Biomedical subjects

M Iino

Publications and source records attributed to M Iino.

At least 19 recordsLinked to original sources

PubMed2005

Influence of arthroscopically observed fibrous adhesions before and after joint irrigation on clinical outcome in patients with chronic closed lock of the temporomandibular joint.

The aim of this study was to investigate the changes in the state of arthroscopically observed fibrous adhesions (FA) after visually guided irrigation (VGIR) and the influence of FA on clinical outcome in patients with chronic closed lock of the temporomandibular joint (TMJ). Forty-eight TMJs of 48 patients with unilateral chronic closed lock were enrolled in this study. All 48 joints underwent VGIR twice. After the first VGIR (immediately before the second VGIR), clinical outcome was assessed as regards maximal interincisal opening (MIO) and self-evaluated TMJ pain (VAS). Thirty patients were symptom-free (good outcome group) and the remaining 18 patients had symptoms (poor outcome group). In each group, the changes of the MIO, VAS and severity of FA (FA score) after the first VGIR were studied. The influence of FA score in the first and second VGIR on clinical outcome was analyzed by logistic regression analysis. There was no joint with disappearance or reduction of FA after the first VGIR. In both groups, MIO and VAS were significantly improved after the first VGIR even though the state of FA became significantly worse. The multivariate logistic regression analysis showed that the risk of poor outcome for FA scores in the first and second VGIR were 0.89-times (95% CI: 0.33-2.40, P=0.82) and 1.76-times (95% CI: 0.54-5.73, P=0.35), respectively. The dose-response relationships between FA scores in the first or second VGIR were not significant. In conclusion, our results indicate that the presence of FA or a postoperative worsening of FA (including postoperative new FA formation) seems not to affect the clinical outcome as regards MIO and VAS in patients with chronic closed lock of the TMJ.

Adult↗

PubMed2004

Immunohistochemical localization of cyclooxygenase-1 and -2 in synovial tissues from patients with internal derangement or osteoarthritis of the temporomandibular joint.

This study examined the immunohistochemical expression and localization of cyclooxygenase-1 and -2 (COX-1 and COX-2) in synovial tissues from patients with internal derangement (ID) or osteoarthritis (OA) of the temporomandibular joint (TMJ). Synovial tissues from patients with condylar fractures of the mandible were studied as control. Synovial tissues from 13 TMJs of 10 patients with ID or OA and from 5 TMJs of 4 patients with fractures were examined for COX-1 and COX-2 expression by immunohistochemical staining using two monoclonal antibodies. In addition, whether the COX-2 expression grade correlated with the synovitis score and clinical findings was assessed. COX-2 was expressed in the synovial lining, infiltrating mononuclear cells, fibroblast-like cells, and blood vessels, including CD31-positive endothelial cells, in the synovium of patients with ID or OA. Expression levels of COX-1 in synovial lining cells and endothelial cells were similar in the specimens obtained from the patients with ID or OA and those obtained from the controls. The expression of COX-2 positively correlated with arthroscopic findings of synovitis (p = 0.55, P = 0.023) and with joint pain (p = 0.56, P = 0.021). These results suggest that up-regulation of COX-2 in synovium may play a part in the pathogenesis of synovitis in patients with ID or OA of the TMJ.

Adult↗

PubMed2003

Suppression of the morphine-induced rewarding effect and G-protein activation in the lower midbrain following nerve injury in the mouse: involvement of G-protein-coupled receptor kinase 2.

The present study was designed to investigate whether a state of neuropathic pain induced by sciatic nerve ligation could alter the rewarding effect, antinociception, and G-protein activation induced by a prototype of mu-opioid receptor agonist morphine in the mouse. The sciatic nerve ligation caused a long-lasting and profound thermal hyperalgesia. Under this neuropathic pain-like state, an i.c.v. morphine-induced place preference was observed in sham-operated mice but not in sciatic nerve-ligated mice. However, no differences in the antinociceptive effect of i.c.v.-administered morphine were noted between the groups. The increases in the binding of guanosine-5'-o-(3-[(35)S]thio)triphosphate induced by morphine in lower midbrain membranes including the ventral tegmental area, which contributes to the expression of the rewarding effect of opioid, were significantly attenuated in sciatic nerve-ligated mice. On the other hand, there were no differences in the stimulation of guanosine-5'-o-(3-[(35)S]thio)triphosphate binding to pons/medulla membranes, which plays an important role in the antinociception of mu-opioid receptor agonists, between the groups. In addition, no changes in levels of guanosine-5'-o-(3-[(35)S]thio)triphosphate binding by either the selective delta- or kappa-opioid receptor agonists were noted in membrane of the lower midbrain and limbic forebrain membranes obtained from sciatic nerve-ligated mice. Reverse transcription-polymerase chain reaction analysis showed that sciatic nerve ligation did not alter the mRNA product of mu-opioid receptors in the lower midbrain, indicating that a decrease in some mu-opioid receptor functions may result from the uncoupling of mu-opioid receptors from G-proteins. We found a significant increase in protein levels of G-protein-coupled receptor kinase 2, which causes receptor phosphorylation in membranes of the lower midbrain but not in the pons/medulla, obtained from mice with nerve injury, whereas there were no changes in the protein level of phosphorylated-protein kinase C in the lower midbrain. These results suggest that the uncoupling of mu-opioid receptors from G-proteins by G-protein-coupled receptor kinase 2 in the lower midbrain may, at least in part, contribute to the suppression of the rewarding effect of morphine under neuropathic pain.

Analgesics, Opioid↗

PubMed2003

Dentoalveolar reconstruction of a missing premaxilla using bone graft and endosteal implants.

We report here on a patient with bilateral cleft lip and palate (BCLP) and a missing premaxilla, who underwent dentoalveolar reconstruction of the cleft and premaxillary alveolus using endosteal implants after bone grafting. The patient, whose maxillary incisors and premaxilla were missing, had corticocancellous bone grafting from the iliac crest, followed by excellent bone formation at the anterior alveolus. After the placement of the endosteal implants and the completion of the pre-surgical orthodontic alignment, orthognathic surgery was performed for the restoration of a Class III open bite. After post-operative orthodontic preparation, the final fixed prostheses were completed. This treatment procedure offers an option of dentoalveolar reconstruction for BCLP patients with an excised premaxilla.

Adult↗

PubMed2002

Partial inferior turbinectomy during secondary alveolar bone grafting.

This study examined the characteristics and outcome of patients undergoing partial inferior turbinectomy during secondary alveolar bone grafting. Thirty-three of 55 patients with cleft lip and palate or cleft lip and alveolus who underwent secondary alveolar bone grafting concurrently received partial inferior turbinectomy to ensure that the height of the nasal floor was similar on the cleft side and non-affected side. At the time of surgery, patients who underwent turbinectomy were significantly older than those who did not undergo the procedure. The proportion of patients who underwent turbinectomy was significantly higher among patients with cleft lip and palate than among those with cleft lip and alveolus. These differences apparently reflected the developmental stage of the inferior turbinate and the relative severity of alveolar and palatal defects. In most patients who underwent partial inferior turbinectomy, postoperative X-ray films revealed excellent bone formation at the graft site. Our findings suggest that partial inferior turbinectomy during secondary alveolar bone grafting is a very useful procedure that facilitates dissection to the height of the nasal floor, reconstruction of the mucosal nasal floor, and formation of a sufficient bone bridge. It also promotes alveolar cleft closure, especially in patients with wide bone defects.

Adolescent↗

PubMed2001

Mapping of heme-binding domains in soluble guanylyl cyclase beta1 subunit.

Soluble guanylyl cyclase (sGC) is activated upon the interaction of NO with heme in the sGC beta1 subunit. To identify the domains contributing to heme-binding, we constructed a series of deletion mutants of the beta1 subunit, and evaluated their heme-binding capability. Deletion mutants consisting of residues 1-120 [beta1(1-120)] and 80-385 [beta1(80-385)] were the shortest mutants exhibiting heme binding among the C-terminal and N-terminal-truncated mutants, respectively. The region common to both beta1(1-120) and beta1(80-385), i.e., residues 80-120, is therefore essential for heme binding, although the residues 341-385 play an auxiliary role in heme binding. Two deletion mutants, beta1(80-195) and beta1(60-195), which include only the essential region, exhibited strong heme binding and spectral properties similar to those of the nitrosyl complex of native sGC. Thus, these heme-binding core proteins may serve as model proteins for future studies on the tertiary structure of the nitrosyl complex of sGC.

Animals↗

PubMed2001

Visualization of IP(3) dynamics reveals a novel AMPA receptor-triggered IP(3) production pathway mediated by voltage-dependent Ca(2+) influx in Purkinje cells.

IP(3) signaling in Purkinje cells is involved in the regulation of cell functions including LTD. We have used a GFP-tagged pleckstrin homology domain to visualize IP(3) dynamics in Purkinje cells. Surprisingly, IP(3) production was observed in response not only to mGluR activation, but also to AMPA receptor activation in Purkinje cells in culture. AMPA-induced IP(3) production was mediated by depolarization-induced Ca(2+) influx because it was mimicked by depolarization and was blocked by inhibition of the P-type Ca(2+) channel. Furthermore, trains of complex spikes, elicited by climbing fiber stimulation (1 Hz), induced IP(3) production in Purkinje cells in cerebellar slices. These results revealed a novel IP(3) signaling pathway in Purkinje cells that can be elicited by synaptic inputs from climbing fibers.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

PubMed2001

Reduced IP3 sensitivity of IP3 receptor in Purkinje neurons.

The inositol 1,4,5-trisphosphate receptor (IP3R) is highly expressed in Purkinje neurons (PNs) and is thought to be essential for the induction of long-term depression at parallel-fiber-PN synapses. Here, by imaging the fluorescence intensity of the low-affinity Ca2+ indicator inside the Ca2+ stores in the permeabilized single PNs, we analyzed the kinetics of Ca2+ release via the IP3R in controlled cytoplasmic environments. The rate of Ca2+ release is dependent on the IP3 concentration with an EC50 of 25.8 microM, which is > 20-fold greater than that of the IP3R in the isolated preparations or in peripheral cells. This property would be advantageous in inducing the release of Ca2+ in a localized space adjacent to the site of synaptic inputs.

Animals↗

PubMed2001

Glia-synapse interaction through Ca2+-permeable AMPA receptors in Bergmann glia.

Glial cells express a variety of neurotransmitter receptors. Notably, Bergmann glial cells in the cerebellum have Ca2+-permeable alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type glutamate receptors (AMPARs) assembled without the GluR2 subunit. To elucidate the role of these Ca2+-permeable AMPARs, we converted them into Ca2+-impermeable receptors by adenoviral-mediated delivery of the GluR2 gene. This conversion retracted the glial processes ensheathing synapses on Purkinje cell dendritic spines and retarded the removal of synaptically released glutamate. Furthermore, it caused multiple innervation of Purkinje cells by the climbing fibers. Thus, the glial Ca2+-permeable AMPARs are indispensable for proper structural and functional relations between Bergmann glia and glutamatergic synapses.

Adenoviridae↗

PubMed2001

Ca(2+)-sensor region of IP(3) receptor controls intracellular Ca(2+) signaling.

Many important cell functions are controlled by Ca(2+) release from intracellular stores via the inositol 1,4,5-trisphosphate receptor (IP(3)R), which requires both IP(3) and Ca(2+) for its activity. Due to the Ca(2+) requirement, the IP(3)R and the cytoplasmic Ca(2+) concentration form a positive feedback loop, which has been assumed to confer regenerativity on the IP(3)-induced Ca(2+) release and to play an important role in the generation of spatiotemporal patterns of Ca(2+) signals such as Ca(2+) waves and oscillations. Here we show that glutamate 2100 of rat type 1 IP(3)R (IP(3)R1) is a key residue for the Ca(2+) requirement. Substitution of this residue by aspartate (E2100D) results in a 10-fold decrease in the Ca(2+) sensitivity without other effects on the properties of the IP(3)R1. Agonist-induced Ca(2+) responses are greatly diminished in cells expressing the E2100D mutant IP(3)R1, particularly the rate of rise of initial Ca(2+) spike is markedly reduced and the subsequent Ca(2+) oscillations are abolished. These results demonstrate that the Ca(2+) sensitivity of the IP(3)R is functionally indispensable for the determination of Ca(2+) signaling patterns.

Adenosine Triphosphate↗

PubMed2001

Retrovirus insertion and transcriptional activation of the multidrug-resistance gene in leukemias treated by a chemotherapeutic agent in vivo.

To understand the molecular basis for multidrug-resistant (MDR) cancer cells in vivo, this study analyzed molecular changes of the mdr1a gene region in leukemia cells in mice during continuous treatment with vincristine. An inverse insertion of murine leukemia retrovirus (MuLV) into the 5'-flanking region of the mdr1a gene was found. This insertion was concomitantly accompanied by up-regulation of the mdr1a gene and the loss of chemosensitivity. Deletion of long-terminal repeat (LTR) sequences dramatically decreased the mdr1a promoter-driven reporter activity. The MuLV LTR insertion appears to exert its enhancer activity on mdr1a transcription during the appearance of MDR leukemia cells. Two mechanisms were postulated to explain the mdr1a gene activation by retrovirus insertion during in vivo chemotreatment: de novo insertion of MuLV induced by vincristine treatment and selection of a small fraction of pre-existing cells carrying MuLV insertion during vincristine treatment. No rearranged sequence was detected by polymerase chain reaction in parental cells. This result argued for the first mechanism. The randomly altered distribution of MuLV during repetitive chemotreatment might also be consistent with this hypothesis. On the other hand, the retrovirus insertion was detected at the same site of the mdr1a promoter region in 2 independent experiments, which suggests the second mechanism. It should be noted that in vivo chemotreatment using vincristine could generate the mdr1a-overexpressing cells through retrovirus insertion and the enhancer effect of the LTR.

ATP Binding Cassette Transporter, Subfamily B↗

PubMed2001

Synthesis and structure--activity analysis of novel dihydropyridine derivatives to overcome multidrug resistance.

The structure activity relationships were studied on newly synthesized 1,4-dihydropyridine derivatives possessing a 1-pentyl group at the 4-position, and 3-pyridylpropylester was found to be one of the effective fragments for overcoming P-glycoprotein mediated multidrug-resistance (MDR) in cultured human cancer cells, in vitro. 3-Pyridylpropylester was also found to be one of the effective fragments for increasing the life span of P-glycoprotein overexpressing MDR P388 leukemia-bearing mice, in vivo. All compounds had weak calcium antagonistic activities, but there appeared no relationship between MDR reversing effect and calcium antagonistic activity.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed2001

Negative control of store-operated Ca2+ influx by B cell receptor cross-linking.

An increase in the intracellular Ca(2+) concentration by B cell receptor (BCR) cross-linking plays important roles in the regulation of B cell functions. [Ca(2+)](i) is regulated by Ca(2+) release from the Ca(2+) store as well as store-operated Ca(2+) influx (SOC). Protein tyrosine kinases downstream of BCR cross-linking were shown to regulate the mechanism for Ca(2+) release. However, it remains elusive whether BCR cross-linking regulates SOC or not. In this study, we examined the effect of BCR cross-linking on thapsigargin-induced SOC in the DT40 B cells. We found that the SOC-mediated increase in intracellular Ca(2+) concentration was inhibited by BCR cross-linking. Using a membrane-potential-sensitive dye, we found that BCR cross-linking induced depolarization, which is expected to decrease the driving force of Ca(2+) influx and SOC channel conductance. When membrane potential was held constant by the transmembrane K(+) concentration gradient in the presence of valinomycin, the BCR-mediated inhibition of SOC was still observed. Thus, the BCR-mediated inhibition of SOC involves both depolarization-dependent and depolarization-independent mechanisms of SOC inhibition. The depolarization-independent inhibition of the SOC was abolished in Lyn-deficient, but not in Bruton's tyrosine kinase-, Syk- or SHIP (Src homology 2 domain containing phosphatidylinositol 5'-phosphatase)-deficient cells, indicating that Lyn is involved in the inhibition. These results show novel pathways of BCR-mediated SOC regulations.

Animals↗

PubMed2001

Osmolality dependence of erythrocyte sedimentation and aggregation in a strong magnetic field.

In order to specify the major determinant of the magnetic enhancement of erythrocyte sedimentation observed previously, the dependence of erythrocyte sedimentation rate (ESR) on osmolality was measured under a strong magnetic field. Even at hypotonic osmolality, an increase in ESR due to aggregation was observed in plasma solution as compared with that without aggregation in saline solution. However, the magnetic field did not enhance ESR at hypotonic osmolality, when the cell shape was an isotropic sphere (spherocyte). Thus, we narrowed our search to a mechanism that would explain the enhanced ESR found specifically in anisotropic erythrocytes. It was concluded that the major determinant can only work for anisotropic erythrocytes and is a magnetic field-induced increase in an intermembrane adhesive area due to magnetic orientation of anisotropic erythrocytes.

Adult↗

PubMed2001

IGF-I-induced apoptosis in LM2d6 cultured at a low concentration of fetal bovine serum.

We examined the effects of IGF-I (1-1000 ng/ml) on cell proliferation in LM2d6 mouse fibroblast cells at 0.1, 1.0 and 5.0% fetal bovine serum (FBS). In medium containing 0.1% FBS, treatment of LM2d6 cells with IGF-I significantly reduced the cell number in a dose- and time-dependent manner, whereas no effects were seen at 1 or 5% FBS. Treatment of the cells with 0.1% FBS for 72 h caused DNA laddering and nuclear condensation. However, Scatchard analysis for IGF-I binding sites on the cells revealed that both the number and the affinity of IGF-I receptors were not greater than that of Balb/3T3 cells. Furthermore, the apoptotic action of Long (R(3))-IGF-I, an analogue of IGF-I that has a reduced affinity for IGF binding proteins, was not greater than that of IGF-I. Taken together, we conclude that IGF-I reduces cell proliferation at low levels of FBS due to the induction of apoptosis. This effect is probably not caused by an excess production of IGF binding proteins in LM2d6 cells.

Animals↗

PubMed2001

Small molecule-based laser inactivation of inositol 1,4,5-trisphosphate receptor.

BACKGROUND: Chromophore-assisted laser inactivation (CALI) is a powerful method for the study of in situ protein function in cellular processes. By using CALI, it is possible to abrogate the function of a target protein with unprecedented spatial and temporal resolution. However, CALI has some limitations, which restrict wider biological application, owing mainly to the use of antibody for target recognition. To circumvent the limitations, we have developed small molecule-based CALI (smCALI). RESULTS: The inositol 1,4,5-trisphosphate receptor (IP3R) was selected as the target protein and a malachite green-conjugated IP3 analog, MGIP3, was used as a small-molecular probe. We examined the effect of MGIP3-based CALI on Ca2+ release via IP3R using permeabilized smooth muscle cells. When the cells were treated with MGIP3 followed by laser irradiation, the IP3-induced Ca2+ release rate was decreased in a concentration- and irradiation time-dependent manner. The effect was specific for IP3R, because the Ca2+ uptake function of the co-localized sarco/endoplasmic reticulum Ca2+-ATPase was not affected. CONCLUSIONS: IP3R was specifically inactivated by smCALI using MGIP3. The efficiency of inactivation was calculated to be substantially greater than that of antibody-based CALI. The efficient and specific inactivation of IP3R would allow us to obtain an insight into spatiotemporal roles of IP3R in various cell functions. Our results may be considered to be a first step for a wider application of smCALI as a useful method to study spatiotemporal protein functions.

Animals↗

PubMed2001

Sequential-replenishment mechanism of exocytosis in pancreatic acini.

Here we report exocytosis of zymogen granules, as examined by multiphoton excitation imaging in intact pancreatic acini. Cholecystokinin induces Ca 2+ oscillations that trigger exocytosis when the cytosolic Ca 2+ concentration exceeds 1 microM. Zymogen granules fused with the plasma membrane maintain their Omega-shaped profile for an average of 220 s and serve as targets for sequential fusion of granules that are located within deeper layers of the cell. This secondary exocytosis occurs as rapidly as the primary exocytosis and accounts for most exocytotic events. Granule-granule fusion does not seem to precede primary exocytosis, indicating that secondary fusion events may require a plasma-membrane factor. This sequential-replenishment mechanism of exocytosis allows the cell to take advantage of a large supply of fusion-ready granules without needing to transport them to the plasma membrane.

Animals↗

PubMed2001

Dendritic spine geometry is critical for AMPA receptor expression in hippocampal CA1 pyramidal neurons.

Dendritic spines serve as preferential sites of excitatory synaptic connections and are pleomorphic. To address the structure-function relationship of the dendritic spines, we used two-photon uncaging of glutamate to allow mapping of functional glutamate receptors at the level of the single synapse. Our analyses of the spines of CA1 pyramidal neurons reveal that AMPA (alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid)-type glutamate receptors are abundant (up to 150/spine) in mushroom spines but sparsely distributed in thin spines and filopodia. The latter may be serving as the structural substrates of the silent synapses that have been proposed to play roles in development and plasticity of synaptic transmission. Our data indicate that distribution of functional AMPA receptors is tightly correlated with spine geometry and that receptor activity is independently regulated at the level of single spines.

Animals↗