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Biomedical subjects

S Tabata

Publications and source records attributed to S Tabata.

At least 127 records · Page 7Linked to original sources

[A case of bronchial stenosis due to postoperative inflammation treated with expandable metallic stent].

We presented a 61-year-old man who had undergone a left sleeve upper resection because of a squamous cell carcinoma of the upper lobe of the left lung. At 5 weeks after the operation, the anastomosis between the left main bronchus and the left lower bronchus became stenotic, therefore pneumonitis occurred below the anastomosis. Because of the inflammatory granulo stenosis for short time, we used an expandable metallic stent to save a residual lung function of the operated side. The anastomosis between the left main bronchus and the left lower bronchus was kept open satisfactorily, and in the late postoperative periods the residual lung function recovered until the levels of predicted residual lung function.

Anastomosis, Surgical↗

Compilation of all genes encoding bacterial two-component signal transducers in the genome of the cyanobacterium, Synechocystis sp. strain PCC 6803.

Bacteria have devised sophisticated signaling systems for eliciting a variety of adaptive responses to their environment, which are generally referred to as the "two-component regulatory system." The widespread occurrence of the two-component systems in both prokaryotes and eukaryotes implies that it is a powerful device for a wide variety of adaptive responses of cells to their environment. The two-component signal transducers contain one or more of three conserved and characteristic phosphotransfer signaling domains, named the "transmitter, receiver, and alternative transmitter." The recently determined entire genomic sequence of Synechocystis sp. strain PCC 6803 allowed us to compile systematically a complete list of genes encoding such two-component signal transduction proteins. The results of such an effort, made in this study, revealed that at least 80 ORFs were identified as members of the two-component signal transducers in this single species of cyanobacteria.

Amino Acid Sequence↗

Sequence analysis of the genome of the unicellular cyanobacterium Synechocystis sp. strain PCC6803. II. Sequence determination of the entire genome and assignment of potential protein-coding regions.

The sequence determination of the entire genome of the Synechocystis sp. strain PCC6803 was completed. The total length of the genome finally confirmed was 3,573,470 bp, including the previously reported sequence of 1,003,450 bp from map position 64% to 92% of the genome. The entire sequence was assembled from the sequences of the physical map-based contigs of cosmid clones and of lambda clones and long PCR products which were used for gap-filling. The accuracy of the sequence was guaranteed by analysis of both strands of DNA through the entire genome. The authenticity of the assembled sequence was supported by restriction analysis of long PCR products, which were directly amplified from the genomic DNA using the assembled sequence data. To predict the potential protein-coding regions, analysis of open reading frames (ORFs), analysis by the GeneMark program and similarity search to databases were performed. As a result, a total of 3,168 potential protein genes were assigned on the genome, in which 145 (4.6%) were identical to reported genes and 1,257 (39.6%) and 340 (10.8%) showed similarity to reported and hypothetical genes, respectively. The remaining 1,426 (45.0%) had no apparent similarity to any genes in databases. Among the potential protein genes assigned, 128 were related to the genes participating in photosynthetic reactions. The sum of the sequences coding for potential protein genes occupies 87% of the genome length. By adding rRNA and tRNA genes, therefore, the genome has a very compact arrangement of protein- and RNA-coding regions. A notable feature on the gene organization of the genome was that 99 ORFs, which showed similarity to transposase genes and could be classified into 6 groups, were found spread all over the genome, and at least 26 of them appeared to remain intact. The result implies that rearrangement of the genome occurred frequently during and after establishment of this species.

Bacterial Proteins↗

In vitro bone formation by rat marrow cell culture.

Fresh marrow cells were obtained from the femora Fischer rats and cultured in a medium containing 15% fetal calf serum (FCS) to leach confluent. After trypsinization, cells were subcultured at a cell density of 100 x 10(3)/35 mm well in the presence of FCS, 10 mM beta-glycerophosphate, 82 micrograms/mL ascorbic acid phosphate, and 10(-8)M dexamethasone (Dex). Osteoblastic cells and microscopic mineralized nodules began to appear at about 1 week after the subculture, and at 2 weeks many macroscopic nodules that showed high alkaline phosphatase activity (ALP) and appearance of bone Gla protein (BGP) mRNA were evident. As demonstrated by in situ hybridization, the mRNA was manifested by cuboid-shaped cells (osteoblastic cells). X-ray diffraction (XRD) and Fourier-transform infrared spectroscopy (FTIR) showed the mineralization of fine crystals of hydroxyapatite comparable to natural rat bone mineral. In contrast to these findings, subculture done under the same conditions except for the lack of Dex did not show mineralized nodules, nor did they show the osteoblastic phenotype expression. These analyses indicate that Dex-induced mineralization using rat bone marrow cell culture is an in vitro counterpart of bone formed in vivo. Such a culture is useful for investigating materials/ osteogenic cells interactions.

Alkaline Phosphatase↗

Osteogenic differentiation of cultured marrow stromal stem cells on the surface of bioactive glass ceramics.

To investigate the significance of apatite-wollastonite-containing glass ceramic (AW ceramic) surfaces and the biological apatite layer formed on these surfaces, rat marrow cell culture, which shows osteogenic differentiation, was carried out on four different culture substrata (control culture dish, two AW ceramics, each having a different surface roughness, and a ceramic on which an apatite layer was formed. A culture period of 2 weeks in the presence of beta-glycerophosphate, ascorbic acid, and dexamethasone resulted in abundant mineralized nodule formations that were positive for alkaline phosphatase (ALP) stain on all substrata. The stain on the apatite-formed AW ceramic was the most intense, the enzyme activity being about twice that of the control culture dish, which had the lowest stain and activity of the four substrata. Northern blot analysis of bone Gla protein (BGP) showed the same tendency, that is, the amount of BGP mRNA from cultured cells on the apatite-formed AW ceramics was the highest and the mRNA on the control dish was the lowest. These data indicate that the glass ceramic surface promotes osteoblastic differentiation and that the promotion can be further enhanced by the formation of a biological apatite layer on the ceramic surface.

Alkaline Phosphatase↗

Localization of mouse Rad51 and Lim15 proteins on meiotic chromosomes at late stages of prophase 1.

BACKGROUND: In meiosis, eukaryotic chromosomes show a series of morphological changes, during which chromosomes synapse and recombine. To understand the mechanisms of the morphological changes and recombination of chromosomes, we examined stage-specific localization of the Rad51 and Lim15 proteins on the chromosomes in meiotic prophase 1. These proteins are homologous with the RecA protein and have general properties of searching and pairing of homologous DNA sequences. We used mouse chromosomes whose small sizes allow us to identify the locations of these proteins on the entire structures of the chromosomes. RESULTS: In the leptotene and zygotene stages, the Rad51 protein was present on chromatin loops of mouse testis chromosomes then the protein left the loops. In the pachytene stage, the Rad51 protein was present almost exclusively along the core of the synaptonemal complexes (SC). When the stage proceeded to diplotene, the protein was present in the synaptic regions of chromosomes, in particular, in the chiasma regions. The protein was not present on separated homologous SC cores. On the other hand, the Lim15 protein that was found on chromatin loops in early prophase 1, was present almost exclusively at both ends of the SC cores throughout the late stages of prophase 1. CONCLUSION: The Rad51 and Lim15 proteins are present in chromatin loops when chromosomes form SC. The proteins may promote pairing of homologous DNA sequences that would lead formation of SC. The Rad51 protein in the SC cores may be involved in chiasma formation in late stages. The Lim15 protein, instead, may be involved in recombination in the telomeric region or in cohesion of sister chromatids for segregation.

Adenosine Triphosphatases↗

Thyroid cell proliferation-inhibiting activity in serum of patients with chronic renal failure on hemodialysis.

To investigate the possible humoral factor(s) influencing thyroid cell activity in chronic renal failure, we measured serum activity which stimulates or inhibits the [3H]thymidine incorporation by using a cultured functioning rat thyroid cell line (FRTL-5 cells) in 17 patients on hemodialysis and 19 healthy controls. Polyethylene glycol-treated serum was centrifuged and FRTL-5 cells were cultured with the supernatant. Thyroid stimulating activity was determined by [3H]thymidine incorporation after incubation for 72 h. There was no significant difference in [3H]thymidine incorporation between cultures incubated with patient and normal serum, suggesting the absence of the stimulating activity. But when patient serum was added to cultures together with 20 or 50 microU/ml of TSH, the TSH-stimulated increase in [3H]thymidine incorporation was significantly decreased, indicating the presence of thyroid inhibiting activity, which possibly inhibits the thyroid cell growth. This activity was not significantly altered by hemodialysis. No significant correlation was observed between this activity and serum levels of thyroid hormones or the iodine concentration. Patients on hemodialysis therefore have serum thyroid inhibiting activity which is nondialysable, differs from iodine, and could influence the thyroid cell growth.

Adult↗

Computer survey for likely genes in the one megabase contiguous genomic sequence data of Synechocystis sp. strain PCC6803.

Using the computer program GeneMark, the open reading frames (ORFs) previously assigned within the one megabase sequence data of the genome of the cyanobacterium, Synechocystis sp. strain PCC6803 (Kaneko et al., DNA Res. 2: 153-166, 1995), were re-examined. Matrices required by GeneMark for its statistical calculation were generated and modified by running a script termed GeneMark-Genesis that performed recursive application of GeneMark against the Synechocystis data and evaluated the probability scores for optimization. Based on the matrices thus generated, 752 of the 818 previously assigned ORFs (92%) were supported by GeneMark as likely coding sequences, of which 26 were predicted to start at more internal positions than previously assigned. In addition, 50 ORFs were newly identified as likely coding sequences, most of them being shorter than 300 bp. Thus, the procedure was proven to be very powerful to locate likely coding regions within the genomic sequence data of Synechocystis without having prior information concerning their similarity to the genes of other organisms. However, GeneMark did not predict 66 previously assigned ORFs as likely genes: 14 of them showed significant degrees of similarity to known genes and 10 others were found within IS-like elements. It seems that these genes, many of which appear to be exogenous origin, escaped detection by GeneMark as in the case of "class 3 (horizontally transferred) genes" of E. coli, which in turn suggests that genes of different phylogenetic origins might also be detected as such by modifying the matrices.

Base Sequence↗

Sequence analysis of the genome of the unicellular cyanobacterium Synechocystis sp. strain PCC6803. I. Sequence features in the 1 Mb region from map positions 64% to 92% of the genome.

The contiguous sequence of 1,003,450 bp spanning map positions 64% to 92% of the genome of Synechocystis sp. strain PCC6803 has been deduced. Computer analysis of the sequence predicts that this region contains at least 818 potential ORFs, in which 255 (31%) were either genes that had already been identified or their homologues, 84 (10%) were homologues to registered hypothetical genes, and 149 (18%) showed weak similarities to reported genes. The remaining 330 ORFs showed no apparent similarity to any reported genes or carried no significant protein motifs. The potential ORFs as a whole occupied 86% of the sequenced region, implying compact arrangement of genes in the genome. As to the structural RNA genes, one rRNA operon consisting of 5,028 bp and at least 11 species of tRNA genes were identified. It is noteworthy that 10 out of the 11 tRNA species showed significant sequence similarities to tRNAs reported in plant chloroplasts. As other notable unique sequences, three classes of IS-like elements each with characteristics typical of IS elements were identified, and a typical unit of WD(Trp-Asp)-repeats which have only been detected in the regulatory proteins of eukaryotes was identified within the large 5,079-bp ORF located at map position 69%.

Base Sequence↗

Expression profiles of a human gene identified as a structural homologue of meiosis-specific recA-like genes.

Using the cDNA clone from mouse testis which carries the conserved sequences among meiosis-specific recA-like genes, a highly homologous cDNA clone was isolated from a cDNA library of human testis. The clone had the coding capacity of a protein consisting of 340 amino acid residues, which coincides with the average size of putative eukaryotic recA-like proteins. When expression of the corresponding gene, named HsLIM15, in various tissues was examined by reverse transcription-PCR, products of two different sizes were detected in testis: While the longer was predominantly seen in the testis, the shorter was commonly to all the tissues including the testis. Analysis of the sequences indicated that the longer product corresponded to the above cDNA clone, and the shorter one was its deletion product missing an internal 165 bp portion. The result strongly suggests that the mRNA species coding for the putative meiosis-specific RecA-like protein in human is predominantly expressed in testis possibly as an alternative splicing product of a ubiquitously expressed gene.

Adenosine Triphosphatases↗

Assignment of 82 known genes and gene clusters on the genome of the unicellular cyanobacterium Synechocystis sp. strain PCC6803.

We have previously constructed the physical map of a cyanobacterium, Synechoystis sp. strain PCC6803 on the basis of restriction and linking clone analysis. Since a total of 82 genes and gene clusters have been isolated from this strain, most of which are involved in oxygenic photosynthesis, portions of their sequences were amplified by the PCR method and assigned on the physical map of the genome by hybridization with restriction fragments, ordered clones, which were obtained from cosmid and lambda libraries, and long PCR-products. An exception was the gene psbG2 which was mapped on an extra-chromosomal unit of 45 kb. Since genetic maps of some of genes assigned above, especially those for photosynthesis, have been reported for two other cyanobacterial strains, Anabaena sp. PCC7120 and Synechococcus sp. PCC7002, gene organizations were compared among the three strains. However, no significant correlation was observed, suggesting that rearrangement of genes occurred in the respective strains during or after establishment of the species.

Bacteriophage lambda↗

Characterization of a mouse recA-like gene specifically expressed in testis.

Using the sequence information on a class of recA-like genes, which have been isolated from meiosis-specific cDNA libraries of Lilium and budding yeast and identified in various plant cells, a new gene was screened from a mouse testis cDNA library. The putative product of the gene, termed MmLim15, was 37.8 kDa consisting of 340 amino acid residues. The predicted amino acid sequence showed 72-78% similarity to those of Lim15 from Lilium and other meiosis-specific RecA-like proteins. Similarity was also found to another class of recA-like proteins, Rad51 and its homologues, which have been detected in both budding and fission yeasts and various animal cells, but to a lesser extent and the similarity patterns were somewhat different from those for Lim15 type proteins, revealing that the two classes of recA-like genes are phylogenetically separate. Reverse transcription-PCR using poly(A)+ RNAs from various tissues as templates indicated that transcription of MmLim15 occurred specifically in the testis, suggesting that the MmLim15 product participates in meiotic recombination.

Adenosine Triphosphatases↗

Characterization of a protein tyrosine phosphatase (RIP) expressed at a very early stage of differentiation in both mouse erythroleukemia and embryonal carcinoma cells.

From our previous studies, several protein tyrosine phosphatases (PTPase) are implicated in the early events leading to in vitro differentiation of both mouse erythroleukemia (MEL) and embryonal carcinoma (F9) cells. Among the PTPases, recent experiments suggest that a new PTPase (RIP) plays a critical role in differentiation processes, particularly at their early stages. We isolated cDNA clones for RIP from a RNA preparation isolated from differentiating MEL cells, and determined the total 7932 bp base sequence for RIP cDNA. The cDNA codes for a putative 269.8 kDa (2450 amino acids) protein with a PTPase catalytic domain. We have demonstrated that the transcripts exist in multiple forms, and among mouse tissues they were found predominantly in kidney and, to a lesser extent, in lung, heart, brain and testis. The RIP gene was mapped between D5Mit90 and D5Mit25 on mouse chromosome 5.

Amino Acid Sequence↗