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C Nakamura

Publications and source records attributed to C Nakamura.

At least 37 records · Page 2Linked to original sources

Single nucleotide polymorphisms in an STS region linked to the Ncc-tmp1A locus are informative for characterizing the differentiation of chromosome 1A in wheat.

Homoeoalleles of Ncc confer nucleus-cytoplasm (NC) compatibility on NC hybrids of wheat with the D plasmon of Aegilops squarrosa. To dissect the chromosomal region containing Ncc, a RAPD marker linked to the Ncc-tmplA locus, which is located on chromosome 1A of T timopheevi, was sequenced and converted to a PCR-based sequence-tagged-site (STS) marker. Five single nucleotide polymorphisms (SNPs) between T timopheevi and T turgidum. were detected in a 509-bp genomic DNA fragment. Based on the SNPs, the STS alleles in 164 accessions from emmer wheat, timopheevi wheat and two einkorn wheats, T. urartu and T. boeoticum were surveyed by PCR-RFLP analysis. The sequence comparisons and PCR-RFLP analyses revealed nine alleles based on six SNPs. These SNPs were highly conserved within each group of wheat, and all groups could be distinguished by particular combinations of the SNPs. All accessions of T. urartu had one unique STS allele as compared with the others. Our results indicate that the SNPs in the STS marker linked to the Ncc-tmplA locus would be informative for studies of the differentiation of chromosome 1A in wheat.

Alleles↗

Effects of antihypertensive agents on blood pressure during exercise.

The relationship between blood pressure (BP) and cardiovascular morbidity has been appreciated for many years. Casual BP may not be representative of the pressure at other times. It is recognized that BP during exercise may be a more accurate predictor than casual BP. There is, however, little information about the effects of antihypertensive drugs on the BP during exercise. This study was designed to investigate the effects of various antihypertensive agents on BP during exercise. Sixty-four patients (age, 49+/-10 years) with untreated essential hypertension (WHO I, II) were studied during a supine ergometric exercise regimen. A graded exercise test was started at a workload of 50 W, and the load was increased by 25 W every 3 min. The hemodynamic responses to exercise were evaluated by changes in systolic and diastolic BP (SBP, DBP) and heart rate (HR). Plasma norepinephrine (NE) levels were measured at rest and during submaximal exercise, and before and after 4 weeks of treatment with metoprolol (METO), doxazosin (DOXA), trichlormethiazide (TCTZ), nifedipine (NIFE), amlodipine (AMLO) and temocapril (TEMO) between left ventricular mass index (LVMI) and BP values at rest, during exercise, and during the recovery period after exercise were assessed by multiple regression analysis. The stepwise selection (forward conditional) method showed that LVMI was significantly associated with SBP during submaximal exercise and during the recovery period. All antihypertensive treatments decreased SBP and DBP (p<0.01) at rest. METO, AMLO and TEMO significantly lowered SBP (p<0.05) during exercise, whereas DOXA, TCTZ and NIFE induced no change in SBP. The exercise-induced increase of plasma NE was further enhanced by METO and NIFE but not by AMLO, DOXA, or TCTZ, and it was significantly suppressed by TEMO (p<0.01). These results suggest that BP during exercise is more highly associated with the progression of left ventricular hypertrophy (LVH) than is casual BP. Because antihypertensive agents differ in their effects on exercise hemodynamics, we recommend that hemodynamic factors during exercise be considered when selecting the optimal antihypertensive medication for highly active patients.

Adult↗

Molecular cloning of three homoeologous cDNAs encoding orthologs of the maize KNOTTED1 homeobox protein from young spikes of hexaploid wheat.

The plant knotted1 (kn1)-like homeobox genes are known to play important roles in the maintenance of shoot apical meristem (SAM), determination of cell fate and differentiation of vegetative tissues. To study structural diversity of the three homologous loci encoding a KN1-like homeobox protein in the hexaploid wheat genome, we isolated clones from a cDNA library of young spikes of Japanese common wheat cultivar 'Norin 26'. Three different but highly homologous cDNAs were isolated and their sequences were determined. The mean homology of the deduced amino acid sequences was 96% as compared to the barley ortholog KNOX3. The wheat kn1-like homeobox proteins named WKNOX1 are encoded by a single set of homologous genes on the homologous group 4 chromosomes in the three component genomes of common wheat, i.e. 4A, 4B and 4D. The nucleotide sequence data and the Southern blot pattern suggested that the three homologous loci of wknox1 genes are highly conserved through polyploid evolution of wheat. They were expressed in SAM-containing shoots and young spikes but not in developed leaves, glumes and lemmas and callus tissues. The ectopic expression of the wknox1 was observed in lemma of wheat Hooded (Hd) mutants. The result suggested that the Hd gene is a dominant allele of the wknox1 locus on chromosome 4A.

Amino Acid Sequence↗

Identification of paternal mitochondrial DNA sequences in the nucleus-cytoplasm hybrids of tetraploid and hexaploid wheat with D and D2 plasmons from Aegilops species.

Mitochondrial (mt) DNA structures were studied in 12 nucleus-cytoplasm (NC) hybrids of tetraploid and hexaploid wheat in which nuclear genomes of the recurrent paternal wheat parents were combined with D and D2 plasmons from Aegilops species. RFLP analysis of 15 mtDNA regions indicated the presence of the paternal sequences in six regions encompassing 11 structural genes in all the NC hybrids. PCR-RFLP analysis showed that one region (a nad3-orf156 region) consisted of a mixture of the maternal, paternal and novel paternal-like sequences. The presence of unexpectedly high levels of the paternal sequences was confirmed by random PCR cloning and sequencing of this region. PCR-RFLP analysis of the random clones further showed that the relative stoichiometry of the maternal and paternal sequences varied depending on the plasmons from the maternal parents and the nuclear backgrounds of the paternal parents. Our results suggest that the differential amplification of the paternal mtDNA sequences is under the control of NC interaction in these NC hybrids.

Base Sequence↗

Homoeoallelic gene Ncc-tmp of Triticum timopheevii conferring compatibility with the cytoplasm of Aegilops squarrosa in the tetraploid wheat nuclear background.

A nuclear gene, Ncc-tmp1A, of Triticum timopheevii is required for the nucleus-cytoplasm (NC) compatibility in tetraploid NC hybrids with the cytoplasm of Aegilops squarrosa. A euploid NC hybrid of T. durum was previously produced by introgressing the gene from chromosome 1A of T. timopheevii. To examine the possible presence of a functional homoeoallele in the G genome of T. timopheevii, segregation of seed viability was studied as a marker phenotype in BC1s involving the two types of NC hybrids, (Ae. squarrosa)-T. timopheevii and (Ae. squarrosa)-T. turgidum. The result of these test crosses suggested that the G genome possesses a functional homoeoallele Ncc-tmp1G. Segregation of two RAPD (random amplified polymorphic DNA) markers that were closely linked to Ncc-tmp1A was further studied among the viable BC1s obtained from a test cross of (Ae. squarrosa)-T. timopheevii x T. turgidum. Some viable BC1 segregants without the markers were obtained, suggesting a limited degree of transmission of chromosome 1G carrying Ncc-tmp1G. However, a similar RAPD analysis of BC1s obtained after backcrosses of reciprocal F1s of T. timopheevii/T. turgidum with T. turgidum showed random marker segregation. Thus, it was concluded that Ncc-tmp1A is not required for compatibility with its own cytoplasm. Southern blot analysis of the euploid NC hybrid using RFLP (restriction fragment length polymorphism) markers on the homologous group 1 chromosomes showed that Ncc-tmp1A locates in the centromeric region.

Alleles↗

New members of a cold-responsive group-3 Lea/Rab-related Cor gene family from common wheat (Triticum aestivum L.).

A Cor (cold-responsive) cDNA that belongs to the group-3 Lea (late embryogenesis abundant)/Rab (responsive to abscisic acid, ABA) family was isolated from a winter-hardy cultivar of common wheat (Triticum aestivum L.). Screening of a cold-acclimated cDNA library was performed using an ABA- and other stress-responsive barley cDNA clone, Hva1, as a probe. A wheat cDNA clone (designated as Wrab19) putatively encoded a basic (pI = 10.3) and hydrophobic protein with 179 amino acids. The deduced protein showed characteristics of the group-3 LEA/RAB protein family. In contrast to the single copy barley Hva1, Wrab19 belonged to a multigene family in the hexaploid wheat genome and six loci were assigned to the homoeologous group 1 chromosomes. Using Wrab19 as a probe, four homologous cDNAs (designated as Wrab17) were isolated that encoded acidic (pI = 4.6-4.7) and hydrophobic proteins, all with 166 amino acids. The deduced proteins showed high homology (a mean of 84% identity) with a barley gibberellic acid (GA3)-inducible protein, ES2A, and several other group-3 LEA/RAB proteins. Wrab17 was considered to be a three-copy gene and each copy was assigned to chromosome 5A, 4B or 4D of hexaploid wheat. Transcripts of both Wrab19 and Wrab17 accumulated within 1 day of cold acclimation at 4 degrees C. They were responsive to ABA and/or GA3, but showed some cultivar differences in their response to these plant hormones. We conclude that the two genes are new members of the group-3 Lea/Rab-related Cor gene family in wheat.

Abscisic Acid↗

Nicotiana tabacum cDNAs encoding alpha and beta subunits of a heterotrimeric GTP-binding protein isolated from hairy root tissues.

Heterotrimeric GTP-binding proteins (G-proteins) play important roles in signal transduction pathways in eukaryotic cells. Through differential screening of a hairy root cDNA library of tobacco (Nicotiana tabacum L.) against transcripts from non-root tissues of normal cuttings, we obtained a partial cDNA clone that showed abundant expression and high homology to the alpha subunit gene of plant G-protein. After RACE-PCR, a full-length cDNA clone was obtained, which was 1,677-bp in length and contained an open reading frame encoding a protein of 384 amino acids. A cDNA clone encoding a beta subunit of G-protein was also isolated from the same cDNA library based on PCR amplification and library screening. The clone was 1,600-bp in length and contained an open reading frame encoding 377 amino acids. The deduced amino acid sequences of these clones showed high homology (75.5 to 99.8% amino acid identity) with alpha and beta subunits of other plant G-proteins. Genomic Southern blot analysis showed that the amphidiploid tobacco genome possessed two major copies of both alpha and beta subunit genes and some minor homologous copies. Northern blot analysis showed that the transcript of alpha subunit gene was abundant in the root tissues, particularly in the hairy root tissues. In contrast, the level of expression of the beta subunit gene was equivalent in all the tissues studied. Possible function of tobacco G-protein was discussed.

Amino Acid Sequence↗

A cold-responsive wheat (Triticum aestivum L.) gene wcor14 identified in a winter-hardy cultivar 'Mironovska 808'.

A cDNA library was constructed from a cold-acclimated winter-hardy common wheat (Triticum aestivum L.) cultivar 'Mironovska 808'. Using this library and a cold- and light-responsive barley cDNA clone cor14b as a probe, cDNAs of a homologous wheat gene wcor14 were isolated. Two identical cDNAs designated as wcor14a had an open reading frame encoding an acidic (pI = 4.71) and hydrophobic polypeptide with 140 amino acids (MW = 13.5 kDa). The deduced WCOR14a polypeptide showed 70% identity with the barley chloroplast-imported COR14b and had a nearly identical N-terminal, putative chloroplast transit peptide of 51 amino acid residues. Another cDNA clone wcor14b was assumed to encode a polypeptide WCORb which had 5 substitutions and a frame shift in the C-terminal region as compared with WCOR14a. RACE PCR, genomic PCR and Southern blot analyses suggested that wcor14 and its related sequences constitute a small multigene family with and without an intron in the hexaploid wheat genome. Northern blot analysis showed that transcripts of wcor14 accumulated within 3-6 hours of cold acclimation at 4 degrees C and the level reached a maximum at day 3. The transcripts became non-detectable within 3 hours after de-acclimation at room temperature. Contrary to the barley cor14b, a similar level of wcor14 transcripts was detected under the continuous darkness. Neither treatment with NaCl, ABA nor dehydration induced its expression. Based on these results we conclude that wcor14 is a wheat orthologue of the barley cor14b and specifically induced by low temperature.

Amino Acid Sequence↗

Self-assembling photosynthetic reaction centers on electrodes for current generation.

Photosynthetic reaction centers (RCs) made from photosynthetic organisms can be used in solar batteries because their molecules cause light-induced charge separation. We present a simple immobilization system of the intact RCs from Rhodobacter sphaeroides on an electrode that uses nickel ligand binding by the hexameric histidine tag on H subunit (HHisRC). The binding constant of HHisRC to the nickel-nitrilotriacetic acid (Ni-NTA) chip measured with a surface plasmon resonance instrument was 1.6 x 10(8) M-1. HHisRCs were immobilized on an indium tin oxide electrode overlaid with an Ni-NTA gold substrate. The photoinduced displacement current of this electrode was measured to estimate the orientation of HHisRC on the electrode, and the detachability of HHisRC from the electrode was determined by using an imidazole solution wash. The direction of the flash-light-induced displacement current suggested that the H subunit side of the immobilized HHisRC faced the surface of the electrode. The photoinduced current disappeared after the electrode was washed in the imidazole solution. This simple immobilization and detachment of HHisRC to the electrode might be useful for making a reproducible photocurrent device.

Biosensing Techniques↗

Fabrication of an electrode-viologen-hydrogenase heterogeneous system and the electrochemical hydrogen evolution.

An indium tin oxide (ITO) electrode was chemically modified by one layer of viologen (VIO) derivative, which possessed a persistent and reproducible electrochemical response. A monolayer of a thermal stable hydrogenase from Thiocapsa roseopersicina was stabilized on a synthesized poly-L-lysine subphase surface and transferred onto the electrode for fabrication of an ITO-VIO-hydrogenase heterogeneous system. Electrochemical properties of both the ITO-VIO monolayer and the heterogeneous ITO-VIO-hydrogenase system have been investigated. Hydrogen evolution could be measured by potentiostating the VIO-hydrogenase-covered ITO electrode to "electroplate" [(VIO+)n]surf, and a large increase in hydrogen evolution was observed when using an electrolyte solution containing sodium dithionite. We discuss the possible electron transfer process.

Electrochemistry↗

[A successfully repaired case of methicillin-resistant Staphylococcus aureus infective endocarditis in a girl with VSD].

We report a successful case of active infective endocarditis due to Methicilin-Resistant Staphylococcus aureus (MRSA). A 2-year-old girl who had a ventricular septal defect (VSD) complained of persistent fever. Echocardiography showeda large vegetation on the tricuspid valve and a small VSD. She underwent vegetectomy, tricuspid valvoplasty and direct closure of VSD. Vancomycin treatment was also effective to abolish infection. She was discharged without any complication.

Child, Preschool↗

Staphylococcal scalded skin syndrome with prosthetic valve endocarditis.

We report staphylococcal scalded skin syndrome (SSSS) in a 67-year-old man. He showed diffuse erythema with erosion on his face and erythema with giant desquamation on his neck, axilla, genitalia, chest and abdomen 39 days after a coronary artery bypass graft and aortic valve replacement. He died of cardiac rupture caused by myocardial necrosis, and autopsy findings demonstrated prosthetic valve endocarditis due to a strain of exfoliative toxin-B producing methicillin-resistant Staphylococcus aureus. To the best of our knowledge, this is the first case of SSSS caused by prosthetic valve endocarditis.

Aged↗

Gastric antiulcerogenic effects of Stryphnodendron adstringens in rats.

The antiulcer activity of the total extract and the fractions of Stryphnodendron adstringens was studied in rats and compared with that of cimetidine. Ulcers were induced in rats by means of three experimental models: acute stress, acidified-ethanol and indomethacin. The total extract and the fractions were found to have significant antiulcer activity in the case of the acute stress and acidified-ethanol models. These findings support the use of S. adstringens extracts in the treatment of gastric lesions.

Animals↗

Ac-mediated trans-activation of the Ds element in rice (Oryza sativa L.) cells as revealed by GUS assay.

A method using particle bombardment and beta-glucuronidase (GUS) assay was applied to rice callus for detecting the excision of the maize Ds element trans-activated by the Ac transposase source. Excision of Ds biolistically introduced into rice callus resulted in the restoration of the interrupted gus gene expression, allowing visual detection of trans-activation two days after bombardment. Only the transgenic callus lines expressing the Ac transposase gene and the wild-type callus co-transformed with Ac and Ds revealed GUS activity. Frequency of excision, estimated based on the relative GUS activity, ranged from 0.3% to 2.2%. Callus lines showing different levels of Ac transcripts revealed varying excision frequencies. At the later stages of callus growth after selection for the Ac/Ds transformed lines, excision events were detected by GUS assay and confirmed by PCR and sequence analyses of the excision sites in individual colonies. GUS activity was also demonstrated in the primary regenerants from the Ac/Ds-transformed callus colonies. The method described in this study may be used as an approach for rapid detection of excision events and assessment of various factors limiting Ac/Ds activity in rice cells.

Biolistics↗

Variations in the maize Ac transposase transcript level and the Ds excision frequency in transgenic wheat callus lines

To investigate the excision of a maize transposable element in wheat cells, plasmid DNAs containing a Dissociation (Ds) element located between a rice actin 1 gene promoter and a beta-glucuronidase (GUS) gene (gus) were introduced into wheat callus lines by microprojectile bombardment, and transient GUS expression was assayed. The gus-expressing cells after Ds excision were detected only when the Activator (Ac) transposase gene was co-transformed. To further examine a relationship between the amount of Ac mRNA and the Ds excision frequency, the Ds-containing plasmids were introduced into 15 independent transgenic callus lines transformed with the Ac transposase gene. Ten lines expressed the Ac transposase gene under the control of either the cauliflower mosaic virus 35S promoter or the Ac native promoter. The gus gene expression that indicated the Ds excision was observed only in the transgenic callus lines stably expressing the Ac transposase gene. The number of blue spots reflecting the frequency of Ds excision was variable among them. Northern-blot analysis also showed a large variability in the amount of Ac transposase transcripts among the lines. It was however noted that the excision frequency was decreased at a high level of the Ac transposase transcripts, supporting the hypothesis that Ds excision is inhibited above a certain level of the Ac transposase as observed in maize and transgenic tobacco.

Journal Article↗

Effects of incorporation of various amphiphiles into recipient liposome membranes on inter-membrane protein transfer.

To obtain information about the factors governing spontaneous inter-membrane protein transfer, we examined the effects of incorporation of various amphiphilic compounds in dimyristoylphosphatidylcholine (DMPC) liposomes on protein transfer from influenza virus-infected cells to the liposomes, and analyzed the physical properties of these liposome membranes. The incorporation of amphiphilic compounds, negatively charged dicetylphosphate (DCP), dipalmitoylphosphatidylserine (DPPS) or positively charged dimethyldipalmitoylammonium (DMDPA), into DMPC liposomal membranes enhanced protein transfer. The liposomes containing DCP, DPPS or DMDPA were unaffected by osmotic shock caused by external addition of glucose, suggesting a decrease in lipid packing in the liposomal membranes. Furthermore, calorimetric study of these liposomes showed that a phase separation occurred partially in the liposomal membranes. Accordingly, the membranes of DMPC liposomes containing DCP, DPPS and DMDPA should be distorted due to the coexistence of two phases, gel and liquid crystalline, in the membranes. Consequently, the membrane distortion could be responsible for the enhancement effects of the amphiphiles on the inter-membrane protein transfer from influenza virus-infected cells to the liposomes.

Cells, Cultured↗