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Biomedical subjects

H Bito

Publications and source records attributed to H Bito.

At least 19 recordsLinked to original sources

Citron, a Rho-target, interacts with PSD-95/SAP-90 at glutamatergic synapses in the thalamus.

Proteins of the membrane-associated guanylate kinase family play an important role in the anchoring and clustering of neurotransmitter receptors in the postsynaptic density (PSD) at many central synapses. However, relatively little is known about how these multifunctional scaffold proteins might provide a privileged site for activity- and cell type-dependent specification of the postsynaptic signaling machinery. Rho signaling pathway has classically been implicated in mechanisms of axonal outgrowth, dendrogenesis, and cell migration during neural development, but its contribution remains unclear at the synapses in the mature CNS. Here, we present evidence that Citron, a Rho-effector in the brain, is enriched in the PSD fraction and interacts with PSD-95/synapse-associated protein (SAP)-90 both in vivo and in vitro. Citron colocalization with PSD-95 occurred, not exclusively but certainly, at glutamatergic synapses in a limited set of neurons, such as the thalamic excitatory neurons; Citron expression, however, could not be detected in the principal neurons of the hippocampus and the cerebellum in the adult mouse brain. In a heterologous system, Citron was shown to form a heteromeric complex not only with PSD-95 but also with NMDA receptors. Thus, Citron-PSD-95/SAP-90 interaction may provide a region- and cell type-specific link between the Rho signaling cascade and the synaptic NMDA receptor complex.

Animals

The effect of fentanyl on sevoflurane requirements for somatic and sympathetic responses to surgical incision.

BACKGROUND: Fentanyl produces a reduction in the minimum alveolar concentration (MAC) of isoflurane and desflurane needed to blockade adrenergic response (BAR) to surgical incision in 50% of patients (MAC-BAR). MAC-BAR of sevoflurane and the reduction in MAC-BAR of sevoflurane by fentanyl have not been described previously. The purpose of this study was to determine the MAC and MAC-BAR reduction of sevoflurane by fentanyl with and without nitrous oxide (N2O). METHODS: Two hundred twenty-six patients were randomly assigned to one of two groups: a sevoflurane group and a sevoflurane/N2O group. Patients in each group were randomly assigned to one of five different fentanyl concentration subgroups. Patients were anesthetized with sevoflurane and fentanyl in the sevoflurane group and with sevoflurane, fentanyl, and N2O (66 vol%) in the sevoflurane/N2O group. Somatic and sympathetic responses to surgical incision were observed for MAC and MAC-BAR assessment at predetermined concentrations of sevoflurane. RESULTS: Fentanyl produced an initial steep reduction in the MAC and MAC-BAR of sevoflurane, with 3 ng/ml resulting in a 61% reduction in MAC and an 83% reduction in MAC-BAR. A ceiling effect was observed for MAC and MAC-BAR, with 6 ng/ml fentanyl providing only an additional 13% and 9% reduction in MAC and MAC-BAR, respectively. In the presence of 66 vol% N2O, MAC and MAC-BAR of sevoflurane were reduced with increasing concentrations of fentanyL A ceiling effect was not observed for reduction in MAC and MAC-BAR in the presence of N2O. CONCLUSIONS: MAC and MAC-BAR decreased similarly with increasing concentrations of fentanyl in plasma, showing an initial steep reduction followed by a ceiling effect. In the presence of N2O, MAC and MAC-BAR decreased similarly but did not exhibit a ceiling effect.

Adult

Role of citron kinase as a target of the small GTPase Rho in cytokinesis.

During mitosis, a ring containing actin and myosin appears beneath the equatorial surface of animal cells. This ring then contracts, forms a cleavage furrow and divides the cell, a step known as cytokinesis. The two daughter cells often remain connected by an intercellular bridge which contains a refringent structure known as the midbody. How the appearance of this ring is regulated is unclear, although the small GTPase Rho, which controls the formation of actin structures, is known to be essential. Protein kinases are also thought to participate in cytokinesis. We now show that a splice variant of a Rho target protein, named citron, contains a protein kinase domain that is related to the Rho-associated kinases ROCK14 and ROK, which regulate myosin-based contractility. Citron kinase localizes to the cleavage furrow and midbody of HeLa cells; Rho is also localized in the midbody. We find that overexpression of citron mutants results in the production of multinucleate cells and that a kinase-active mutant causes abnormal contraction during cytokinesis. We propose that citron kinase regulates cytokinesis at a step after Rho in the contractile process.

Alternative Splicing

Different regions of Rho determine Rho-selective binding of different classes of Rho target molecules.

Based on their Rho binding motifs several Rho target molecules can be classified into three groups; class I includes the protein kinase PKN, rhophilin, and rhotekin, class II includes the protein kinases, Rho-associated coiled-coil containing protein kinases, ROCK-I and ROCK-II, and class III includes citron. Taking advantage of the selectivity in recognition by these targets between Rho and Rac, we examined the regions in Rho required for selective binding of each class of Rho target molecules. Yeast two-hybrid assays were performed using Rho/Rac chimeras and either rhophilin, ROCK-I, or citron. This study showed the existence of at least two distinct regions in Rho (amino acids 23-40 and 75-92) that are critical for the selective binding of these targets. The former was required for binding to citron, whereas the latter was necessary for binding to rhophilin. On the other hand, either region showed affinity to ROCK-I. This was further confirmed by ligand overlay assay using both recombinant ROCK-I and ROCK-II proteins. Consistently, Rho/Rac chimeras containing either region can induce stress fibers in transfected HeLa cells, and this induction is suppressed by treatment with Y-27632, a specific inhibitor of ROCK kinases. These results suggest that the selective binding of different classes of Rho targets to Rho is determined by interaction between distinct Rho-binding motifs of the targets and different regions of Rho.

Adaptor Proteins, Signal Transducing

Molecular dissection of the Rho-associated protein kinase (p160ROCK)-regulated neurite remodeling in neuroblastoma N1E-115 cells.

A critical role for the small GTPase Rho and one of its targets, p160ROCK (a Rho-associated coiled coil-forming protein kinase), in neurite remodeling was examined in neuroblastoma N1E-115 cells. Using wild-type and a dominant-negative form of p160ROCK and a p160ROCK-specific inhibitor, Y-27632, we show here that p160ROCK activation is necessary and sufficient for the agonist-induced neurite retraction and cell rounding. The neurite retraction was accompanied by elevated phosphorylation of myosin light chain and the disassembly of the intermediate filaments and microtubules. Y-27632 blocked both neurite retraction and the elevation of myosin light chain phosphorylation in a similar concentration-dependent manner. On the other hand, suppression of p160ROCK activity by expression of a dominant-negative form of p160ROCK induced neurites in the presence of serum by inducing the reassembly of the intermediate filaments and microtubules. The neurite outgrowth by the p160ROCK inhibition was blocked by coexpression of dominant-negative forms of Cdc42 and Rac, indicating that p160ROCK constitutively and negatively regulates neurite formation at least in part by inhibiting activation of Cdc42 and Rac. The assembly of microtubules and intermediate filaments to form extended processes by inhibitors of the Rho-ROCK pathway was also observed in Swiss 3T3 cells. These results indicate that Rho/ROCK-dependent tonic inhibition of cell process extension is exerted via activation of the actomysin-based contractility, in conjunction with a suppression of assembly of intermediate filaments and microtubules in many cell types including, but not exclusive to, neuronal cells.

3T3 Cells

Effects of the water content of soda lime on compound A concentration in the anesthesia circuit in sevoflurane anesthesia.

BACKGROUND: Sevoflurane anesthesia is usually performed with fresh gas flow rates greater than 2 l/min due to the toxicity of compound A in rats and limited clinical experience with sevoflurane in low-flow systems. However, to reduce costs, it would be useful to identify ways to reduce compound A concentrations in low-flow sevoflurane anesthesia. This goal of this study was to determine if compound A concentrations can be reduced by using soda lime with water added. METHODS: Low-flow sevoflurane anesthesia (fresh gas flow of 1 l/min) was performed in 37 patients using soda lime with water added (perhydrated soda lime) or standard soda lime as the carbon dioxide (CO2) absorbent. The soda lime was not changed between patients, but rather was used until CO2 rebreathing occurred. The perhydrated soda lime was prepared by spraying 100 ml distilled water onto 1 kg fresh soda lime, and water was added only when a new bag of soda lime was placed into the canister. Compound A concentrations in the circle system, soda lime temperatures, inspired and end-tidal CO2 and end-tidal sevoflurane concentrations, and CO2 elimination by the patient were measured during anesthesia. RESULTS: Compound A concentrations were significantly lower for the perhydrated soda lime (1.9 +/- 1.8 ppm; means +/- SD) than for the standard soda lime (13.9 +/- 8.2 ppm). No differences were seen between the two types of soda lime with regard to the temperature of the soda lime, end-tidal sevoflurane concentrations, or CO2 elimination. Compound A concentration decreased with the total time of soda lime use for both types of soda lime. The CO2 absorption capacity was significantly less for perhydrated soda lime than for standard soda lime. CONCLUSIONS: Compound A concentrations in the circuit can be reduced by using soda lime with water added. The CO2 absorption capacity of the soda lime is reduced by adding water to it, but this should not be clinically significant.

Anesthesia, Closed-Circuit

[A comparison of the grade of laryngeal visualisation;--the McCoy compared with the Macintosh and the Miller blade in adults].

Effectiveness in visualization of the vocal cord during orotracheal intubation with McCoy (McC) compared with Macintosh (Min) and Miller (Mil) blades were investigated. After an institutional review board approval, 117 patients for elective surgery under general anesthesia requiring tracheal intubation were investigated. Five board certified anesthesiologists tried to visualize the vocal cord of a patient three times with the three different types of laryngoscope. Total of 351 intubation attempts were studied. The view obtained at laryngoscopy with each of the three blades was recorded as follows. Grade 1. If most of the glottis is visible. Grade 2. If only the posterior extremity of the glottis is visible. Grade 3. If no part of the glottis can be seen. Grade 4. If not even the epiglottis can be exposed. Eight-two Grade 1 views were obtained with McC, 72 with Mil and 47 with Min, respectively. Thirty-three Grade 2 views were obtained with McC, 36 with Min and 24 with Mil. Two Grade 3 views with McC, 34 with Min and 14 with Mil were obtained. Seven Grade 4 views were obtained with Mil. The grades of laryngeal visualization with McC were significantly lower than those with Min and Mil.

Adolescent

[Determination of the distance between the upper incisors and the laryngoscope blade during laryngoscopy: comparisons of the McCoy, the Macintosh, the Miller, and the Belscope blades].

We compared the distance between the upper central incisors and the laryngoscope blade with the four different types of laryngoscope blade (McCoy, Macintosh, Miller, Belscope). Twenty-three patients scheduled for general anesthesia were studied. The tooth-blade distance was measured when optimum visibility of the glottis was obtained. The visibility was determined according to the Cormack and Lehane grading. The distance with the McCoy and the Belscope was greater than that with the Macintosh or the Miller. The visibility grade was significantly worse with the Macintosh than with the other types of laryngoscope. The results indicate that the McCoy and the Belscope provide less incidence of upper dental injuries and greater visibility than either with the Macintosh or the Miller. Furthermore, the force applied to the handle is thought to be smaller with the McCoy than with the Belscope.

Adult

Dendritic Ca2+ channels characterized by recordings from isolated hippocampal dendritic segments.

Dendritic arbors are critical for the information processing capability of central neurons, but quantitative analysis of their membrane properties has been hampered by their geometrical complexity. Here, we have focused on an important source of Ca2+ entry in dendrites, the voltage-gated Ca2+ channels, by applying the whole-cell voltage-clamp technique to isolated dendritic segments ("dendrosomes") from rat hippocampal neurons. We found that low voltage-activated T-type Ca2+ channels provide a significantly larger fraction of the Ca2+ influx in dendrites than their counterparts in cell bodies. Surprisingly, 60%-70% of the high voltage-activated Ca2+ current in dendrosomes was N and P/Q type, and these channels were susceptible to neurotransmitter inhibition, suggesting a novel physiological role for G protein-regulated Ca2+ channel modulation in controlling dendritic excitability and Ca2+ signaling.

Animals

Ca2+-dependent regulation in neuronal gene expression.

Ca2+ is an important signal-transduction molecule that plays a role in many intracellular signaling pathways. Recent advances have indicated that in neurons, Ca2+-controlled signaling mechanisms cooperate in order to discriminate amongst incoming cellular inputs. Ca2+-dependent transcriptional events can thereby be made selectively responsive to bursts of synaptic activity of specific intensity or duration.

Animals

Does nitrous oxide antagonize sevoflurane-induced hypnosis?

We have studied 64 ASA I and II patients (aged 20-60 yr) to determine if nitrous oxide affects sevoflurane requirement for achieving 50% probability of no movement in response to verbal commands (MACawake). Patients were allocated randomly to one of four nitrous oxide concentration groups (0, 20, 40 and 60 vol.%). Patients in each group received sevoflurane at two different end-tidal concentrations according to a predetermined randomization table. After steady state sevoflurane and nitrous oxide concentrations had been maintained for at least 15 min, patients were assessed as being awake or asleep. The MACawake for sevoflurane was 0.63% and this was reduced significantly in a non-linear manner by increasing nitrous oxide concentration. A 50% reduction in MACawake was produced by a nitrous oxide concentration of 45%. The reduction in MACawake by nitrous oxide was non-linear; the interaction coefficient between nitrous oxide and sevoflurane being significantly less than zero (P = 0.0238), indicating that the reduction in MACawake by nitrous oxide was smaller than would be expected from simple additivity and that nitrous oxide antagonized the effects of sevoflurane in preventing response to verbal commands.

Adult

Partly exhausted soda lime or soda lime with water added, inhibits the increase in compound A concentration in the circle system during low-flow sevoflurane anaesthesia.

We performed low-flow sevoflurane anaesthesia at a flow rate of 1 litre min-1 in three groups (n = 8 each) using 600 g of fresh soda lime (control group), 600 g of soda lime with 60 ml of water added (water group) or 600 g of soda lime saturated with carbon dioxide, that is partly exhausted soda lime (carbon dioxide group). Degradation products in the system were measured hourly. Inspired and end-tidal carbon dioxide and sevoflurane concentrations, carbon dioxide and temperature of the soda lime were monitored. CF2 = C(CF3)-O-CH2F (compound A) was the only sevoflurane degradation product detected. The mean maximum concentration of compound A was significantly higher in the control group (mean 16.0 (SD 5.0) ppm) than in the water (1.4 (1.0) ppm) or carbon dioxide (4.0 (1.8) ppm) group, and the maximum temperature of the soda lime was significantly lower in the carbon dioxide group (30.7 (3.5) degrees C) than in the control (43.4 (1.8) degrees C) or water (40.8 (1.8) degrees C) group (P < 0.05). The use of partly exhausted soda lime or soda lime with water added reduced compound A concentrations in the system during low-flow sevoflurane anaesthesia.

Adult

Effects of low-flow sevoflurane anesthesia on renal function: comparison with high-flow sevoflurane anesthesia and low-flow isoflurane anesthesia.

BACKGROUND: The safety of low-flow sevoflurane anesthesia, during which CF2=C(CF3)-O-CH2F (compound A) is formed by sevoflurane degradation, in humans has been questioned because compound A is nephrotoxic in rats. Several reports have evaluated renal function after closed-circuit or low-flow sevoflurane anesthesia, using blood urea nitrogen (BUN) and serum creatinine as markers. However, these are not the more sensitive tests for detecting renal damage. This study assessed the effects of low-flow sevoflurane anesthesia on renal function using not only BUN and serum creatinine but also creatinine clearance and urinary excretion of kidney-specific enzymes, and it compared these values with those obtained in high-flow sevoflurane anesthesia and low-flow isoflurane anesthesia. METHODS: Forty-eight patients with gastric cancer undergoing gastrectomy were studied. Patients were randomized to receive sevoflurane anesthesia with fresh gas flow of 1 l/min (low-flow sevoflurane group; n = 16) or 6-10 l/min (high-flow sevoflurane group; n = 16) or isoflurane anesthesia with a fresh gas flow of 1 l/min (low-flow isoflurane group; n = 16). In all groups, the carrier gas was oxygen/nitrous oxide in the ratio adjusted to ensure a fractional concentration of oxygen in inspired gas (FiO2) of more than 0.3. Fresh Baralyme was used in the low-flow sevoflurane and low-flow isoflurane groups. Glass balls were used instead in the high-flow sevoflurane group, with the fresh gas flow rate adjusted to eliminate rebreathing. The compound A concentration was measured by gas chromatography. Gas samples taken from the inspiratory limb of the circle system at 1-h intervals were analyzed. Blood samples were obtained before and on days 1, 2, and 3 after anesthesia to measure BUN and serum creatinine. Twenty-four-hour urine samples were collected before anesthesia and for each 24-h period from 0 to 72 h after anesthesia to measure creatinine, N-acetyl-beta-D-glucosaminidase, and alanine aminopeptidase. RESULTS: The average inspired concentration of compound A was 20 +/- 7.8 ppm (mean +/- SD), and the average duration of exposure to this concentration was 6.11 +/- 1.77 h in the low-flow sevoflurane group. Postanesthesia BUN and serum creatinine concentrations decreased, creatinine clearance increased, and urinary N-acetyl-beta-D-glucosaminidase and alanine aminopeptidase excretion increased in all groups compared with preanesthesia values, but there were no significant differences between the low-flow sevoflurane, high-flow sevoflurane, and low-flow isoflurane groups for any renal function parameter at any time after anesthesia. CONCLUSIONS: The only difference between the low-flow and high-flow sevoflurane groups was compound A formation, and postanesthesia laboratory data showed no significant effects of compound A formation during sevoflurane anesthesia on renal function. No significant effects on renal function were observed in either the low-flow or high-flow sevoflurane groups compared with the low-flow isoflurane group.

Aged

[Cervical spine movement during orotracheal intubation using the McCoy laryngoscope compared with the Macintosh and the Miller laryngoscopes].

The movement of cervical spine during orotracheal intubation was compared using the McCoy, Macintosh or Miller laryngoscope blade. Twenty ASA 1-2 patients requiring tracheal intubation were studied. Following induction of anesthesia and obtaining muscle relaxation, the cross-table lateral X-ray was taken before and during laryngoscopy using three types of laryngoscopes. Degree of cervical spine movement was evaluated by measuring the distance between the spinous processes of C1 and the occiput, and the amount of displacement of C1 and C5 against C3 by tracing on each films. The results indicated that delta C1-occiput was larger and delta C1 + C5 smaller with the McCoy laryngoscope compared with the others. The use of the McCoy laryngoscope results in less cervical spine movement during laryngoscopy and therefore should be of particular benefit in the presence of cervical spine instability as well as in the normal patients.

Adolescent

[Perioperative management of a patient with severe anemia caused by rare Jr (a-) blood].

We experienced the anesthetic management of a patient with severe anemia as low as 1.8 g.dl-1 of hemoglobin and 6.5 per cent of hematocrit before surgery. His blood had been determined as type B, anti-Jra antibody positive and the proper blood was unavailable. We gave mainly colloid fluids and partially blood products of different blood groups to maintain the circulating volume with monitoring central venous pressure peri-operatively. The operation finished relatively uneventfully and the patient was transferred to the common ward from the intensive care unit on the fifth postoperative day. We conclude that maintaining the patient's circulating volume to avoid shock should be given priority in a case of severe anemia.

Aged