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Biomedical subjects

O Tanaka

Publications and source records attributed to O Tanaka.

At least 19 recordsLinked to original sources

Opaque eyes developed in transgenic mice with T-cell receptor delta gene.

PURPOSE: During the generation of transgenic mice (TGs) introduced with mouse T-cell receptor delta (TCR delta) gene, the authors found a TG line with corneal opacity that coincided with the presence of the transgene. The authors investigated the pathogenesis and molecular mechanisms of this corneal opacity in this line. METHODS: The pathologic features and pathogenesis of the corneal opacity in TGs were examined histologically using transmission and scanning electron microscopy, as well as light microscopy. DNA and RNA blot analyses were performed to examine the copy number and the expression of the transgenes, respectively. RESULTS: Histologically, edema of the corneal epithelium and adhesion of the iris to the cornea were observed in adult TGs. In the developmental analysis, the authors first observed relative hypoplasia of the ciliary body on day 18 of gestation and dysgenesis of the anterior chamber angle from postnatal day 2. Corneal opacity was observed from postnatal day 8, coinciding with the histologic vesicular change of the epithelium. No inflammation was observed through its life. In the sublines that have different copy numbers of the transgene, the occurrence of the opacity depended on the copy number of the transgene. Expression of the transgene in the thymus was consistent with the number of the introduced transgene. CONCLUSION: In a TCR delta TG line, the overexpression of transgenes coincided with abnormal development of the ocular anterior segment and the corneal opacity. Pathogenesis is described, and possible molecular mechanisms are discussed.

Animals

Different characteristics of hepatoid and non-hepatoid alpha-fetoprotein-producing gastric carcinomas: an experimental study using xenografted tumors.

The characteristics, including metastatic potential, of 5 xenografts of alpha-fetoprotein (AFP)-producing gastric carcinomas in nude mice, designated TSG1, TSG3, TSG11, TSG17 and TSG20, were examined. Of these xenografts, TSG1, TSG11 and TSG20 were regarded as hepatoid adenocarcinomas based on their morphological resemblance to hepatocellular carcinoma, frequent immunoreactivity for liver-cell markers, and excessive production of AFP with a high concanavalin A (Con-A)-binding property of hepatic type. On the other hand, TSG3 and TSG17 tumors showed the features of poorly differentiated medullary adenocarcinoma with scattered AFP-positive cells consistent with low AFP levels in mouse sera, and negative immunoreactivity for other liver-cell markers. Ultrastructurally, these tumors were composed of undifferentiated cells with a little adenocarcinomatous differentiation. Moreover, the AFP produced by TSG3 and TSG17 tumors had an extremely high Con-A nonbound fraction (80% to 90%), which was different from that of the hepatic or yolk-sac types. Therefore, both TSG3 and TSG17 tumors were regarded as non-hepatoid, poorly differentiated adenocarcinomas which could be differentiated from any types of AFP-producing gastric carcinoma. Furthermore, cells from hepatoid adenocarcinoma strains (TSG1, TSG11 and TSG20) injected into the spleens of nude mice produced liver metastases in all the mice examined, whereas cells from non-hepatoid carcinoma strains (TSG3 and TSG17) produced few or no liver metastases. Our data show that some non-hepatoid AFP-producing gastric carcinomas have lower liver-metastasizing potential than hepatoid AFP-producing gastric carcinomas.

Adenocarcinoma

Transgenic mouse model of hemifacial microsomia: cloning and characterization of insertional mutation region on chromosome 10.

The 643 transgenic mouse line carries an autosomal dominant insertional mutation that results in hemifacial microsomia (HFM), including microtia and/or abnormal biting. In this paper, we characterize the transgene integration site in transgenic mice and preintegration site of wildtype mice. The locus, designated Hfm (hemifacial microsomia-associated locus), was mapped to chromosome 10, B1-3, by chromosome in situ hybridization. We cloned the transgene insertion site from the transgenic DNA library. By using the 5' and 3' flanking sequences, the preintegration region was isolated. The analysis of these regions showed that a deletion of at least 23 kb DNA occurred in association with the transgene integration. Evolutionarily conserved regions were detected within and beside the deleted region. The result of mating between hemizygotes suggests that the phenotype of the homozygote is lethality in the prenatal period. These results suggest that the Hfm locus is necessary for prenatal development and that this strain is a useful animal model for investigating the genetic predisposition to HFM in humans.

Animals

Stage-specific expression of cancer-associated type 1 and type 2 chain polylactosamine antigens in the developing pancreas of human embryos.

Expression of type 1 and type 2 chain carbohydrate antigens during the course of morphogenesis of human embryonic pancreas was investigated using specific monoclonal antibodies and compared with the carbohydrate antigen profiles of human pancreatic cancers. The type 2 chain antigens, such as stage-specific embryonic antigen 1 (Le(x)) and I-antigens, appeared much earlier than the type 1 chain antigens; the epithelial cells of primitive foregut were Le(x)+I-antigen- in the embryos at Carnegie stages 16-23, while the pancreatic primordial cells, which had differentiated from the Le(x)+ gut epithelial cells, were Le(x)-I-antigen+ at Carnegie stages 22-23. The type 1 chain antigens, such as Le(a), Le(b), Le(c), and their sialylated derivatives, were not expressed in any cells at these stages and appeared much later in the pancreas of the 10-12-week embryos, when the primitive pancreatic ductal cells in the primordia exhibited an extensive budding of the daughter cells. At this stage, Le(a) appeared and was expressed strongly in the epithelial cells of primitive pancreatic ducts as well as in the daughter cells that were destined to differentiate into future centroacinar cells; Le(b) was localized in the daughter cells which were to become future acinar cells; and Le(c) was specifically expressed in the daughter cells which were to form future Langerhans islets. With regard to the sialylated derivatives of Le(a), expression of the 2-3 sialyl Le(a) antigen was limited to the epithelial cells of the primitive pancreatic ducts, while the 2-6 sialyl Le(a) antigen was strongly expressed in the future centroacinar cells, which had differentiated from the corresponding daughter cells. Among these antigens, the Le(a) and 2-3 sialyl Le(a) antigens showed the highest incidence in human pancreatic cancer tissues. These results indicate that the expression of these carbohydrate antigens in embryonic pancreas is differentiation dependent and cell lineage specific and that most human pancreatic cancer cells mimic the carbohydrate antigen profile of the epithelial cells of the primitive pancreatic ducts in human embryos.

Adult

Supra-neuroectodermal cells and fibers on the primary nasal cavity and in the fourth ventricle of mouse and human embryos: scanning and transmission electron microscopic studies.

Neuroectoderm-derived epithelia of the primary nasal cavity and the fourth ventricular floor and roof were observed by scanning (SEM) and transmission electron microscopy (TEM) and SEM-TEM correlative views in mouse embryos of 9th to 13th days of gestation, and in 38 externally normal human embryos ranging at Carnegie stages from 13 to 18 (about 5 to 7 weeks of gestation). Smooth-surfaced spindle-shaped cells with one or more cytoplasmic processes and cord-like cytoplasmic structures were observed by SEM on the wall of the primary nasal cavity of both species. They had morphological features similar to those of neuronal type 1 supraependymal (SE) cells and SE fibers on the floor and roof of the fourth ventricle in both species. Type 1 SE cells, SE fibers, and corresponding structures in the primary nasal cavity were localized in relation to the underlying developing nerve and vascular systems. Furthermore, their processes and fibers ran roughly parallel to these underlying structures and they penetrated the epithelial layer at the ends, suggesting a connection with underlying structures. From TEM and SEM-TEM correlative observations, SE fibers in the fourth ventricle and cord-like structures in the primary nasal cavity, both with a larger diameter, were deduced as single axon-like processes or bundles of processes. Those fibers and cord-like structures of smaller diameters were interpreted as elongated telophase bridges; both contained parallel packed microtubules and connected distant cells. Since these processes and fibers were generally longer and became fewer at later developmental stages, they appeared to be transient neuronal structures. They may play a development-related role in such morphogenetic cell movements as in the developing nerve and vascular systems in the epithelial and/or subepithelial layers, but not as direct rudiments of adult nerve tissues.

Animals

Oleanane glycosides from Glycyrrhiza yunnanensis roots.

From the roots of Glycyrrhiza yunnanensis, collected in Yunnan, China, six new oleanane-type triterpene glycosides named yunganosides A1, B1, C1, D1, E2 and F2 were isolated together with hypaphorine. The structures of these glycosides were established by spectroscopic and chemical means.

Carbohydrate Sequence

Timing and sequence of the events in the development of extraocular muscles in staged human embryos: ultrastructural and histochemical study.

The ultrastructure and the appearance of glycogen were studied in the extraocular muscles of 14 externally normal human embryos (Carnegie stages 13-21). At stage 16, myofibrils with an immature Z line and glycogen granules appeared in the cytoplasm of the myoblast. The myoblasts came into cluster at stage 18, and fusion between the myotubes was observed at stage 20. At this stage, an M line appeared in the myofibrils. At stage 21, an A band with a Z line and an H band with an M line were observed, the sarcoplasmic reticulum appeared in the cytoplasm of the muscle fibers and glycogen increased in volume in the cytoplasm. In the previous study, we showed that the muscle-specific isoenzymes, such as creatine kinase, beta-enolase and glycogen phosphorylase, appeared from stage 18 to 20 in the extraocular muscles. The previous findings and the present results suggest that the fusion of the muscle cells occurs in the period when some molecular markers of muscle differentiation are expressed in vivo.

Glycogen

Immunohistochemical study of carbonic anhydrase III in the extraocular muscles of human embryos.

The differentiation of extraocular muscles was studied immunohistochemically in externally normal human embryos (Carnegie stages 13-23), using antibodies to carbonic anhydrase (CA) III and beta-enolase as the markers of type 1 and type 2 muscle fibers, respectively. At stage 18, some myoblasts were immunoreactive to beta-enolase antibodies, however, CA-III immunoreactivity was not observed around the optic vesicle. At stage 20, CA-III immunoreactivity appeared in some muscle fibers of extraocular muscles. From stage 21 to stage 23, CA-III-immunoreactive fibers increased and almost equalled the number of beta-enolase-immunoreactive fibers. These findings suggest that CA-III-immunoreactive type 1 fibers appear in the late stage of myogenesis compared with beta-enolase-immunoreactive type 2 fibers.

Carbonic Anhydrases

Studies on single-dose toxicity of hydrophobically modified hydroxypropyl methylcellulose in rats.

Single-dose toxicological studies of hydrophobically modified hydroxypropyl methylcellulose (HM-HPMC, hydroxypropyl methylcellulose modified with stearylglycidylether) were conducted. A dispersion of HM-HPMC was administered to rats orally or by dermal application at doses up to 900 mg/kg. After the oral administration, the mean body weight of the 900 mg/kg group on the first day after administration was slightly but significantly lower (P less than 0.05) than that of the control group, and one rat had loose stools at 30 min. after the administration. No other abnormalities were noted. In the case of dermal application, no abnormalities were observed. No rats died, and no abnormalities in their organs were found by either route. In conclusion, there was no observed toxicity of HM-HMPC after oral or dermal administration at single dose up to 900 mg/kg under the conditions of these studies.

Administration, Cutaneous

Primary dermal and eye irritability tests of hydrophobically modified hydroxypropyl methylcellulose in rabbits.

Primary dermal and eye irritation tests of hydrophobically modified hydroxypropyl methylcellulose (HM-HPMC, hydroxypropyl methylcellulose modified with stearylglycidylether), a new cellulose derivative used as a thickener for topical pharmaceuticals and cosmetics, were conducted in rabbits. A dispersion of HM-HPMC (3%) was applied to intact and abraded skins and reactions were observed. A very slight erythema was observed in both skins and this polymer was categorized as a "mild irritant". In the eye irritation test, with a dispersion of the same concentration, it was categorized as "marginal" in unrinsed eyes and "negative" in rinsed eyes.

Administration, Cutaneous

[Effects of sex hormones and propylthioracil on growth of transplantable rat thyroid tumor with estrogen receptor (ER)].

UNLABELLED: Effects of hormones (E2: estradiol, TP: testosterone) and propylthiouracil (PTU) on the growth of transplantable rat thyroid tumor (F2D1) having estrogen receptors were studied. Rat thyroid neoplasms, induced by N-bis (2-hydroxypropyl) nitrosamine, were inoculated subcutaneously from donor to recipient rats, in order to establish 16 transplantable rat thyroid tumor lines. The grafts were used for histological studies and for the assay of estrogen receptor (ER) and androgen receptor (AR). Of these lines, we designated papillary carcinoma, which was positive for ER (N: 12.5fmol/mg protein, Kd: 0.4nM) but negative for AR, as F2D1. For studies on the effects of sex hormones and PTU on the growth of transplantable tumors, the rats which had been inoculated with tumors were divided into the following 8 groups; (1) Intact, (2) PTU, (3) ovariectomy (OV), and (4) OV + E2 for female, and (5) Intact, (6) PTU, (7) castration (CA), and (8) CA + TP for male. RESULTS: The growth rate of F2D1 in female rats was decreased by OV, but no change was observed in OV + E2 as compared with Intact. CA and CA + TP in male rats did not influence the growth rate. PTU produced a significant increase in the growth rate in both sexes. These results demonstrate that estrogen and PTU act on the growth of ER-positive rat thyroid tumors.

Animals

[Flow cytometric analysis of DNA ploidy in nasosinal papilloma].

Using paraffin embedded specimens taken from 32 patients with histologically benign nasosinal papillomas, we conducted nuclear DNA analysis by flow cytometry and studied the biological degree of malignancy in this disease. Aneuploidy, which is frequently observed in malignant tumors was not seen in any of these nasosinal papilloma cases. Age did not affect either S+G2M % or polyploid %, two parameters that reflect cell proliferation capacity. Both parameters, S+G2M % and polyploid %, were higher in inverted papillomas which are more likely to become malignant than epithelial papillomas. In recurrent cases of nasosinal papilloma both S+G2M % and polyploid % were higher than in nonrecurrent cases. Moreover, the polyploid % was significantly different, supporting speculation that this can be used as a parameter for predicting recurrence of nasosinal papilloma.

Cell Nucleus

[A case of renal hemangiopericytoma].

A 27-year-old female was referred with an abdominal mass. Examination revealed a non tender firm mass in the right flank and hypertension (200/100 mmHg). An angiomyolipoma was suspected on computed tomography and arteriography and a radical nephrectomy was performed. On cut section, the kidney was occupied by a well-capsulated, grayish tumor measuring 10 x 9 cm. Pathological diagnosis was a renal hemangiopericytoma without involvement of the capsule. Her blood pressure has normalized after the operation. She has no evidence of recurrence after 18 months' follow-up.

Adolescent

[Flow cytometric BrdU/DNA assay for anticancer agent sensitivity test].

The significance of BrdU/DNA double staining method using flow cytometry as a chemosensitivity test of anticancer agents is herein reported. Experimentally, CDDP, ADM and MMC yielded a significant increase of G2 phase fraction at 24, 48 and 72 hours, and caused a significant decrease of BrdU labeling index at 48 hours. The changes of G2 phase fraction and BrdU labeling index were correlated well with the result of colony forming test, and, therefore, were considered as a useful index of lethal effect of anticancer agents. For the clinical application, enzymatic preparation and discontinuous Ficoll density gradient method might be advantageous to collect viable tumor cells from solid specimen. Flow cytometric BrdU/DNA assay can be clinically applied as a chemosensitivity test in selecting effective anticancer drugs.

Antineoplastic Agents

[Studies on chemical constituents of the roots of Lantana camara].

Six oligosaccharides (I-VI) and six iridoid glucosides (VII-XII) isolated from the ethanolic extract of Lantana camara roots were identified as stachyose (I), verbascose (II), ajugose (III), verbascotetracose (IV), alpha-D-galac-(1-[-6)-alpha-D-galac(-1](3)-6-D-gluc(V ) , alpha-D-galac-(1-6)-alpha-D-galac(-1]-(4)6-D-)gluc(VI) , theveside (VII), 8-epiloganin (VIII), shanzhsid methyl ester (IX), theviridoside (X), lamiridoside (XI) and geniposide (XII), on the basis of spectral analysis (1H-NMR, 13C-NMR, FD-MS, GC-MS), physico-chemical constants and preparation of derivatives. V and VI were new compounds named lantanose A and lantanose B, respectively. The others were isolated from this plant for the first time.

Drugs, Chinese Herbal

[Management of patients with impaired glucose tolerance following esophagectomy in carcinoma of the esophagus].

We demonstrated the effectiveness of the "sliding scale" insulin infusion in diabetic patients undergoing esophagectomy. Fifty-eight patients were followed after esophagectomy to clarify differences in energy expenditure and caloric contributions of substrates. Energy expenditure was measured by indirect calorimetry on the 1st, 3rd, 5th, and 7th days after esophagectomy. Out of 58 patients, 7 were divided into diabetic (D), 30 into borderline (B), 21 into normal (N), according to a 75 g oral glucose tolerance test, preoperatively. Forty-four patients underwent esophagectomy by means of right thoracotomy, and blunt esophagectomy was done on 14 patients. The results were as follows: 1) No relation was found between postoperative morbidity and severity of glucose intolerance. 2) There was no significant difference in changes in energy expenditure following operation among groups D, B, and N. No difference was found in ratio of caloric intake to energy expenditure among groups D, B, and N. Caloric contributions of substrates seemed to be comparable among groups D, B, and N. These results suggest that "sliding scale" insulin infusion with total parenteral nutrition enable us to control not only blood glucose level but energy metabolism following esophagectomy in diabetic patients.

Diabetes Complications