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Biomedical subjects

K Takehana

Publications and source records attributed to K Takehana.

17 recordsLinked to original sources

Histochemistry of glandular metaplasia at the trigone of the urinary bladder in cows.

The epithelium at the trigone of the urinary bladder showed intestinal metaplasia of a colonic type in three Holstein-Friesian cows affected with chronic polypoid cystitis. Except for Ricinus communis (RCA-I), almost all goblet cells in the whole crypt were positive to periodic acid-Schiff (PAS), alcian blue (AB) pH 2.5, AB pH 1.0, periodate borohydride-potassium hydroxide-PAS (PB-KOH-PAS), Ulex europaeus (UEA-I), Triticum vulgare (WGA) and Arachis hypogaea (PNA) after neuraminidase digestion. This result indicated that most goblet cells contained acidic and neutral glycoconjugates as O-acetylated sialomucin, L-fucose, N-acetyl-glucosamine, neuraminic acid residues and sialic acid-galactose dimers and were devoid of beta-D-galactose. The goblet cells at the surface in the upper half of the crypt contained both sulpho- and sialo-mucins with N-acetyl-galactosamine residues by AB (pH 2.5)-PAS, high iron diamine (HID)-AB (pH 2.5), Dolichos biflorus (DBA) and Glycine maximus (SBA) reactions. On the other hand, the lower goblet cells were found to contain predominantly sulphated mucins with D-mannose and D-glucose residues by AB-PAS, HID-AB and Concanavalia ensiformis (Con A) reactivities. This suggested that mucin secreted from these cells was similar to that secreted from the goblet cells of the large intestine in cattle.

Alcian Blue

Cytoplasmic filaments in the endothelial cells of the sheathed capillary: an ultrastructural and immunocytochemical study in the pig spleen.

Cytoplasmic filaments of the endothelial cells of sheathed capillaries in the pig spleen were identified and their ultrastructure was studied. Two types of cytoplasmic filaments were found: intermediate filaments (diameter: 10 nm) which filled most of the interior of the cells, and thin filaments (diameter: 5 nm) which were located just beneath the cell membrane and filled the lateral cytoplasmic processes. In immunocytochemical preparations, the intermediate filaments were positive for vimentin and desmin, and were negative for keratin. Staining of the thin filaments with heavy meromyosin resulted in arrowhead formations. These observations suggest that the intermediate filaments maintain the cytoarchitecture, possibly protecting the cell from structural alterations induced by blood pressure changes. Concurrently, thin filaments may facilitate the passage of red blood cells and blood platelets through the interendothelial fenestrae of the sheathed endothelial cell to the reticular meshwork in the capillary sheath.

Animals

Diastolic time and left ventricular performance during one- and two-handed weight holding.

To evaluate the difference in DT and systolic time intervals during one-handed and two-handed weight holding with the same total load, the DT-heart rate relationship was studied in 12 patients and ten normal volunteers. Heart rate, blood pressure, and pressure-rate product were significantly higher in one-handed compared with two-handed weight-holding exercise in patients with MI. Although there were no significant differences in QS2 and DT between the two types of weight-holding exercise in normal volunteers, QS2 was significantly longer and DT was significantly shorter in one-handed compared with two-handed weight-holding exercise in patients with MI. Disproportionate shortening of DT with higher pressure-rate product during one-handed compared with two-handed weight-holding exercise can initiate an imbalance of myocardial oxygen supply and demand in patients with MI.

Analysis of Variance

Interaction of nuclear factors with the regulatory region of the N-myc gene during differentiation of human embryonal carcinoma cells.

The human embryonal carcinoma cell line, NEC14, can be induced to differentiate by the addition of 10(-2) M N,N'-hexamethylene-bis-acetamide (HMBA). During the early stage of HMBA-induced differentiation, the level of N-myc expression decreased steeply and transiently, and then quickly returned to its original level after reaching a minimal level at 18 h after addition of HMBA. Nuclear run-on experiments indicated that this transient decrease is regulated at the transcription start point. To investigate the mechanism of this down-regulation, the 5'-flanking region of the human N-myc gene was cloned and sequenced. Computer analysis of the sequence revealed high homology with the 5'-flanking region of the mouse N-myc gene, especially (greater than 80%) in the region of nt positions -1777 to -1732, nt positions -763 to -501 and nt positions -260 to + 1. The patterns of protein binding to the upstream region during the early stage of NEC14 cell differentiation were analyzed by gel retardation assay. The DNA fragments VIII and X, containing the sequences of nt positions -1437 to -1237 and nt positions -1863 to -1710, respectively, formed the DNA-protein complexes which were greatly reduced in quantity in the cell extract prepared 18 h after the addition of HMBA. This reduction, however, was not observed with an extract similarly prepared from the NEC14 derivative cell line, H10, expressing the N-myc gene constitutively. These results suggest a causal connection between the complex formation and the high-level transcription of the N-myc gene.

Acetamides

A transient decrease in N-myc expression and its biological role during differentiation of human embryonal carcinoma cells.

The human embryonal carcinoma (EC) cell line, NEC14 can be induced to morphologically differentiate by the addition of 10(-2) M N,N'-hexamethylene-bis-acetamide (HMBA) in vitro. The expression of several cellular oncogenes (c-onc) in NEC14 cells was examined after induction of differentiation by HMBA. The level of N-myc expression was the highest in undifferentiated cells but decreased transiently to less than 1/10 of the original level shortly after the induction of differentiation. To investigate the role of the transient decrease in N-myc level on NEC14 cell differentiation, a chimeric human N-myc gene in which transcription is initiated at the human beta-actin gene promoter was constructed and introduced into NEC14 cells. Several transformants expressing the exogenous N-myc gene constitutively were established. These transformants showed 10- to 70-fold increases in plating efficiency and shorter population doubling times as compared with the parental NEC14 cells. The transformants were hard to induce, spontaneously differentiated cells on the periphery of cell clusters in culture, unlike parental NEC14 cells, and took longer for HMBA-induced morphological differentiation. The populations of the cells expressing HLA and SSEA-1 antigens increased from 10%-20% to nearly 100% in NEC14 cells after the induction of differentiation, while the populations expressing these antigens increased only to 50%-60% in one of the transformants, S11. The transformants gained an increased tumorigenic potential in nude mice, and the tumors produced consisted exclusively of EC stem cells. These results suggest that the additional expression of the exogenous N-myc gene (increased about two-fold) confers the more transformed state on the cells.

Antigens, Surface

Duodenal glands of the pony (Equus caballus).

The ultrastructure and histochemistry of the duodenal glands of the pony (Equus caballus) was examined in four horses. Unlike that of most species except for the rabbit, the submucous glands of the horse contain two distinct cell types, serous and mucous. These cells are described.

Animals

Ultrastructure of the red pulp in spleen innervation in horse and pig.

The innervation of the red pulp in the spleen of horse and pig was investigated by electron microscopy. In addition, the neurilemma was studied by immunohistochemistry specific for S-100 protein. In the pig, a large population of smooth-muscle cells extending from the smooth-muscle trabeculae was present in the red pulp. The cytoplasmic processes of reticular cells enwrapped the smooth-muscle cells, and nerve fibres were distributed between the smooth-muscle cells and the reticular cells. The nerve terminals clustered toward the facing of the smooth-muscle cells. Nerve fibres and terminals were not disclosed within the sheathed artery. Immunohistochemically the neurilemma showed a reaction positive for S-100 protein. In the horse, no smooth-muscle cells were noted in the red pulp. The nerve fibres terminated around the cytoplasmic processes of the reticular cells. Nerve fibres and terminals were disclosed within the sheathed artery, and the terminals contained both large and small dense-cored vesicles. Immunohistochemically the neurilemma showed a reaction negative for S-100 protein. These findings support the presence of the axon-bearing reticular cells described earlier in the horse spleen.

Animals

Carbohydrate histochemistry of bovine duodenal glands.

The complex carbohydrates in the bovine duodenal glands were examined histochemically using 5 peroxidase-labeled lectins to achieve a better knowledge of their glycoprotein profiles. The presence of at least 2 different cell types and secretion of neutral mucin were observed in the bovine duodenal glands. The duodenal gland cells located in the central area of the lobules contained neutral glycoproteins with different saccharide residues as beta-D-gal-(1-3)-D-galNAc, alpha-D-galNAc, beta-(1-4)-D-glcNAc, NeuNAc and D-galNAc. The other cell types located in the peripheral area of the lobules contained alpha-L-Fuc, alpha-D-galNAc, beta-(1-4)-D-glcNAc, NeuNAc and D-galNAc. These findings seemed to be characteristic of a unique digestive process of the ruminant.

Animals

Anomalous caudal vena cava passing through the vertebral canal in a calf.

A 15-day-old female Holstein-Friesian calf with an anomalous caudal vena cava was examined macroscopically, roentgenologically, and histologically. The calf, weighing 43 kg, had severe scoliosis. A common renal vein merged into a single venous trunk formed by the union of the left and right common iliac veins. The trunk entered the vertebral canal through the left intervertebral foramen formed by the last (13th) thoracic and the first lumbar vertebrae. The trunk continued along the ventral side of the narrowing spinal cord inside the canal, and then ran out the left intervertebral foramen formed by the 8th and 9th thoracic vertebrae and emptied via the right azygos vein into the cranial vena cava. In contrast, the hepatic vein passed through the foramen vena cava independently of the trunk and entered the right atrium directly. The pathogenesis of the present anomaly may be explained as follows: The right subcardinal vein, failing to make connection with the liver, shunted directly into the right azygos vein derived from the right supracardinal vein. The body axis began to curve before ossification of the vertebrae occurred. Consequently, the developing right supracardinal vein, located close to the spinal cord, is thought to have become enclosed in the vertebrae with the spinal cord during the early fetal stages.

Animals

Seven-legged calf--dipygus with an extra foreleg at the pelvic region.

A male Holstein-Friesian calf with seven legs was examined macroscopically and radiographically. External features included two normal forelimbs, two normal hindlimbs (lateral hindlimbs), and two abnormal hindlimbs (medial hindlimbs) which were underdeveloped. Also, a rudimentary forelimb, which was attached to the pelvic region, was observed between both the medial hindlimbs. It consisted of an underdeveloped humerus, a duplicated ulna, several carpal bones, a partially duplicated metacarpal bone and three digits with three hoofs. This leg was connected with two sets of coxae by a irregular-shaped bone considered the vestigial vertebrae and ribs. Two penises and scrotums, three kidneys and testes were also observed. This calf is the first case of dipygus associated with pygopagus parasiticus in cattle. Based on these findings, the pathogenesis of this rare anomaly was briefly discussed from an embryological point of view.

Abnormalities, Multiple

Scanning electron microscopic studies on the red pulp of the mink spleen.

In order to obtain three-dimensional information on the fine architecture of the red pulp of the mink spleen, especially the circulation pattern in the red pulp, perfusion-fixed and freeze-cracked specimens were examined with a scanning electron microscope. A latticework was formed by elongated endothelial cells (rod cells) with side processes and the spongy reticular tissue. The outer surface of the sinus was covered with fine processes of reticular cells. Numerous sheathed arteries were found in the splenic cord. The sheath was composed of a few layers of flat reticular cells. The arterial capillaries of the red pulp opened directly into the cordal space. No evidence could be detected to prove or suggest any direct continuity between arterial capillaries and splenic sinuses. These results strongly support the concept of "open circulation", at least in the red pulp of the mink spleen, with the possibility of a "functionally closed circulation" under some physiological conditions.

Animals

Histochemistry of complex carbohydrates in the horse duodenal gland.

Complex carbohydrates were examined in glandular cells of the horse duodenal gland by using lectin histochemical techniques. In the horse, the duodenal gland was distributed in the area from the uppermost part of the small intestine to a point about 6m caudal to the pylorus. It consisted of two types of cells, mucous and serous cells. The former was found in glands distributed almost all over this part, but the latter was present in glands distributed restrictedly to the uppermost part of the small intestine at a point about 10 cm caudal to the pylorus. The cytoplasm of the mucous cell contained neutral glycoproteins with different saccharide residues as alpha-D-mannose, N-acetyl-beta (1----4)-D-glucosamine, galactose, alpha-galactose, alpha-N-acetylglucosamine, beta-D-Gal (1----3)-D-GalNAc, alpha-L-fucose and sialic acid. On the other hand, the serous cell contained neutral and acid glycoproteins with different residues such as alpha-D-mannose, alpha-D-glucose, beta-D-galactose(1----3)-D-N-acetylgalactose, alpha-L-fucose, N-acetyl-alpha-D-galactosamine, N-acetyl-beta (1----4)-D-glucosamine, galactose, alpha-galactose, alpha-N-acetylglucosamine and sialic acid. It is also elucidated in the present study that lipase, an enzyme for digestion, is contained in the serous cell of the equine duodenal gland.

Animals

Small-square (SS) net exists in trout muscle.

The small-square-net form (with the large-square-net form) was clearly observed in brown trout muscle in cross-square-net-section. Fresh adductor mandibularis and dorsal muscle strips, isometrically clamped to prevent contraction, were prefixed in 3% glutaraldehyde, 0.1 M cacodylate, pH 7.4, and post-fixed in 1% osmium tetroxide in veronal acetate buffer at 2 degrees C. The samples were dehydrated, embedded in epon-araldite, cross-sectioned, and stained with uranyl acetate and lead citrate for electron microscopic observation. The sections provided the first clear evidence of the small-square-net form, coexisting with the basket-weave pattern, in fish muscle. In previous experiments, we could not demonstrate the small-square-net pattern convincingly with guppy tissue (Yamaguchi et al., 1985a) which may be more susceptible to chemical or physical stress than is trout tissue. The existence of the diverse structural patterns in cross-sections of some, if not all, vertebrate striated muscle must be derived from a basic unit which although it may vary in size among species is structurally identical. The findings agreed with our unified model of narrow and wide Z-line structure (Yamaguchi et al., 1980; Yamaguchi et al., 1985a; Yamaguchi et al., 1985b).

Animals

[A possible circadian rhythm of susceptibility to thiamylal].

A circadian rhythm of the susceptibility to thiamylal was evaluated in 30 patients. They were divided into two groups; 15 patients were induced around 9 AM (Morning-group) and the rest were in the early afternoon (Afternoon-group). The induction was performed by intravenous administration of thiamylal sodium. Parameters measured were the time and doses of thiamylal sodium to obtain the loss of consciousness and the loss of eyelash reflex. The results were as follows; the doses of thiamylal sodium to obtain the loss of consciousness were 3.5 +/- 0.6 mg.kg-1 (mean +/- SD) in Morning-group, and 4.1 +/- 0.4 mg.kg-1 in Afternoon-group, respectively. Thus, larger doses of thiamylal were necessary to obtain the loss of consciousness in Afternoon-group than in Morning-group (P less than 0.01). The doses of thiamylal sodium to obtain the loss of eyelash reflex were 4.4 +/- 0.8 mg.kg-1 in Morning-group and 5.0 +/- 0.6 mg.kg-1 in Afternoon-group, respectively. Also, larger doses of thiamylal were necessary to obtain the loss of eyelash reflex in Afternoon-group than in Morning-group (P less than 0.05). This study suggests that a circadian rhythm of the susceptibility to thiamylal exists in humans.

Adult

Molecular cloning and characterization of cellular genes whose expression is repressed by the adenovirus E1a gene products and growth factors in quiescent rat cells.

Several cDNA clones of cellular genes, whose expression is repressed by the adenovirus type-12 E1a gene products, were isolated from a rat 3Y1 cell cDNA library by differential plaque hybridization with labeled cDNA probes prepared from 3Y1 and the derivative cell line expressing the E1a gene constitutively. The changes in the levels of these gene transcripts during cell-cycle progression from G0 to G1 to S phase were analyzed with 3Y1 cells and gMA cell lines, derived from 3Y1 cells, in which the expression of the E1a gene or its 13S, 12S cDNA can be switched on by the addition of dexamethasone. Quantitation of the transcripts by Northern-blot hybridizations and by nuclear run-on experiments revealed the following. (i) The level of clone-53 mRNA (which turned out to be the fibronectin (FN)-coding mRNA) is very high in resting gMA cells and decreased rapidly after switching on of the E1a gene or its 13S, or 12S cDNA. (ii) The addition of serum or platelet-derived growth factor to resting 3Y1 cells also resulted in a rapid decrease in the FN mRNA level, but the addition of epidermal growth factor (EGF) had little or no effect. (iii) The level of clone-56 mRNA in gMA cells was not affected by the induction of the E1a gene expression; however, the addition of EGF to resting gMA or 3Y1 cells resulted in a decrease of this mRNA after a 12- to 16-h lag period. Induction of the E1a gene expression in gMA cells treated with EGF shortened the lag period. The addition of serum to resting 3Y1 cells decreased the clone-56 mRNA level without a significant lag period.

Adenoviridae