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Biomedical subjects

O Tanaka

Publications and source records attributed to O Tanaka.

At least 181 records · Page 10Linked to original sources

Immunohistochemical study of creatine kinase isozymes in the human upper limb bud.

Creatine kinase (CK) is involved in the production of ATP and is composed of two immunologically distinct subunits, B (CK-B) and M (CK-M). In the differentiation of myoblasts, the isozyme of CK changes from CK-B to CK-M. In the present study, the expression of CK subunits was studied immunohistochemically in the upper limb bud of human embryos (Carnegie stages 13-21). It was found that CK-B and CK-M immunoreactive cells appeared at stage 15 and at stage 18, respectively.

Arm↗

Chromosome analysis of postimplantation stage embryos for studying possible causes of developmental abnormalities in nonobese diabetic mice.

The chromosomes of postimplantation stage embryos of nonobese diabetic (NOD) mice were analyzed to investigate the causal mechanism of congenital anomalies in diabetic pregnancies. Postimplantation stage embryos (day 12 of gestation) in diabetic (NOD-DM) and nondiabetic (NOD-N) NOD mice had either a high or low incidence of chromosomal abnormalities. A large majority of externally normal embryos from NOD-DM and NOD-N mice had low incidences. A high incidence of chromosomal abnormalities was found in externally abnormal embryos of NOD-DM and NOD-N mice, and in a smaller number of externally normal NOD-N and NOD-DM embryos. No control ICR embryo manifested a high incidence of chromosomal abnormalities. In the NOD-DM embryos, the chromosomes appeared to be influenced by long-term maternal diabetic conditions, while high incidences of chromosomal abnormalities in the NOD-N embryos suggested a probable cause by other factor(s) (e.g. a genetic predisposition) or by a very mild diabetic condition because the NOD-N mice were prediabetic.

Animals↗

Studies on Balanites aegyptiaca fruits, an antidiabetic Egyptian folk medicine.

An aqueous extract of mesocarps of the fruits of Balanites aegyptiaca exhibited a prominent antidiabetic activity by oral administration in streptozotocin induced diabetic mice. From one of the active fractions of this extract, two new steroidal saponins were isolated, and their structures were determined as 26-O-beta-D-glucopyranosyl-(25R)-furost-5-ene-3 beta,22,26-triol 3-O-[alpha-L-rhamnopyranosyl-(1----2)]-[beta-D-xylopyranosyl-(1---- 3)]-[alpha-L-rhamnopyranosyl-(1----4)]-beta-D-glucopyranoside and its 22-methyl ether. In addition, two known saponins, 26-O-beta-D-glucopyranosyl-(25R)-furost-5-ene-3 beta,22,26-triol 3-O-(2,4-di-O-alpha-L-rhamnopyranosyl)-beta-D-glucopyranoside and its methyl ether were isolated and identified. It was revealed that the individual saponins did not show antidiabetic activity, while the recombination of these saponins resulted in significant activity. From an ethanolic extract of the epicarps, two known flavonol glycosides, isorhamnetin-3-O-robinobioside and isorhamnetin-3-O-rutinoside were isolated and identified.

Animals↗

Transfructosylation of rebaudioside A (a sweet glycoside of Stevia leaves) with Microbacterium beta-fructofuranosidase.

It was found that a beta-fructofuranosidase produced by Microbacterium sp. H-1 has potent trans-beta-fructofuranosylation activity from sucrose (donor). By means of this enzyme system, rebaudioside A (RA), the second major sweet steviol glycoside of the leaves of Stevia rebaudiana, was subjected to transfructosylation, affording a mono-beta-fructofuranosylated product (RA-F) in a high yield. The structure of RA-F was elucidated as beta-D-fructofuranosyl-(2----6)-beta-D-glucopyranosyl ester of steviol-13-O-[beta-D-glucopyranosyl-(1----2)]- [beta-D-glucopyranosyl-(1----3)]-beta-D-glucopyranoside. Some improvement in the quality of sweetness was observed for RA-F.

Actinomycetales↗

Solubilization of steviolbioside and steviolmonoside with gamma-cyclodextrin and its application to selective syntheses of better sweet glycosides from stevioside and rubusoside.

1,4-alpha-Glucosylation at the 13-O-glycosyl moiety of stevioside (S) and rubusoside (RU) results in a significant increase of sweetness. Saponification of the 19-COO-beta-glucosyl linkage of S and RU yielded steviolbioside (SB) (= 13-O-beta-sophorosyl-steviol) and steviolmonoside (SM) (= 13-O-beta-glucosyl-steviol), respectively, both of which are poorly soluble in an acetate buffer. It was found that the solubilities of SM and SB in the buffer solution were remarkably increased in the presence of gamma-cyclodextrin (gamma-CD). SB was solubilized in the buffer solution with the aid of gamma-CD, and the solution was subjected to 1,4-alpha-transglucosylation by using a cyclodextrin glucanotransferase-starch system to give a mixture of products which were glucosylated at the 13-O-glycosyl moiety. This mixture was acetylated, and the acetate was subjected to chemical beta-glucosylation of 19-COOH followed by deacetylation to afford compounds which have superior sweetness to S. In the same way, derivatives with superior sweetness were selectively prepared from RU through SM.

Cyclodextrins↗

Diabetic environment and genetic predisposition as causes of congenital malformations in NOD mouse embryos.

Congenital malformations such as neural tube defects and a kinky or waved vertebral column were observed at higher incidence in embryos from nonobese diabetic (NOD) female mice with overt diabetes (NOD-D; 40.3%, P less than 0.005) or without overt diabetes (NOD-N; 8.4%, P less than 0.05) than in control Institute of Cancer Research (ICR) mouse embryos (1%) at day 13 of gestation. In vivo and in vitro preimplantation development of NOD-N, NOD-D, and ICR embryos did not differ in rate of development, size, or morphology. Embryos cultured from one-cell to early blastocyst stage were mutually transferred to uterine horns of pseudopregnant females between NOD-D and ICR mice and examined at day 13 of gestation. There were significant decreases in ratios of implantation and of viable embryos in ICR embryos transferred to NOD-D recipients (52%, P less than 0.001 and 14%, P less than 0.001, respectively) compared with those ratios in ICR embryos transferred to ICR uteri (79.2 and 56.2%) or those in NOD-D embryos transferred to ICR uteri (70.3 and 33.1%). Furthermore, 18 of 45 viable ICR embryos transferred to NOD-D dams had malformations, whereas there were no malformations in 73 viable ICR embryos transferred to ICR recipients, suggesting deleterious effects of maternal diabetic environment to embryos. On the other hand, 8 of 58 viable NOD-D embryos that were cultured in vitro and transferred to ICR uteri had malformations such as neural tube defects.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Skeletal malformations in rat offspring. Long-term effect of maternal insulin-induced hypoglycemia during organogenesis.

We studied the effect of maternal hypoglycemia on skeletal growth in the offspring of nondiabetic and diabetic rats. Female Wistar rats were injected with streptozocin (30 mg/kg i.v.) 2-3 wk before mating, and diabetes was confirmed by an intraperitoneal glucose tolerance test. On postconception day 9.5 or 10.5, both control and diabetic dams received saline or Actrapid human insulin (400 mU/rat i.p.). Hypoglycemia (approximately 2.8 mM) was induced for 120 min in the insulin-treated mothers. Pregnancy was terminated on gestational day 20. Fetal bones and cartilage were double-stained with alizarin red S and alcian blue 8GS. Insulin-induced hypoglycemia caused delayed ossification in the fetuses of the control dams. The number of malformations, e.g., costal fusion waves, increased greatly. These effects were more striking in the fetuses of dams that had received insulin on day 10.5 rather than on day 9.5 of embryo development. This type of insulin-induced hypoglycemia further delayed ossification of the fetal bones in diabetic dams. The influence of maternal hypoglycemia on skeletal malformations and/or variations was greater in the fetuses of diabetic dams than in the fetuses of control dams. These data suggest that maternal hypoglycemia in early pregnancy has a striking effect on skeletal growth and malformations in fetuses. In addition, mild glucose intolerance in dams may amplify these hypoglycemic effects.

Animals↗

[Flow cytometric analysis of DNA content in paraffin-embedded tissue in head and neck cancer (the second report)--primary lesions of laryngeal cancer].

Flow cytometric analysis of DNA content using paraffin-embedded materials has become an important diagnostic, as well as prognostic, method for clinical pathology and investigative oncology. DNA content as measured in paraffin-embedded materials is closely related with that obtained from fresh specimens. This method also permits retrospective analysis on a lot of cases and studying different specimens of a tumor for intratumor heterogeneity. Flow cytometry was used to investigation of the DNA distribution in biopsy specimens from 30 patients with squamous cell carcinoma of the larynx. These patients had initially treated in our institute in the period from 1986 to 1988 and followed for two to four years. A modification of Hedley's method was used to prepare paraffin-embedded materials. DNA histogram were assessed in terms of DNA index (DI). The coefficient of variation for determination of DI ranged 4.2 to 13.7% (mean 7.8 +/- 2.4%). Using infiltrated lymphocytes and mesenchymal cells as an internal standards, aneuploid DNA histogram were found in 13 of 30 specimens (43.3%). DI ranged from 1.05 to 1.22. There was no significant correlation between DNA content and sex, age, tumor size, staging or treatment. The aneuploidy was found in 2 of 17 cases with well differentiated tumor (11.8%), 10 of 12 with moderately differentiated tumor (83.3%), one with poorly differentiated tumor (100%). 11.8% of patients with diploid tumor cells had recurrence during the interval of observation, compared to 38.5% of those with aneuploid tumor cells (p less than 0.05). In T1 lesions of glottic cancer aneuploid group had a significantly lower local control rate than diploid group (p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Further study on the 1,4-alpha-transglucosylation of rubusoside, a sweet steviol-bisglucoside from Rubus suavissimus.

Rubusoside (the beta-D-glucosyl ester of 13-O-beta-D-glucosyl-steviol), which is the major sweet principle of leaves of Rubus suavissimus S. Lee, was subjected to 1,4-alpha-transglucosylation by the cyclodextringlucanotransferase-starch system (the CGTase system). The tri- and tetra-glucosylated products were isolated together with the mono- and di-glucosylated products, which had already been isolated. A prominent increase in intensity of the sweetness was observed for the compounds which were di- and tri-glucosylated at the 13-O-glucosyl moiety. This result further substantiated the structure-sweetness relationship for 1,4-alpha-glucosylated compounds of steviol-glycosides reported previously. For protection of the 19-COO-glucosyl moiety against glucosylation by the CGTase system, the 4-hydroxyl group of the 19-COO-glucosyl moiety was beta-galactosylated by the beta-galactosidase-lactose system. This galactosylated compound was subjected to a regio-selective glucosylation of the 13-O-glucosyl moiety by the CGTase system, which was followed by enzymic elimination of the galactosyl group to furnish an exclusive preparation of the improved sweeteners just mentioned.

Carbohydrate Sequence↗

Enzymic production of sweet stevioside derivatives: transglucosylation by glucosidases.

For the purpose of improving sweetness and a further study on the structure-sweetness relationship of steviol glycosides, transglycosylation of stevioside by a variety of commercial glucosidases was investigated. It was revealed that two alpha-glucosidases gave glucosylated products. Transglucosylation of stevioside by Pullulanase and pullulan exclusively afforded three products, 13-O-[beta-maltotriosyl-(1----2)-beta-D-glucosyl]-19-O-beta-D-g luc osyl- steviol (1), 13-O-[beta-maltosyl-(1----2)-beta-D-glucosyl]-19-O-beta-D-glucosyl- steviol (2) and 13-O-beta-sophorosyl-19-O-beta-maltotriosyl-steviol (3). All of these products have already been obtained by trans-alpha-1,4-glucosylation of stevioside by the cyclodextrin glucanotransferase starch system, and 1 and 2 have been proven to be tasty and potent sweeteners. Transglucosylation of stevioside by Biozyme L and maltose afforded three new products, 4, 5 and 6, the structures of these compounds being elucidated as 13-O-beta-sophorosyl-19-O-beta-isomaltosyl-steviol (4), 13-O-[beta-isomaltosyl(1----2)-beta-D-glucosyl]-19-O-beta-D-glucosyl- steviol (5) and 13-O-[beta-nigerosyl-(1----2)-beta-D-glucosyl]-19-O-beta-D- glucosyl-steviol (6). A significantly high quality of taste was evaluated for 4.

Carbohydrate Sequence↗

Microtia as an autosomal dominant mutation in a transgenic mouse line: a possible animal model of branchial arch anomalies.

Microtia was found in a transgenic mouse 643 and all offspring with microtia had the transgene. No anomalies, other than occasional low set ear and abnormal biting, were identified in other tissues and organs. In the developmental analysis, on the 9th and 10th days of gestation, hypoplasia of the second branchial arch was observed, while various kinds of malformed hillocks were noted on the 12th day. All of these anomalous embryos were transgenic. Histologically, hemorrhage and subsequent phagocytosis were noted at the second branchial arch. Left sided anomalies were predominant and in bilaterally defective ones asymmetry existed. These findings closely resembled to those in experimental animals with a phenocopy of the first and second branchial arch syndrome in humans. Since all other transgenic mouse lines with the same transgene as 643 appeared normal, this dysmorphic phenotype may be caused by an insertional mutation of a host gene, although inappropriate expression of the transgene should be examined further as a possible cause. These results suggest that this transgenic mouse line 643 may be useful as an animal model of branchial arch anomalies in humans.

Animals↗

Variabilities in prenatal development of orofacial system.

Reliable information on embryonic and fetal development of the human oro-facial system is meager. Much of the data available at present is not entirely reliable, because it was derived from a small number of specimens. An embryological approach with human materials is important for establishing a normal standard of development including individual variabilities as well as clarifying the embryogenesis and etiology of defective development (Nishimura et al. 1977). It is important in human craniofacial embryology to know the variabilities, that is, individual differences in developmental phenomena of the oro-facial region during human prenatal life. In recent times the importance of morphologic investigations of human development has received less emphasis. Yet, without thorough knowledge of the basic facts of prenatal human development, erroneous assumptions can be made in more dynamic approaches and lead investigators astray. Knowledge of prenatal development of human orofacial structures and some of their deviations will therefore be welcomed by many basic scientists and clinicians in the field of facial clefts and other craniofacial malformations. The author was engaged in the collection and systematic study of human embryos and fetuses with Dr. Hideo Nishimura, Emeritus Professor of Kyoto University, Kyoto, Japan, and has been studying the normal and abnormal development during prenatal life. Several results obtained from the study of a large number of specimens are presented laying stress on the orofacial development.

Cleft Lip↗

Differentiation-dependent expression of I and sialyl I antigens in the developing lung of human embryos and in lung cancers.

The localization of two carbohydrate antigens, I and sialyl I antigens, in the lungs of developing human embryos was investigated using specific monoclonal antibodies and compared with the distribution patterns of the known embryonic antigen, stage-specific embryonic antigen-1 (Lex hapten). When the future bronchi were actively developing from the bronchial buds in the lungs of 50- to 53-day-old embryos, the immature bronchial bud cells were I-, Lex+, while the fully differentiated epithelial cells of the larger bronchus were I+, Lex-. When the bronchiolar bud cells matured into bronchiolar epithelial cells in the lung of a 12-week-old embryo, the immature bronchiolar bud cells were I-,Lex+, while the fully differentiated epithelial cells of the bronchioles were I+,Lex-. Sialylated forms of the antigens finally appeared in the lungs of 18-week-old embryos, when the terminal bud cells actively proliferated and underwent the differentiation process into epithelial cells of alveoli and alveolar ducts. The immature terminal bud cells at this stage were I-, sialyl I-, Lex+, sialyl Lex-i+, while the fully differentiated alveolar epithelial cells were I+, sialyl I+, Lex-, sialyl Lex-i-. After 8 months, the flattened mature alveolar epithelial cells were strongly positive for both I and sialyl I antigens, the strong expression of which continued after birth and even into the adult stage. These distribution patterns indicate that the I and sialyl I antigens are specific markers for the differentiated type cells in each stage of development, while Lex and related embryonic antigens were specific to the immature bud cells in every stage. The above-described differentiation-dependent expression patterns of these antigens seem to be reflected in the distribution of these antigens in human lung cancers, i.e., I and sialyl I antigens were expressed in lung cancer cells more weakly than in normal lung cells, while the Lex and sialyl Lex-i were expressed in cancer cells much more strongly than in normal lung cells. This was further reflected in the serum levels of these antigens in the patients with respiratory disorders. The distribution pattern of the serum levels of these antigens in patients with lung cancers showed sialyl Lex-i greater than sialyl I, indicating that these serum antigens originated from the lung cancer lesion where sialyl Lex-i is much more dominant than sialyl I antigen.(ABSTRACT TRUNCATED AT 400 WORDS)

Adenocarcinoma↗

Ca2(+)-ATPase cytochemistry in the spinal cord microvasculature of prenatal rats.

Membrane-bound Ca2(+)-ATPase activity was localized cytochemically in the blood vessels of the spinal cord of rat embryos to obtain a better understanding of the membrane activities of vascular cells. The cytochemical method revealed a growth of the parenchymal vasculature. In the parenchyma, reaction product was dense over the entire plasma membrane of voluminous endothelial cells provided with large nuclei and enriched cytoplasmic organelles, suggesting that the endothelial cells may be of a vascular sprout. The parenchymal vessels with a wide lumen were frequently associated with pericytes, and the Ca2(+)-ATPase activity was diminished in intensity on the luminal surface of the flattened endothelial cells. On the other hand, the endothelium of extraparenchymal capillaries exhibited Ca2(+)-ATPase activity primarily on the luminal surface of the plasma membrane. Quercetin, a Ca2(+)-transporting ATPase inhibitor, considerably decreased the abluminal activity in the voluminous endothelial cells with slit-like vascular lumen and the luminal activity of functioning capillary endothelium as well. Thus, a dual activity of Ca2(+)-ATPase, postulating for the activities of Ca2(+)-transporting ATPase and ecto-ATPase, was closely correlated with the maturation processes of the capillary endothelium.

Animals↗

Morphological study of normal and tumor cells in the head and neck region using irradiated collagen gel embedding technique: morphological study using collagen gel.

Fourteen specimens of normal, benign, or malignant tumor cells from the head and neck region were subjected for culture using 0.2% of irradiated collagen gel embedding technique. Re-differentiation of glands within the gel in serum-free medium was observed. There were marked differences in the growth patterns within the gel between normal or benign and malignant cells. Four normal glands and 5 out of 6 benign tumors showed branch-like growth patterns. On the other hand, malignant tumors showed no branch-like pattern, or could not grow at all. The results showed that the collagen gel technique could be useful for the differential diagnosis of malignancy in head and neck tumors.

Cell Differentiation↗

A New Minor Saponin from the Leaves of Panax ginseng.

Seventeen compounds were isolated from the leaves of PANAX GINSENG C. A. Meyer. Among them, a new minor saponin was established as 3beta,6alpha,12beta-trihydroxy-dammar-20(22), 24-diene-6- O-alpha- L-rhamno-pyranosyl-(1-->2)-beta- D-glucopyranoside ( 2). Fourteen compounds were identified as 20( R)-protopanaxadiol ( 1), 20( R)-protopanaxatriol, ginsenoside-Rh (3), 20( R)-ginsenoside-Rh (2), 20( S)-ginsenoside-Rh (2), ginsenoside-Rh (1), -Rg (3), -Rg (2), -Rg (1), -Re, -Rd, -Rc, -Rb (2), -Rb (1); the others are still under investigation.

Journal Article↗

A flower-inducing substance of high molecular weight from higher plants.

The flower-inducing activities of aqueous extracts of several plants were fractionated by gel filtration. Three major peaks, corresponding to molecular weights of about 120, 20 to 30, and 5 to 10 kilodaltons, were detected in extracts of Lemna, Pharbitis, and Brassica. The latter two peaks may be degradation products generated during the extraction procedure. In extracts of soybean seeds, only the peak of material of 120 kilodaltons was detected. This is the first published report of a high molecular mass substance with florigenic activity in Lemna plants. The florigenic substance had some properties associated with proteins (or polypeptides), but the activity was unaffected by treatment with proteinase K.

Journal Article↗