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Biomedical subjects

M Iino

Publications and source records attributed to M Iino.

At least 37 records · Page 2Linked to original sources
PubMed2001

Sindbis viral-mediated expression of Ca2+-permeable AMPA receptors at hippocampal CA1 synapses and induction of NMDA receptor-independent long-term potentiation.

Gene manipulation in order to artificially express a particular gene in neurons in the central nervous system is a powerful tool for the analysis of brain function. Sindbis viral vectors have been developed to express high levels of foreign genes in postmitotic brain neurons with little transfection of glial cells. In this study, we expressed the gene encoding the unedited GluR2 (GluR-B) subunit of the AMPA-type glutamate receptor that forms inwardly rectifying and Ca2+-permeable channels, in rat CA1 hippocampal neurons in slice cultures using Sindbis viral vectors. The pyramidal cell layer of the CA1 region was injected with recombinant Sindbis viruses encoding both enhanced green fluorescent protein (GFP) and unedited GluR2. The GFP fluorescence from CA1 neurons could be detected as early as 6 h and reached a maximal level about 48 h postinfection. The inwardly rectifying and Ca2+-permeable AMPA receptors were expressed in most CA1 pyramidal cells expressing GFP. These AMPA receptors expressed by gene transfer were involved in fast excitatory neurotransmission elicited by electrical stimulation of the Schaffer collaterals in the stratum radiatum. Tetanic stimulation of Schaffer collaterals induced NMDA receptor-independent, long-term potentiation due to Ca2+ influx through the newly expressed AMPA receptors in the area densely stained with GFP. Thus, the combined use of Sindbis viral vectors with the GFP reporter allowed physiological examination of the roles of a specific gene product in synaptic function in well-characterized brain neurons.

Animals↗
PubMed2001

Auxin- and abscisic acid-dependent osmoregulation in protoplasts of Phaseolus vulgaris pulvini.

Protoplasts isolated from the laminar pulvinus of Phaseolus vulgaris and bathed in a medium containing KCl as the major salt were found to swell in response to IAA and to shrink in response to ABA. The protoplasts of flexor cells and those of extensor cells responded similarly. The results indicate that the cellular content of osmotic solutes is enhanced by IAA and reduced by ABA. The IAA-induced swelling was abolished when either the K(+) or the Cl(-) of the bathing medium was replaced by an impermeant ion or when the medium was adjusted to neutral pH (instead of pH 6). The response was inhibited by vanadate. It is concluded that the swelling is caused by enhanced influxes of K(+) and Cl(-), which probably occur through K(+) channels and Cl(-)/H(+) symporters, respectively. The ABA-induced shrinking was inhibited by 5-nitro-2-(3-phenylpropylamino)-benzoic acid, an anion-channel inhibitor, suggesting that it is caused by Cl(-) efflux through anion channels and charge-balancing K(+) efflux through outward-rectifying K(+) channels. It appears that the two plant hormones act on pulvinar motor cells to regulate their turgor pressure, as they do in stomatal guard cells. The findings are discussed in relation to the pulvinar movements induced by environmental stimuli.

Abscisic Acid↗
PubMed2001

Blue-light-dependent osmoregulation in protoplasts of Phaseolus vulgaris Pulvini.

Blue light was found to induce shrinkage of the protoplasts isolated from first-leaf lamina pulvini of 18-day-old Phaseolus vulgaris. The response was transient following pulse stimulation, while it was sustainable during continuous stimulation. No apparent difference was found between flexor and extensor protoplasts. Protoplasts of the petiolar segment located close to the pulvinus showed no detectable response. In the plants used, the pulvinus was fully matured and the petiole was ceasing its elongation growth. When younger, 12-day-old, plants were used, however, the petiolar protoplasts did respond to blue light. The pulse-induced response was similar to that in pulvinar protoplasts, although the response to continuous stimulation was transient and differed from that in pulvinar protoplasts. No shrinkage was induced in pulvinar protoplasts when the far-red-light-absorbing form of phytochrome was absent for a period before blue-light stimulation, indicating that the blue-light responsiveness is strictly controlled by phytochrome. Inhibitors of anion channels and H(+)-ATPase abolished the shrinking response, supporting the view that protoplasts shrink by extruding ions. The response of pulvinar protoplasts is probably involved in the blue-light-induced, turgor-based movement of pulvini. The blue-light responding system in pulvini is suggested to have evolved from that functioning in other growing organs.

Adaptation, Physiological↗
PubMed2001

Light-dependent osmoregulation in pea stem protoplasts. photoreceptors, tissue specificity, ion relationships, and physiological implications.

Light-induced changes in the volume of protoplasts bathed in a medium of constant osmolarity are useful indications of light-dependent cellular osmoregulation. With this in mind, we investigated the effect of light on the volume of protoplasts isolated from the elongating stems of pea (Pisum sativum) seedlings raised under red light. The protoplasts were isolated separately from epidermal peels and the remaining peeled stems. Under continuous red light, the protoplasts of peeled stems swelled steadily, but those of epidermal peels maintained a constant volume. Experiments employing far-red light and phytochrome-deficient mutants revealed that the observed swelling is a light-induced response mediated mainly by phytochromes A and B with a little greater contribution by phytochrome A. Protoplasts of epidermal peels and peeled stems shrank transiently in response to a pulse of blue light. The blue light responsiveness in this shrinking response, which itself is probably mediated by cryptochrome, is under the strict control of phytochromes A and B with equal contributions by these phytochromes. We suggest that the swelling response participates in the maintenance of high tissue tension of elongating stems and that the shrinking response is involved in stem growth inhibition. Other findings include the following: The swelling is caused by uptake of K+ and Cl-. The presence of Ca2+ in the bathing medium is required for phytochrome signaling in the swelling response, but not in the response establishing blue light responsiveness. Phytochrome A mediates the two responses in a totally red/far-red light reversible manner, as does phytochrome B.

Calcium↗
PubMed2001

Vestibuloplasty after secondary alveolar bone grafting.

OBJECTIVE: This paper introduces a surgical technique for vestibuloplasty after secondary alveolar bone grafting of patients with cleft lip and palate (CLP). This paper also reports on the patients who underwent this modified vestibuloplasty. SURGICAL PROCEDURE: The vestibuloplasty technique described in this paper consists of: (1) reduction of submucosal scar tissue of the upper lip, (2) V-Y plasty of the superficial mucosa, (3) placement of horizontal mattress sutures between nostril floor skin and freed marginal mucosa, (4) application of artificial skin to cover the exposed periosteal surface, and (5) use of a removable retention splint. CONCLUSION: This surgical procedure appears to be very useful for patients with CLP. The technique enables the surgeon to obtain an adequate sulcus depth around the graft area. In addition, this technique releases the mucosal scar contraction and improves the shape and mobility of the upper lip.

Adolescent↗
PubMed2001

Evaluation of distal and proximal axonal degeneration in patients with carpal tunnel syndrome.

In patients with carpal tunnel syndrome, varying degrees of demyelination and axonal degeneration occur in the median nerve. Only a few studies have examined axonal degeneration produced at proximal to the lesion. In this study proximal axonal degeneration was evaluated and compared with other parameters. In 40 consecutive CTS patient hands, distal latency (DL), compound muscle action potential amplitude (CMAP) and motor conduction velocity (MCV) were analyzed by conventional motor nerve conduction studies. Intrafascicular compound nerve action potential amplitude (N-CNAP) at the elbow after wrist simulation and its nerve conduction velocity (NCV) between wrist and elbow were also analyzed. The negative correlation of DL with CMAP was statistically significant (r = 0.577, p < 0.001). CMAP was correlated with either MCV (r = 0.537, p < 0.001) or N-CMAP (r = 0.710, p < 0.001). A significant correlation of MCV with NCV (r = 0.517, p < 0.001) was also indicated. There were no any other significant correlation among the parameters. In CTS the degree of demyelination and axonal degeneration influence the prognosis for nerve recovery after decompressive surgery. DL is mainly influenced by demyelination that results in conduction block and slowing at the carpal tunnel. CMAP and N-CNAP indicate the degree of axonal degeneration at distal and proximal to the compression site. As in electrophysiologic evaluation of mononeuropathies, proximal axonal degeneration is best assessed by both stimulation and recording electrode locationing proximal to the lesion. Recording of intrafascicular nerve action potential was a little invasive method, but it provided important informations. The negative correlation between DL and CMAP implies that distal axonal degeneration can occur in proportion to the conduction disturbance. Moreover, N-CNAP had a higher correlation with CMAP. The greater the distal axonal degeneration, the more the proximal axonal degeneration. Conduction velocity represents the velocity of the fastest conduction fiber, not the degree of axonal degeneration.

Action Potentials↗
PubMed2001

[Prevalence of Martin-Gruber anastomosis on motor nerve conduction studies].

Prevalence of median to ulnar anastomosis in the forearm(Martin-Gruber anastomosis; MGA) to the first dorsal interosseous(FDI), abductor digiti quinti (ADQ) and adductor pollicis(AP) was investigated. Subjects contained 106 patients with normal nerve conduction or patients with various neuropathies. Recording electrodes were placed on the motor point of FDI, ADQ and AP. Supramaximal stimulations were given to the median and ulnar nerves at the wrist or above the elbow. The diagnosis of MGA was made by the following criteria; amplitude of compound muscle action potential(CMAP) increased after elbow stimulation as compared with the wrist stimulation in median nerve conduction studies. The corresponding decrease in CMAP amplitude was found after above elbow stimulation as compared with the wrist stimulation in ulnar nerve conduction studies. No MGA was found in 80(75%) out of 106 patients. MGA to FDI was found in all 26 patients who had MGA. MGA to ADQ and AP was found in 11% and 10% of the patients, respectively. Only 8 out of 26 patients had MGA to all 3 muscles. In the presence of MGA median motor nerve conduction studies demonstrate larger CMAP, with a small initial positivity, after elbow stimulation than after wrist stimulation. And moreover, ulnar motor nerve conduction studies reveal a conduction block-like finding in the forearm. In this study MGA was found in 25% of the patient to FDI, in 11% to ADQ and in 10% to AP. Although a very small MGA might be overlooked in our method, such a small MGA doesn't mislead us into erroneous interpretation of motor nerve conduction studies.

Aged↗
PubMed2000

Antibody studies of factor VIII inhibitor in a case with Waldenström's macroglobulinemia.

We report a case of Waldenström's macroglobulinemia with prominent bleeding tendency; laboratory investigation revealed an elevated activated partial thromboplastin time. Further laboratory evaluation showed circulating factor VIII anticoagulant, deemed polyclonal IgG, with a titer of 700 Bethesda Units/ml. The factor VIII inactivation kinetics of the patient plasma were identical to those of a type II inhibitor, and the inhibitor was found to recognize the A2 domain of the factor VIII heavy chain. Apparently, paraprotein is not always the cause of reduced activity of coagulation factors in neoplastic dysproteinemias.

Aged↗
PubMed2000

Junctophilins: a novel family of junctional membrane complex proteins.

Junctional complexes between the plasma membrane (PM) and endoplasmic/sarcoplasmic reticulum (ER/ SR) are a common feature of all excitable cell types and mediate cross-talk between cell surface and intracellular ion channels. We have identified the junctophilins (JPs), a novel conserved family of proteins that are components of the junctional complexes. JPs are composed of a carboxy-terminal hydrophobic segment spanning the ER/SR membrane and a remaining cytoplasmic domain that shows specific affinity for the PM. In mouse, there are at least three JP subtypes: JP-1, -2, and -3. JP-2 is abundantly expressed in the heart, and mutant mice lacking JP-2 exhibited embryonic lethality. Cardiac myocytes from the mutant mice showed deficiency of the junctional membrane complexes and abnormal Ca2+ transients. Our results suggest that JPs are important components of junctional membrane complexes.

Amino Acid Motifs↗
PubMed2000

A fluorescent indicator for visualizing cAMP-induced phosphorylation in vivo.

We have developed a method for visualizing phosphorylation of proteins in living cells using a novel fluorescent indicator composed of two green fluorescent protein (GFP) variants joined by the kinase-inducible domain (KID) of the transcription factor cyclic adenosine monophosphate (cAMP)-responsive element binding protein (CREB). Phosphorylation of KID by the cAMP-dependent protein kinase A (PKA) decreased the fluorescence resonance energy transfer (FRET) among the flanking GFPs. By transfecting COS-7 cells with an expression vector encoding this indicator protein (termed ART for cAMP-responsive tracer), we were able to visualize activation dynamics of PKA in living cells.

Amino Acid Sequence↗
PubMed2000

Molecular basis of spatio-temporal dynamics in inositol 1,4,5-trisphosphate-mediated Ca2+ signalling.

Inositol 1,4,5-trisphosphate (IP3)-mediated Ca2+ signalling regulates many important cell functions, and the spatio-temporal dynamics of the Ca2+ signalling is a crucial factor for its versatility. The molecular mechanisms that control Ca2+ signalling are now being investigated, and I here describe the subtypes of IP3 receptors that have distinct functional properties and contribute to the diversity of Ca2+ signalling patterns. I also discuss the spatio-temporal dynamics of intracellular IP3 concentration, describing recent methodological advances in monitoring intracellular IP3 concentration. These findings highlight the potential importance of the spatio-temporal information of any signalling molecule.

Animals↗
PubMed2000

Histological evaluation of autogenous iliac particulate cancellous bone and marrow grafted to alveolar clefts--a preliminary report of five young adult cases.

OBJECTIVE: We histologically evaluated iliac particulate cancellous bone and marrow (PCBM) grafted to alveolar clefts in five young adults with cleft lip and palate. PATIENTS AND METHODS: Five young adults with cleft lip and palate underwent secondary alveolar bone grafting. The mean age of the patients was 21.2 years (range = 17 to 27 years). Bone specimens were taken from the graft site 5 to 10 months after the surgery simultaneously with implant fixture placement (two cases), vestibuloplasty (two cases), and rebone grafting (one case). These five bone specimens were prepared and stained with hematoxylin and eosin and examined microscopically. RESULT: One specimen, taken 5 months after surgery, showed immature trabecular bone partially lined by osteoblasts. The remaining four specimens showed well-mineralized trabeculae and fatty change in the marrow space. CONCLUSION: In young adult cases, PCBM remodeled after grafting and became mature bone about 5 to 6 months after the surgery.

Adolescent↗
PubMed2000

[Analysis of segmental motor conduction in the median and the ulnar nerves: comparison between normal and diabetic individuals].

We investigated characteristics of segmental motor conduction in the median and the ulnar nerves. Subjects were 55 individuals with normal conduction of the upper extremity and 71 patients with diabetes mellitus. Mean polyneuropathy index (PNI), which was determined as a mean percentage of the normal for 6 indices concerning to the conduction velocity in the upper limb, was 99.0% in the normal group and 85.6 % in the diabetic group on the mean. In the normal group distal latency was longer in the median nerve than in the ulnar nerve, and the conduction time between Erb's point and the wrist was longer in the ulnar nerve than the median nerve both in men and women. In the diabetic group these differences were accentuated; that means the distal latency was relatively more prolonged in the median nerve and the conduction time between Erb's point and the wrist was much longer in the ulnar nerve. Prolonged distal latency in the median nerve of women and conduction delay between Erb's point and the wrist in the ulnar nerve of men altogether resulted in the gender difference in the median minus ulnar F-wave latency after wrist stimulation in the diabetic group. Carpal tunnel segment of the median nerve and the elbow segment of the ulnar nerve are known to be common entrapment sites. This phenomenon of accentuated conduction delay in these common entrapment sites might be considered as a sort of 'double crush syndrome'.

Diabetes Mellitus↗
PubMed2000

[Sequential nerve conduction studies in a patient with ulnar neuropathy at the elbow treated by night athletic supporter].

Ulnar nerve can be stretched with the elbow flexed position. To avoid elbow flexed position in patients with ulnar neuropathy at the elbow we used an athletic elbow supporter. We herein demonstrate a 31-year-old man with right ulnar neuropathy at the elbow whose neuropathy was resolved by using this supporter only at night. He had complained of weakness and paraesthesia in the ulnar side of his right hand. Nerve conduction studies of right ulnar nerve revealed decrease in the amplitude of compound nerve action potentials and a severe motor nerve conduction block with apparent conduction delay around the ulnar groove. A diagnosis of ulnar neuropathy at the elbow was done and we recommended him to wear an athletic elbow supporter at night. Paraesthesia of his right hand improved in a few days after starting this therapy. Three months later paraesthesia was resolved. One year later grip power of his right hand increased to 35 kg from 20 kg, and the conduction block at the elbow completely disappeared. Compound nerve action potentials, recorded at the segment of wrist to above elbow and wrist to finger, were improved equally. These observations suggest that the conduction block at the elbow entrapment site and the distal axonal degeneration gradually recovered together.

Elbow↗
PubMed1999

Abnormal features in skeletal muscle from mice lacking mitsugumin29.

Physiological roles of the members of the synaptophysin family, carrying four transmembrane segments and being basically distributed on intracellular membranes including synaptic vesicles, have not been established yet. Recently, mitsugumin29 (MG29) was identified as a novel member of the synaptophysin family from skeletal muscle. MG29 is expressed in the junctional membrane complex between the cell surface transverse (T) tubule and the sarcoplasmic reticulum (SR), called the triad junction, where the depolarization signal is converted to Ca(2+) release from the SR. In this study, we examined biological functions of MG29 by generating knockout mice. The MG29-deficient mice exhibited normal health and reproduction but were slightly reduced in body weight. Ultrastructural abnormalities of the membranes around the triad junction were detected in skeletal muscle from the mutant mice, i.e., swollen T tubules, irregular SR structures, and partial misformation of triad junctions. In the mutant muscle, apparently normal tetanus tension was observed, whereas twitch tension was significantly reduced. Moreover, the mutant muscle showed faster decrease of twitch tension under Ca(2+)-free conditions. The morphological and functional abnormalities of the mutant muscle seem to be related to each other and indicate that MG29 is essential for both refinement of the membrane structures and effective excitation-contraction coupling in the skeletal muscle triad junction. Our results further imply a role of MG29 as a synaptophysin family member in the accurate formation of junctional complexes between the cell surface and intracellular membranes.

Amino Acid Sequence↗
PubMed1999

Potentiating effects of 4-[2-(phenylsulfonylamino)ethylthio]-2,6-difluoro-phenoxyaceta mide (PEPA) on excitatory synaptic transmission in dentate granule cells.

A novel sulfonylamino compound, 4-[2-(phenylsulfonylamino)-ethylthio]-2,6-difluoro-phenoxyaceta mide (PEPA) has been shown to selectively potentiate glutamate-induced currents in Xenopus oocytes expressing recombinant AMPA receptor subunits, GluR1-GluR4, by attenuation of desensitization. Here, we examined the effects of PEPA on responses to excitatory amino acids as well as on excitatory synaptic transmission in dentate granule cells of rat hippocampal slices using the whole-cell patch clamp technique. PEPA at 100 microM produced a 3-4-fold increases in the peak amplitude of current responses to AMPA and glutamate applied iontophoretically in the dentate granule cells, whereas it showed no effect on NMDA-induced currents. Excitatory postsynaptic currents (EPSCs) evoked in these neurons by stimulation of the perforant path had fast and slow components mediated by AMPA and NMDA receptors, respectively. PEPA at concentrations between 10 and 100 microM potentiated only the AMPA component of the EPSC (AMPA EPSC) in a dose-dependent manner without affecting the NMDA component. Although the potentiating effect of PEPA on the amplitude of the AMPA EPSC was weaker than that on the AMPA-induced current, it clearly prolonged the duration of the EPSC. PEPA at 100 microM increased the peak amplitude of the AMPA EPSC by 17%, and increased the area enclosed by the AMPA EPSC by 72%.

Animals↗