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Biomedical subjects

Y Yamauchi

Publications and source records attributed to Y Yamauchi.

At least 127 records · Page 7Linked to original sources

[Massive swelling of the tongue in two patients following the repair of cleft palate].

Massive swelling of the tongue occurred following the repair of cleft palate in a one-year-old girl and a one-year-old boy. The duration of operation was 285 min and 225 min respectively. Massive swelling of the tongue became prominent within 30 min after the operation in both infants. The trachea was intubated immediately. Swelling of the tongue recovered slowly, and the trachea was extubated 6 and 7 days after the operation, respectively. The compression of the tongue by the retractor for a long time during the operation probably caused the reperfusion edema of the tongue. Early recognition of the swelling and prompt maintenance of the airway are essential to prevent the serious outcomes resulting from the swelling of the tongue.

Airway Obstruction↗

In vivo implantation of human osteosarcoma cells in nude mice induces bones with human-derived osteoblasts and mouse-derived osteocytes.

Two human osteosarcoma cell lines, Hu09 and OST, were suspended in Matrigel (Becton Dickinson Labware, Bedford, Massachusetts) and implanted subcutaneously in the backs of nude mice. To study phenotypic changes of tumor cells and host cells, expression of mRNA for osteopontin (OPN), osteocalcin (OC), and osteonectin (ON) was analyzed by in situ hybridization. Bone tissue was formed in the tumors derived from Hu09 cells. OPN mRNA was transcribed predominantly in osteocyte-like cells within the bone, whereas OC mRNA was transcribed in osteoblast-like cells that surrounded the bone. ON mRNA was detected in both types of cells. The similarity of the expression pattern of OPN, OC, and ON during osteogenesis of Hu09 cells to that of normal skeletal development suggests that the bone formed in Hu09-implanted mice is the same as normal bone tissue. By DNA-DNA in situ hybridization using a human-specific Alu probe and a mouse-specific m-L1 probe, osteoblast-like cells in Hu09 tumorous bone were, however, of human origin, whereas osteocyte-like cells were of mouse origin. In the tumors derived from OST cells, no osteogenesis was observed during the experimental period, and the expression of OPN, OC, and ON was not detected in tumor cells. An endochondral bone formation was not evident when these cells were simply implanted into muscle tissue. An endochondral bone was, however, reactively induced in the host mUscle tissue either when 1 alpha-hydroxyvitamin D3 and all-transretinoic acid were administered to OST-implanted mice or when Hu09 cells were pretreated with dexamethasone before implantation. Hu09 implantation seems to be a useful tool not only for the study of the differentiation of osteosarcoma cells but also for the investigation of the mechanism of bone formation. This system, using Hu09 and OST, may provide us with a new tool for the isolation of the unidentified factors that induce or inhibit osteogenesis in vivo.

Animals↗

Prostaglandin E2 at priming of naive CD4+ T cells inhibits acquisition of ability to produce IFN-gamma and IL-2, but not IL-4 and IL-5.

We investigated the effect of prostaglandin E2 on the acquisition of cytokine-producing ability by naive CD4+ T cells in human cord blood. Naive CD4+ T cells were stimulated for 3 days with mouse monoclonal anti-CD3 Ab, then washed and expanded in IL-2-containing medium for 3 more days. These activated T cells produced IL-2, IL-4, IL-5, and IFN-gamma upon stimulation with PMA and ionomycin. PGE2 added at priming of naive T cells inhibited the production of IL-2 and IFN-gamma, but not of IL-4 and IL-5, in a dose-dependent manner. This change in the cytokine production profile induced by PGE2 was maintained in T cells restimulated with anti-CD3 in the absence of PGE2, expanded by IL-2, and stimulated with PMA and ionomycin. The mRNA expression of IFN-gamma and IL-2, but not that of IL-4, was also decreased in these cells. Forskolin and dibutyryl cAMP had a similar effect. PGE2 must exist at an early stage of T cell activation to inhibit priming for IL-2 and IFN-gamma production. PGE2 also showed this effect, even in the presence of exogenous IFN-gamma, at the primary stimulation. These results indicate that PGE2 inhibits the acquisition of the ability to produce IL-2 and IFN-gamma by acting directly on naive T cells. Our results suggest that PGE2 plays a role in facilitating the development of the Th2-type cytokine production profile.

Animals↗

Suppression of age-related changes in mouse hippocampal CA3 nerve cells by a free radical scavenger.

This study was designed to evaluate the relationship between oxygen free radicals and age-related morphological changes in hippocampal nerve cells using K-7259 (N,N' bis[4-(3,4,5-trimethoxyphenyl)butyl] homopiperazine dihydrochloride), a known neuro-protective agent. A chemiluminescence assay has shown that this agent is a potent free radical scavenger with an IC50 of 1.6 x 10(-5)M. Mice fed diets containing 10, 20, 40 mg/kg/day of K-7259, for periods ranging from 25 to 40 or 50 weeks of age were used as test groups, and 10-, 20-, 30-, 40-, and 50-week-old mice fed a standard diet were used as controls. We measured the number and area of pyramidal nerve cells within a defined frame in the hippocampal CA3 field using an image analyzer and the density of nerve cells by the disector method. These values decreased gradually in controls as expected, and the number and area yielded a significant difference between control mice at the ages of 10 and 30 weeks. As compared with the corresponding controls, all test groups had greater cell numbers (statistical significance at 40 weeks in the 40 mg/kg/day group) and density, while cell areas were greater in all but a 10 mg/kg/day group (statistical significance at 50 weeks). In summary, the free radical scavenger K-7259 forestalled an age-related decrease in the number and size of hippocampal CA3 nerve cells, thus suggesting that free radicals play an important role in the cellular morphological changes which appear with age.

Aging↗

Neural activity of chorda tympani mechanosensitive fibers during licking behavior in rats.

The chorda tympani nerve, supplying the anterior two-thirds of the tongue, contains gustatory and mechanosensitive afferent fibers. We have analyzed discharge patterns in rats of various fibers recorded from dissected nerve filaments during licking behavior of which 4 were taste-sensitive and 12 mechanosensitive. The incidence of these two types were estimated electrophysiologically under anesthesia and their conduction velocity measured. Recordings in freely moving animals showed that the mechanosensitive fibers innervating the dorsal part of the tongue gave two burst discharges per lick, suggesting that contact of the tongue with the upper incisors and/or lip occurred during tongue protrusion and retraction. The fibers from the tip of the tongue showed one burst discharge per lick, which was the response to contact with a drinking spout. No rhythmical discharges synchronized with lick signals were observed in the fibers from the lateral part of the tongue or the taste-sensitive fibers. Such mechanoreceptor discharges were difficult to detect in recordings from the whole chorda tympani nerve. This masking of responses was due mainly to activation of a small number of mechanosensitive fibers by licking-induced mechanical stimulation. The lubricating action of saliva also decreased mechanoreceptor sensitivity. Despite their small number, the mechanosensitive fibers had axons with faster conduction velocities (larger diameter) than the taste-sensitive fibers. This was probably the reason why dissected nerve bundles more frequently showed mechanical than taste responses in conscious rats.

Action Potentials↗

Antinuclear antibodies in healthy aging people: a prospective study.

In order to evaluate the expression of antinuclear antibodies (ANA) in normal elderly individuals over time and clinical significance, a cross-sectional ANA testing in healthy Japanese was performed, followed by annual evaluations of ANA positive aged (> or = 65 years) and a control group. ANA was more prevalent in the aged (11.4% vs. 3.8%) and most were persistent after 4 years. Anti-ssDNA and anti-histone antibodies were increased in aged ANA positive as compared to ANA negative controls. Except for a history of spontaneous abortion, there was no differences in clinical findings. HLA DRB1*0901 and the DQB1*0602 + 0302 + 0303 set of alleles were increased in ANA positive. Therefore, ANA in the aged were persistent, apparently directed toward chromatin elements, and shared MHC associations with autoimmune diseases. Longer follow-up may be necessary to improve the evaluation of clinical significance of ANA in the aged.

Adult↗

Physical interaction with monocytes rescues human mature CD4+ T-cell lines from anti-CD3-induced apoptosis.

Crosslinking of the TcR-CD3 complex with immobilized anti-CD3 antibodies without sufficient co-stimulation induced cell death in human mature CD4+ T-cell lines. In these T cells, DNA fragmentation and morphological characteristics of apoptosis were seen. The anti-CD3-induced apoptosis was inhibited by co-culture with monocytes. The rescue signal provided by monocytes does not need to be present simultaneously with signals mediated by anti-CD3. When T cells were precultured with monocytes for 24 h before anti-CD3 stimulation and then the monocytes were removed from the culture, anti-CD3-induced T-cell apoptosis was also inhibited. To determine whether the monocyte-derived rescue signals were transduced by soluble factors or by direct cell-to-cell interaction with monocytes, we precultured T cells with monocytes separated by a micropore membrane which prevented T cell-monocyte physical interaction but not the diffusion of secreted molecules. In this system, rescue signals could not reach the T cells. To further assess the importance of physical interaction, we precultured T cells with fixed monocytes. T cells could not be rescued from apoptosis under these experimental conditions, either. The results considered collectively suggest that sufficient physical interaction with viable monocytes is important for the rescue of anti-CD3-induced apoptosis of CD4+ T cells.

Antibodies, Monoclonal↗

Transient expression assay for antisense RNAs using episomal replication of plasmids: effective reduction of retinoblastoma gene (Rb-1) product by its antisense RNA complementary to 3'-untranslated region.

We have developed a transient expression assay for selection of effective antisense RNAs using episomal replication of plasmids in COS-7 cells, an African green monkey kidney-derived cell line expressing SV40 large T antigen. The transient expression assay was enabled by a liposome-mediated DNA transfection method, by which about 70% of the cells were reproducibly transfected with exogenous DNAs. Plasmids expressing antisense RNAs for the retinoblastoma gene (Rb-1) mRNA and harboring SV40 ori were constructed and introduced into COS-7 cells to examine their inhibitory effect on the accumulation of endogenous Rb protein (pRb). Only the antisense RNA complementary to the 3'-untranslated region (UTR) in Rb-1 mRNA was expressed stably at high levels for 3 days after the transfection. This antisense RNA reduced by 73% the content of endogenous pRb 70 h after transfection. A similar inhibition was detected in mouse mammary carcinoma cells (FM3A) that were stably transfected with the antisense RNA expressing vector directed to 3'UTR. In contrast, no obvious change in pRb was observed with antisense RNAs complementary to the coding region of Rb-1 mRNA. The cellular content of these antisense RNAs was lowered by degradation; thus these RNAs did not affect the levels of pRb in COS-7 and FM3A cells. These results, taken together, suggest that the expression levels and the stability of antisense RNAs are involved in their repressive activity, and our transient expression assay provides a rapid and easy system for evaluation of ectopic antisense RNA activity in COS-7 cells.

Animals↗

Selective induction of interleukin-4- and interferon-y-producing T cells from cord blood naive T cells. Effects of costimulatory signaling through CD28.

We investigated the effect of costimulation through CD28 and CD11a on the differentiation of human naive CD4+ T cells with restricted cytokine production profiles. Interleukin (IL)-4 and interferon-gamma (IFN-gamma) were measured by ELISA and IL-2 was detected by a bioassay. Naive CD4+ T cells proliferated and produced IL-2 upon cross-linking of CD3, and costimulation through CD28 enhanced IL-2 production. After repeated stimulation, CD4+ T cells which were stimulated in the absence of costimulation through CD28 lost their ability to secrete IL-2 and started secreting IL-4 and IFN-gamma. Instead in the presence of costimulation through CD28, they secreted IL-2, IL-4 and IFN-gamma. Blocking of endogenous IL-4 activity with anti-IL-4 Ab suppressed the IL-4 secretion and proliferation of T cells.

CD28 Antigens↗

Temperature-sensitive mutation of DNA polymerase alpha induces growth-suppressive phenotypes involving retinoblastoma protein and cyclin D1.

Temperature-sensitive (ts) cell cycle mutant mouse cell, tsFT20, is deficient in DNA polymerase alpha activity to initiate DNA replication at replicon origins. Here, we analyzed phenotypes concerning growth control genes in the arrested tsFT20 cells. Analysis of cyclins showed that expression levels of cyclin D1, which is essential for G1/S transition, remarkably decreased in the mutant cells after temperature up-shift. Further we examined phosphorylation states of retinoblastoma protein (pRB) in the cells. Though the tsFT20 cells arrested in G1/S-S phase at nonpermissive temperature (Eki et al., (1990) J. Biol. Chem. 265 26-33), a large proportion of pRB was found as an underphosphorylated growth-suppressive form in the arrested cells. In revertant cell lines of tsFT20, pRB was not underphosphorylated even at nonpermissive temperature. The pRB underphosphorylation occurred later than the decrease of mRNA levels of cyclin D1, thus the underphosphorylation may be caused by the decrease in amount of cyclin D1 protein. These results indicated that the mutational inactivation of DNA polymerase alpha evokes phenotypes in which the inhibitory machinery of G1/S transition has been turned on.

Animals↗

CJ-12,371 and CJ-12,372, two novel DNA gyrase inhibitors. Fermentation,isolation, structural elucidation and biological activities.

A fermentation broth of an unidentified fungus (N983-46) was found to produce DNA gyrase inhibitors, CJ-12,371 (1) and CJ-12,372 (2). Following isolation by solvent extraction and silica gel and ODS (reverse phase) chromatographies, the structures were determined to be novel spiro-ketal compounds with S-configuration at position C-1. CJ-12,371 and CJ-12,372 inhibit both DNA supercoiling and relaxation mediated by Escherichia coli DNA gyrase. The interaction of these compounds with DNA gyrase appears to be novel in that the compounds inhibit supercoiling and relaxation without blocking religation; thus, no cleavage intermediate of double strand DNA is observed. Both compounds have antibacterial activity against several species of pathogenic Gram-positive bacteria, with MICs between 25 and 100 micrograms/ml. These results suggest that the antibacterial potency of CJ-12,371 and CJ-12,372 is attributed to the inhibition of DNA gyrase. However, the compounds did not inhibit DNA gyrase selectively, as they also inhibited eukaryotic topoisomerase II-mediated relaxation. Semi-synthetic modifications to the dihydroxy motif in CJ-12,371 altered both gyrase- and topoisomerase II-inhibitory activities, but did not enhance selectivity.

Anti-Bacterial Agents↗

Generation of novel rapamycin structures by microbial manipulations.

A new rapamycin producing culture was isolated and designated as Actinoplanes sp. N902-109. The addition of a cytochrome P-450 inhibitor and precursor feeding using this culture, and biotransformation approaches generated new rapamycin analogs with modifications at C-4, C-9, C-16, C-27, C-28 and/or C-39. The immunosuppressive activity of the resulting analogs was established in the mixed lymphocyte reaction assay.

Actinomycetales↗

[Whole mouth gustatory test (Part 2)--Effect of aging, gender and smoking on the taste threshold].

The whole mouth test method was employed for examining changes in gustatory senses due to aging. Participants were 314 male and 356 female healthy volunteers not complaining of taste disorders. Simultaneously, the effects of smoking were examined, comparing 71 male smokers to 142 male non-smokers, ranging from 20 to more than 60 years of age. A multiple comparison procedure (Tukey-Kramer) was applied to age groups, revealing a significant increase in the threshold with age; high threshold values were observed in the "70s and older" group in comparison to the younger group for all tastes other than the sweet taste. Concerning the differences between males and females, the gustatory thresholds of males were found to be higher than those of females for some in the "20s and older" group and for all tastes other than the sweet taste, whereas the thresholds of males were lower than those of females for all taste qualities in the late teen group. As to male subjects in whom the effects of smoking could be studied, smokers in their 20s demonstrated a slight rise in the threshold only for the bitter taste.

Adolescent↗

Valid method to evaluate the slope of Fourier transformed spectrum for the analysis of biological rhythm fluctuation.

Influences of lower frequency components of power spectrum calculated from biological rhythm data on the spectral slope were examined and the validity of conventional method to calculate the spectral slope was tested. Heart-beat-period data obtained from the electrocardiogram of rats were cut into two successive data sets, each of them was converted into power spectrum density by the fast Fourier transform, and the power spectra were compared with each other. The results showed that two spectra were overlapped only in the frequency range higher than a frequency about 15 times that of the dominant one, which means that the lower frequency components are not always the same and thus, the regression line by which the spectral slope is determined is considerably affected by the components. To evaluate the spectral slope, regression lines of spectra were obtained in three methods and their slopes were compared with each other. The methods were: "conventional" method, in which the weight of a frequency component was independent of the frequency (i.e., equal-weight = 1); "weight" method, in which the weight was the wave number included in the rhythm data; and "limited-data" method, where the weight was 1 and lower-frequency components were excluded. The results showed that there was not a large difference between the slopes in cases where the spectrum looked rather linear. However, in cases of considerably curved spectra the difference was large and the "weight" method may be the most reasonable to use, and in cases of short data length the "limited-data" method should be avoided.

Animals↗

[Whole mouth gustatory test (Part 1)--basic considerations and principal component analysis].

The whole mouth method gustatory test is a simple gustatory test. It can be applied easily, and can be used to assess overall taste which a subject is supposed to be feeling. In the present study, a new series of taste solutions for the use in the whole mouth gustatory test was prepared. In order to determine the normal range of gustation and relationships between each taste solution, gustatory threshold tests were conducted on 123 healthy volunteers (17-22 years old). The series of taste solutions represented 4 tastes, i.e., sweet, salty, sour, and bitter, and was prepared using sucrose, salt, tartaric acid and quinine, respectively. Recognition thresholds measured for each taste solution yielded normal ranges i.e., 0.0165 mol/l for the sweet solution, 0.0316 mol/l for the salty solution, 0.000734 mol/l for the sour solution, and 0.0000203 mol/l for the bitter solution which were almost the same as those published. The average thresholds obtained for all solutions were at almost always at the middle concentration level, i.e., level 6. Thus, this method can serve as a standard method of performing the whole mouth taste test. Principal component analysis of the thresholds obtained revealed that the primary component mainly demonstrated taste detection function, and that it could more or less be represented by the simple sum of each detection threshold. The analysis also showed that approximately 90% of taste threshold variation is explained by the 4 principal components. These findings provide further evidence that these four tastes comprise the basic elements of taste.

Adolescent↗

A new method for continuous measurement of regional cerebral blood flow using laser Doppler flowmetry in a conscious rat.

We have developed a new system to continuously measure regional cerebral blood flow (rCBF) in the cortex of a conscious animal. For this purpose, we used rats and laser Doppler flowmetry. Under pentobarbital anesthesia, the animal's skull was opened making a small square hole 3 mm x 3 mm in size. A transparent acrylic plate was placed over the hole in the skull. A polyethylene cannula (inner diameter 1.0 mm, length 5.0 mm) was fixed on the plate as a guide for the laser Doppler flowmeter (LDF) probe (outer diameter 1.0 mm, length 5.5 mm). Both the plate and guide cannula were fixed to the skull by dental cement. Every day for the following two weeks after surgery, the conscious animal was placed in a hammock for recording rCBF. A LDF probe was freely attachable to the plate above the cortex via the guide cannula during measurement of rCBF. The rats were kept in a hammock with their legs firmly touching the floor during measurement of rCBF. It was possible to measure rCBF every day for about two weeks, and rCBF responded consistently to inhalation of 7% CO2 when the responses were expressed as percentages of the prestimulus control rCBF values. This system is recommended for the continuous measurement of rCBF in a conscious animal.

Animals↗