Biomedical subjects

Y Yamagata

Publications and source records attributed to Y Yamagata.

At least 109 records · Page 6Linked to original sources

PubMed1990

Convergence of cardiovascular and vestibular inputs on neurons in the medullary paramedian reticular formation.

Extracellular recordings were made from 47 neurons in the caudal medullary paramedian reticular formation of chloralose-urethan anesthetized cats. The firing rate of all of these neurons was affected by stimulation of the contralateral vestibular nerve. The onset latency of the response to vestibular nerve stimulation was accurately determined for 43 of these neurons. The large majority (36 of 43) received short-latency (onset latency less than 5 ms) inputs; 7 were driven within 2 ms and were classified as receiving disynaptic inputs. Twenty of the 47 neurons also received inputs from the ipsilateral carotid sinus nerve. The paramedian reticular formation thus appears to be an important site for the integration of cardiovascular afferent and vestibular signals. However, only a small fraction of the neurons (2 of 13 tested) which received such convergent inputs could be antidromically driven from the upper thoracic spinal cord.

Action Potentials↗

PubMed1990

Concanavalin A affects beta-tubulin mRNA expression during neuritic processes of mouse neuroblastoma N18TG2 cells in a different manner from colchicine.

Addition of concanavalin A (Con A) to mouse neuroblastoma N18TG2 cells cultured with dibutyryl-cAMP which can stimulate neurite outgrowth, stopped the neuritic processes effectively. The extended neurites showed a gradual retraction for at least 8 hrs after addition of Con A, while addition of colchicine caused rapid retraction of the neurites. Immunocytochemistry showed that the addition of Con A did not disorganize the microtubules but the addition of colchicine did. The increase in beta-tubulin mRNA expression which was observed after cell culture and after stimulation by dB-cAMP was suppressed by the addition of Con A. Con A did not affect the beta-tubulin mRNA expression when the cells had already been cultured, while colchicine drastically decreased it. Thus, Con A appeared to affect the beta-tubulin mRNA expression in a different manner from colchicine, probably through inhibition of cell movement.

Animals↗

PubMed1990

Cervical afferent pathway to inferior oblique motoneuron in the cat.

The neuronal pathway implicated in the vertical cervico-ocular reflex (COR) was investigated electrophysiologically in chloralose anesthetized cats. The effect of bilateral C2 dorsal root afferent stimulation on inferior oblique motoneurons (IO-MN) was investigated by intracellular recording. Control disynaptic excitatory postsynaptic potentials elicited in IO-MNs following stimulation of the contralateral anterior semicircular canal nerve (ACN) were invariably facilitated by conditioning stimulation to both ipsilateral and contralateral C2 dorsal roots (DR) in all motoneurons tested. This result indicates that inputs from the C2 DR of both sides and the contralateral ACN converge onto secondary vestibular neurons ('common interneurons') which project directly to the IO-MN. Common interneurons mediating vestibular and cervical excitation to the IO-MNs were studied in the vestibular nuclei on the side opposite to the motoneurons by extracellular recording. Nineteen vestibular neurons were identified as common interneurons; they were distributed in the caudal half of the lateral nucleus and the rostral half of the descending nucleus. The present experiment provides electrophysiological evidence of the projection of upper cervical afferents to the ipsilateral vestibular nuclei. The difference in neuronal organization between the horizontal and vertical COR is also briefly discussed.

Action Potentials↗

PubMed1990

Diketopiperazine-mediated peptide formation in aqueous solution.

Though diketopiperazines (DKP) are formed in most experiments concerning the prebiotic peptide formation, the molecules have not been paid attention in the studies of chemical evolution. We have found that triglycine, tetraglycine or pentaglycine are formed in aqueous solution of glycine anhydride (DKP) and glycine, diglycine or triglycine, respectively. A reaction of alanine with DKP resulted in the formation of glycylglycylalanine under the same conditions. These results indicate that the formation of the peptide bonds proceeds through the nucleophilic attack of an amino group of the amino acids or the oligoglycines on the DKP accompanied by the ring-opening. The formation of glycine anhydride, di-, tri- and tetraglycine was also observed in a mixed aqueous solution of urea and glycine in an open system to allow the evaporation of ammonia. A probable pathway is proposed for prebiotic peptide formation through diketopiperazine on the primitive Earth.

Amino Acid Sequence↗

PubMed1990

Prebiotic synthesis of orotic acid parallel to the biosynthetic pathway.

By heating an aqueous solution of aspartic acid and urea, carbamylaspartic acid is first formed and then the molecule is cyclized to dihydroorotic acid (DHO) with loss of water. Irradiation of an aqueous solution of DHO with a tungsten lamp yields orotic acid by photo-dehydrogenation of the molecule. This pathway of orotic acid formation is quite similar to that of biosynthesis of the molecule.

Aspartic Acid↗

PubMed1990

Response of vestibular neurons to head rotations in vertical planes. III. Response of vestibulocollic neurons to vestibular and neck stimulation.

1. To compare the properties of the vestibulocollic reflex (VCR) with those of vestibular neurons projecting to the neck [vestibulocollic (VC) neurons], we have studied the behavior of the latter in the decerebrate cat. Neurons were identified by their antidromic responses to stimulation in C1-C2, but not C5. Responses to stimulation of vestibular and neck receptors were produced by rotation of the body and head in vertical planes. 2. We determined the plane of whole body (vestibular) or body with head counter-rotated (neck) rotation, which produced the maximal modulation of each neuron (response vector orientation). Neuron dynamics were then studied with sinusoidal (0.02-2 Hz) stimuli aligned with this orientation. 3. On the basis of dynamics and vector orientation, the neuron was assigned a vestibular input classification of otolith, vertical canal, otolith + canal, or spatial-temporal convergence (STC). 4. The properties of this sample of VC neurons are similar to those of a larger population of vestibular neurons whose projection was not identified. For example, the distributions of cells with different types of vestibular inputs were roughly the same; in particular, few cells showed STC responses. In addition, there was no evidence of significant convergence of like canals across the midline (e.g., right anterior + left anterior). 5. Also similar to the larger unidentified population, 80% of VC neurons tested for neck input received such an input. The neck and vestibular responses tended to be antagonistic; the vector orientations were usually opposite, and the response gains and phases similar.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1990

Calpain and calpastatin in rabbit corneal epithelium.

The purpose of this study was to provide a direct assay for calpain and its endogenous inhibitor calpastatin in normal rabbit epithelium. Corneal epithelial extracts were fractionated by DEAE (1) chromatography on HPLC. Fractions were analyzed for calpain by ELISA, immunoblotting, and caseinolytic enzyme activity with FITC-labeled casein. Results demonstrated immunoreactive peaks for calpains I and II. Calpain II from the soluble fraction of corneal epithelium eluted at a similar NaCl concentration (260 mM) as calpain II from other tissues, was inhibited by both E64 and the removal of Ca, contained an 80 kDa subunit in immunoblots, and was present at specific activity of 220 units/g protein (in a crude homogenate). Calpain antigen was also present in the EDTA/EGTA washed insoluble fraction of corneal epithelium. Calpastatin in corneal epithelium eluted at 130 - 160 mM NaCl on DEAE, coeluted with calpain I, and was present at 330 units/g protein (crude homogenate). The results demonstrated a calpain/calpastatin system in corneal epithelium, where it is speculated to play a role in epithelial cell turnover and wound healing.

Animals↗

PubMed1990

Gene expression for calpain isozymes in human hematopoietic system cells.

Calpain (EC 3.4.22.17; Ca2(+)-dependent cysteine endopeptidase) is known to exist in two forms of isozyme. Calpain I requires low (or microM)-Ca2+ for activation and calpain II requires high (or mM)-Ca2+. Both isozymes consist of one heavy (approx.80 kDa) and one light (approx. 30 kDa) subunit each. The heavy subunits of isozymes I and II are different genetic products, while the light subunits are identical. Antibodies respectively specific for the heavy subunits of pig calpains I and II were raised in rabbits, and the affinity-purified IgG proteins were used for Western blot analysis. When 23 human hematopoietic system cells were examined for the degree of their expression of the genes for calpains I and II, all of them were found to contain calpain I of detectable amounts in their cytosolic fluid. By contrast, only nine cell-line cells were positive in calpain II, and they were, without exception, the lineage which had been infected with HTLV-I, the retrovirus responsible for human adult T-cell leukemia. The enhanced production of calpain II in HTLV-I infected T-cells was also confirmed by running chromatographic analyses on the homogenates of these cells, and comparing them with those of uninfected T-cells. When YT-C3 cell, which is an uninfected, natural killer-like cell, was transfected with HTLV-I gene, the resulting transformed stable cells, YT-4 and YT-5.1, were found to produce increased amounts of calpain II concomitant with that of interleukin (IL)-2 receptor protein. These results suggest that the gene expression for calpain isozymes may vary during the course of differentiation of T-lymphocytes. The mechanism of regulation of calpain isozyme genes and the biological significance of the variation in expression during differentiation still remain unanswered.

Animals↗

PubMed1989

Neurite outgrowth from mouse neuroblastoma and cerebellar cells induced by the protein kinase inhibitor H-7.

The protein kinase inhibitor H-7 (1-(5-isoquinolinesulfonyl)-2-methylpiperazine dihydrochloride) or staurosporine was found to induce neurite outgrowth from mouse neuroblastoma N18TG2 cells and cultured cerebellar cells by light microscopy. Examination by electron microscopy revealed some morphological differences between the neurites of N18TG2 cells induced by H-7 and dB-cAMP, in which H-7 formed longer and thinner neurites and more abundant localized varicose structures along neurites than dB-cAMP. These results indicate that protein kinase inhibitor itself can act as a potent stimulus for inducing neurite outgrowth in a different manner from dB-cAMP.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

PubMed1989

Effects of spinal cord compression on repetitive impulse conduction of the ascending fibers of the dorsal column.

The effects of spinal cord compression on conduction of compound action potentials (CAPs) in the dorsal column fibers at various stimulus frequencies were analysed in pentobarbital anesthetised cats. The L 6 posterior root was given 1 to 500 Hz stimuli, and the CAPs were recorded from the L 2 posterior column. The 100th CAP was recorded at 100, 200 and 500 Hz. The L 4 cord segment was compressed by stepped increments (0.5 mm/5 min.) with a plastic plate (4 x 10 mm) until the responses at 1 Hz disappeared, then the compression was released. Before compression, each CAP at all frequencies showed almost uniform amplitudes and latencies. During compression, the CAPs were not altered significantly at any frequency until the spinal cord was compressed 2.5-4.0 mm. Further compression produced a progressive decrease in amplitude and increase in latency of the CAPs. At 500 Hz, however, the conduction block (amplitude loss, latency increase) was much severer than that produced at lower frequencies. After release of compression, the CAPs at all frequencies recovered progressively, and became almost stable after about 1 hour. The recovery rate of amplitude was only about 30% compared to the amplitude before compression at 500 Hz, and about 50% at 1 and 200 Hz. However, the recovery rate of amplitude at 100 Hz (65%) exceeded those at 1 and 200 Hz. To observe the difference in recovery rates among them, all the trains of CAPs were analyzed. Before compression, the amplitudes were almost uniform at stimulus frequencies as high as 500 Hz.(ABSTRACT TRUNCATED AT 250 WORDS)

Action Potentials↗

PubMed1989

Biomechanical effect of anteriorly directed extraoral forces on the craniofacial complex: a study using the finite element method.

This study was designed to investigate the biomechanical effect of protractive maxillary orthopedic forces on the craniofacial complex by use of the three-dimensional finite element method (FEM). The three-dimensional FEM model was developed on the basis of a dry skull of a young human being. The model consisted of 2918 nodes and 1776 solid elements. Eighteen cranial and facial sutural systems were integrated in the model. An anteriorly directed 1.0-kg force was applied on the buccal surfaces of the maxillary first molars in both a horizontal parallel direction and a 30 degree obliquely downward direction to the functional occlusal plane. The nasomaxillary complex showed a forward displacement with upward and forward rotation in a horizontal protraction case, whereas a downward force produced almost translatory repositioning of the complex in an anterior direction. High stress levels were observed in the nasomaxillary complex and its surrounding structures. However, the pattern of stress distributions within the complex was different in two force systems. A downward protraction force produced relatively uniform stress distributions, indicating the importance of the force direction in determining the stress distributions from various orthopedic forces.

Biomechanical Phenomena↗

PubMed1989

Neck muscle spindle activity in the decerebrate, unparalyzed cat: dynamics and influence of vestibular stimulation.

1. Using floating electrodes, we recorded from neck-muscle spindle afferents in the C2 dorsal root ganglion of the decerebrate cat. Nerves to dorsal neck muscles were cut so that the afferents presumably originated mainly from ventral and ventrolateral perivertebral muscles and sternocleidomastoid. One goal of our experiments was to study possible vestibular influence exerted on these spindles via the fusimotor system. Unparalyzed preparations were therefore used. 2. Stimuli consisted of sinusoidal rotations in vertical planes. Neck tilt stretched neck muscles, whereas whole-body tilt stimulated vestibular receptors. 3. For each afferent we first determined the most effective direction of neck tilt, then used stimuli oriented close to this direction to study response dynamics, particularly gain of responses to stimuli of different amplitudes (0.5-7.5 degrees). 4. Three-quarters of the afferents failed to respond to 0.5 degrees, 0.2-Hz neck rotations. Stimuli that were effective usually elicited responses that had low gain and were linear over the whole range of amplitudes. Only a few afferents had behavior typical of spindle primary afferents: high-gain responses to small sinusoidal stimuli, gain decreasing as stimulus amplitude increases. This prevalence of static spindle responses in the unparalyzed cat is in striking contrast to results obtained on neck-muscle spindles in paralyzed, decerebrate cats, and on hindlimb extensor muscle spindles in decerebrate, unparalyzed cats. 5. Paralysis produced by injection of Flaxedil changed the behavior of 2/4 spindle afferents tested, causing the appearance of high-gain responses to 0.5 degrees stimuli and of nonlinear behavior.(ABSTRACT TRUNCATED AT 250 WORDS)

Afferent Pathways↗

PubMed1989

Crystal and molecular structures of carbon-bridged pyrimidine cyclonucleosides. Substrate analogs of ribonuclease A.

The crystal and molecular structures of carbon-bridged 6,5'-cyclo-5'-deoxy-4-thiouridine (6,5'-Cs4U), 6,5'-cyclo-5'-deoxy-2',3'-O-isopropylideneuridine (6,5'-CiU) and 6,6'-cyclo-5',6'-dideoxy-allofuranosyluracil (6,6'-CU) have been determined by X-ray diffraction. The molecular conformations of 6,5'-Cs4U and 6,5'-CiU are very similar; the conformation about the glycosidic bond is anti (low region), the torsion angle O(4')-C(1')-N(1)-C(2) being -150.0 degrees for 6,5'-Cs4U and -145.5 degrees for 6,5'-CiU, and the sugar puckering being both O(4')-exo. On the other hand, 6,6'-CU takes the glycosidic torsion angle of -116.9(4) degrees (middle anti region) and the sugar conformation of C(4')-endo. The cyclization causes little alteration in the geometry of the base moiety. 6,5'-Cs4U and 6,5'-CiU exhibit the similar base-base interactions between adjacent molecules, although their molecular packings are quite different; the 4-thiouracil or uracil moiety interacts with adjacent base moieties through hydrogen bonding and stacking interactions. In 6,6'-CU, cyclonucleosides were connected by hydrogen bondings between the hydroxyl and sugar ring oxygen atoms and between the hydroxyl groups and the base nitrogen and oxygen atoms. As the 2',3'-cyclic phosphates of these carbon-bridged cyclonucleosides are hydrolyzed by ribonuclease A, it is suggested that the conformers found in these cyclonucleosides are recognized by the enzyme.

Chemical Phenomena↗

PubMed1988

Crystallization and preliminary X-ray diffraction studies of 3-methyladenine-DNA glycosylase II from Escherichia coli.

Single crystals of 3-methyladenine-DNA glycosylase II produced from the alkA gene of Escherichia coli have been obtained by the method of vapour diffusion with polyethylene glycol 6000 as precipitant at pH 8.5. The crystals belong to the monoclinic space group C2, with a = 58.6 A, b = 76.8 A, c = 62.2 A, beta = 110.3 degrees. They contain one molecule per asymmetric unit and diffract to 2.7 A resolution.

Crystallization↗

PubMed1988

Structural studies of O6-methyldeoxyguanosine and related compounds: a promutagenic DNA lesion by methylating carcinogens.

O6-Methylation of guanine residues in DNA can induce mutations by formation of base mispairing due to the deprotonation of N(1). The electronic, geometric and conformational properties of three N(9)-Substituted O6-methylguanine derivatives, O6-methyldeoxyguanosine (O6mdGuo), O6-methylguanosine (O6mGuo) and O6, 9-dimethylguanine (O6mdGua), were investigated by X-ray and/or NMR studies. O6mdGuo crystallizes in the monoclinic space group P2(1) with cell parameters a = 5.267(1), b = 19.109(2), c = 12.330(2) A, beta = 92.45(1) degrees, V = 1239.8(3) A3, z = 4 (two nucleosides per asymmetric unit), and O6mGua in the monoclinic space group P2(1)/n with cell parameters a = 10.729(2), b = 7.640(1) c = 10.216(1) A, beta = 92.17(2) degrees, V = 836.7(2) A3, z = 4. The geometry and conformation of O6-methylguanine moieties observed in both crystals and are very similar. Furthermore, the molecular dimensions of the O6methylguanine residue resemble more closely those of adenine than those of guanine. The methoxy group is coplanar with the purine ring, the methyl group being cis to N(1). The conformation of O6-methylguanine nucleosides is variable. The glycosidic conformation of O6mdGuo is anti for molecule (a) and high-anti for molecule (b) in the crystal, while that of O6mGuo is syn [Parthasarathy, R & Fridey, S. M. (1986) Carcinogenesis 7, 221-227]. The sugar ring pucker of O6mdGuo is C(2')-endo for molecule (a) and C(1')-exo for molecule (b). The C(4')-C(5') exocyclic bond conformation in O6mdGuo is gauche- for molecule (a) but trans for molecule (b), in contrast with gauche+ for O6mGuo. The hydrogen bonds exhibited by O6-methylguanine derivatives differ from those in guanine derivatives; the amino N(2) and ring N(3) and N(7) atoms of O6-methylguanine residues are involved in hydrogen bonding. 1H-NMR data for O6mdGuo and O6mdGuo reveal the predominance of a C(2')-endo type sugar puckering. In O6mdGuo, however, a contribution of a C(1')-exo sugar puckering is significant. The NOE data also indicate that O6mdGuo molecules exist with nearly equal population for anti (including high anti) and syn glycosidic conformations. These observations and their biological implications are discussed.

Alkylating Agents↗

PubMed1988

Glycolipid-directed FH6 monoclonal antibody recognizes high molecular weight glycoprotein antigen carrying sialyl Lex-i determinant in the culture supernatant of PC-9 cells.

The properties of the antigen recognized by monoclonal antibody FH6 have been analyzed. FH6 was originally generated against a glycolipid, i.e. a difucoganglioside isolated from human colonic adenocarcinoma, and specifically reacts with sialyl Lex-i determinant. Several culture supernatants of human carcinoma cell line cells were found to have high levels of FH6-reactive antigen, and PC-9, a human lung carcinoma cell line was used for the analysis. A solid-phase sandwich radioimmunoassay was performed to detect the antigen. The antigenic activity was extractable in 0.6 M PCA or 7% TCA, and was sensitive to mild alkaline treatment and to Pronase digestion. Most of the antigen was eluted in the void volume of a Sepharose CL-2B column, which indicates that its molecular weight is greater than several million. It was eluted from a DEAE-cellulose column at a NaCl concentration in the range of 0.2-0.25 M. The immunoaffinity-purified antigen has a high carbohydrate content of more than 80%. These data indicate that the antigen recognized by FH6 in the culture supernatant of PC-9 is not a glycolipid, but a high molecular weight glycoprotein which could be referred to as a mucin, or a proteoglycan, which contains keratan-sulfate like glycosaminoglycan chains, as judged from the results of the glycosidase treatments.

Antibodies, Monoclonal↗

PubMed1988

Common interneurones mediating cortical and tectal excitation of abducens motoneurones in the cat.

Tectal and cortical effects on abducens motoneurones were examined with intracellular recording techniques in cats under chloralose anaesthesia. Abducens motoneurones exhibited disynaptic EPSPs after stimulation of the contralateral superior colliculus and cerebral peduncle. The tectal disynaptic EPSPs were observed invariably in all motoneurones tested, while the peduncular EPSPs were observed only in 40% of motoneurones after stimulation of the contralateral cerebral peduncle. However, the tectal disynaptic EPSPs were consistently facilitated by conditioning peduncular stimulation in all motoneurones tested. These results indicated that the disynaptic excitatory tecto-abducens and cortico-abducens pathways shared common premotor interneurones. The common interneurones which mediated the tectal and cortical disynaptic excitation of abducens motoneurones were explored in the pons. These interneurones were identified by the criteria that they were fired monosynaptically from both the tectum and the cerebral peduncle and were activated antidromically from the abducens nucleus. Systematic threshold mapping for the antidromic activation in and around the abducens nucleus indicated that they gave off many collateral branches in the nucleus. Such neurones were found in the nucleus reticularis pontis caudalis, being distributed in the area extending 0.8-3 mm rostral to the rostral pole of the abducens nucleus, 1.3-2.7 mm deep from the dorsal surface of the brain stem, and 0.8-1.8 mm lateral from the midline. The present experiments strongly suggest that a group of neurones in the paramedian pontine reticular formation make direct excitatory connexions with abducens motoneurones and play a role of common interneurones that transmit both tectal and cortical commands.

Abducens Nerve↗

PubMed1988

Condensation of oligoglycines with trimeta- and tetrametaphosphate in aqueous solutions.

The dehydration condensation of glycine with trimetaphosphate in aqueous solution has been reinvestigated. Although it has been reported that the condensation of glycine under the alkaline conditions was brought about through the formation of cyclic acylphosphoramidate and hence the condensation of polyglycines could not occur, we found that the condensation of oligoglycines with trimeta- and tetrametaphosphate in aqueous solution are possible through the formation of their acylphosphates under the neutral or weak acidic conditions. Aqueous solutions of 1.0 M glycylglycine and 1.0 M trimetaphosphate in the various pH from 4.0 to 9.0 were incubated at 38 degrees C. The solutions were analyzed by HPLC with ninhydrin reaction system. Tetraglycine and hexaglycine were detected and their maximum yields were given in the reaction carried out around pH 7. They are approximately 15% and 4% after 30 days, respectively. Analogous experiments were performed with tetrametaphosphate. The results showed a similar pH dependence for the condensation, but the yields were about one-tenth of those of corresponding experiments with trimetaphosphate. Relative rates of dimerization of glycine, diglycine and triglycine in the equimolar concentration were also investigated at pH 6.0 at 38 degrees C. The rates for diglycine and triglycine were approximately twice and four times as large as that for glycine. Relevance of the experiments to chemical evolution is discussed.

Chemical Phenomena↗