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Biomedical subjects

T Ohyama

Publications and source records attributed to T Ohyama.

At least 19 recordsLinked to original sources

Bone mineral density of the mandible in ovariectomized rats: analyses using dual energy X-ray absorptiometry and peripheral quantitative computed tomography.

INTRODUCTION: Although previous studies have shown that maxillary molar extraction in ovariectomized (OVX) animals causes mandibular loss of bone, it is still questionable as to whether estrogen deficiency affects mandibles with functional occlusion. MATERIALS AND METHODS: To answer this question, 13-week-old female Sprague-Dawley rats were bilaterally OVX or sham-operated. After 109 days, the bone mineral density (BMD) of the femurs and mandibles was measured using dual-energy X-ray absorptiometry (DEXA) and peripheral quantitative computed tomography (pQCT). RESULTS: In DEXA analysis, although the BMD of the total mandible of the OVX rats was similar to that of the sham-operated rats, the BMD of the condylar region in the OVX rats had decreased by 12.5%. In pQCT analysis, decrease in trabecular BMD of the mandibular bone was detectable but low in the molar region (maximal 13%), whereas no difference was seen in cortical BMD. In the femurs, the trabecular bone prominently decreased in OVX rats (30% decrease in pQCT analysis) as previously reported. CONCLUSION: This study revealed regional differences in the mandibular bone decrease in OVX rats. Although the mechanism of low susceptibility of the mandible to estrogen-deficient conditions remains unknown, it is likely that mechanical stress derived from functional occlusion is preventing bone loss in this pathological condition. Furthermore, this study demonstrated the advantage of pQCT in analyzing rat mandibular bone.

Absorptiometry, Photon↗

A new and simple method for evaluating masticatory function using newly developed artificial test food.

The aim of this study was to develop an objective evaluation system for the masticatory function. This system used paraffin wax cubes as a test food, which had six red- and green-coloured layers so that each of the six surfaces showed a pseudo-checkered pattern. A total of 100 paraffin cubes were chewed by 37 subjects and the images of these samples were captured and analysed using a digital image analyzer. With regard to the colour and the shape of each sample, five parameters were obtained. Furthermore, an independent examiner graded the degree of colour mixing in the chewed samples into three groups (poor, medium and good) by visual inspection. A discriminant analysis was performed using the five variables as predictors of two groups (good and poor). Mixing Ability Index (MAI) was calculated from the discriminant function and using this index, 97% of the samples from these two groups were classified correctly. This system needed only a few minutes to complete and is easy to use. Therefore, it has high potentials for clinical use.

Adult↗

Validity and reliability of a newly developed method for evaluating masticatory function using discriminant analysis.

The objective of the study was to assess validity and reliability of a newly developed method for evaluating mixing ability of masticatory function. A two-coloured paraffin wax cube was used as a test food in the mixing ability test. Eleven full dentate and 33 denture wearers participated in the validity assessment. They were divided into three groups according to their dental state. Validity was assessed by the Spearman's coefficient of correlations between the mixing ability test and the sieving method used as a 'gold standard' test of the masticatory performance. The subjects were asked to masticate the test foods for 5, 7, 10, 15, 20 and 30 strokes in each test. Significant correlations (P < 0.001) were obtained in natural dentition group (r(s) = 0.66), in denture-natural dentition group (r(s) = 0.72) and in denture-denture group (r(s) = 0.56). In the reliability assessment, three full dentate subjects were asked to repeat the mixing ability test a number of times. The intraclass correlation coefficients were 0.94 in interexaminer consistency, 0.98 in intra-examiner consistency, 0.89 in test-retest consistency and 0.99 in measuring consistency. These results indicated that the mixing ability test had the validity and the reliability for evaluation of the masticatory performance.

Adult↗

Influence of working side contacts on masticatory function for mandibular distal extension removable partial dentures.

The aim of this study was to clarify the influence of working side contacts for masticatory function for a distal extension removable partial denture. Five subjects who had edentulous arches from second premolar to second molar opposing natural teeth participated. Working side contacts were altered by using three types of artificial teeth attached to the denture base. A-form teeth made contact with opposite teeth while acting as the working side in a lateral excursion. The cusp angles of B- and C-form teeth were decreased by 10 degrees and 20 degrees, respectively compared with A-form teeth. B-and C-form teeth discluded with opposite teeth on the working side. Subjects were asked to perform two kinds of masticatory performance test. Obtained data were evaluated by the repeated-measures anova and the Tukey test for multiple comparisons. Results revealed that there was a significant difference (P=0.001) in the three types of teeth in the mixing ability test and the value of A-form teeth was statistically less value than those of the other teeth. However, there was no significant difference in the comminuting ability test. These results suggested that working side contacts between artificial teeth and opposite natural teeth influenced mixing ability, but not comminuting ability.

Adult↗

3-D Finite element analysis of all-ceramic posterior crowns.

The purpose of this study was to evaluate the stress distribution under various loading conditions within posterior all-ceramic crowns. A three-dimensional finite element model representing a lower first molar was constructed. Variations of the model had two types of single layer all-ceramic crowns (Dicor and Empress) and two types of double layer all-ceramic crowns (In-Ceram and Empress2) cemented. A load of 600 N, simulating the maximum bite force, was applied vertically to the crowns. Loads of 225 N, simulating masticatory force, were applied from three directions (vertically, at a 45 degrees angle, and horizontally). In the test simulating maximum bite force, the maximum tensile stresses on all crowns (17.4-19.4 MPa) concentrated around the loading points. In the masticatory force simulation test, the specimens experienced maximum tensile stresses of 19.7-27.0 MPa under a horizontal load and 10.8-10.9 MPa under a vertical load. When the load was applied horizontally, the maximum tensile stress was observed around the loading points on the surface in the case of the single layer crowns, and of the cervical area of the inner core of the double layer crowns. Within the limitation of this study, it was found that the strength of occlusal contact points is important to the integrity of posterior all-ceramic crowns and that bite forces applied from the horizontal direction are a critical factor.

Bite Force↗

Enhancement of osteogenesis on hydroxyapatite surface coated with synthetic peptide (EEEEEEEPRGDT) in vitro.

Some dental implants are coated with hydroxyapatite (HA), which preferentially binds to bone. Several matrix proteins have an arginine-glycine-aspartic acid (RGD) sequence where cells attach via an integrin receptor. We hypothesized that coating an HA surface with an RGD-containing peptide might enhance the attachment and differentiation of osteoblasts. The HA disks (diameter 34 mm, thickness 1 mm) were treated with a solution (50 mM Tris/HCl and 150 mM NaCl, pH 7.4) containing the peptide EEEEEEEPRGDT, in which the E repetition exerts a high affinity to HA. After washing with phosphate-buffered saline, KUSA/A1 mouse osteoblastic cells were inoculated onto the HA surface and cultured. After 30 min, the number of cells attached to the surface was counted. The DNA content and alkaline phosphatase (ALP) activity were measured after 10 days in culture. Expression of bone matrix proteins was also examined by means of reverse transcriptase-polymerase chain reaction at 7 days; the mineralized area of the culture was also evaluated by staining with Alizarin Red S after 10 days. Treatment with the peptide stimulated cell attachment and increased DNA content and ALP activity. Furthermore, matrix protein expression and mineralized nodule formation were enhanced to a greater extent on the peptide-treated surface than on the nontreated surface. Our results indicate that coating an HA surface with RGD-containing peptide enhances osteoblast attachment and differentiation. This peptide treatment of HA-coated implants may stimulate the osseointegration of the implants.

Animals↗

Testing a symptom-based surveillance system at high-profile gatherings as a preparatory measure for bioterrorism.

We tested symptom-based surveillance during the G8 conference in 2000 as a means of detecting outbreaks, including bio-terrorism attacks, promptly. Five categories of symptoms (skin and haemorrhagic, respiratory, gastrointestinal, neurological and unexplained) were adopted for the case definition of the surveillance. The surveillance began I week before the conference, and continued until 1 week after the conference ended. We could not detect any outbreaks during this surveillance. Compared to the existing diagnosis-based surveillance system, symptom-based surveillance has the advantages of timeliness and simplicity. However, poor specificity and difficulties in determining epidemic threshold were important limitations of this system. To increase the specificity of surveillance, it is essential to incorporate rapid laboratory diagnoses into the system.

Bioterrorism↗

Vibration analysis of maxillary removable partial denture frameworks.

The purpose of this study was to investigate the vibratory characteristics of three designs of the Class I Kennedy maxillary removable partial denture frameworks as the basic study. Their major connectors comprised a U-shaped palatal connector (UPC), single palatal bar (SPB), and anterior-posterior palatal bars (APB). Frequency response functions were measured when the framework was impacted. The modal shape was observed and the decay rate was calculated using modal analysis software. The results showed that the vibratory properties of each framework differed from each other. Within the range of frequencies from 10 to 2000 Hz, the UPC type had seven natural frequencies, while the SPB and the APB types had six. The UPC type had a greater number of natural modes accompanied by elastic deformation, including fluttering and twisting, than the other type, and the UPC type was considered to be unfavourable. The decay rate of the APB type was significantly higher than those of the UPC and the SPB types (P < 0.01).

Analysis of Variance↗

Digital acoustic analysis of five vowels in maxillectomy patients.

The aim of the study was to characterize the acoustics of vowel articulation in maxillectomy patients. Digital acoustic analysis of five vowels, /a/, /e/, /i/, /o/ and /u/, was performed on 12 male maxillectomy patients and 12 normal male individuals. A simple set of acoustic descriptions called the first and second formant frequencies, F1 and F2, were employed and calculated based on linear predictive coding. The maxillectomy patients had a significantly lower F2 for all five vowels and a significantly higher F1 for only /i/ vowel. From the data plotted on an F1-F2 plane in each subject, we determined the F1 range and the F2 range, which are the differences between the minimum and the maximum frequencies among the five vowels. The maxillectomy patients had a significantly narrower F2 range than the normal controls. In contrast, there was no significant difference in the F1 range. These results suggest that the maxillectomy patients had difficulty in controlling F2 properly. In addition, the speech intelligibility (SI) test was performed to verify the results of this new frequency range method. A high correlation between the F2 range and the score of SI test was demonstrated, suggesting that the F2 range is effective in evaluating the speech ability of maxillectomy patients.

Acoustics↗

Fracture resistance of pressable glass-ceramic fixed partial dentures.

The aim of the present study was to evaluate the mechanical strength of the Empress2 system, which is based on the use of a high-strength glass--ceramic core of lithium disilicate, and the fracture resistance of fixed partial dentures fabricated with this material. To evaluate mechanical strength, four types of ceramic materials were tested for four-point flexural strength and diametral tensile strength: Empress2 core material, Empress2 layering porcelain, conventional Empress material and Dicor. Then, using Empress2, conventional Empress and Dicor, actual clinical type anterior fixed partial dentures were fabricated for fracture testing. The results showed that the Empress2 core material, at 329 MPa, has more than twice the flexural strength of conventional materials and at 271 MPa, more than four times the diametral tensile strength of conventional materials. Furthermore, fixed partial dentures fabricated with Empress2 had a fracture resistance of 1424 N. That is, they were more than twice as fracture resistant as fixed partial dentures made with conventional materials.

Ceramics↗

The influence of altered occlusal guidance on condylar displacement.

This study investigated mandibular displacement and masticatory muscle activity during clenching in lateral occlusal position in relation to the lateral occlusal pattern. Twenty healthy human subjects (mean = 26.5 years) volunteered for this study. Metallic occlusal overlays were fabricated for the lower working side canine to second molar and non-working side second molar in order to simulate a canine protected occlusion, group function occlusion and bilateral balanced occlusion (balancing contact). Three-dimensional displacements of the bilateral condyle and electromyography (EMG) activities in the bilateral masseter, anterior and posterior temporalis were recorded during maximal clenching. The experimental occlusal pattern was revealed to have statistically significant effects on both condylar displacement and EMG activity (P < 0.001 and P < 0.001, respectively). When compared to the simulated group function occlusion, the simulated canine protected occlusion and balancing contact caused statistically significant smaller superior displacements of the non-working side condyle, which suggests that they may result in reduced temporomandibular joint (TMJ) loading. Furthermore, the simulated canine protected occlusion was associated with the lowest EMG activity, which suggests that this occlusal pattern has the capacity to reduce the level of parafunctional activity.

Adult↗

Role of C-terminal region of HA-33 component of botulinum toxin in hemagglutination.

Using SDS-PAGE, we found that one subcomponent, hemagglutinin (HA-33), from the Clostridium botulinum progenitor toxin of type D strain 1873 and type C strain Yoichi had slightly smaller molecular sizes than those of type C and D reference strains, but other components did not. Based on N- and C-terminal sequence analyses of HA-33, a deletion of 31 amino acid residues from the C-terminus at a specific site was observed in the HA-33 proteins of both strains. The progenitor toxins from both strains showed poor hemagglutination activities, titers of 2(1) or less, which were much lower than titers from the reference strains (2(6)), and did not bind to erythrocytes. These results suggest strongly that the short C-terminal region of the HA-33 plays an essential role in the hemagglutination activity of the botulinum progenitor toxin. Additionally, a sequence motif search predicted that the C-terminal region of HA-33 has a carbohydrate-recognition subdomain.

Amino Acid Sequence↗

Linearization and integration of DNA into cells preferentially occurs at intrinsically curved regions from human LINE-1 repetitive element.

A bent DNA library was constructed from human genomic DNA, from which a new clone belonging to the human LINE-1 sequence family was isolated and characterized. This clone, with a length of 378 base pairs and termed HBC-1 (human bent clone-1), contained an intrinsically occurring curved DNA structure. By permutation analysis, the center of curvature of this fragment was mapped onto the nucleotide position 886 from the 5' terminus of the complete LINE-1 sequence. Reporter plasmids, which contain HBC-1, were effectively integrated into human chromosome, indicating that the bent DNA structure provides a preferential donor site for the integration of human LINE-1 sequences. The present finding may provide an explanation as to why some inactivated LINE-1 sequences on human chromosomes carry the deletion at their 5' termini.

3T3 Cells↗

Cu-metallothioneins (Cu(I)8-MTs) in LEC rat livers 13 weeks after birth still act as antioxidants.

Redox properties of metallothioneins (MTs) and Cu in the cytosol from Long-Evans Cinnamon (LEC) rat livers 13 weeks after birth were investigated. MTs from LEC rat livers contain 8 g atoms of Cu and 1 g atom of Zn per mole of protein (Cu(I)8-MTs). Titration of Cu(I)8-MTs with CuCl2 indicates that Cu(I)8-MTs were able to reduce further 2-g atoms of cupric ions per mole MTs as bound form. Hg2+-induced hydroxyl radical generation from Cu(I)8-MTs was demonstrated by ESR using the spin trap, 5,5-dimethyl-1-pyrroline N-oxide (DMPO). The intensity of DMPO-OH signal from Cu-loaded MTs was increased with the increasing number of Cu in MTs. The used cytosol fraction contained 1.37 mM total Cu and 5 mM DTNB titrable-SH groups has a potential to reduce 2 mM CuCl2. No ESR signal due to Cu2+ was also detected with LEC rat liver cytosol, whereas strong Cu2+ signal appeared by the addition of HgCl2. The rate constants for the reaction of Cu(I)8-MTs with superoxide and hydroxyl radicals were estimated to be 2 x 10(6) and > or = 10(12) M(-1)s(-1), respectively, from competition kinetics. Cu2+-catalyzed oxidation of DNA was strongly inhibited both in the presence of Cu-unsaturated MTs and GSH. The results suggest that Cu(I)8-MTs from LEC rat livers just before hepatitis still act as antioxidants.

Animals↗

MST, a physiological caspase substrate, highly sensitizes apoptosis both upstream and downstream of caspase activation.

The human serine/threonine kinase, mammalian STE20-like kinase (MST), is considerably homologous to the budding yeast kinases, SPS1 and STE20, throughout their kinase domains. The cellular function and physiological activation mechanism of MST is unknown except for the proteolytic cleavage-induced activation in apoptosis. In this study, we show that MST1 and MST2 are direct substrates of caspase-3 both in vivo and in vitro. cDNA cloning of MST homologues in mouse and nematode shows that caspase-cleaved sequences are evolutionarily conserved. Human MST1 has two caspase-cleavable sites, which generate biochemically distinct catalytic fragments. Staurosporine activates MST either caspase-dependently or independently, whereas Fas ligation activates it only caspase-dependently. Immunohistochemical analysis reveals that MST is localized in the cytoplasm. During Fas-mediated apoptosis, cleaved MST translocates into the nucleus before nuclear fragmentation is initiated, suggesting it functions in the nucleus. Transiently expressed MST1 induces striking morphological changes characteristic of apoptosis in both nucleus and cytoplasm, which is independent of caspase activation. Furthermore, when stably expressed in HeLa cells, MST highly sensitizes the cells to death receptor-mediated apoptosis by accelerating caspase-3 activation. These findings suggest that MST1 and MST2 play a role in apoptosis both upstream and downstream of caspase activation.

3T3 Cells↗

Latent acquisition of timed responses in cerebellar cortex.

Evidence indicates that rabbit eyelid conditioning is mediated by plasticity in the interpositus cerebellar nucleus and in cerebellar cortex. Although the relative contributions of these sites are not fully characterized, evidence suggests that plasticity in the cerebellar cortex influences conditioned response amplitude and timing, whereas plasticity in the interpositus nucleus is necessary or permissive for conditioned response expression. Recent empirical and computational analyses suggest that, during training, plasticity is initially established in the cerebellar cortex, whereas conditioned response expression begins later as plasticity is induced in the interpositus nucleus. We used the dependence of response timing on the interstimulus interval (ISI) to test this latent learning hypothesis. Rabbits were initially trained using a tone conditioned stimulus (CS) with a relatively long ISI to a low-criterion threshold. The relative absence of plasticity in the interpositus nucleus was then examined via reversible disconnection of the cerebellar cortex. Later, to induce plasticity in the interpositus nucleus, subjects were trained to robust levels of conditioned response expression using a shorter ISI. Reversible disconnection of the cerebellar cortex at this time confirmed the presence of robust interpositus nucleus plasticity after the second phase. Subsequent probe trials with the long CS alone then revealed double-peaked responses whose peaks were appropriately timed to the two ISIs. The results are consistent with the hypothesis that temporally specific learning occurs first in the cerebellar cortex before the appearance of conditioned responses. This latent learning is expressed only after plasticity is induced in the interpositus nucleus.

Acoustic Stimulation↗

Crystal structure of a depsipeptide, Boc-(Leu-Leu-Lac)3-Leu-Leu-OEt.

A sequential polydepsipeptide, Boc-(Leu-Leu-Lac)3-Leu-Leu-OEt (1) (Lac = L-lactic acid residue) has been synthesized by the segment condensation method. The sequential unit of 1, -Leu-Leu-Lac-, is consisted of two amino acid residues and one hydroxy acid residue. X-ray diffraction measurement with an imaging plate detector and a direct-methods procedure of Shake-and-Bake successfully revealed the crystal structure of 1. In the solid state, the 11-mer depsipeptide, 1, have clear alpha-helical conformation even with the three ester linkages.

Crystallization↗

Intrinsic DNA bends: an organizer of local chromatin structure for transcription.

DNA with a curved trajectory of its helix axis is called bent DNA, or curved DNA. Interestingly, biologically important DNA regions often contain this structure, irrespective of the origin of DNA. In the last decade, considerable progress has been made in clarifying one role of bent DNA in prokaryotic transcription and its mechanism of action. However, the role of bent DNA in eukaryotic transcription remains unclear. Our recent study raises the possibility that bent DNA is implicated in the "functional packaging" of transcriptional regulatory regions into chromatin. In this article, I review recent progress in bent DNA research in eukaryotic transcription, and summarize the history of bent DNA research and several subjects relevant to this theme. Finally, I propose a hypothesis that bent DNA structures that mimic a negative supercoil, or have a right-handed superhelical writhe, organize local chromatin infrastructure to help the very first interaction between cis-acting DNA elements and activators that trigger transcription.

Chromatin↗