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Biomedical subjects

H Mitani

Publications and source records attributed to H Mitani.

At least 37 records · Page 2Linked to original sources

Prevention of the photodamage in the hairless mouse dorsal skin by kojic acid as an iron chelator.

Kojic acid, a fungal metabolic product, has been used as a skin-depigmenting agent in skin care products marketed in Japan. Iron in the skin is known to be involved in wrinkling as a result of chronic photodamage. Kojic acid was expected to have anti-wrinkling activity, since it possesses iron-chelating activity. We now evaluated the anti-wrinkling activity of kojic acid by using hairless mice exposed to chronic solar-simulating ultraviolet (UV) irradiation as model animal. At the end of a 20-week irradiation period, topical application of kojic acid before UV irradiation was observed to dramatically prevent: (1) the wrinkling, (2) hyperplasia of the epidermis, (3) fibrosis of the lower dermis, and (4) the increase of extracellular matrix components in the upper dermis. These findings indicate that kojic acid is a typical agent preventing wrinkling of the skin due to chronic photodamage.

Animals↗

Efficient ex vivo generation of human dendritic cells from mobilized CD34+ peripheral blood progenitors.

We tried to efficiently generate human dendritic cells (DCs) from CD34+ peripheral blood hematopoietic progenitor cells mobilized by high-dose chemotherapy and subsequent administration of granulocyte colony-stimulating factor, using a liquid suspension culture system. Among various combinations, the combination of c-kit ligand, flt-3 ligand, c-mpl ligand (TPO), and interleukin (IL)-4 most potently generated the number of CD1a+CD14- DCs in cultures containing granulocyte-macrophage colony-stimulating factor (GM-CSF) and tumor necrosis factor alpha (TNF-alpha). The delayed addition of IL-4 on day 6 of culture gave rise to an additional increase in the yield of CD1a+CD14-DCs that were characterized by the expression of HLA-ABC, HLA-DR, CD80, CD86, and CD83. The majority of the sorted CD1a-CD14+ cells derived from 6-day culture of CD34+ cells gave rise to CD1a+CD14- DCs and CD1a-CD14+ macrophages on day 12 of culture in the presence and absence of IL-4, respectively. These findings suggest that IL-4 promotes the differentiation of CD1a- CD14+ cells derived from mobilized CD34+ peripheral blood hematopoietic progenitors to CD1a+ CD14- DCs. The majority of these DCs expressed CD68 but not the Langerhans-associated granule antigen, a finding that suggests they emerge through the monocyte differentiation pathway. The addition of TPO and IL-4 to cultures did not affect the potential of DCs to stimulate the primary allogeneic T-cell response. These findings demonstrated that the combination of c-kit ligand plus flt-3 ligand plus TPO with GM-CSF plus TNF-alpha, followed by IL-4, is useful for ex vivo generation of human DCs from mobilized CD34+ peripheral blood progenitors.

Antigens, CD1↗

Differences in recombination frequencies during female and male meioses of the sex chromosomes of the medaka, Oryzias latipes.

In the medaka, Oryzias latipes, sex is determined chromosomally. The sex chromosomes differ from those of mammals in that the X and Y chromosomes are highly homologous. Using backcross panels for linkage analysis, we mapped 21 sequence tagged site (STS) markers on the sex chromosomes (linkage group 1). The genetic map of the sex chromosome was established using male and female meioses. The genetic length of the sex chromosome was shorter in male than in female meioses. The region where male recombination is suppressed is the region close to the sex-determining gene y, while female recombination was suppressed in both the telomeric regions. The restriction in recombination does not occur uniformly on the sex chromosome, as the genetic map distances of the markers are not proportional in male and female recombination. Thus, this observation seems to support the hypothesis that the heterogeneous sex chromosomes were derived from suppression of recombination between autosomal chromosomes. In two of the markers, Yc-2 and Casp6, which were expressed sequence-tagged (EST) sites, polymorphisms of both X and Y chromosomes were detected. The alleles of the X and Y chromosomes were also detected in O. curvinotus, a species related to the medaka. These markers could be used for genotyping the sex chromosomes in the medaka and other species, and could be used in other studies on sex chromosomes.

Animals↗

Aggressive neoplastic plasma cell growth with MLL gene rearrangement after high-dose therapy with autologous stem cell support for multiple myeloma.

We report a case of a patient with IgA kappa multiple myeloma (MM) mobilized with etoposide and subsequently receiving high-dose melphalan (HDM) with stem cell support. She relapsed rapidly post transplantation. Southern blot and fluorescent in situ hybridization analysis showed MLL gene rearrangement in the myeloma cells, which was not detected in the sample at diagnosis or in the PBSC harvested with etoposide plus G-CSF. These observations suggest that clonal rearrangement of the MLL gene is caused by etoposide. Patients with MM undergoing HDM with stem cell rescue may be at an increased risk of not only secondary leukemia, but also secondary genetic abnormalities in myeloma cells, especially those receiving priming with etoposide for peripheral blood stem cell collection.

Antineoplastic Agents, Phytogenic↗

The influences of molar intrusion on the inferior alveolar neurovascular bundle and root using the skeletal anchorage system in dogs.

The skeletal anchorage system (SAS) was developed as intraoral rigid anchors for open-bite correction by intrusion of molars. Since the application of SAS is a new modality in orthodontic treatment, the influences of radical molar intrusion on the root and the inferior alveolar neurovascular bundle were unknown. The purpose of this research is to verify the effect of molar intrusion on the neurovascular bundle, the level of osseointegration of bone screws, and root resorption. The results of this study showed mandibular molars were intruded 3.4 mm on the average over 7 months in dogs. The miniplates were well stabilized with osseointegrated bone screws and the peri-implant soft tissues showed slight inflammatory changes. Neither nerves nor blood vessels were damaged. Root resorption was observed but was repaired with new cementum. We concluded that the SAS utilizing transmucosal titanium miniplates as an immovable orthodontic anchorage could provide a new modality for molar intrusions without serious iatrogenic problems.

Alveolar Process↗

Efficient ex vivo generation of dendritic cells from CD14+ blood monocytes in the presence of human serum albumin for use in clinical vaccine trials.

Dendritic cells (DC) with the potential to induce anti-tumour immunity represent one of the promising candidates for cancer vaccines. Efficiency of ex vivo DC generation depends on culture conditions, especially protein components in the plasma or serum used. Using human serum albumin (HSA), we devised a constant and reproducible culture method for DC generation from peripheral blood CD14+ cells. The number of DC obtained with 2% HSA-supplemented cultures containing granulocyte-macrophage colony-stimulating factor and interleukin 4 were consistently higher than in cultures with various concentrations of autologous plasma or serum. The concentrations and time points tested for plasma or serum considerably affected the number of DC recovered. DC prepared with HSA acquired the ability to uptake dextran, and expressed high levels of major histocompatibility (MHC) and co-stimulatory molecules similar to DC cultured with autologous plasma or serum. Although DC cultured with autologous plasma or serum consisted of CD1a+ and CD1a- populations, DC differentiated in the presence of HSA expressed CD1a. DC obtained with HSA primed and induced immunogenic peptide-specific cytotoxic T lymphocytes against a tumour rejection antigen, HER2. These findings suggest that our method for preparation of DC with HSA should prove valuable in DC generation for immunotherapy.

Cancer Vaccines↗

P300 abnormalities in patients with severe sleep apnea syndrome.

We investigated the mechanism of abnormal P300 findings in the patients with severe sleep apnea syndrome (SAS). Compared with controls, SAS patients showed significantly longer latency of P300. Not the mean sleep latency in multiple sleep latency test but the percentage of total periods in which percentage desaturation time fell below 90% to total sleep time correlated with P300 latency in the patients. During nasal CPAP treatment, P300 latency was significantly shortened in the patients under 45 years, while the elderly patients did not show statistical change. We speculated that P300 latencies might be prolonged due to nocturnal hypoxia with SAS and the abnormality might be irreversible, especially in elderly patients.

Adult↗

An accurate method of predicting mandibular growth potential based on bone maturity.

Mandibular growth prediction provides important information for planning treatment and for evaluating occlusal stability after treatment. At present, several methods can predict mandibular growth, but it is not clear which method is the most accurate. This study compared the predictive error of several methods by using skeletal maturity indicators. Twenty-two longitudinal cephalograms and hand-wrist radiographs of female subjects (average initial age, 8.3 years; final age, 18.4 years) were collected to construct the prediction formula. Another 22 female subjects (initial age, 10.8 years; final age, 18.6 years) were examined to compare differences between the predicted values and the actual values. Mandibular total length (condylion-gnathion) at the final stage can be accurately predicted by (1) the ossification events of the third middle phalanx and the radius, (2) the growth potential method, (3) the growth percentage method, (4) the multiple regression method, and (5) the growth chart method. Bone age as a parameter was calculated by both the Tanner-Whitehouse 2 (TW2) method and the computer-aided skeletal maturity assessment system (CASMAS) that we developed. The average error between the predicted length of condylion to gnathion and the actual length at the final stage for each method was (1) 3.0 mm; (2) 2.1 mm (TW2), 2.4 mm (CASMAS); (3) 2.3 mm (TW2), 2.6 mm (CASMAS); (4) 4.3 mm (TW2), 4.9 mm (CASMAS); and (5) 3.6 mm (TW2), 3.7 mm (CASMAS). The growth potential method and the growth percentage method were the most accurate predictors of mandibular growth potential.

Adolescent↗

Tooth movement and changes in periodontal tissue in response to orthodontic force in rats vary depending on the time of day the force is applied.

The purpose of this study was to investigate whether there are any differences in tooth movement or in the response of periodontal tissue to orthodontic force when the force is applied at different times of the day. One hundred 6-week-old male Wistar rats were divided into one control group without force application and three experimental groups based on the time of day the force was applied to the upper first molars. Animals in the whole-day group received force continuously throughout the experimental period, while animals in the light- and dark-period groups received force only during the light (07:00-19:00) or dark period (19:00-07:00), respectively. Tooth movement was measured using the occlusal view of a precise plaster model with a profile projector. Periodontal tissues were evaluated histologically. The time course of tooth movement varied among the groups. Tooth movement over 21 days in the whole-day and light-period groups was about twice that as in the dark-period group. The formation of new bone on the tension side in the whole-day and light-period groups was more than twice that as in the dark-period group. On the pressure side, more osteoclasts appeared on the alveolar bone in the whole-day and light-period groups than in the dark-period group. The light-period group showed less extensive hyalinization of the periodontal ligament (PDL) than the whole-day group. The area of root resorption on day 21 also varied among the groups. Interference by masticatory forces did not seem to be a principal cause of the decreased tooth movement in the dark-period group. These results indicate that there are considerable variations in tooth movement and in the response of periodontal tissue to orthodontic force when the force is applied at different times of the day in rats. The results suggest that diurnal rhythms in bone metabolism have important implications in orthodontic treatment.

Alveolar Process↗

Determination of pyrimidine dimers in Escherichia coli and Cryptosporidium parvum during UV light inactivation, photoreactivation, and dark repair.

UV inactivation, photoreactivation, and dark repair of Escherichia coli and Cryptosporidium parvum were investigated with the endonuclease sensitive site (ESS) assay, which can determine UV-induced pyrimidine dimers in the genomic DNA of microorganisms. In a 99.9% inactivation of E. coli, high correlation was observed between the dose of UV irradiation and the number of pyrimidine dimers induced in the DNA of E. coli. The colony-forming ability of E. coli also correlated highly with the number of pyrimidine dimers in the DNA, indicating that the ESS assay is comparable to the method conventionally used to measure colony-forming ability. When E. coli were exposed to fluorescent light after a 99.9% inactivation by UV irradiation, UV-induced pyrimidine dimers in the DNA were continuously repaired and the colony-forming ability recovered gradually. When kept in darkness after the UV inactivation, however, E. coli showed neither repair of pyrimidine dimers nor recovery of colony-forming ability. When C. parvum were exposed to fluorescent light after UV inactivation, UV-induced pyrimidine dimers in the DNA were continuously repaired, while no recovery of animal infectivity was observed. When kept in darkness after UV inactivation, C. parvum also showed no recovery of infectivity in spite of the repair of pyrimidine dimers. It was suggested, therefore, that the infectivity of C. parvum would not recover either by photoreactivation or by dark repair even after the repair of pyrimidine dimers in the genomic DNA.

Animals↗

Lyme disease Borrelia spp. in ticks and rodents from northwestern China.

In May 1999, field surveys of Lyme disease spirochetes were conducted around the Tianshan Mountains in Xinjiang Uygur Autonomous Region in northwestern People's Republic of China. Ixodes persulcatus ticks were obtained in a Tianchi Lake valley with primary forest, while the tick fauna was poor in the semidesert or at higher altitudes in this region. Species identities were confirmed by molecular analysis in which an internal transcribed spacer sequence was used. Of 55 adult ticks, 22 (40%) were positive for spirochetes as determined by Barbour-Stoenner-Kelly culture passages. In addition, some rodents, including Apodemus uralensis (5 of 14 animals) and Cricetulus longicaudatus (the only animal examined), and some immature stages of I. persulcatus (4 of 11 ticks) that had fed on A. uralensis were positive for spirochetes. Based on 5S-23S rRNA intergenic spacer restriction fragment length polymorphism analysis and reactivity with monoclonal antibodies, 35 cultures (including double isolation cultures) were identified as Borrelia garinii (20 isolates, including 9 Eurasian pattern B isolates and 11 Asian pattern C isolates), Borrelia afzelii (10 pattern D isolates), and mixed cultures (5 cultures, including isolates that produced B. garinii patterns B and C plus B. afzelii pattern D). These findings revealed that Lyme disease pathogens are distributed in the mountainous areas in northwestern China even though it is an arid region, and they also confirmed the specific relationship between I. persulcatus and genetic patterns of Borrelia spp. on the Asian continent.

Animals↗

Dual regulation of osteoclast differentiation by periodontal ligament cells through RANKL stimulation and OPG inhibition.

Periodontal ligament (PDL) cells play an important role in maintaining the homeostasis of periodontal tissues. However, it is not known how PDL cells contribute to osteoclastogenesis. In this study, we examined the consequences of cell-to-cell interactions between peripheral blood mononuclear cells (PBMCs) and PDL cells during osteoclastogenesis. PBMCs were co-cultured directly or indirectly with PDL cells for two to four weeks. PBMCs that were directly co-cultured with PDL cells formed significantly more resorption pits on dentin slices than did PBMCs that were cultured alone. However, soluble factor(s) produced from PDL cells inhibited the formation of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells. Furthermore, PDL cells expressed both receptor activator nuclear factor kappa B ligand (RANKL) and osteoprotegerin (OPG) mRNA. In conclusion, PDL cells support osteoclastogenesis through cell-to-cell contact. PDL cells might regulate osteoclastogenesis by opposing mechanisms--stimulation of resorptive activity by RANKL and inhibition by OPG--thus affecting processes such as periodontitis and orthodontic tooth movement.

Acid Phosphatase↗

Identification and characterization of two distinct GnRH receptor subtypes in a teleost, the medaka Oryzias latipes.

We report the identification and characterization of two distinct GnRH receptor (GnRH-R) subtypes, designated GnRH-R1 and GnRH-R2, in a model teleost, the medaka Oryzias latipes. These seven-transmembrane receptors of the medaka contain a cytoplasmic C-terminal tail, which has been found in all other nonmammalian GnRH-Rs cloned to date. The GnRH-R1 gene is composed of three exons separated by two introns, whereas the GnRH-R2 gene has an additional intron and therefore consists of four exons and three introns. The GnRH-R1 and GnRH-R2 genes, both of which exist as single-copy genes in the medaka genome, were mapped to linkage groups 3 and 16, respectively. Inositol phosphate assays using COS-7 cells transfected with GnRH-R1 and GnRH-R2 demonstrated that they had remarkably different ligand sensitivities, although both receptors showed highest preference for chicken-II-type GnRH. Phylogenetic analysis showed the presence of three paralogous lineages for vertebrate GnRH-Rs and indicated that neither GnRH-R1 nor GnRH-R2 is the medaka ortholog to mammalian GnRH-Rs that lack a cytoplasmic tail. This, together with an observation that medaka-type GnRH had low affinity for GnRH-R1 and GnRH-R2, suggests that a third GnRH-R may exist in the medaka.

Amino Acid Sequence↗

Compressive force promotes chondrogenic differentiation and hypertrophy in midpalatal suture cartilage in growing rats.

Midpalatal suture cartilage (MSC) is secondary cartilage located between the bilateral maxillary bones and has been utilized in the analysis of the biomechanical characteristics of secondary cartilage. The present study was designed to investigate the effects of compressive force on the differentiation of cartilage in midpalatal suture cartilage in rats. Forces of various magnitudes were applied to the midpalatal suture cartilage in 4-week-old male Wistar rats for 1, 2, 4, 7, or 14 days, mediated through the bilateral 1st molars using orthodontic wires. The differentiation pathways in the MSC cells were examined by immunohistochemistry for the differentiation markers type I, type II and type X collagen, and glycosaminoglycans (GAGs), chondroitin-4-sulfate, chondroitin-6-sulfate and keratan sulfate. Histologically and immunohistochemically, the midpalatal suture cartilage in control rats had the characteristic appearance of secondary cartilage. In the experimental groups, the center of the midpalatal suture cartilage that contained osteo-chondro progenitor cells seemed to become mature cartilage and its immuno-reaction to type II and X collagen and GAGs increased as the experiment progressed. This differentiation was dependent upon the magnitude and duration of the force applied to the midpalatal suture cartilage; i.e., cartilaginous differentiation progressed more rapidly as the applied force increased. The present results suggest that the differentiation of osteo-chondro progenitor cells into mature and hypertrophic chondrocytes in the precartilaginous cell layer is promoted by compressive force.

Animals↗

Age-specific effects of noradrenergic alpha-2 agonist clonidine on the development of amygdaloid kindling in developing rats.

The effects of clonidine on the development of amygdaloid kindling were studied in rats of various ages (14, 21, 28 and 70 postnatal days). Administration of clonidine (0.2, 0.5 mg/kg i.p.) caused a significant retardation of kindling development in the 28-day-old rats as well as in the adult rats, whereas, in the 14-day-old rats, the development of kindling was significantly facilitated by clonidine. No significant effect of clonidine was observed in the 21-day-old rats. These results indicate that in rats the effects of clonidine on the development of amygdaloid kindling vary during development.

Adrenergic alpha-Agonists↗

Increases of vascular endothelin-converting enzyme activity and endothelin-1 level on atherosclerotic lesions in hyperlipidemic rabbits.

The aim of this study was to investigate vascular endothelin-converting enzyme activity and the tissue level of endothelin-1 in the aorta related to atherosclerotic lesions in high cholesterol diet-fed rabbits. Rabbits were fed two atherogenic diets, 0.5% and 1.5% cholesterol, and a normal diet for 16 weeks. Vascular endothelin-converting enzyme activity in the aortic arch and thoracic aorta was significantly increased (2.0-4.4 times) by the atherogenic diet as compared with the normal diet group as well as the levels of lipids and lipid peroxide in plasma were significantly increased. Tissue endothelin-1 levels in both aortas were also elevated (2.3-6.8 times), corresponding well to the increased tissue enzyme activity. In contrast, plasma endothelin-1 levels increased only in the 1.5% cholesterol diet group (2.7 times). These results indicate that the endothelin-converting enzyme activity and the corresponding endothelin-1 level in the vascular walls increase in association with the development of atherosclerotic lesions.

Animals↗