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Biomedical subjects

T Hamano

Publications and source records attributed to T Hamano.

At least 19 recordsLinked to original sources

Inhibitory role of regucalcin in the regulation of Ca2+ dependent protein kinases activity in rat brain neurons.

The role of regucalcin in the regulation of protein kinase activity in rat brain neuronal cells obtained from primary culture was investigated. Protein kinase activity was assayed using the 5500 g supernatant fraction of the cell homogenate. Protein kinase activity was significantly raised by the addition of calmodulin (5 microg/ml) or dioctanoylglycerol (5 microg/ml) in the presence of CaCl2 (10(-4) M), indicating that Ca2+/calmodulin-dependent protein kinase and protein kinase C is present in the neuronal cells. The addition of regucalcin (10(-9)-10(-7) M) in the enzyme reaction mixture caused a significant decrease in protein kinase activity in the absence of calmodulin or dioctanoylglycerol without Ca(2+) addition. Moreover, regucalcin completely prevented the activation of protein kinase by the addition of calmodulin or dioctonoylglyceral in the presence of CaCl(2) (10(-4) M). The presence of anti-regucalcin monoclonal antibody (25 or 50 ng/ml) caused a significant elevation of protein kinase activity without CaCl2 addition. Such an effect was significantly inhibited by the addition of trifluoperazine (2x10(-5) M), an antagonist of calmodulin, or staurosporine (10(-6) M), an inhibitor of protein kinase C. The present study demonstrates that endogenous regucalcin in rat brain neuronal cells has an inhibitory effect on Ca2+ dependent protein kinase activity.

Animals↗

Epithelial pigment slide in contact lens wearers: a possible marker for contact lens-associated stress on corneal epithelium.

PURPOSE: To compare the incidence of epithelial pigment slide among wearers of various types of contact lenses. METHODS: Prospectively, we studied 432 eyes of 432 patients. The patients were separated into 6 groups: hard contact lens (HCL) wearers (n = 166), conventional soft contact lens (CSCL) wearers (n = 30), extended disposable lens (EDCL) wearers (n = 46), frequent replacement SCL (FRCL) wearers (n = 60), daily disposable SCL (DDCL) wearers (n = 65), and normal controls (n = 65). The incidence of prominent pigment slide, defined as spike-like epithelial opacities in the corneal limbus and longer than 1 mm from the base to the apex of the wedges-shaped process detected by slit-lamp examination, was compared in these 6 groups. The relationship between the incidence of prominent pigment slide and the length of contact lens wear was examined. RESULTS: The overall incidence of prominent pigment slide in the CSCL, EDCL, HCL, FRCL, DDCL, and normal groups was 63.3, 23.9, 13.9, 8.3, 7.7, and 4.6%, respectively. The incidence of prominent pigment slide in the CSCL and EDCL groups was significantly higher than that in the control group. A higher incidence of prominent pigment slide was correlated with longer wearing period in each group. CONCLUSIONS: The presence of epithelial pigment slide may be a marker for contact lens-associated stress to the corneal epithelium.

Biomarkers↗

Expression of Ca(2+)-binding protein regucalcin in rat brain neurons: inhibitory effect on protein phosphatase activity.

The expression of Ca(2+)-binding protein regucalcin and its role in the regulation of protein phosphatase activity in rat brain neuronal cells obtained with primary culture was investigated. The expression of regucalcin mRNA was demonstrated by reverse transcription-polymerase chain reaction (RT-PCR) analysis in brain neuronal cells using rat regucalcin-specific primers. Moreover, regucalcin protein in brain neuronal cells was detected by Western blot analysis using a polyclonal rabbit anti-regucalcin antibody. The presence of anti-regucalcin monoclonal antibody (20 or 50 ng/ml) in the enzyme reaction mixture caused a significant increase in protein phosphatase activity toward phosphotyrosine, phosphoserine and phosphothreonine in the reaction mixture containing the cytosol of neuronal cell homogenates. This increase was completely prevented by the addition of regucalcin (10(-8) M). Protein phosphatase activity toward three phosphoaminoacids was significantly elevated by the addition of Ca(2+) (100 microM) and calmodulin (5 microg/ml). This elevation was completely blocked by the addition of regucalcin (10(-8) M). The present study demonstrates that regucalcin is expressed in rat brain neuronal cells, and that it has an inhibitory effect on protein phosphatase activity in the cells.

Animals↗

Phosphoglycerate kinase deficiency: an adult myopathic form with a novel mutation.

The authors report a 36-year-old man with exertional myoglobinuria and muscle cramp without hemolytic anemia or CNS symptoms. They found a deficiency of phosphoglycerate kinase (PGK) activity in muscle and erythrocytes and a 4-base pair deletion in exon 6 of the PGK gene. This mutation may cause a frameshift, yielding an abnormal stop codon in exon 6 by which a truncated PGK protein was produced. This phenotype is caused by a novel mutation of the PGK gene.

Adult↗

Decreased phosphorylation levels of TrkB neurotrophin receptor in the spinal cords from patients with amyotrophic lateral sclerosis.

Amyotrophic lateral sclerosis (ALS) is characterized by the selective degeneration of specific populations of cranial and spinal motor neurons. In this study, we examined the expression of the high affinity functional receptor for BDNF, TrkB, and assessed the functional state of TrkB by examining the level of phosphorylation on tyrosine residues in ALS spinal cords. The data showed that TrkB-immunoprecipitates prepared from cell-free lysates of ALS spinal cords by use of an anti-TrkB antibody contained much more TrkB protein than from controls. These TrkB proteins expressed in ALS spinal cords, however, are much less phosphorylated on tyrosine residues than those of controls. Moreover, RT-PCR analysis of TrkB mRNA in ALS spinal cords demonstrated that the expression of Trk B mRNA is also upregulated in ALS spinal cords compared with those of controls. These data strongly suggest that there exists an abnormality in TrkB-mediated intracellular signaling in ALS spinal cords and shed a light on the possibility of the therapeutic intervention by normalizing this intracellular signaling.

Amyotrophic Lateral Sclerosis↗

Graft-versus-host-disease-associated donor cell engraftment in an F1 hybrid model is dependent upon the Fas pathway.

The graft-versus-host disease (GVHD) generated in BDF1 mice by the injection of spleen cells from the C57BL/6 parental strain induces a direct cell-mediated attack on host lymphohaematopoietic populations, resulting in the reconstitution of the host with donor cells. We examined Fas-Fas ligand (FasL) interactions in donor and host haematopoietic cells over a prolonged period of parental-induced GVHD. Fas expression on bone marrow cells of both donor and host origin increased at 2 weeks. Host cell incubation with anti-Fas antibody induced apoptosis, and the number of haematopoietic progenitor cells decreased. Fas-induced apoptosis by the repopulating donor cells, however, did not increase until 12 weeks, when more than 90% of the cells were donor cells. The expression of various cytokines, such as interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF-alpha), and FasL gene expression in the bone marrow increased concomitantly. To examine directly whether FasL has a major role in the development of donor cell engraftment, FasL-deficient (gld) mice were used as donors. Injection of B6/gld spleen cells induced significantly less host lymphohaematopoietic depletion, resulting in a failure of donor cell engraftment. Furthermore, injection of IFN-gamma gene knockout (gko) B6 spleen cells failed to augment Fas and FasL expression in recipient mice, resulting in a failure of donor cell engraftment. This suggests that the induction of apoptosis by Fas-FasL interactions in host cells may contribute to a reconstitution of the host with donor cells and that donor-derived IFN-gamma plays a significant role for Fas-FasL interactions in host cells during parental-induced GVHD.

Animals↗

Impression pressures against teeth in a partially edentulous model with a mobile tooth: influence of impression tray design.

The purpose of the present study was to examine the effect of custom tray designs on local pressures against teeth during the impression procedure. In a previous study, a partially edentulous simulation model with a mobile tooth was used, and the effect of custom tray designs on the displacement of the mobile tooth was examined during the impression procedure. Based on that study's results, we have assumed that the differences in impression pressures between the labial and the lingual sides of a mobile tooth could either cause or affect displacement. The present study was undertaken to determine the local impression pressures against each side of three anterior teeth, including one mobile tooth, using the same simulation model and the same custom trays as in the previous study. It was found that the local pressures exerted against teeth during the impression procedure were affected by the custom tray designs and varied according to the coronal shape, axis inclination and location of the teeth.

Analysis of Variance↗

IgD multiple myeloma preceding the development of extensive extramedullary disease without medullary involvement.

We present a unique case of IgD multiple myeloma (MM) preceding the development of extensive extramedullary disease without medullary involvement. A 63-year-old man was diagnosed with IgD-lambda MM when he developed anemia. After 3 months of chemotherapy, he was in complete remission as evidenced by the disappearance of bone marrow (BM) plasmacytosis, monoclonal IgD protein in his serum, and Bence Jones proteinuria. Six months after diagnosis, his disease took an unusual course with the development of plasmacytomas in the skin, without medullary involvement. He then received chemotherapy, resulting in the complete disappearance of the subcutaneous plasmacytomas. Two years after the initial diagnosis, his disease took an aggressive clinical course with retroperitoneal relapse, leading to the patient's death within 1 month. The two separate episodes of extramedullary disease were associated with elevated serum lactic dehydrogenase levels and the absence of plasma cells in the BM. This case provides evidence of two separate transformations of the original malignant MM clone.

Antineoplastic Combined Chemotherapy Protocols↗

Cortical potentials associated with voluntary mandibular movements.

Movement-related cortical potentials (MRCPs) are negative potentials over the scalp, which gradually increase prior to voluntary movements, and might be applied to elucidate the cortical efferent function of the mandibular movements. We compared the MRCPs accompanying various mandibular movements to study the motor control mechanism underlying these movements. Electroencephalograms (EEGs) were recorded from 11 electrodes placed over the scalp (F3, Fz, F4, T3, C3, Cz, C4, T4, P3, Pz, and P4), according to the International 10-20 System, and electromyograms (EMGs) were obtained from surface electrodes over the masseter muscle and the anterior belly of the digastric muscle. Ten healthy subjects were requested to make brisk and self-paced mandibular movements in 4 different directions (mouth-opening and -closing, and left and right lateral movements). We obtained MRCPs by averaging the EEG, using the visually determined EMG onset as a trigger signal. In all the movements, a slowly increasing, bilaterally widespread negativity starting 1.5 to 2.0 sec before the EMG onset (Bereitschaftspotential, or BP proper) was observed, with the maximum over the vertex region. The negative slope (NS') occurred about 300 to 700 msec before the EMG onset. The cortical maps of BP/NS' (BP and NS' combined), immediately prior to the mouth-opening and closing, showed a symmetrical distribution, whereas that for the lateral movements showed a tendency of predominance over the hemisphere ipsilateral to the direction of the movement. BP/NS' amplitudes at the onset of movement differed significantly or tended to do so between open, close, and lateral movements, suggesting that MRCP recordings may thus provide a means to explore the role of the cerebral cortex in the control of mandibular movements.

Adult↗

[Dopamine transporter SPECT in patients with Parkinson's disease].

The major neuropathological feature in Parkinson's disease (PD) is severe degeneration of the dopamine (DA) neurons in the substantia nigra. Dopamine transporter (DAT) is an important protein in the regulation of DA neurotransmission. It has been reported that PD patients show a loss of DAT in striatum. We report here the findings of single photon emission computed tomography (SPECT) of the DAT with 2 beta-carboxymethoxy-3 beta-(4[123I]iodophenyl)tropane ([123I] beta-CIT) to investigate striatal DAT in 10 patients with PD, one patient with vascular parkinsonism (VP), and one patient with dystonia syndrome. Patients were evaluated using the Webster rating scale. Specific/nondisplaceable striatal binding ratio (V3") was obtained in each case. In PD patients, the uptake of [123I] beta-CIT was reduced, especially in the tail of putamen compared with caudate nucleus. Even in the early stage of PD, the uptake of beta-CIT was reduced not only in the severely affected side, but also in the mildly disturbed side of the brain. Putamen caudate ratio was generally low in PD patients. In VP patient, the uptake was reduced, but putamen caudate ratio was not decreased. V3" values showed significant correlation with the severity of clinical symptoms such as self-care, facies, posture, gait, speech, and Hoehn-Yahr's stage. On the other hand, V3" values were not significantly correlated with the degree of tremor, seborrhea, and duration of the illness. In conclusion, we found that SPECT of the [123I] beta-CIT is a useful method for the diagnosis in the patients presenting parkinsonism, and for the clinico-physiological estimation of parkinsonian symptoms such as self-care, facies, posture, gait, and speech.

Adult↗

Effect of graft-versus-host disease (GVHD) on host hematopoietic progenitor cells is mediated by Fas-Fas ligand interactions but this does not explain the effect of GVHD on donor cells.

The acute graft-versus-host disease (GVHD) generated in BDF1 mice by the injection of spleen cells from the C57BL/6 parental strain induces a direct cell-mediated attack on host lymphohematopoietic populations, resulting in the reconstitution of the host with donor hematopoietic stem cells. We examined the effect of GVHD on the donor and host hematopoiesis in parental-induced acute GVHD. The bone marrow was hypoplastic and the number of hematopoietic progenitor cells significantly decreased at 4 weeks after GVHD induction. However, extramedullary splenic hematopoiesis was present and the number of hematopoietic progenitor cells in the spleen significantly increased at this time. Fas expression on the host spleen cells and bone marrow cells significantly increased during weeks 2 to 8 of GVHD. Host cell incubation with anti-Fas Ab induced apoptosis, and the number of hematopoietic progenitor cells decreased during these weeks. A significant correlation between the augmented Fas expression on host bone marrow cells and the decreased number of host bone marrow cells by acute GVHD was observed. Furthermore, the injection of Fas ligand (FasL)-deficient B6/gld spleen cells failed to affect host bone marrow cells. Although Fas expression on repopulating donor cells also increased, Fas-induced apoptosis by the repopulating donor cells was not remarkable until 12 weeks, when more than 90% of the cells were donor cells. The number of hematopoietic progenitor cells in the bone marrow and the spleen by the repopulating donor cells, however, decreased over an extended time during acute GVHD. This suggests that Fas-FasL interactions may regulate suppression of host hematopoietic cells but not of donor hematopoietic cells. Hematopoietic dysfunctions caused by the reconstituted donor cells are independent to Fas-FasL interactions and persisted for a long time during parental-induced acute GVHD.

Animals↗

MRI findings of benign monomelic amyotrophy of lower limb.

We report here magnetic resonance imaging (MRI) findings of two patients with benign monomelic amyotrophy of lower limb. Both subjects showed unilateral amyotrophy of the lower limb with a benign clinical course, and the affected muscles demonstrated neurogenic changes. On T1- and T2-weighted MRI, marked atrophy and increased signal intensity were found mainly in gastrocnemius and soleus muscles. Moreover, MRI examination also revealed that thigh muscles including semitendinosus, semimembranosus, and vastus intermedius and lateralis muscles were involved in one of the patients. We concluded that muscle MRI is very useful for detecting affected muscles, especially deep skeletal muscles in patients with benign monomelic amyotrophy of lower limb.

Adult↗

Cortical distribution of Bereitschaftspotential and negative slope potential preceding mouth-opening movements in humans.

Cortical potentials associated with the voluntary movement of various body parts are known as movement-related cortical potentials, which allow evaluation of the cortical efferent function and other higher functions controlling voluntary movement. The cortical potentials have three main components: the Bereitschaftspotential (or 'readiness potential'), the negative slope and the motor potential. Here, the cortical potentials preceding mouth-opening movements were recorded to investigate which of these components could be observed. Electroencephalograms (EEGs) were recorded from 11 electrodes placed over the scalp (F3, Fz, F4, T3, C3, Cz, C4, T4, P3, Pz and P4) according to the international 10-20 system. Electromyograms (EMGs) were recorded from the anterior belly of the digastric and the masseter. The 10 healthy participants were requested to make brisk and self-paced mouth-opening movements. All data were digitized with a sampling frequency of 200 Hz and stored for off-line analysis. Movement-related cortical potentials were obtained by averaging the EEG, using the digastric EMG onset as a trigger signal. The Bereitschaftspotential was recorded as a gradually increasing, bilaterally widespread negativity starting 1.7 s (mean) +/-0.23 s (S.D.) before the onset of the opening movement. The amplitude, which was measured at movement onset, was maximum at Cz (3.8 +/- 1.3 microV). The negative slope was observed 500-600 ms before the movement onset, and the motor potential was not clearly identified, because of postmotion artefacts. The cortical maps of the Bereitschaftspotential and negative slope combined together before to the mouth opening showed a symmetrical distribution with a maximum at the vertex region. The study reveals that the Bereitschaftspotential and the negative slope, two components of the movement-related cortical potentials, can be observed preceding mouth-opening movements and are similar to those associated with other voluntary movements. Findings such as distribution, amplitude and onset of the potentials could provide important information for studying the cortical control of mandibular movements.

Adult↗

Inhibitory effect of regucalcin on Ca(2+)-dependent protein kinase activity in rat brain cytosol: involvement of endogenous regucalcin.

The effect of regucalcin, a Ca(2+)-binding protein, on Ca(2+)-dependent protein kinase activity in the brain cytosol of rats with different ages (5 and 50 weeks old) was investigated. The addition of calmodulin (10 microg/ml) or dioctanoylglycerol (5 microg/ml) in the enzyme reaction mixture caused a significant increase in protein kinase activity in the presence of CaCl2 (1 mM), indicating that Ca2+ calmodulin or protein kinase C is present in the cytosol. Such an increase was completely prevented by the addition of regucalcin (10(-7) M). Moreover, regucalcin (10(-7) M) significantly inhibited cytosolic protein kinase activity without Ca2+/calmodulin or dioctanoylglycerol addition. Meanwhile, the presence of anti-regucalcin monoclonal antibody (10-50 ng/ml) in the enzyme reaction mixture caused a significant elevation of protein kinase activity, suggesting an inhibitory effect of endogenous regucalcin. Brain cytosolic protein kinase activity was significantly elevated by increasing age (50-week-old rats). Also, regucalcin (10(-7) M) significantly decreased protein kinase activity without Ca(2+) addition in the brain cytosol of aged rats. However, the effect of anti-regucalcin monoclonal antibody (50 ng/ml) in elevating protein kinase activity was not seen in the brain cytosol of aged rats. These results suggest that regucalcin has an inhibitory effect on Ca(2+)-dependent protein kinase activity in rat brain cytosol, and that the effect of endogenous regucalcin may be weakened in the brain cytosol of aged rats.

Animals↗

Abnormal contingent negative variation in writer's cramp.

OBJECTIVE: To investigate the physiological abnormality in writer's cramp, a focal dystonia which specifically affects writing. METHODS: We recorded brain potentials that precede hand and neck movements (contingent negative variation or CNV) in 11 patients and 11 age-matched normal subjects. A 1000 Hz tone burst (S1) was delivered to the right or left ear in random sequence, and 2 s after, a 2000 Hz tone burst (S2) was delivered to both ears simultaneously. For the response task to S2, the subjects were instructed to extend their fingers ipsilateral to the ear to which S1 was given in one experiment or to rotate the head to the side of the S1 presentation in another. All the patients had symptoms in the right hand only, and performed both tasks normally. CNV amplitudes were compared between normals and patients using unpaired t test. RESULTS: They showed normal CNV for neck movement but significantly decreased CNV amplitudes for movements both in the affected and unaffected hands. CONCLUSIONS: Our findings suggest that motor programming is specifically abnormal for the affected body part, including the asymptomatic contralateral limb, and that the clinical symptom may result from a deficient compensatory mechanism for abnormal motor programs or subroutines.

Adult↗