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Biomedical subjects

H Mitani

Publications and source records attributed to H Mitani.

At least 109 records · Page 6Linked to original sources

Effects of expansive force on the differentiation of midpalatal suture cartilage in rats.

In an attempt to clarify the effects of biomechanical tensional force on chondrogenic and osteogenic differentiation of secondary cartilage, the midpalatal sutures of 4-week-old Wistar male rats were expanded by orthodontic wires which applied 20 g force for 4, 7, 10, and 14 days. The differentiation pathways in the midpalatal suture cartilage were examined by immunohistochemistry for osteocalcin, type I and type II collagen, and von Kossa histochemistry. Although the midpalatal sutures of the control animals consisted mainly of two separate secondary cartilages with mesenchyme-like cells at their midlines, type I collagen-rich fibrous tissue began to appear at day 4 and increased at the midline of the cartilage with days of experiment. At the end of the experiment, type I collagen-rich and calcified bone matrix appeared at the boundary between the precartilaginous and the cartilaginous cell layers. Most of the cartilaginous tissues were separated from each other and the midpalatal suture was replaced by osteocalcin-positive intramembranous bone and fibrous sutural tissue. These results strongly suggest that tensional force changed the phenotypic expression of collagenous components in secondary cartilage, which may reflect the differentiation pathway of osteochondro progenitor cells.

Animals↗

A pressure-distribution sensor (PDS) for evaluation of lip functions.

The purpose of this study was to develop and to test a pressure-distribution sensor (PDS) for evaluating lip functions. The PDS is fabricated as a disposable cartridge and is based on the principle of optical-pressure conversion used in tactile sensors of robot arms. Its advantages are in measuring sealing forces, contact area, and pressure-distribution patterns of the lips at maximum effort. We used the PDS to evaluate pressure-distribution patterns of long-face subjects with extremely large interlabial distances. The results suggest that the PDS is a useful device for evaluating lip sealing functions and their changes after orthodontic therapy or orthognathic surgery.

Disposable Equipment↗

Inhibitory effects of fluvastatin, a new HMG-CoA reductase inhibitor, on the increase in vascular ACE activity in cholesterol-fed rabbits.

1. The effects of fluvastatin, a new 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor, on the vascular angiotensin converting enzyme (ACE) activity in hyperlipidaemic rabbits were compared with those of enalapril, an ACE inhibitor. 2. Rabbits were fed a 1.5% cholesterol containing diet or normal diet for 16 weeks and treated with either fluvastatin or enalapril in the diet at the respective doses of 2 and 10 mg kg-1 day-1. The total cholesterol, triglyceride and phospholipid levels in serum were significantly increased in rabbits fed the high cholesterol diet. Treatment with fluvastatin but not enalapril resulted in a decrease in serum lipids. 3. The vascular ACE activities assessed via the cleavage rate from synthetic substrate in the aortic arches and upper thoracic aortae were increased by 8 to 10 times when the rabbits were made hyperlipidaemic. Fluvastatin as well as enalapril significantly lowered the tissue ACE in the aortae. 4. The ACE activities in serum did not alter in hyperlipidaemic rabbits either in the presence or absence of fluvastatin. The serum ACE activity was lowered by enalapril. 5. The lipid peroxide in serum as well as the plaque area in the thoracic aorta was significantly increased in the cholesterol diet-fed rabbits. Treatment with fluvastatin or enalapril reduced both serum lipid peroxide and plaque formation. The relaxant responses to acetylcoholine (ACh) were significantly suppressed in the cholesterol-fed rabbits. Treatment with fluvastatin or enalapril significantly reversed the suppression of ACh-induced relaxation. 6. It seems that the reduction of vascular ACE is not coupled to lipids and ACE activity in serum, but rather to lipid peroxidation. Thus, the decrease in vascular ACE activity by fluvastatin as well as the lipid-lowering effect may reduce the risk of atherosclerosis progression in the vasculature.

Animals↗

Localization of glycosaminoglycans in periodontal ligament during physiological and experimental tooth movement.

Localization of chondroitin sulphates in periodontal ligaments (PDL) of rat molar roots during physiological and experimental tooth movement were analysed immunohistochemically with the use of monoclonal antibodies, 3B3 and 2B6, specific to chondroitin 6-sulphate (CH-6S) and chondroitin 4-sulphate/dermatan sulfate (CH-4S/DS), respectively. The maxillary first molars of experimental animals were forced to move laterally with a 10 g weight by U-shaped wires for 3 and 7 d. In control animals, 3B3 epitope was seen in the PDL near to the bone surface facing the distal half of roots, which corresponded to the compressive side during physiological tooth movement. Immunoreactivity for 2B6 was weak or negative in the PDL. Both epitopes were present at osteoid, precementum, lacunae and canaliculli of osteocytes and cementocytes. In 3-d-treated animals, the early stage of hyalinization characterized with visible cells and fibres was observed in the PDL at the buccal side of the mesial root, which showed intense immunoreactivity for 3B3. Further 3B3 positive area seen in control animals changed its position from the distal to the buccal side of the PDL. Immunoreactivity for 2B6 did not change in the PDL of 3-d-treated animals. In 7-d-treated animals, the typical hyalinization characterized with no visible cells and fibres was seen in the PDL at the buccal sides of both mesial and disto-buccal roots, where both epitopes were present at the peripheral part of the tissue. Observation of serial sections suggested that the 3B3-positive area was present at the peripheral part of the 2B6-positive area. The present results suggest that the expression of CH-6S is related to the compressive force in non-hyalinized and hyalinized PDL, whereas that of CH-4S/DS is not influenced by the mechanical stress.

Alveolar Process↗

Induction of cyclobutane pyrimidine dimer photolyase in cultured fish cells by UVA and blue light.

The cyclobutane pyrimidine dimer (CPD) photolyase in fish cells is known to be regulated by environmental factors, such as light, hydrogen peroxide and growth inhibition. The induction of CPD photolyase by light in cultured goldfish cells was dependent on the wavelength of the light, and UVA and blue light had high inductive activity. The spectrum for CPD photolyase activity was different from that for the induction. Treatment with blue or yellow light for a short time, which did not induce any CPD photolyase, induced high CPD photolyase activity in the presence of the photosensitizers, TPPS (monosulfonated meso-tetraphenyl porphine) and ALPS (aluminum phthalocyanine tetrasulfonate), respectively. These results suggest that the induction of CPD photolyase might be triggered by active oxygen produced by light and cellular photosensitizers. We also found that immediately after treatment with UVA, blue light or a photosensitizer in combination with light, cellular attachment to the substratum was enhanced, as was the CPD photolyase activity. Pretreatment with a flavonoid, quercetin, inhibited both photoinduction of CPD photolyase and enhancement of cellular attachment. Vitamin E inhibited only photoinduction of CPD photolyase activity. Treatment with H7, a strong inhibitor for protein kinase C, after light treatment inhibited photoinduction of CPD photolyase activity, but an analogue of H7, Ha1004, which is a weak inhibitor of protein kinase C, did not have such an effect.

Animals↗

Overexpression of Medaka (Oryzias latipes) photolyase gene in Medaka cultured cells and early embryos.

To study the role and the regulation of the photolyase gene in the Medaka (small teleost), we constructed a eukaryotic expression plasmid of the Medaka photolyase gene and introduced it into Medaka cells in vivo and in vivo. The expression plasmid contains a cytomegalovirus enhancer and a thymidine kinase promoter to overexpress the photolyase gene of the Medaka. First, we transfected this construct into cultured Medaka cells and established several lines of transfectant. Every transfectant showed enhanced ability of pyrimidine dimer repair in the presence of fluorescent light. In the transfectant that showed the most enhanced ability of photorepair, the augmented transcription of photolyase gene was observed compared with that of progenitor OL32 cells. In this transfectant, we also observed an enhanced rate of UV survival with 20 min of fluorescent light treatment after irradiation with a 400 J/m2 UV sunlamp. Next, the expression construct was microinjected into the embryos of the Medaka at the one cell stage. Compared with the nontreated counterparts, the overexpression of a photolyase gene was detected in the microinjected embryos, but we failed to detect a significant increase in photo-reactivability of death at the midblastula stage.

Animals↗

Increased activity of vascular ACE related to atherosclerotic lesions in hyperlipidemic rabbits.

The relationship between vascular angiotensin-converting enzyme (ACE) activity in the aorta and atherosclerotic lesions was investigated in rabbits fed two atherogenic diets, 0.5 and 1.5% cholesterol, for 17 wk. Tissue ACE activity was assessed by the cleavage rate of hippuric acid from a synthetic substrate. Total cholesterol, triglycerides, and phospholipids in the serum were significantly increased by the diet. Vascular ACE activity in the femoral artery, abdominal aorta, and aortic arch was significantly increased (2-5 times) by the atherogenic diet in a high-cholesterol-related manner compared with the rabbits fed a normal diet. The increase was associated with vascular heterogeneity of regions. There was a significant correlation between plaque area and vascular ACE activity in the aortic arch isolated from rabbits fed the higher (1.5%)-cholesterol diet. Contractile responses of the femoral artery to angiotensin I and II in the atherogenic diet-fed rabbits did not differ from those in animals fed the normal diet. It is conceivable that increased ACE in the vascular walls is involved in hyperlipidemia-induced atherogenesis, presumably through production of growth or stimulating factor(s) contributing to plaque formation.

Animals↗

Inhibitory effect of the topical administration of a bisphosphonate (risedronate) on root resorption incident to orthodontic tooth movement in rats.

Root resorption associated with tooth movement is an unsolved problem in orthodontics. If such root resorption could be prevented, it would be an important contribution toward reducing risk factors in orthodontic treatment. The purpose of this study was to examine the effects of the topical administration of a bisphosphonate, risedronate, which is known to be a potent blocker of bone resorption, on root resorption during tooth movement and on the repair of the resorbed root surface after tooth movement in rats. In the first experiment, both the right and left upper first molars were moved buccally with a standardized expansion spring under administration of risedronate. After day 7, extensive root resorption had occurred on the control side, and the area of root resorption reached a maximum on day 14. The topical administration of risedronate caused a significant and dose-dependent inhibition of root resorption after the orthodontic force was applied. In the second experiment, the right and left upper molars were first moved buccally for 3 weeks. Risedronate treatment began on the day the spring was removed. After the force was withdrawn, the resorbed root surfaces on both the control and risedronate-treated sides were gradually restored by apposition of repair cementum (cementoid). The topical administration of risedronate did not appear to inhibit the repair process of root resorption. These results suggest that the topical administration of risedronate may be useful in preventing root resorption of teeth during orthodontic treatment.

Administration, Topical↗

Efficacy of direct injection of ethanol into the myocardium to control aconitine-induced ventricular tachycardia in anesthetized dogs.

The effect of injecting ethanol directly into the myocardium to control aconitine-induced ventricular tachycardia (VT) was evaluated in anesthetized dogs. In 17 dogs, VT was induced by injecting aconitine (1.0 microgram, median dose) directly into the epicardium. After inducing persistent VT for up to 3 min, 0.6 ml (median volume) of 96% ethanol was injected into the same epicardial region. Regular sinus rhythm reappeared in 15 dogs with no change in systolic blood pressure; the other 2 dogs died of ventricular fibrillation (VF). In another 13 dogs, VT was induced by injecting aconitine directly into the endocardium using a Variocath needle catheter. After persistent VT for up to 3 min, a regular sinus rhythm was restored in 7 dogs by injecting 2.0 ml (median volume) of 96% ethanol; the remaining 6 dogs died of VF. Histology showed no transmural necrosis and the subendocardial necrotic areas were essentially the same in the dogs that recovered from VT as in those that died. There was no statistically significant relationship between doses of ethanol and VT duration. These preliminary results suggest that the injection of ethanol into the myocardium may efficiently terminate VT when other techniques fail.

Aconitine↗

[Prognostic factors of lateral wall oropharyngeal squamous cell carcinoma--retrospective study of 79 patients].

We reviewed 79 patients with squamous cell carcinoma of the lateral wall of the oropharynx who received treatment at the Cancer Institute Hospital. Tokyo, between 1971 and 1990. There were 67 men and 12 women, aged 31 to 81 years (average 59.2 years). The tumors could be staged by TNM classification (UICC 1987) as follows: Stage 10 Stage II 12, Stage III 27, Stage IV 40. It must be noted that more than 50% of the tumors were classified as stage IV. As the initial treatment, 50 patients underwent radical radiotherapy, and the remaining 29 were treated mainly by surgery after pre-operative radiation. Some patients underwent adjuvant chemotherapy, but the role of chemotherapy was not considered in this study. The purpose of the present study was to determine the prognostic factors and to determine the survival rate after each treatment modality. The results were as follows: i) Patients with either T4, over N2b or a stage IV tumor had a significantly poor prognosis. ii) Cox multivariate analysis revealed that age, local extension (base of tongue, anterior pillar, soft palate and mid-line), pathological findings (poorly, moderately or well differentiated), tumor type (exophytic or endophytic) were not useful prognostic factors. iii) Radiosensitivity was not reflected in the survival rate judging from the high recurrence rate (44%) after radical radiotherapy. On the other hand, local recurrence after radical resection was seen in 18% of the patients. iv) Analysis of the data supported the recommendation of surgery after preoperative radiation as the treatment for stage III patients. It seems difficult to find the best treatment modality for stage IV patients in order to improve the survival rate, and this is an issue that still remains to be resolved. v) From the study of the surgical specimens, we are unable to demonstrate a significant survival advantage of the patients with a high grade radiation effect, or patients with no vessel involvement.

Adult↗

Surgical simulation of facial paralysis.

We are developing a prototype of a surgical simulation system for restoring facial expression lost in cases of facial paralysis. Correcting facial paralysis requires careful planning of surgery procedures and accurate prediction of the surgical results. So far techniques to perform this task largely relied on the physician's experience, and there were few quantitative data to refer to. The system we propose is finally aimed at helping the surgeon plan the surgery and predict the results. The 3D model of the face used in our system is generated directly from the CT scan of the patient. The face model consists of skin, skull and selected facial muscles based on anatomy. The mass and spring lattice approach is used to give physical attributes to the face model. The present prototype system lets the user directly manipulate graphical representations of facial expression.

Computer Graphics↗

Effects of lateral pterygoid muscle hyperactivity on differentiation of mandibular condyles in rats.

BACKGROUND: The effects of biomechanical stress on the growth and development of the mandibular condyle have been studied by many investigators. However, the role of the lateral pterygoid muscle in this development is not clear. METHODS: Hyperfunction of the lateral pterygoid muscles of male 3-week-old Sprague-Dawley rats was induced by electrical stimulation, and the responses of the mandibular condyles were compared to control tissues by a double-fluorescent staining technique using polyclonal antibodies against type I and type II collagen. Electrical stimulation consisted of repeated application (5 seconds on/5 seconds off) of a 5 Hz current for up to 7 days. RESULTS: In the first 2 days, cartilaginous tissues rich in type II collagen disappeared in the anterior and posterior areas, which were loaded by tensional force due to direct and indirect attachment of the lateral pterygoid muscles. Tissues in these areas were replaced by intramembranous bone that was reactive for type I collagen at 7 days. By the end of the experiment, the trabecula of the condyle was remodeled more perpendicularly, thus resisting the compressive force due to hyperfunction of the lateral pterygoid muscles. CONCLUSIONS: These results suggest that the activity of the lateral pterygoid muscle might play a significant role in the differentiation of progenitor cells and in the maturation and calcification of chondrocytes in mandibular condyles.

Animals↗

Molecular genetic basis of renal carcinogenesis in the Eker rat model of tuberous sclerosis (Tsc2).

We have recently identified on rat chromosome 10q a germline mutation in the tuberous sclerosis gene (Tsc2), the gene predisposing to renal carcinoma (RC) in the Eker rat. The homozygous mutant condition is lethal at around the 13th day of fetal life. In heterozygotes, RCs invariably develop in the first year of life. Histologically, RCs develop through multiple stages from early preneoplastic lesions (i.e., phenotypically altered tubules) to adenomas. The wild-type allele mutation has been found even in the earliest preneoplastic lesions, fitting Knudson's two-hit hypothesis and supporting the hypothesis that Tsc2 is a tumor suppressor gene. In this study, homozygous deletion of the Ink4 homologue on rat chromosome 5q was observed in 14 of 24 (58%) RC-derived cell lines. This may represent involvement of a second tumor suppressor gene, contributing to tumor progression. Considering previous results of studies of homozygous deletion of the Ifn alpha gene in five of 24 cases (21%) and the Ifn beta gene in one of 24 cases (4%), the order of the genes may be Ink4-Ifn alpha-Ifn beta. Microsatellite instability was not observed in 26 Eker rat tumors.

Amino Acid Sequence↗

Colonic and peritoneal tuberculosis associated with coloduodenal fistula.

We report a very rare case of tuberculous colitis that showed relatively long-segment involvement of the colon near the hepatic flexure with coloduodenal fistula that caused severe malnutrition. The formation of fistula in abdominal tuberculosis is very rare. This is the eighth reported case of abdominal tuberculosis with fistula and the first reported case with a coloduodenal fistula.

Adult↗

Allelic loss at the tuberous sclerosis (Tsc2) gene locus in spontaneous uterine leiomyosarcomas and pituitary adenomas in the Eker rat model.

Hereditary renal carcinomas (RCs) develop in virtually all Eker rats by the age of one year. Investigation of extra-renal primary tumors co-occurring in Eker rats late in life (at 2 years) additionally revealed enhanced development of hemangiosarcomas of the spleen, uterine leiomyosarcomas and pituitary adenomas, although the demonstrated predilection for these extra-renal tumors was not as complete as with RCs. We identified the germline mutated tuberous sclerosis (Tsc2) gene as the predisposing Eker gene and revealed the tumor suppressor nature of Tsc2 gene function in renal carcinogenesis. In the present study, we examined allelic loss at the Tsc2 gene locus in uterine leiomyosarcomas and pituitary adenomas developing in hybrid F1 rats carrying the Eker mutation as well as in pituitary adenomas from non-carrier rats. We detected loss of heterozygosity in 4 of 11 uterine leiomyosarcomas (36%) and 11 of 31 pituitary adenomas (35%) from Eker rats but in none of 9 pituitary adenomas from non-carrier rats (P < 0.05), suggesting that inactivation of the Tsc2 gene is also a critical event in the pathogenesis of these extra-renal tumors. Our present data indicate that there might be different pathways for tumorigenesis of pituitary adenomas between Eker and

Adenoma↗

Induction of cyclobutane pyrimidine dimer photolyase in cultured fish cells by fluorescent light and oxygen stress.

The expression of a gene for photolyase in RBCF-1 cells, a line of cultured goldfish cells, is known to be enhanced by fluorescent light. We have now found that H2O2 is another strong inducer of cyclobutane pyrimidine dimer photolyase. Northern blot analysis suggested that regulation by H2O2 occurs at the transcriptional level and the time course of induction of photolyase by H2O2 was similar to that by fluorescent light. Treatment with fluorescent light in the presence of riboflavin, which is known as an endogenous photosensitizer, also enhanced the induction of photolyase. These results suggest the involvement of oxygen stress in the induction of photolyase by fluorescent light. A cell clone with high-level expression of the goldfish gene for photolyase was obtained by transfection with plasmids that expressed the goldfish photolyase in OL32 cells derived from another fish, the medaka (Oryzias latipes). The induction of the medaka gene for photolyase was not affected by the high-level expression of the goldfish gene for this enzyme.

Animals↗

Cloning and characterization of cDNAs for 70-kDa heat-shock proteins (Hsp70) from two fish species of the genus Oryzias.

cDNA corresponding to two hsp70-related genes (OLHSC70 and CEHSC70) were isolated from two lines of cultured fish cells derived from the genus Oryzias. OLHSC70 was 2,261 bp in length and encoded a protein of 686 amino acids with a predicted molecular mass of 76,120 daltons. CEHSC70 was 2,114 bp in length and it lacked the 5' region found in OLHSC70. Two-dimensional electrophoresis revealed that Oryzias latipes has at least three heat-inducible proteins with molecular masses of about 70,000 daltons. One of these proteins (Hsp70.1) was barely expressed under normal conditions but its high-level expression was induced by hyperthermia. The other two proteins (Hsc70.1, and Hsc70.2) were constitutively expressed under normal conditions and only slightly enhanced levels were induced by hyperthermia. Transfection with the cloned sequence, RNA dot-blot analysis and the two-dimensional electrophoresis of proteins showed that OLHSC70 encoded Hsc70.1.

Amino Acid Sequence↗