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Biomedical subjects

S Miyachi

Publications and source records attributed to S Miyachi.

At least 37 records · Page 2Linked to original sources

Molecular structure, localization and function of biliproteins in the chlorophyll a/d containing oxygenic photosynthetic prokaryote Acaryochloris marina.

We investigated the localization, structure and function of the biliproteins of the oxygenic photosynthetic prokaryote Acaryochloris marina, the sole organism known to date that contains chlorophyll d as the predominant photosynthetic pigment. The biliproteins were isolated by means of sucrose gradient centrifugation, ion exchange and gel filtration chromatography. Up to six biliprotein subunits in a molecular mass range of 15.5-18.4 kDa were found that cross-reacted with antibodies raised against phycocyanin or allophycocyanin from a red alga. N-Terminal sequences of the alpha- and beta-subunits of phycocyanin showed high homogeneity to those of cyanobacteria and red algae, but not to those of cryptomonads. As shown by electron microscopy, the native biliprotein aggregates are organized as rod-shaped structures and located on the cytoplasmic side of the thylakoid membranes predominantly in unstacked thylakoid regions. Biochemical and spectroscopic analysis revealed that they consist of four hexameric units, some of which are composed of phycocyanin alone, others of phycocyanin together with allophycocyanin. Spectroscopic analysis of isolated photosynthetic reaction center complexes demonstrated that the biliproteins are physically attached to the photosystem II complexes, transferring light energy to the photosystem II reaction center chlorophyll d with high efficiency.

Amino Acid Sequence↗

Corticostriatal projections from distal and proximal forelimb representations of the monkey primary motor cortex.

Corticostriatal projections from one distal and two proximal subregions in the forelimb representation of the primary motor cortex (MI) were examined in the macaque monkey. The distal and proximal subregions in the anterior bank of the central sulcus (distal and proximal-bank subregions) and the proximal subregion in the surface of the precentral gyrus (proximal-surface subregion) of the MI were identified using intracortical microstimulation. Different anterograde tracers were then injected into two of these three forelimb subregions of the MI. In the ipsilateral putamen, the distribution areas of corticostriatal fibers from the distal, proximal-bank and proximal-surface subregions were arranged from ventrolateral to dorsomedial in this order. These corticostriatal input zones were largely segregated from one another.

Animals↗

Fluorescence properties of chlorophyll d-dominating prokaryotic alga, acaryochloris marina: studies using time-resolved fluorescence spectroscopy on intact cells

Antenna components and the primary electron donor of the photosystem (PS) II in the Chlorophyll (Chl) d-dominating prokaryote, Acaryochloris marina, were studied using time-resolved fluorescence spectroscopy in the ps time range. By selective excitation of Chl a or Chl d, differences in fluorescence properties were clearly resolved. At physiological temperature, energy transfer was confirmed by a red shift of emission maximum among PS II antenna components, and the equilibrium of energy distribution among Chl a and Chl d was established within 30 ps. A fluorescence component that can be assigned to delayed fluorescence (DF) was observed at 10 ns after the excitation; however, it was not necessarily resolved by the decay kinetics. At -196 degrees C, a red shift of emission maximum was reproduced but the equilibrium of energy distribution was not detected. DF was resolved in the wavelength region corresponding to Chl a by spectra and by decay kinetics. The lifetime of the DF was estimated to be approx. 15 ns, and the peaks were located at 681 and 695 nm, significantly shorter wavelengths than those of Chl d. These findings strongly suggest that an origin of DF is Chl a, and Chl a is most probably the primary electron donor in the PS II reaction center (RC). These results indicate that the constitution of PS II RC in this alga is essentially identical to that of other oxygenic photosynthetic organisms.

Journal Article↗

Purification and characterization of hydrogenase from the marine green alga, Chlorococcum littorale.

Hydrogenase from the marine green alga, Chlorococcum littorale, was purified 1485-fold, resulting in a specific activity for hydrogen evolution of 75.7 micromol/min/mg of protein at 25 degrees C, using reduced methyl viologen as an electron donor. The K(m) value for methyl viologen was 0.5 mM. The purity of the enzyme was judged by native PAGE. The molecular weight was estimated to be 55 kDa by SDS-PAGE, and 57 kDa by gel filtration. The optimum temperature and pH value for hydrogen evolution were 50 degrees C and 7.5, respectively. The partially purified hydrogenase catalyzed hydrogen evolution from ferredoxin that had been isolated from the same cells, but not from NADH or NADPH. The K(m) value for ferredoxin was 0.68 microM. The enzyme was extremely oxygen sensitive, losing over 95% of its activity upon exposure to air within minutes, even at 4 degrees C. Two peptide fragments were obtained from the hydrogenase protein digested enzymatically, and their amino acid sequences were determined. No significant homology was found to any other known sequences of hydrogenases.

Amino Acid Sequence↗

Intra-arterial vein bypass using a vein-loaded stent system to occlude wide-necked aneurysms: an experimental study in dogs.

We treated 13 experimental wide-necked aneurysms of lateral type produced in dog carotid arteries with a vein-loaded stent system consisting of an autologous vein graft, a vein-carrier stent, and two other self-expanding Z-shaped stents. The vein-loaded stent, made by connecting the autologous vein graft to the vein-carrier stent, was implanted in the common carotid artery to seal the aneurysm's orifice, after which two booster stents were deployed. The distal stent was placed to fix the distal end of the vein graft, and the proximal stent was placed at the proximal venous portion of the vein-loaded stent to reinforce its proximal end. In this manner an intra-arterial bypass was established across the aneurysm. Of 13 aneurysms seven were completely occluded, preserving the parent artery. The remaining six procedures resulted in parent artery occlusion because of failures in stent deployment. Angiograms performed in within 1 week six successfully treated dogs revealed the parent artery to be patent in four but thrombosed in two. Histological study of successful cases showed as smooth intima with slight endothelial hypertrophy and partially organised thrombus. The intraluminal features were also observed with a vascular endoscope at the time of treatment in four dogs and at the time of follow-up angiography in two. The appearances corresponded well to the macroscopic findings in the specimens. Our vein-loaded stent system offers complete closure of the orifice, which does not occur with conventional stent placement. The method is believed to show clinical promise for treating wide-necked aneurysms not curable by embolisation of the sac.

Aneurysm↗

A dural arteriovenous fistula of the tentorium successfully treated by intravascular embolization.

BACKGROUND: Dural arteriovenous fistulas of the tentorium are rare lesions that often present with intracranial hemorrhage. Definitive treatment is therefore necessary, but transarterial embolization has rarely been curative. CASE DESCRIPTION: A 59-year-old man presenting with sudden onset of severe headache had subarachnoid hemorrhage demonstrated by computed tomography. Left carotid angiography showed a tentorial dural arteriovenous fistula fed by a tentorial branch from the internal carotid artery and by a middle meningeal artery; the fistula drained to the marginal sinus via a dilated varicosity. Transarterial embolization successfully obliterated the fistula, and the patient was discharged with no neurologic deficit. CONCLUSION: This tentorial fistula, that showed extremely rare angiographic features, particularly venous drainage, was embolized successfully. The literature concerning tentorial dural arteriovenous fistulas is discussed in terms of effective therapeutic choice.

Arteriovenous Fistula↗

Parallel neural networks for learning sequential procedures.

Recent studies have shown that multiple brain areas contribute to different stages and aspects of procedural learning. On the basis of a series of studies using a sequence-learning task with trial-and-error, we propose a hypothetical scheme in which a sequential procedure is acquired independently by two cortical systems, one using spatial coordinates and the other using motor coordinates. They are active preferentially in the early and late stages of learning, respectively. Both of the two systems are supported by loop circuits formed with the basal ganglia and the cerebellum, the former for reward-based evaluation and the latter for processing of timing. The proposed neural architecture would operate in a flexible manner to acquire and execute multiple sequential procedures.

Animals↗

Rubisco activase transcript (rca) abundance increases when the marine unicellular green alga Chlorococcum littorale is grown under high-CO2 stress.

cDNA and the corresponding genomic DNA region encoding Rubisco activase were isolated from the unicellular green alga Chlorococcum littorale. The deduced amino acid sequence encoded by the cDNA was 403 amino acids long and exhibited important homology with those of other known Rubisco activases. Its N-terminal sequence was similar to the chloroplastic transit peptides in Chlamydomonas reinhardtii. The mature protein had a predicted molecular mass of 42 kDa. Five introns were located inside the genomic gene encoding Rubisco activase (rca). Genomic Southern blots indicated that two copies of the rca gene were present in the genome of C. littorale. The level of rca messenger RNA increased when cells of C. littorale were subjected to high-CO2 stress (i.e. grown under at least 20% CO2). Hsp70 heat-shock protein was also induced under high-CO2 conditions and, as expected, was also induced at 35 degrees C. The rca gene, in contrast, was not induced at 35 degrees C, indicating that this gene was induced in response to the high CO2 concentration and not to general stress. A search of the promoter-binding proteins by a gel retardation assay showed that, under the high-CO2 conditions, a protein(s) which was probably an activator of the rca transcription was synthesized.

Amino Acid Sequence↗

A photosystem I reaction center driven by chlorophyll d in oxygenic photosynthesis.

A far-red type of oxygenic photosynthesis was discovered in Acaryochloris marina, a recently found marine prokaryote that produces an atypical pigment chlorophyll d (Chl d). The purified photosystem I reaction center complex of A. marina contained 180 Chl d per 1 Chl a with PsaA-F, -L, -K, and two extra polypeptides. Laser excitation induced absorption changes of reaction center Chl d that was named P740 after its peak wavelength. A midpoint oxidation reduction potential of P740 was determined to be +335 mV. P740 uses light of significantly low quantum energy (740 nm = 1.68 eV) but generates a reducing power almost equivalent to that produced by a special pair of Chl a (P700) that absorbs red light at 700 nm (1.77 eV) in photosystem I of plants and cyanobacteria. The oxygenic photosynthesis based on Chl d might either be an acclimation to the far-red light environments or an evolutionary intermediate between the red-absorbing oxygenic and the far-red absorbing anoxygenic photosynthesis that uses bacteriochlorophylls.

Journal Article↗

Characteristics of a long-term procedural skill in the monkey.

The purpose of this study was to characterize the nature and structure of procedural memory. We have previously studied the process of learning sequential behavioral procedures using monkeys. The monkey's task was to press five consecutive pairs of buttons (indicated by illumination) in the correct order for every pair, which he had to find by trial-and-error in a block of trials. The whole sequence was called a "hyperset"; each pair was called a "set". We first examined whether monkeys learned to perform a hyperset as a single sequence or learned the order of button-presses individually for each set. To answer this question, we generated hypersets that were the same as the hypersets that had been extensively learned except that the order of the sets was reversed. The performance of these "reversed hypersets" was much worse than the performance of the original learned hypersets and was similar to the performance of new hypersets, as regards both the number of errors and the performance time. The result suggests that monkeys learned a hyperset as a sequence. To examine whether the learned performance was specific to the hand used for practice, we had monkeys use the same hand throughout the long-term practice of each hyperset, and then tested the opposite hand. The performance using the opposite hand was worse than the performance using the trained hand, but was better than the performance for new hypersets. This indicates that the memory for the sequential procedure is only partially accessible to the hand that was not used for the practice.

Animals↗

Cloning and characterization of high-CO2-specific cDNAs from a marine microalga, Chlorococcum littorale, and effect of CO2 concentration and iron deficiency on the gene expression.

Two cDNA clones exclusively induced under an extremely high-CO2 concentration (20%) were isolated from Chlorococcum littorale by differential screening and named HCR (high-CO2 response) 1 and 2, respectively. The amino acid sequence of the protein encoded by HCR2 exhibited homology to the gp91-phox protein, a critical component of a human phagocyte oxidoreductase, and to the yeast ferric reductases, Saccharomyces cerevisiae FRE1 and FRE2 and Schizosaccharomyces pombe Frp1. The induction of both HCR mRNAs required extremely high-CO2 conditions and iron deficiency, being suppressed under air conditions and by iron sufficiency, suggesting that the expression of these two HCR genes required extremely high-CO2 conditions and iron deficiency in combination. The HCR2 protein was detected in the membrane fractions of cells grown under conditions which would favor the induction of HCR2-mRNA and the protein level was lowered when the cells were transferred from iron deficient to 10 microM FeSO4 conditions (with 20% CO2).

Algal Proteins↗

Role of monkey cerebellar nuclei in skill for sequential movement.

To examine whether the cerebellum is involved in learning and memory of visuomotor sequences, we trained two monkeys on a sequential button press task and inactivated different portions of the cerebellar nuclei by injecting a small amount of muscimol (gamma-aminobutyric acid agonist). Before the injection experiments started, the monkeys had learned a set of sequences (n = 21 and 12) extensively. After each injection, we had the monkeys perform the learned sequences and, in addition, learn new sequences. We found deficits in learning/memory by the injections into the dorsal and central part of the dentate nucleus. The number of errors increased significantly for the learned sequences but not for the new sequences. This effect was present only when the hand ipsilateral to the muscimol injection was used. Consistent with this result, anticipatory saccades, the occurrence of which is correlated closely with motor skill, also became less frequent particularly when the ipsilateral hand was used. No effect on learning/memory was observed after injections into the ventral or lateral parts of the dentate nucleus, interpositus nucleus, or fastigial nucleus. In contrast, hand movements became slower after ipsilateral injections at all of the injection sites. These results suggest that, among the cerebellar nuclei, the dentate nucleus, especially its dorsal and central regions, is related to the storage and/or retrieval of long-term memory for motor skill.

Animals↗

Light-harvesting in Acaryochloris marina--spectroscopic characterization of a chlorophyll d-dominated photosynthetic antenna system.

Oxygenic photosynthesis of the prokaryote Acaryochloris marina involves chlorophyll d (Chl d) as the major pigment [Miyashita et al. (1996) Nature 383, 402]. Four spectral forms of Chl d (peak wavelengths: 694, 714, 726 and 740 nm) are resolvable by low-temperature absorption spectroscopy on intact cells. Based on fluorescence spectra (at 290 K and 77 K) and on analysis of fluorescence induction curves we conclude: (1) excitation energy is efficiently transferred between the various spectral forms of Chl d and the PS II reaction center; (2) Chl d serves as a light-harvesting pigment for both, Photosystem II (PS II) and PS I; (3) excitation energy transfer between PS II units occurs.

Chlorophyll↗

Isolation and characterization of biliprotein aggregates from Acaryochloris marina, a Prochloron-like prokaryote containing mainly chlorophyll d.

Phycobiliprotein aggregates were isolated from the prokaryote Acaryochloris marina, containing chlorophyll d as major pigment. In the electron microscope the biliprotein aggregates appear as rod-shaped structures of 26.0 x 11.3 nm, composed of four ring-shaped subunits 5.8 nm thick and 11.7 nm in diameter. Spectral data indicate that the aggregates contain two types of biliproteins: phycocyanin and an allophycocyanin-type pigment, with very efficient energy transfer from the phycocyanin- to allophycocyanin-type constituent. The chromophore-binding polypeptides of the pigments have apparent molecular masses of 16.2 and 17.4 kDa. They crossreact with antibodies against phycocyanin and allophycocyanin from a red alga.

Bacterial Proteins↗

Ribulose-1,5-bisphosphate carboxylase/oxygenase from thermophilic red algae with a strong specificity for CO2 fixation.

Strongly carboxylase-specific ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) was found in Galdieria partita and Cyanidium caldarium (Cyanidiophyceae). The relative specificity, VcKo/VoKc, of Galdieria and Cyanidium RuBisCO was 238 and 222, respectively; 2.4 to 2.5-fold that of higher plant RuBisCOs. The apparent Km of RuBisCO from the thermophilic red algae for CO2 was 6 to 7 microM and the smallest of the values reported so far for other RuBisCOs. The pre-rhodophyte Porphiridium purpureum, which lives at moderate temperatures, had RuBisCO with the relative specificity value of 144. A large difference (5.2 kcal x mol(-1)) in the activation energies between the carboxylase and oxygenase activities in Galdieria RuBisCO was a cause of the strong specificity for the carboxylase activity.

Carbon Dioxide↗

Differential roles of monkey striatum in learning of sequential hand movement.

To study the role of the basal ganglia in learning of sequential movements, we trained two monkeys to perform a sequential button-press task (2x5 task). This task enabled us to examine the process of learning new sequences as well as the execution of well-learned sequences repeatedly. We injected muscimol (a GABA agonist) into different parts of the striatum to inactivate the local neural activity reversibly. The learning of new sequences became deficient after injections in the anterior caudate and putamen, but not the middle-posterior putamen. The execution of well-learned sequences was disrupted after injections in the middle-posterior putamen and, less severely, after injections in the anterior caudate/putamen. These results suggest that the anterior and posterior portions of the striatum participate in different aspects of learning of sequential movements.

Animals↗