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Biomedical subjects

A Naito

Publications and source records attributed to A Naito.

At least 73 records · Page 4Linked to original sources
PubMed1998

Interaction of Doc2 with tctex-1, a light chain of cytoplasmic dynein. Implication in dynein-dependent vesicle transport.

Doc2 has one Munc13-interacting domain at the N-terminal region and two C2-like domains interacting with Ca2+ and phospholipid at the C-terminal region. Doc2 consists of two isoforms, Doc2alpha and -beta. Doc2alpha is specifically expressed in neuronal cells and implicated in Ca2+-dependent neurotransmitter release, whereas Doc2beta is ubiquitously expressed and its function is unknown. We show here that both Doc2alpha and -beta interact with rat tctex-1, a light chain of cytoplasmic dynein, in both cell-free and intact cell systems. Overexpression of the N-terminal fragment of Doc2 containing the tctex-1-interacting domain induces changes in the intracellular localization of cation-independent mannose 6-phosphate receptor and its ligand, cathepsin D, which are transported from trans-Golgi network to late endosomes. Overexpression of the C-terminal fragment containing two C2-like domains shows the similar effect, but to a lesser extent, whereas overexpression of full-length Doc2 or the C-terminal fragment of rabphilin3 containing two C2-like domains does not show this effect. Because dynein is a minus-end-directed microtubule-based motor protein, these results suggest that Doc2, especially Doc2beta, plays a role in dynein-dependent intracellular vesicle transport.

Amino Acid Sequence↗
PubMed1998

Evidence of local conformational fluctuations and changes in bacteriorhodopsin, dependent on lipids, detergents and trimeric structure, as studied by 13C NMR.

We examined how the local conformation and dynamics of [3-13C]Ala-labeled bacteriorhodopsin (bR) are altered as viewed from 13C NMR spectra when the natural membrane lipids are partly or completely replaced with detergents. It turned out that the major conformational features of bR, the alphaII-helices, are generally unchanged in the delipidated or solubilized preparations. Upon partial delipidation or detergent solubilization, however, a significant conformational change occurs, ascribed to local conversion of alphaII-->alphaI-helix (one Ala residue involved), evident from the upfield displacement of the transmembrane helical peak from 16.4 ppm to 14.5 ppm, conformational change (one or two Ala residues) within alphaII-helices from 16.4 to 16.0 ppm, and acquired flexibility in the loop region (especially at the F-G loop) as manifested from suppressed peak-intensities in cross-polarization magic angle spinning (CP-MAS) NMR spectra. On the other hand, formation of monomers as solubilized by Triton X-100, Triton N-101 and n-dodecylmaltoside is characterized by the presence of a peak at 15.5 ppm and a shifted absorption maximum (550 nm). The size of micelles under the first two conditions was small enough to yield 13C NMR signals observable by a solution NMR spectrometer, although 13C CP-MAS NMR signals were also visible from a fraction of large-sized micelles. We found that the 16.9 ppm peak (three Ala residues involved), visible by CP-MAS NMR, was displaced upfield when Schiff base was removed by solubilization with sodium dodecyl sulfate, consistent with our previous finding of bleaching to yield bacterioopsin.

Bacteriorhodopsins↗
PubMed1998

A novel brain-specific isoform of beta spectrin: isolation and its interaction with Munc13.

Munc13 is a component of the neurotransmitter release machinery which is specifically expressed in brain. Munc13 interacts with Doc2 and syntaxin which are also implicated in the neurotransmitter release process. Here we isolated another Munc13-interacting molecule from a rat brain cDNA library by use of the yeast two-hybrid system, identified it to be a novel type of beta spectrin, and named it beta SpIII sigma 1. beta SpIII sigma 1 was specifically expressed in brain, where it was enriched in the synaptic vesicle and plasma membrane fractions. Because spectrin has been shown to interact with the actin cytoskeleton which is involved in the exocytotic process, the present results suggest that the Munc13-beta SpIII sigma 1 interactions play a role in neurotransmitter release.

Amino Acid Sequence↗
PubMed1998

Quantitation of telomerase activity in hepatocellular carcinoma: a possible aid for a prediction of recurrent diseases in the remnant liver.

Because telomerase activity is necessary for cell immortality and probably associated with tumor progression, we have evaluated a possible aid for quantitation of the activity to predict intrahepatic recurrences after surgery in patients with hepatocellular carcinoma (HCC). HCC tissues obtained by surgical resection from 20 patients were studied. Telomerase activity was expressed as peaks with a periodicity through a fluorescence-based telomeric repeat amplification protocol using an autosequencer, and the quantity of activity was calculated from peak areas. A ratio of fluorescence intensity depending on telomerase to that of an internal standard was used as a value of relative telomerase activity (RTA). RTA in serially diluted S100 extracts from HepG2 cells was well correlated with the amount of the extracts. The mean RTA value of 36.4 +/- 27.8 (mean +/- SD, 3.21 to 105) in 9 patients suffering from early recurrences after surgery was significantly higher than that (9.84 +/- 7.65; mean +/- SD, 3.00 to 29.0) in 11 patients without intrahepatic recurrences during the early period (P = .004). These results indicate that RTA value can be a useful predictor for intrahepatic recurrences during the early period after surgical resection of HCC.

Adult↗
PubMed1998

Inhibitory projections from pronator teres to biceps brachii motoneurones in human.

Neural projections from the pronator teres (PT) muscle to biceps brachii (BB) motoneurones were studied in three healthy human subjects using a post-stimulus time histogram method. In 25 BB motor units, electrical stimulation to the PT nerve with intramuscular needle electrodes induced inhibition in nine units (36%), whereas facilitation was produced in 18 units (72%) by stimulation to the median nerve trunk with surface electrodes at the distal end of the intermuscular septum of the arm or in the cubital fossa. Six motor units (24%) received both inhibition (PT nerve stimulation) and facilitation (median nerve trunk stimulation). In the six, the latency of the inhibition was, on average, 1.2 ms longer than that of the facilitation. The stimulation site for the inhibition was, on average, 4.8 cm distal to that for the facilitation. The inhibition was evoked with an intensity well below the motor threshold. These findings suggest that BB motoneurones receive oligosynaptic inhibition of group I afferents from PT in human.

Adult↗
PubMed1998

Purification and characterization of four catechol 1,2-dioxygenase isozymes from the benzamide-assimilating bacterium Arthrobacter species BA-5-17.

When Arthrobacter sp. BA-5-17 was grown on benzamide, the bacterium synthesized four different catechol 1,2-dioxygenase (CD, EC 1.13.11.1) isozymes (CD-I, II, III-1, and III-2). We purified each CD to homogeneity by a series of column chromatography. The molecular masses of the four CDs were between 68 and 72 kDa. The enzymes were made up of two identical subunits each with the molecular mass of 33 kDa. CD-I and II were indistinguishable in enzymatic properties tested. Most properties of CD-III-1 were similar to those of CD-III-2. However, CD-III-1 had a marked adsorption peak at 325 nm, which disappeared in CD-III-2 as well as in CD-I and II. CD-III-1 and III-2 were much more resistant to heating and inhibitors than CD-I and II.

Amino Acid Sequence↗
PubMed1998

Perikaryal myelination of cultured chick embryo statoacoustic ganglion cells: an electron microscopic study.

Perikaryal myelin formation on cultured chick embryo statoacoustic ganglion (SAG) cells was studied using electron microscopy. SAGs were dissected from 13-day embryos and cultured for 1 to 6 weeks. Myelinated perikarya, which were completely encircled by two or more layers of loose and/or compact myelin lamellae like those in vivo in the chick, were first observed in 3-week-cultured SAGs; myelinated axons appeared in 2-week-cultured SAGs. The perikaryal myelination progressively increased loose and compact lamellae and the axonal myelination increased compact lamellae. In 4- to 6-week-cultured SAGs, 11 to 12% of SAG cells had the myelinated perikaryon of which perikaryal myelin lamellae terminated at the axon hillock in the same manner as those in vivo in the chick. The number of layers of myelin lamellae around the myelinated perikaryon in 5- to 6-week-cultured SAGs and around the myelinated axon in 4- to 6-week-cultured SAGs ranged between 2 and 15, and between 12 and 26, respectively. Since these numbers were consistent with those in vivo in chicks 3 days after hatching, it is suggested that the culture provides perikaryal myelin sheaths, which are equivalent to those in vivo in structure, in more than 10% of SAG cells, while the myelination process in vitro is carried out much more slowly than that in vivo.

Animals↗
PubMed1998

Stability of the C-terminal alpha-helical domain of bacteriorhodopsin that protrudes from the membrane surface, as studied by high-resolution solid-state 13C NMR.

We have recorded 13C NMR spectra of [1-(13)C]Ala- and [3-(13)C]Ala-bacteriorhodopsin (bR), [1-(13)C]Ala- and [3-(13)C]Ala-papain-cleaved bR, and [3-(13)C]Ala-labeled R227Q bR mutant by cross polarization-magic angle spinning (CP-MAS) and dipolar decoupled-magic angle spinning (DD-MAS) methods. The pH and temperature were varied, and Arg 227 was replaced with Gln (R227Q), in order to clarify their effects on the stability of the alpha-helical domain of the C-terminus that protrudes from the membrane surface. The comparative 13C CP- and DD-MAS NMR study of [3-(13)C]Ala-bR, rather than [1-(13)C]Ala-bR, turned out to be the best means to distinguish the 13C NMR signals of the C-terminus from those of the rest of the transmembrane helices or loops. The inner segment of the C-terminus, from Ala 228 to Ala 235, forms an alpha-helical domain (resonated at 15.9 ppm) either at neutral pH and/or at 10 to -10 degrees C. The alpha-helical peak was not seen, however, after either cleavage of the C-terminus with papain or lowering the pH to 4.25. This alpha-helical structure, and a part of the random coil which was produced from the helix at pH 4.25, were further converted to a low-temperature-type alpha-helix, as indicated by an upfield displacement of the 13C NMR signal, when the temperature was lowered to 10- -10 degrees C. Surprisingly, the corresponding helical structure in R227Q is more stable than in the wild type at the acidic pH. This alpha-helical peak was classified as an alphaII-helix from the 13C chemical shifts of Cbeta carbon, although it was ascribed to an alphaI-helix on the basis of the carbonyl shifts. This is in contrast to Ala 53 which adopts the alphaII-helix as judged from the 13C chemical shifts of Cbeta and the carbonyl carbons. Therefore, this discrepancy might be caused by differential sensitivity of the two types of carbon signals to conformation and to modes of hydrogen bonding when motional fluctuation is involved. It is likely that the alphaII-helix form present at the C-terminus is not always the type originally proposed but should be considered as a form undergoing large-amplitude conformational fluctuation around alpha-helix.

Amino Acid Sequence↗
PubMed1998

Two statoacoustic ganglion (SAG) cells holding a common perikaryal myelin sheath in between in the chick.

Ganglion cells of the eighth cranial nerve in vertebrates have perikaryal myelin sheaths. The present electron microscopic study showed a case of two statoacoustic ganglion (SAG) cells holding a common perikaryal myelin sheath in between in the chick. Observations of numbers and terminals of myelin lamellae suggested that the sheath would be composed of an abortive lamella sandwiched between respective perikaryal myelin sheaths of the two SAG cells. The outer surface of the two perikaryal sheaths was covered by a continuous basal lamina. The process of common myelin sheath formation is discussed.

Animals↗
PubMed1998

Regulation of promoter and intron enhancer activity in immunoglobulin heavy-chain genes during B-cell differentiation.

Chloramphenicol acetyltransferase (CAT) transgenic mice, in which the transgene is regulated by the VH promoter and heavy-chain intron enhancer (E mu), were examined to investigate the variation of activity of these cis-acting elements during the late stage of B-cell development. CAT enzyme activity decreased when resting B cells were stimulated through B-cell receptors (BCR) with goat anti-mouse IgM antibodies in vitro. On the other hand, when these B cells were stimulated by lipopolysaccharide (LPS) in vitro, they showed enhanced CAT activity, accompanied by an increase in the number of CD43+ B220+ cells (pro-plasma and plasma cells). In addition, the CAT activities in CD43+ B220+ and PNAhi B220+ cells from immunized mice were higher than those in CD43- B220+ and PNAlo B220+ cells, respectively. These results suggest that the activity of E mu in the context of VH promoter was transiently down-regulated by stimulation through the BCR but enhanced at the pro-plasma and plasma stages.

Animals↗
PubMed1998

Electromyographic study of the elbow flexors and extensors in a motion of forearm pronation/supination while maintaining elbow flexion in humans.

Activities of the elbow flexors (biceps brachii, BB; brachialis, B; brachioradialis, BR) and extensors (triceps brachii, TB) in a motion of forearm pronation/supination with maintenance of elbow flexion (PS-movement) in nine healthy human subjects were studied by electromyography (EMG). The subject performed the PS-movement slowly or quickly with or without a load extending the elbow. In the slow PS-movement, an increase and decrease of EMG activities during supination and pronation, respectively, were seen in BB and the reverse was in B. A clear increment of EMG activities in BB accompanied with a reduction of EMG activities in B and/or BR, and the reverse were often observed. The contraction level and gain with the forearm supine were higher and larger than those with the forearm prone, respectively, in BB and the reverse was in B and BR. In a series of the quick PS-movement, alternating increases of EMG activities between BB and the other flexors (B and BR) were seen. Since TB showed no EMG activities throughout the experiment, it is suggested that reciprocal contractions between BB and the other flexors, which produce a complementary force in flexion direction, enable motions of pronation/supination with maintenance of flexion. Contraction properties of the flexors were discussed.

Adult↗
PubMed1998

[Terminal care of the cancer patient at home--patient and family spending the last moments together].

Some 129 cancer patients were taken care of at home by the visiting nurse team of our hospital since 1994 to 1997; 116 patients have already died, 50 of them at home (group A), and 66 at the hospital (group B). We analyze the differences the between two groups in nutritional assistance, pain control and manpower in looking after a patient. Twice as many patients in group A took nutritional medication (drip infusion or intravenous hyperalimentation, for example) than group B. More patients in group A used oral and suppository morphine than group B. In group A 70% of patients (56% in group B) had more than two persons looking after at home. The important factors for a good recuperation at home are: 1. Available nutritional assist available. 2. Good control. 3. Less burden and anxiety of the person looked after. To spend pain desirable last moments together, it is important for the visiting nurse team to take proper care and to confirm the wishes of the patient and family, considering the rapidly changing conditions of the individual patient.

Aged↗
PubMed1997

Physical and functional interactions of Doc2 and Munc13 in Ca2+-dependent exocytotic machinery.

Doc2 has two C2 domains that interact with Ca2+ and phospholipid. Munc13 has two C2 domains and one C1 domain that interacts with phorbol ester or diacylglycerol (DAG) and phospholipid. Both Doc2 and Munc13 are implicated in Ca2+-dependent neurotransmitter release, but their modes of action still remain unclear. We show here that Doc2 interacts with Munc13 both in a cell-free system and in intact PC12 cells during the high K+-induced Ca2+-dependent exocytosis. The Doc2-Munc13 interactions are stimulated by phorbol ester through the C1 domain of Munc13. Overexpression of the Doc2-interacting domain of Munc13 reduces the Ca2+-dependent exocytosis from PC12 cells, and co-expression with Doc2 suppresses this reduction. These results, together with the earlier findings that secretagogues produce DAG and elevate cytoplasmic Ca2+, suggest that the DAG-induced Doc2-Munc13 interactions play an important role in Ca2+-dependent exocytotic machinery.

Animals↗
PubMed1997

Molecular cloning of mouse Doc2alpha and distribution of its mRNA in adult mouse brain.

We have previously isolated from a human brain cDNA library, a new protein having two C2-like domains which interact with Ca2+ and phospholipid, and named Doc2alpha. Doc2alpha is abundantly expressed in brain, where it is highly concentrated on the synaptic vesicle fraction, and is implicated in Ca2(+)-dependent exocytosis. We have isolated here a mouse Doc2alpha cDNA and determined the localization of its mRNA in adult mouse brain. The amino acid sequence of the mouse Doc2alpha cDNA is 92% identical with that of the human counterpart. Northern blot analysis and in situ hybridization on adult mouse brain sections have revealed that Doc2alpha is predominantly expressed in mouse brain, where it is expressed in neuronal cells, but not in non-neuronal cells. Doc2alpha is highly expressed in the olfactory bulb, cerebral cortex, hippocampus, amygdaloid complex, and ventromedial hypothalamus nucleus, but not in the cerebellum, caudate-putamen, or ventral thalamus. These results indicate that Doc2alpha is expressed heterogeneously in mouse brain, where it is predominantly expressed in neuronal cells, and suggest that Doc2alpha plays a specific role in the area where it is expressed.

Amino Acid Sequence↗
PubMed1997

Stability of C3 convertase in the rat classical complement pathway.

It has been reported that rat serum complement causes efficient hemolysis of antibody-sensitized sheep erythrocytes (EA) at 20 C but not at 37 C. In connection with this, we demonstrated that C3 convertase of rat complement was significantly unstable at 37 C using purified components of rat complement.

Animals↗
PubMed1997

Electromyographic (EMG) study of cold shivering in the chronic spinal dog.

Cold shivering in six chronic spinal dogs (T9-10), which became capable of weight-bearing standing and/or walking with their hindlimbs, was studied by electromyography (EMG). In the forequarters, EMG activities in the form of grouping discharges (GD) accompanied by visible tremor (VT) were induced at an ambient temperature of 15 degrees C in all the dogs. In the hindquarters, GD with VT were induced at 3 degrees C in one dog. Although GD with VT were not induced in in the remaining dogs, GD without VT were produced by additional cutaneous stimuli to the hindquarters at 3 degrees C. These stimuli never produced GD in the hindquarters at room temperature. Though the mean frequency of GD in the hindquarters was lower than that in the forequarters, the range of the frequency in the hindquarters was much the same as that in the forequarters. During the experiment, the drop in rectal temperature was less than 0.6 degrees C in all of the dogs. These findings suggest that the chronically isolated spinal cord is responsible for cold shivering, while its excitability to cold exposure is less than that of the spinal cord governed by the supraspinal center.

Animals↗
PubMed1997

Cause and repair of flap necrosis over cochlear implant.

OBJECTIVE: The aim of this report was to study the cause and treatment of flap-related complications over cochlear implant. SETTING: The study was performed in an academic tertiary referral center. PATIENT: A 53-year-old Japanese male patient had had two retro-auricular skin incisions for tympanomastoidectomy in the postauricular region before implantation of a cochlear implant. He also had worn a helmet daily during work in his factory. INTERVENTIONS: Corrective surgery was performed for reformation of the local blood supply to the overlying skin flap and reinforcement of the tissue overlying the implant by use of a superiorly based temporal muscle and fascia flap that were sutured with an inferiorly based muscle and fascia flap. RESULTS: We successfully transposed the flap covering the implant without explantation of the implant. CONCLUSIONS: The delayed development of flap necrosis was thought to be due to pressure necrosis produced by the band in the patient's helmet lying on the skin over the implant and to poor local blood supply in the postauricular area stemming from the two previous skin incisions for mastoid surgery performed for cholesteatoma. In those patients who wear helmets, it is important to position the cochlear implant sufficiently behind the ear and to avoid the use of a helmet.

Cochlear Implants↗