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Biomedical subjects

T Tachibana

Publications and source records attributed to T Tachibana.

At least 37 records · Page 2Linked to original sources
PubMed2001

Sporothrix schenckii thermo-intolerant mutants losing fatal visceral infectivity but retaining high cutaneous infectivity.

Four mutants of the Sporothrix schenckii lung isolate IFM 41598 were isolated by their inability to form colonies on a Sabouraud glucose agar plate incubated at 37 degrees C for 4 days. In contrast to the parent IFM 41598, these thermo-intolerant mutants were all defective in producing fatal visceral infections in mice, even though they retained infectivity in footpad tissues with a small fungal inoculum (approximately equals 10 cfu).

Animals↗
PubMed2001

Metabolic effects of 20 kDa and 22 kDa human growth hormones on adult male spontaneous dwarf rats.

BACKGROUND: Two molecular forms of human GH (hGH) have been shown to be biologically active. The 20 kDa form has been reported to have weaker diabetogenic and lipolytic actions than the 22 kDa form. OBJECTIVE: To analyze the carbohydrate metabolism of 20 kDa and 22 kDa hGH, using the adult male spontaneous dwarf rat (SDR), which is GH deficient. DESIGN: SDRs were given 20 kDa or 22 kDa hGH in doses of 125 microg/rat or 500 microg/rat, or saline, for 10 days, and their weight, serum IGF-I, glucose, insulin, leptin and body composition were measured. RESULTS: Weight and serum IGF-I increased both in the 20 kDa and 22 kDa groups, but IGF-I concentrations were significantly lower in the 20 kDa group than in the 22 kDa group. Serum glucose was not increased by either 20 kDa or 22 kDa hGH, whereas insulin was significantly increased after the higher dose of the 22 kDa hGH. Although blood concentrations of leptin were decreased by both 20 kDa and 22 kDa hGH, values were lower in the high-dose 20 kDa group than in the group given the same dose of 22 kDa hGH. Both forms of GH increased the percentage body water and body protein content, and decreased the percentage of body fat by the same degree. The observation that the higher dose of the 22 kDa hGH increased insulin concentrations without changing blood glucose demonstrates that this concentration of the hormone induces insulin resistance, whereas the same dose of 20 kDa hGH does not. CONCLUSIONS: The results can be interpreted to indicate that the higher dose of the 22 kDa hGH has diabetogenic activity, as reported previously, whereas the 20 kDa hGH has lower diabetogenic activity. The 20 kDa form of hGH may therefore be more useful in treating adult GH deficiency, especially those with severe obesity.

Animals↗
PubMed2001

New approach for the establishment of mouse early embryonic stem cells and induction of their differentiation.

Eleven early embryonic stem (EES) cell lines were established using a new novel method. Two cell stage embryos from the ddY mouse strain were cultured in alpha-MEM supplemented with 10% fetal calf serum (FCS) and embryotrophic factors (ETFs) and allowed to develop to the trilaminal germ disc embryonic stage. Only small round cells (EES cells) were isolated by the colony isolating technique and subsequently cultured in the same medium containing the ETFs and leukemia inhibitory factors (LIF-10 ng/ml). The newly established embryonic stem (ES) cells isolated from inner cell mass of blastocysts differentiated from two cell stage embryo in culture. The EES and ES cell lines were maintained in an undifferentiated state using Ham's F12 medium supplemented with 10% FCS and 1 ng/ml of LIF. The EES cells maintained their normal genetic and morphological features as well as their potential to differentiate into a broad spectrum of cell types as well as their ability to contribute to all cell lineages in chimeric mice. Moreover, these cell lines changed and differentiated into various kinds of cells by removing LIF and by the addition of ETFs to the vitro culture system. All 11 EES cell lines and 3 ES cell lines formed embryoid bodies; however, cell line EES-4 formed tube-like structures which extended, anastomosed with each other, and finally formed networks when the LIF were absent. Primitive germ organ-like structures composed of 3 germ layers were recognized in the cultures following the administration of ETFs. In conclusion, the new method devised by us is a novel, easy and reliable technique for establishing EES cell lines.

Animals↗
PubMed2000

Altered nociceptive neuronal circuits after neonatal peripheral inflammation.

Nociceptive neuronal circuits are formed during embryonic and postnatal times when painful stimuli are normally absent or limited. Today, medical procedures for neonates with health risks can involve tissue injury and pain for which the long-term effects are unknown. To investigate the impact of neonatal tissue injury and pain on development of nociceptive neuronal circuitry, we used an animal model of persistent hind paw peripheral inflammation. We found that, as adults, these animals exhibited spinal neuronal circuits with increased input and segmental changes in nociceptive primary afferent axons and altered responses to sensory stimulation.

Afferent Pathways↗
PubMed2000

The small GTP-binding protein TC10 promotes nerve elongation in neuronal cells, and its expression is induced during nerve regeneration in rats.

We have made a rat cDNA library using nerve-transected hypoglossal nuclei. Using this library, we performed expressed-sequence tag analysis coupled with in situ hybridization to identify genes whose expression is altered in response to nerve injury. In this gene screening, a member of Rho family GTPases, TC10, which had not yet been characterized in neuronal cells, was identified. TC10 mRNA expression was very low in normal motor neurons; however, axotomy induced its expression dramatically. Other family members such as RhoA, Rac1, and Cdc42 were moderately expressed in normal motor neurons and showed slight upregulation after axotomy. The expression level of TC10 mRNA was low in the embryonic brain and gradually increased with development. However, the expression of TC10 mRNA in the adult brain was lower and more restricted than that of RhoA, Rac1, and Cdc42. Cultured dorsal root ganglia exhibited dramatic neurite extension secondary to adenovirus-mediated expression of TC10. It can be concluded that although TC10 expression is lower in developing and mature motor neurons compared with other Rho family members, TC10 expression is induced by nerve injury to play a crucial role in nerve regeneration, particularly neurite elongation, in cooperation with other family members.

Adenoviridae↗
PubMed2000

Spontaneous oscillation of oxy- and deoxy- hemoglobin changes with a phase difference throughout the occipital cortex of newborn infants observed using non-invasive optical topography.

We investigated spontaneous changes in the cerebral oxygenation state of infants during quiet sleeping by using a form of multi-channel near-infrared spectroscopy: non-invasive optical topography. Eight infants born at 32-39 weeks were studied at postconceptional term age (38-43 weeks). Spatially synchronized oscillations of changes in the concentration of oxy- and deoxy- hemoglobin ([oxy-Hb] and [deoxy-Hb]) were observed throughout the occipital cortex. Time series analysis based on the theory of non-linear oscillators showed that the mean periods of the oscillation for each infant ranged from 11 to 18 s. The phase lag of [oxy-Hb] relative to [deoxy-Hb] was stable at about 3pi/4. This phase difference may result from interplay between the vasomotion and the oxygen consumption in relation to brain activity.

Female↗
PubMed2000

Characterization of the nuclear transport of a novel leucine-rich acidic nuclear protein-like protein.

We previously reported that the nuclear localization signal (NLS) peptides stimulate the in vitro phosphorylation of several proteins, including a 34 kDa protein. In this study, we show that this specific 34 kDa protein is a novel murine leucine-rich acidic nuclear protein (LANP)-like large protein (mLANP-L). mLANP-L was found to have a basic type NLS. The co-injection of Q69LRan-GTP or SV40 T-antigen NLS peptides prevented the nuclear import of mLANP-L. mLANP-L NLS bound preferentially to Rch1 and NPI-1, but not to the Qip1 subfamily of importin alpha. These findings suggest that mLANP-L is transported into the nucleus by Rch1 and/or NPI-1.

Amino Acid Sequence↗
PubMed2000

Cloning and expression of the chicken immunoglobulin joining (J)-chain cDNA.

The J chain is a component of polymeric immunoglobulin (Ig) molecules and may play an important role in their polymerization and the transport of polymeric Ig across epithelial cells. In this study, the primary structure of the chicken J chain was determined by sequencing cDNA clones. The cDNA had an open reading frame of 476 nucleotides encoding a putative protein of 158 amino acid residues including the signal sequence. The 3' untranslated region consisted of 1216 nucleotides and a poly(A) tail. The deduced amino acid sequence of the chicken J chain had a high degree of homology to that of human, cow, rabbit, mouse, frog, and earthworm, with eight conserved Cys residues identical to the mammalian J chains. Northern blot hybridization performed with total RNA from various chicken tissues revealed high levels of J-chain mRNA expression in spleen, intestine, Harderian gland, and bursa of Fabricius, and low levels in the thymus. The J chain was expressed in the bursa as early as day 15 of embryogenesis. These data indicated that the chicken J-chain gene displays a high degree of homology with that of other species, and is expressed at an early stage of development of the chicken immune system.

Amino Acid Sequence↗
PubMed2000

Postnatal differentiation of Merkel cells in the rat palatine mucosa, with special reference to the timing of peripheral nerve development and the potency of cell mitosis.

The origin and mechanism of the differentiation and proliferation of Merkel cells are enigmatic. A preliminary study in our laboratory showed that Merkel cells in the rat palatine mucosa emerge after birth. This is in contrast to the case of similar cells in the skin that differentiate during the embryonic period prior to the establishment of peripheral nerve innervation. We studied immunohistochemically the developmental timings of Merkel cells and peripheral nerves in the rat palatine mucosa using antibodies to cytokeratins 18 and 20, PGP 9.5, and CGRP using developing palates of prenatal and postnatal rats. We also studied the potency of mitosis in Merkel cells by immunohistochemistry using antibodies for a cell proliferation marker Ki67 and cyclin D-kinase inhibitors p16, p21 and p27. It was shown that Merkel cells in the rat palatine mucosa differentiate postnatally, after the development of peripheral nerve fiber terminals was almost established. The emergence and increase in number of Merkel cells progressed in an anterior-to-posterior wave. Newly appearing Merkel cells were usually negative for anti-cytokeratin 20 antibody but gained affinity for the antibody with progress of maturation. All Merkel cells in the palatine mucosa were negative for anti-Ki67 antibody but positive for anti-p27 antibody. These results indicate that Merkel cells in the rat palatine mucosa are not responsible for the development of peripheral nerve fiber terminals and that these cells differentiate in situ from intraepithelial stem cells.

Animals↗
PubMed2000

Suppression of neuropeptides' mRNA expression by herbal medicines in a rat model of peripheral inflammation.

The traditional Chinese medicines have been used clinically for a long time in some Asian countries, however, very few studies have been done to demonstrate the working mechanisms of these medicines using recently developed biochemical methodologies. In this study, we examined the anti-inflammatory effect of Huang-Lian-Jie-Du-Tang (HLJDT), a combination of herbs used in traditional Chinese medicine, on paw edema, thermal hyperalgesia and the mRNA increase of neuropeptides in spinal dorsal horn and hypothalamic neurons using a rat model of peripheral inflammation and hyperalgesia. The rats that received HLJDT from 3 days before the injection of complete Freund's adjuvant (CFA) into the plantar had significantly less edema and reduced thermal hyperalgesia compared to control rats that received CFA injection. The up-regulation of preprodynorphin mRNA in L4-5 dorsal horn neurons 8 hours after CFA injection that was observed in control rats, was also decreased in the HLJDT-treated rats. Moreover, there was a significant decrease in mRNA level of corticotropin-releasing factor in the paraventricular hypothalamic nucleus in the HLJDT-treated rats. These data demonstrate that HLJDT is anti-inflammatory, and produces changes in mRNA expression in dorsal horn and hypothalamic neurons. This is the first demonstrated that a traditional Chinese medicine can affect the excitability of neurons through an anti-inflammatory action.

Animals↗
PubMed2000

Extracellular norepinephrine in the medial hypothalamus increases during feeding in chicks: a microdialysis study.

Norepinephrinergic function in the medial hypothalamus is important for the regulation of feeding behavior in chicks as well as in rats. This study was conducted to clarify the variation of extracellular norepinephrine (NE) in the medial hypothalamus, including the paraventricular nucleus (PVN) and the ventromedial hypothalamic nucleus (VMN), during feeding behavior of layer-type chicks. To measure extracellular NE and 4-hydroxy-3-methoxyphenylglycol (MHPG), a major metabolite of NE, we used microdialysis and high-pressure liquid chromatography (HPLC) with electrochemical detection. After the collection of baseline samples, food-deprived animals were allowed access to the food for 3 h. Extracellular NE significantly increased during the first hour of access to food, and then returned to baseline levels. MHPG also increased during the feeding, but its increase continued throughout the remainder of the experiment. This study suggests that the variation of NE in the medial hypothalamus may be involved in the control of feeding in layer-type chicks.

Animals↗
PubMed2000

Comparison of the somatogenic action of 20 kDa- and 22 kDa-human growth hormones in spontaneous dwarf rats.

The somatogenic action of the 20 kilodalton human growth hormone (20 K) was studied using the spontaneous dwarf rat (SDR), which has an isolated GH deficiency. Saline or 2.5 microg, 10 microg, or 100 microg/rat/day of recombinant 20 K or 22 K was administered to prepubertal male and female SDRs for 10 days. Their body weights, serum IGF-I, glucose and insulin were measured, and their body composition was determined. Body weights and serum IGF-I increased dose-dependently in both the 20 K- and 22 K-treated groups. There was no significant difference in body weights and serum IGF-I between the 20 K- and 22 K-treated groups except at the dose of 100 microg/rat, in which the IGF-I concentrations were higher in the 22 K-treated male SDRs (P< 0.05: 20 K vs 22 K). Blood glucose was not significantly different between the Spague-Dawley (SD) normal rats and the SDR control groups; however, serum insulin levels of the SDR were higher than those of the SD control group (P< 0.05). Additionally, there was a tendency for serum insulin and glucose levels to increase following 22 K treatment, but the differences were not significant. The percentage of body fat decreased with hGH treatment in both groups (P< 0.01: GH 10, 100 microg/rat group vs SDR control group), however, no significant differences were observed in body composition between the 20 K and 22 K treatment groups. In summary, the 20 K-hGH showed almost the same somatogenic activity as the 22 K-hGH in prepubertal male and female SDRs.

Animals↗
PubMed2000

Rapid improvement of psoriasis vulgaris during drug-induced agranulocytosis.

The role of neutrophils in psoriasis has long been discussed. We report a patient with long-standing psoriasis vulgaris who showed rapid improvement during agranulocytosis caused by ticlopidine. The patient did not develop any new psoriatic lesions for several days, although neutrophils increased daily after the administration of ticlopidine was stopped. The day after the peripheral blood neutrophil count recovered, several psoriatic plaques reappeared. The correlation of psoriatic activity with peripheral blood neutrophil counts suggests that a certain number of neutrophils may be required to initiate and maintain psoriasis.

Aged↗
PubMed2000

Nucleocytoplasmic shuttling of the aryl hydrocarbon receptor.

The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that acts in concert with the AhR nuclear translocator (ARNT), and alters gene expression in response to environmental contaminants such as 2,3,7, 8-tetrachlorodibenzo-p-dioxin (TCDD). We have previously shown that AhR contains both a nuclear localization signal (NLS), AhR(13-39), and a nuclear export signal (NES), AhR(55-75), in its NH(2)-terminal region. In this study, we obtained direct evidence for the nucleocytoplasmic shuttling of AhR and show the biological significance of the shuttling in terms of the transcriptional activation of its target gene, CYP1A1. When AhR(13-75) fused with glutathione S-transferase (GST)-green fluorescent protein (GFP) was microinjected into the nucleus of a polykaryotic of BHK21 cell, the GST-AhR(13-75)-GFP migrated from one nucleus to the other. This event, nucleocytoplasmic shuttling, was completely inhibited in the presence of leptomycin B (LMB). The interaction between chromosome region maintenance 1 (CRM1) and endogenous AhR was shown by immunoprecipitation with antibodies to AhR followed by immunoblot analysis with antibodies to CRM1. The inhibition of the nuclear export of AhR by LMB repressed the transcriptional activation of the CYP1A1 gene. The findings suggest that nuclear-cytoplasmic shuttling of AhR is essential for the inducible expression of the CYP1A1 protein.

Animals↗
PubMed2000

Recycling of importin alpha from the nucleus is suppressed by loss of RCC1 function in living mammalian cells.

We previously reported that the nuclear import of substrates containing SV40 T antigen nuclear localization signal (NLS) was suppressed in a temperature-sensitive RCC1 mutant cell line, tsBN2, at nonpermissive temperature. Moreover, it was shown that import into wild type BHK21 cell-derived nuclei gradually decreased in heterokaryons between the tsBN2 and BHK21 cells, although the BHK21 nuclei retained wild type RCC1 and should contain RanGTP (Tachibana et al., 1994). In this study, it was found that in the heterokaryons cultured at non-permissive temperature, endogenous importin alpha was not detected immunocytochemically in the cytoplasm or BHK21 nuclei but only in the tsBN2 nuclei, suggesting that importin alpha cannot be exported from the RCC1-depleted nuclei. In fact, importin alpha microinjected into the nucleus of tsBN2 cells at non-permissive temperature remained in the nucleus. These results strongly support the hypothesis that the recycling of importin alpha from the nucleus requires nuclear RanGTP. Moreover, it was found that cytoplasmic injection of importin alpha restored the import of SV40 T-NLS substrates in the BHK21 nuclei but not the tsBN2 nuclei in the heterokaryons. This indicates that the decrease of importin alpha from the cytoplasm in the heterokaryons leads to a suppression of the efficiency of nuclear import of the T-NLS substrate and provides support for the view that nuclear RanGTP is essential for the nuclear entry of the substrates.

Animals↗
PubMed2000

Regulation of pituitary growth hormone-secretagogue and growth hormone-releasing hormone receptor RNA expression in young Dwarf rats.

Growth hormone-secretagogue receptor (GHSR) RNA is known to be expressed in the hypothalamus and pituitary. Since endogenous GH secretagogue (GHS) is still unknown, the physiological role of GHS and GHSR in growth is not well understood. In this study, we have determined the effects of growth hormone in GH-releasing hormone receptor (GHRHR) and GHSR RNA expression in spontaneous Dwarf rats (SDRs) which are deficient in GH secretion, with or without GH replacement. Twenty-five-day-old SDRs received daily s.c. injection of human GH (40 microg/kg BW x 2/day) or control solution for two weeks. On day 40, the rats were sacrificed by decapitation and the pituitaries were immediately removed and quickly frozen. Total RNA was extracted from the pituitary, and mRNA coding GHSR was detected and semi-quantitated by competitive RT-PCR. Pituitaries from control SDRs showed strong GHSR RNA expression and the expression level was 5 to 10 times higher in females than in males. When GH was replaced, GHSR RNA expression greatly decreased. Pituitary GHRHR RNA expression, determined by RNase Protection Assay, was similar in male and female control animals; and was also greatly reduced in rats treated with GH when compared to the control. These results suggest that the expression of both GHSR and GHRHR is regulated by growth hormone, presumably via changes in hypothalamic GHRH and/or endogenous GHS. The apparent sexual dimorphism in GHSR indicates different regulatory effects of sex steroid in young growing SDRs.

Animals↗
PubMed2000

Effects of scopolamine on working memory in rats in a delayed matching-to-position task, employing a subject-centered procedure.

The effect of scopolamine hydrobromide on a delayed matching-to-position task was examined while controlling for two confounding factors, i.e., mediating behavior and slow performance on a task. The task was given on the basis of a subject-centered method in which delay intervals are dependent on subjects' performance. The performance of individual subjects, rather than averaged group performance, was taken as the focus of the analysis. The results indicated that scopolamine had an effect not only on speed of performing the task but also on the length of the retention interval. The effects differed considerably among individuals: the effects on both the length of the retention interval and the speed of performance were found for two of the five subjects. An effect on speed of performance alone was found for one subject. No effects on either measure were found in a further two subjects.

Animals↗