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Biomedical subjects

T Kuroda

Publications and source records attributed to T Kuroda.

At least 37 records · Page 2Linked to original sources

Gas6 rescues cortical neurons from amyloid beta protein-induced apoptosis.

Gas6, a product of the growth-arrest-specific gene 6, protects neurons from serum deprivation-induced apoptosis. Neuronal apoptosis is also caused by amyloid beta protein (Abeta), whose accumulation in the brain is a characteristic feature of Alzheimer's disease. Abeta induces Ca(2+) influx via L-type voltage-dependent calcium channels (L-VSCCs), leading to its neurotoxicity. In the present study, we investigated effects of Gas6 on Abeta-induced cell death in primary cultures of rat cortical neurons. Abeta caused neuronal cell death in a concentration- and time-dependent manner. Gas6 significantly prevented neurons from Abeta-induced cell death. Gas6 ameliorated Abeta-induced apoptotic features such as the condensation of chromatin and the fragmentation of DNA. Prior to cell death, Abeta increased influx of Ca(2+) into neurons through L-VSCCs. Gas6 significantly inhibited the Abeta-induced Ca(2+) influx. The inhibitor of L-VSCCs also suppressed Abeta-induced neuronal cell death. The present cortical cultures contained few non-neuronal cells, indicating that Gas6 affected the survival of neurons directly, but not indirectly via non-neuronal cells. In conclusion, we demonstrate that Gas6 rescues cortical neurons from Abeta-induced apoptosis. Furthermore, the present study indicates that inhibition of L-VSCC contributes to the neuroprotective effect of Gas6.

Amyloid beta-Peptides↗

Disturbed tooth development in parathyroid hormone-related protein (PTHrP)-gene knockout mice.

Parathyroid hormone-related protein (PTHrP) is involved in epithelial-mesenchymal cell interactions during development of various tissues and organs. Tooth germ development is a classical model for this interaction. In tooth germs, PTHrP is expressed in the enamel organ (epithelial component), whereas its major receptor, the type I PTH/PTHrP receptor is expressed in cells of the alveolar bone and dental follicle (mesenchymal components). To clarify the role of PTHrP during fetal tooth germ development, PTHrP gene-knockout mice were used for histochemical and ultrastructural analysis. In wild-type mice, osteoclastic cells were aligned predominantly in the inner aspects of the alveolar bone surrounding the developing tooth germs throughout the late embryonic (after embryonic, 17.5 days) and neonatal animals examined. In contrast, osteoblasts were predominant in corresponding areas of fetal homozygous PTHrP-gene knockout mice with only occasional osteoclasts. In such areas, cell-free surfaces showing cement line-like tartrate-resistant acid phosphatase (TRAP) reactions were frequently observed. In neonatal homozygous mice, bone spicules were often shown to penetrate and/or compress the enamel organ and caused partial destruction of the tooth germs. Osteoclasts were few in number in the inner aspects of the alveolar bone, and had poorly developed ruffled border. No morphological abnormality was noted in cells of the tooth germs proper. On bone surfaces away from developing tooth germs, functional osteoclasts with structural features similar to those in wild-type mice were observed in homozygous mice. These observations suggest that PTHrP is required to maintain an appropriate spatiotemporal arrangement of bone cells and osteoclast function, which are necessary for the normal development of tooth germ and alveolar bone encasing the tooth germ. The observation also demonstrates that PTHrP deficiency affects the structure and function of osteoclasts exclusively those located in the vicinity of the growing tooth germ.

Acid Phosphatase↗

Bending property and phase transformation of Ti-Ni-Cu alloy dental castings for orthodontic application.

Bending property of Ti-Ni-Cu alloy castings was investigated in a three-point bending test for orthodontic application in relation to the phase transformation. The compositions of the alloys were Ti-50.8Ni and Ti-40.8Ni-10.0Cu (mol %), and four cross-sectional shapes of the specimens were selected. Heat treatment was performed at 713, 753 or 793 K for 1.8 ks. The bending load changed by the cross-sectional size and shape mainly because of the difference in the moment of inertia of area, but the load-deflection relation did not differ proportionally in the unloading process. The difference between the load values in the loading and the unloading processes was relatively small for Ti-Ni-Cu alloy. With respect to the residual deflection, there was no significant difference between Ti-Ni and Ti-Ni-Cu alloys with the same treatment condition. The load values in the loading and the unloading processes decreased by each heat treatment for Ti-Ni alloy; however, the decrease in the load values for Ti-Ni-Cu alloy was not distinct. It is proved that Ti-Ni-Cu alloy castings produce effective orthodontic force as well as stable low residual deflection, which is likely to be caused by the high and sharp thermal peaks during phase transformation.

Journal Article↗

Breathing modes, body positions, and suprahyoid muscle activity.

AIM: To determine (1) how electromyographic activities of the genioglossus and geniohyoid muscles can be differentiated, and (2) whether changes in breathing modes and body positions have effects on the genioglossus and geniohyoid muscle activities. METHOD: Ten normal subjects participated in the study. Electromyographic activities of both the genioglossus and geniohyoid muscles were recorded during nasal and oral breathing, while the subject was in the upright and supine positions. The electromyographic activities of the genioglossus and geniohyoid muscles were compared during jaw opening, swallowing, mandibular advancement, and tongue protrusion. RESULTS: The geniohyoid muscle showed greater electromyographic activity than the genioglossus muscle during maximal jaw opening. In addition, the geniohyoid muscle showed a shorter (P < 0.05) latency compared with the genioglossus muscle. Moreover, the genioglossus muscle activity showed a significant difference among different breathing modes and body positions, while there were no significant differences in the geniohyoid muscle activity. CONCLUSION: Electromyographic activities from the genioglossus and geniohyoid muscles are successfully differentiated. In addition, it appears that changes in the breathing mode and body position significantly affect the genioglossus muscle activity, but do not affect the geniohyoid muscle activity.

Adult↗

From three-dimensional space vision to prehensile hand movements: the lateral intraparietal area links the area V3A and the anterior intraparietal area in macaques.

The posterior parietal cortex is included in the dorsal cortical visual pathway underlying the three-dimensional (3-D) visual recognition of space and objects. The neurons in the lateral intraparietal area (LIP) respond visually to the three-dimensional objects, whereas those in the anterior intraparietal area (AIP) respond to hand movements to grasp them. LIP receives visual inputs from V3A, whereas AIP projects to the premotor areas; however, it is not known whether the neurons in LIP project to AIP. We herein investigated the connectional substrates that underlie the transformation of three-dimensional vision to prehensile hand movements in the Japanese monkey (Macaca fuscata). After identifying the three-dimensional visually responsive region in the posterior part of LIP by the unit recordings, we injected a bidirectional tracer, wheat germ agglutinin conjugated to horseradish peroxidase, into one of the recording sites. We found that LIP receives neuronal projections from V3A and sends axons to AIP. To confirm our findings, we injected several orthograde tracers into V3A and retrograde tracers into AIP in the same hemispheres. We found that the V3A neurons projecting to LIP terminate in the vicinity of the LIP neurons projecting to AIP. The results suggest that the cortical connections of V3A-LIP-AIP in the lateral bank of the intraparietal sulcus play an important role in the visuomotor transformation for prehensile hand movements.

Animals↗

Lipoprotein sorting signals evaluated as the LolA-dependent release of lipoproteins from the cytoplasmic membrane of Escherichia coli.

Escherichia coli lipoproteins are anchored to either the inner or outer membrane through fatty acyl chains covalently attached to an N-terminal cysteine. Aspartate at position 2 functions to retain lipoproteins in the inner membrane, although the retention is perturbed depending on the residue at position 3. We previously revealed that LolCDE and LolA play critical roles in this lipoprotein sorting. To clarify the sorting signals, the LolA-dependent release of lipoprotein derivatives having various residues at positions 2 and 3 was examined in spheroplasts. When the residue at position 3 was serine, only aspartate at position 2 caused the retention of lipoproteins in spheroplasts. We then examined the release of derivatives having aspartate at position 2 and various residues at position 3. Strong inner membrane retention occurred with a limited number of species of residues at position 3. These residues were present at position 3 of native lipoproteins having aspartate at position 2, whereas residues that inhibited the retention were not. It was also found that a strong inner membrane retention signal having residues other than aspartate at position 2 could be formed through the combination of the residues at positions 2 and 3. These results indicate that the inner membrane localization of native lipoproteins is ensured by the use of a limited number of strong inner membrane retention signals.

Aspartic Acid↗

Contribution of enzymic alpha, gamma-elimination reaction in detoxification pathway of selenomethionine in mouse liver.

The objective of this study was to clarify the detoxification pathways of selenomethionine (SeMet) in mouse liver. It has been postulated that SeMet may be metabolized to selenocysteine (SeCyH) via a pathway similar to methionine (Met). CySeH may be decomposed to H(2)Se, which is consequently methylated to CH(3)SeH, (CH(3))(2)Se, and (CH(3))(3)Se(+). In this study, we estimated that the median lethal single oral dose (LD(50)) was 67.0 mg/kg. We also found that (CH(3))(3)Se(+) was quickly produced in mouse liver after single oral administration of SeMet. This result suggested the existence of a quick alpha,gamma-elimination pathway. We measured the amounts of alpha-ketobutyrate, NH(3), and CH(3)SeH produced by enzymic alpha,gamma-elimination reaction of SeMet in the liver of periodate-oxidized adenosine (PAD) or D,L-propargylglycine (PPG)-treated mice in order to verify the existence of alpha,gamma-elimination enzyme. PAD is an inhibitor of S-adenosylhomocysteinase (EC 3.3.1.1), which is necessary for conversion of SeMet to SeCyH. PPG is an effective inhibitor of the pyridoxal 5'-phosphate (PLP)-containing enzyme bacterial L-methionine gamma-lyase (EC 4.4.1.11) contributing to the alpha,gamma-elimination reaction of SeMet and cystathionine gamma-lyase (EC 4.4.1.1) relating to conversion of SeMet to SeCyH. When SeMet was incubated with the S9 fraction from liver of PAD-treated mice, the formation of alpha-ketobutyrate was much the same as that from nontreated mouse liver. However, the amount of alpha-ketobutyrate formed significantly decreased in the reaction of SeMet with S9 fraction from the liver of PPG-treated mice. In an in vivo experiment using mice treated with PAD before a toxic dosage of SeMet, the amount of SeMet in the liver decreased and the amount of acid-volatile Se derived from CH(3)SeH increased gradually. This phenomenon was not observed in the PPG-pretreated group. Furthermore, the protein fraction that had the alpha,gamma-elimination enzyme activity was found in mouse liver cytosol by gel chromatographic technique. The results of this study indicated that SeMet was directly metabolized to CH(3)SeH by an alpha,gamma-elimination enzyme analogous to bacterial L-methionine gamma-lyase, in addition to the generally acceptable pathway via SeCyH.

Ammonia↗

Na+-driven multidrug efflux pump VcmA from Vibrio cholerae non-O1, a non-halophilic bacterium.

A chromosomal DNA fragment from Vibrio cholerae non-O1 containing a drug resistance determinant was cloned and sequenced. The deduced amino acid sequence suggested that the determinant gene encodes a multidrug efflux pump. We designated the pump VcmA. Escherichia coli cells transformed with a plasmid carrying the vcmA gene showed increased resistance against norfloxacin, ciprofloxacin, ofloxacin, daunomycin, doxorubicin, streptomycin, kanamycin, ethidium bromide, 4',6-diamidino-2-phenylindole dihydrochloride, Hoechst 33342 and acriflavine. Na+ (or Li+)-dependent efflux of ethidium bromide was detected in transformant cells. Efflux of Na+, elicited by ethidium bromide, was observed from transformant cells. Thus, we concluded that the VcmA is a Na+/drug antiporter.

Anti-Bacterial Agents↗

Construction of a series of mutants lacking all of the four major mex operons for multidrug efflux pumps or possessing each one of the operons from Pseudomonas aeruginosa PAO1: MexCD-OprJ is an inducible pump.

We constructed a series of deletion mutants lacking all of the four major mex operons for Mex multidrug efflux pumps or possessing each one of the operons from Pseudomonas aeruginosa PAO1. The drug specificity of MexAB-OprM, MexXY-OprM and MexCD-OprJ was investigated. Surprisingly, we found that the MexCD-OprJ was an inducible pump, inducers of which were tetraphenylphosphonium chloride, ethidium bromide, rhodamine 6G and acriflavine. Fluoroquinolones, chloramphenicol, erythromycin and tetracycline were not inducers although they were substrates of MexCD-OprJ.

Anti-Bacterial Agents↗

Effects of a titratable oral appliance on supine airway size in awake non-apneic individuals.

STUDY OBJECTIVES: To define morphological changes in the upper airway and its surrounding structures after the insertion of a titratable mandibular repositioner. DESIGN: Ten non-apneic adult males participated in this study. A set of supine lateral cephalograms was taken for each subject at the end of expiration with a titratable oral appliance in place in four mandibular positions: most retruded (RP), maximum protrusion (MAX), 33% of MAX (MAX33), and 67% of MAX (MAX67). Changes in the anteroposterior width of the upper airway, positions of the hyoid bone and the third cervical vertebra were compared between the four mandibular positions. An ANOVA was used to test for statistical significance. SETTING: N/A. PATIENTS OR PARTICIPANTS: N/A. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: The anteroposterior width of the velopharynx significantly increased when the mandible was advanced from RP to MAX67 and MAX. However, there were no significant changes in the anteroposterior width of the oropharynx. Significant forward displacement of the hyoid bone and third cervical vertebra together with the mandible was found in MAX67 and MAX compared to RP. CONCLUSION: Especially in MAX67 and MAX, the titratable oral appliance significantly enlarges upper airway size in the velopharynx and results in a forward displacement of the hyoid bone and the third cervical vertebra.

Adult↗

Pneumatosis cystoides intestinalis in a patient with polymyositis.

A 29-year-old Japanese woman with polymyositis (PM) developed pneumatosis cystoides intestinalis (PCI). The patient experienced a gradual onset of mild epigastric abdominal pain and abdominal distension. Radiographs revealed pneumatosis intestinalis involving the small bowel, and colonoscopy revealed submucosal cysts. Treatment with antibiotics and high-flow oxygen resulted in remission. This is an extremely rare case of PCI in a patient with PM.

Adult↗

An association of the polymorphic repeat of tetranucleotide (TCAT) in the first intron of the human tyrosine hydroxylase gene with schizophrenia in a Japanese sample.

Tyrosine hydroxylase (TH) is the rate limiting enzyme in the synthesis of dopamine and norepinephrine. A polymorphic repeat of the tetranucleotide (TCAT) in the first intron of the TH gene may behave as a regulatory element for the gene expression. Allelic fragments of the tandem repeat were typed by a PCR-based process with a pair of primers specific for the polymorphic sequence. The association between the polymorphism and schizophrenia was examined in a Japanese sample. There was a statistically significant association between the polymorphism and schizophrenia in females (chi2 = 26.018, p = 0.010), but not in males (chi2 = 15.995, p = 0.305). The genotype heterozygous for the TH9 and TH6 was significantly decreased in female schizophrenics (chi2 = 5.125, p = 0.0236). These results suggest that TH could be considered as a minor gene contributing to the susceptibility of Japanese female schizophrenia.

Adult↗

Identification of a novel frameshift mutation (383insT) in the RUNX2 (PEBP2 alpha/CBFA1/AML3) gene in a Japanese patient with cleidocranial dysplasia.

Cleidocranial dysplasia (CCD) is an autosomal dominant disorder due to mutations in runt-related gene 2 (RUNX2)/polyomavirus enhancer-binding protein 2alphaA (PEBP2alphaA)/core-binding factor A1 (CBFA1)/acute myeloid leukemia 3 (AML3). To investigate the RUNX2 mutations in a Japanese patient with classic CCD, we analyzed the RUNX2 gene using polymerase chain reaction (PCR)-single-strand conformation polymorphism and PCR-restriction fragment length polymorphism. The patient had hypoplasia of the clavicles, patent fontanelles, short stature, supernumerary teeth, and retention of deciduous dentition. We identified a 1-bp insertion (383insT) at codon 128 of the RUNX2 gene. The 383T insertion affects the conserved residue in the runt domain and results in premature termination in the runt domain.

Adolescent↗

Preserved otoacoustic emission in a child with a profound unilateral sensorineural hearing loss.

We report a case of profound unilateral sensorineural hearing loss with good response of otoacoustic emission. The patient was a 5-year-old boy. who was diagnosed to have unilateral hearing impairment on pure tone audiometry at the first visit. The affected ear showed the absence of auditory brainstem response; however, its transiently evoked otoacoutic emission and distortion product otoacoustic emission were considered to be normal. These findings indicated that the outer hair cell of cochlea was not impaired and that the impairment should be localized between inner hair cells, primary afferent fiber or its synapses, spiral ganglion of the cochlea and acoustic fiber, or at a combination of these areas. That is, evaluation of otoacoustic emission was useful in determining the region of impairment in sensorineural hearing loss. Further follow-up will be necessary to differentiate the present case from auditory neuropathy.

Child, Preschool↗

Optimal stimulus level conditions for measurements of distortion product otoacoustic emissions.

OBJECTIVE: Distortion product otoacoustic emission (DPOAE) is a tool for an objective examination of the cochlea function. While the clinical application of DPOEA test is in progress, the measurements vary depending on the stimulus conditions. We aimed to determine the most appropriate stimulus level in the clinical application of DPOAE test. METHODS: Ninety-seven normal hearing ears and 80 hearing-impaired ears (total 177) were subjected to this study. Two levels of stimulations (L1 and L2) were applied as follows: L1 = L2 = 70 dB; L1 = L2 = 60 dB and L1 = 60/L2 = 50 dB, and DP level was measured at each stimulation, and compared with hearing level. RESULTS: DP level was highest at L1 = L2 = 70 dB. Normal hearing ears and hearing-impaired ears were well differentiated at L1 = L2 = 60 dB. CONCLUSION: In clinical application of DPOAE test, DP levels should be measured at the stimulus level most appropriate for the purposes.

Acoustic Stimulation↗

Effects of spontaneous otoacoustic emissions on distortion product otoacoustic emission.

OBJECTIVE: It has been reported that spontaneous otoacoustic emission (SOAE) can prolong the responses or increase the echo power of transiently evoked otoacoustic emission (TEOAE), yet the effects of SOAE on distortion product otoacoustic emission (DPOAE) have been studied less thoroughly. As most of the previous studies have not paid attention to the patient's age, sex and hearing level, they have not reflected possible effects of those factors. We studied the effects of SOAE specifically on DPOAE in the following subjects. SUBJECTS AND METHODS: The subjects were all females ranging in age from 19 to 24 (average: 21.4) and the 78 ears had a hearing threshold under 15 dB for 1.2, and 4 kHz on pure-tone hearing test. IL088 (Otodynamics) was used for measurement of SOAE and IL092 (Otodynamics) for DPOAE. SOAEs were measured by time-averaging over 100 of the responses, of which those showing a clear peak 3 dB above the noise floor and being reproducible were considered as SOAE-positive. In all the ears. DPOAE responses were measured at L1 = L2 = 70 dB, and in 42 ears also at L1 = L2 = 60 dB and L1/L2 = 60/50 dB. The subjected ears were grouped into two by the presence or the absence of SOAE, and DPOAE amplitudes of 1, 2, and 4 kHz were compared. respectively. RESULTS: Of the total, 39 ears were SOAE-positive and 39 SOAE-negative. Statistically no significant difference was observed in the average hearing level between the SOAE-positive and SOAE-negative groups. The hearing levels did not significantly differ in the frequencies of 1, 2. and 4 kHz, respectively, indicating that influence of the hearing level on DPOAE could be excludable. DPOAE amplitudes at L1 = L2 = 70 dB in the frequencies of 1, 2, and 4 kHz were higher in the SOAE-positive group than in the SOAE-negative group. And DPOAE amplitudes were also higher in SOAE-positive group at L1 = L2 = 60 dB and L1/L2 = 60/50 dB in the frequency of 1.2, and 4 kHz, but significant differences were observed only in the frequencies of 4 kHz. By grouping the ears by the number of SOAE. we revealed the tendency that the larger the number of SOAE, the higher the DPOAE amplitudes. CONCLUSIONS: We evidenced that SOAE has significant effects on DPOAE responses. In clinical application of DPOAE measurement, therefore, the effects should be seriously taken into account.

Adult↗

An anti-mumps IgM antibody level in the serum of idiopathic sudden sensorineural hearing loss.

PURPOSE: Measurement of anti-IgM antibody for mumps enables us to diagnose silent mumps infection. From the viewpoint of prophylactic medicine, we examined the incidence of silent mumps infection in idiopathic sudden sensorineural hearing loss (ISSNHL) by measurement of anti-mumps IgM antibody. MATERIALS AND METHODS: Serum level of anti-mumps IgM antibody was evaluated by EIA method in 69 ISSNHL cases. RESULTS: Of the 69 serum samples examined, five samples were positive, two samples were regarded as quasi-positive and other samples were negative for anti-mumps IgM antibody. The positive rate was 7.2% (5/69). The relationship of anti-mumps IgM and IgG antibody was also described. CONCLUSIONS: From the measurement of anti-IgM antibody for mumps, it was possible to diagnose silent mumps infection in ISSNHL. The positive rate of anti-IgM antibody in ISSNHL was 7.2% suggesting that the silent mumps infection could be considered as one of the causative factors of ISSNHL even though its incidence is not so high. Since mumps often occurs without clinical symptoms, it is still considered to be one of the important causes of profound hearing loss. From the viewpoint of prophylaxis of profound hearing loss and deafness, we should understand the situation of virus epidemiology and vaccination.

Adolescent↗