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J Kusuda

Publications and source records attributed to J Kusuda.

32 records · Page 2Linked to original sources
PubMed1996

Human rasGTPase-activating protein (human counterpart of GAP1m): sequence of the cDNA, primary structure of the protein, production and chromosomal localization.

When cDNA encoding rat rasGTPase-activating protein (rat GAP1m) was used as a probe, two partial cDNA clones of a human counterpart of rat GAP1m were isolated from a cDNA library derived from growth-arrested normal human ectocervical epithelial cells. One clone was found to be a cDNA of premature mRNA with two introns. A complete cDNA of human GAP1m was constructed by a series of reverse transcription-polymerase chain reaction (RT-PCR) using total RNA from human epidermoid carcinoma A431 cells. Human GAP1m shows 87.7% nucleotide identity to rat GAP1m in open reading frame and encodes an 850-amino acid protein that shows 89.2% identity to rat GAP1m. A 100-kDa protein was detected in A431 cells by Western blotting with anti-rat GAP1m antibody. The human GAP1m gene was mapped to chromosome 3q24-q26.

Animals↗
PubMed1996

Identification of the gene-richest bands in human chromosomes.

The human genome is a mosaic of isochores, long DNA segments which are compositionally homogeneous and which can be partitioned into five families, L1, L2, H1, H2 and H3, characterized by increasing GC levels and by increasing gene concentrations. Previous investigations showed that in situ hybridization with a DNA fraction derived from the GC-richest and gene-richest isochores of the H3 family produced the highest concentration of signals on 25 R(everse) bands that include the 22 most thermal-denaturation-resistant T(elomeric) bands, a subset of R bands. Using an improved protocol for in situ hybridization and cloned H3 isochore DNA, we have now shown (i) that the number of bands which are characterized by strong hybridization signals, and which are here called T or H3+, is 28; (ii) that 31 additional R bands, here called T'or H3* bands, also contain H3 isochores, although at a lower concentration than H3+ bands; and (iii) that the remaining R bands (about 140 out of 200, at a resolution of 400 bands), here called R" or H3- bands, do not contain any detectable H3 isochores. H3+ and H3* bands contain all the gene-richest isochores of the human genome. The existence of three distinct sets of R bands is further supported (i) by the different compositional features of genes located in them; (ii) by the very low gene density of chromosomes 13 and 18, in which all R bands are H3- bands; (iii) by the compositional map of a H3* band, Xq28; (iv) by the overwhelming presence of GC-rich and GC-poor long (> 50 kb) DNA sequences in H3+/H3* and in H3-/G bands, respectively; and (v) by the large degree of coincidence of H3+ and H3* bands with CpG island-positive bands. These observations have implications for our understanding of the causes of chromosome banding and provide a classification of chromosomal bands that is related to GC level (and to gene concentration).

Chromosome Banding↗
PubMed1996

Sequence analysis of the cDNA for the human casein kinase I delta (CSNK1D) gene and its chromosomal localization.

A cDNA clone coding for human casein kinase I (CK1) has been isolated and sequenced. The insert of 1911 bp contained an open reading frame of 415 amino acids. The entire amino acid sequence of human CK1 was 97% homologous to that of rat CK1 delta, and their sequences in the kinase domain (284 amino acid residues) were completely identical, predicting that the obtained cDNA is for a human homolog of the CK1 delta isoform (CSNKID). The considerable similarity in the amino acid sequence of the kinase domain of human CK1 delta to the Saccharomyces cerevisiae CK1, HRR25 (66%), and to the Saccharomyces pombe CK1, HHP1 (78%), which are involved in the repair of DNA strand break, supports the speculation that human CK1 delta might also act in DNA metabolism through excision and recombinational repair. The human CK1 delta gene was mapped to chromosome 17q25.2-q25.3 by fluorescence in situ hybridization and polymerase chain reaction analysis of the human/rodent hybrid cell panels.

Amino Acid Sequence↗
PubMed1996

Primary culture of chicken hepatocytes in serum-free medium (pH 7.8) secreted albumin and transferrin for a long period in free gas exchange with atmosphere.

To study liver functions of chicken, we examined the primary culture of chicken hepatocytes, and found an easy method of long-term culture with free atmosphere exchange. Chicken hepatocytes were obtained by collagenase perfusion and cultured at 37 degrees C as a monolayer without substratum in serum-free L-15 medium (pH 7.8) with free atmosphere exchange. The amounts of albumin and transferrin in medium were assayed by ELISA. The culture of chicken hepatocytes was maintained in the serum-free L15-medium )pH 7.) and 37 degrees C with free atmosphere exchange for 20 days. The amount of albumin secreted in the medium decreased to low levels early in culture; however, this was followed by marked increase from day 9 to day 17 of culture. The amount of transferrin was constant until day 6, then it too increased with further culture. We reported an easy method for the simple monolayer culture of chicken hepatocytes in serum-free L12 medium (pH 7.8) with free atmosphere exchange over an extended period. Expression of liver-specific functions, viz. albumin and transferrin synthesis, was observed after 1 week of culture.

Air↗
PubMed1995

Assignment of a human autoimmune antigen, p80-coilin gene to chromosome 17q21-q23 and of its possible pseudogene to chromosome 14.

In order to determine the chromosomal locations of an autoimmune antigen, the coilin gene and its pseudogene, we amplified the segments of the two genes by the polymerase chain reaction (PCR) and screened a panel of somatic cell hybrids for the presence of the gene products. The results indicate that the human coilin gene and its pseudogene can be assigned to chromosome 17 and chromosome 14, respectively. Further analysis of cell hybrids bearing chromosome 17 with various deletions localized the coilin gene to the region q21-q23.

Animals↗
PubMed1995

[Stereotactic radiosurgery using a linear accelerator (LINAC): simulation and positioning].

Stereotactic radiosurgery using a Gamma unit obtains good results for small intracranial diseases, arteriovenous malformation (AVM) and acoustic neurinoma. In stereotactic radiosurgery using a linear accelerator (LINAC), many fundamental problems are to be solved. 1) accuracy of a LINAC, 2) making the collimators for high energy X-ray narrow beams, 3) dosimetry for high energy X-ray narrow beams, 4) irradiation methods for stereotactic radiosurgery, 5) fixation of a patient's head and 6) simulation of a target. The usefulness of our method for simulation of a target and for positioning for radiosurgery was investigated. High energy X-ray narrow beams obtained with the collimators for narrow beams (field sizes: 9mm phi, 18mm phi and 27mm phi) satisfy clinical requirements for stereotactic radiosurgery, as indicated by dose profiles and isodose curves. No dosimetry method for high energy X-ray narrow beams has been established yet. Of the main irradiation methods for stereotactic radiosurgery, the method using multiple non-coplanar converging arcs needs no drastic reconstruction for use with LINAC. A patient's head was completely fixed by the stereotactic frame (Patil stereotaxic system or Leksell micro-stereotactic system). Simulation of a target was performed under CT scan. On CT image, the center of a target was determined and the three-dimensional coodinate on the stereotactic head frame target was settled so that the target would be reached. The three-dimensional coordinate for the target was coincided with the isocenter of a LINA by the laser beams of three-directional pointers. Afterwards, the target position was finely adjusted, by using the target positioner manipulator system.(ABSTRACT TRUNCATED AT 250 WORDS)

Humans↗
PubMed1991

[Fracture of percutaneously inserted IVC filters--two cases of Günther's filter].

During the period from 1988 to 1990, percutaneous insertion of Günther vena caval filters were performed in ten patients in our institution. In two of them, fracture and partial migration of anchoring limbs were observed. Günthter's filter is an attractive device for interventional radiologist because of its technical ease of percutaneous placement and relative stability within the vein. However, our experience suggests that this device is not entirely safe and reliable. The patients should be carefully followed up with a possibility of distal migration in mind. Further structural improvement seems necessary.

Adult↗
PubMed1990

Over 80% of NotI sites are associated with CpG rich islands in the sequenced human DNA.

Clones containing DNA segments linked to NotI sites are not only useful for ordering the NotI fragments fractionated by pulsed field gel, but also valuable in the search of unknown genes, because they often contain the CpG rich islands and genes related with them. To know the probability of association of NotI sites with CpG rich islands, we screened 5,188 sequences accumulated in DNA data base for the presence of NotI site and examined the distribution of CpGs around them. The sequential calculation of G + C content and frequency of CpG occurrence at each nucleotide position identified the CpG rich domains close to NotI sites in 77 sequences, which corresponds to 84% of total number of candidate sequences. This frequency is consistent well with the prediction that 89% of NotI sites in mammalian genome are likely to be present in CpG rich islands and would stress the importance of cloning of NotI linking sequences for direct isolation of desired genes. Furthermore, 63 islands newly identified in this study should provide a clue for understanding the transcriptional regulation of associated genes.

Base Sequence↗
PubMed1987

Organization and nucleotide sequence of a densovirus genome imply a host-dependent evolution of the parvoviruses.

The genome structure of a densovirus from a silkworm was determined by sequencing more than 85% of the complete genome DNA. This is the first report of the genome organization of an insect parvovirus deduced from the DNA sequence. In the viral genome, two large open reading frames designated 1 and 2 and one smaller open reading frame designated 3 were identified. The first two open reading frames shared the same strand, while the third was found in the complementary sequence. Computer analysis suggested that open reading frame 2 may encode all four structural proteins. The genome organization and a part of the nucleotide sequence were conserved among the insect densovirus, rodent parvoviruses, and a human dependovirus. These viruses may have diverged from a common ancestor.

Amino Acid Sequence↗
PubMed1986

The complete nucleotide sequence of the tobacco chloroplast genome: its gene organization and expression.

The complete nucleotide sequence (155 844 bp) of tobacco (Nicotiana tabacum var. Bright Yellow 4) chloroplast DNA has been determined. It contains two copies of an identical 25 339 bp inverted repeat, which are separated by a 86 684 bp and a 18 482 bp single-copy region. The genes for 4 different rRNAs, 30 different tRNAs, 39 different proteins and 11 other predicted protein coding genes have been located. Among them, 15 genes contain introns. Blot hybridization revealed that all rRNA and tRNA genes and 27 protein genes so far analysed are transcribed in the chloroplast and that primary transcripts of the split genes hitherto examined are spliced. Five sequences coding for proteins homologous to components of the respiratory-chain NADH dehydrogenase from human mitochondria have been found. The 30 tRNAs predicted from their genes are sufficient to read all codons if the ;two out of three' and ;U:N wobble' mechanisms operate in the chloroplast. Two sequences which autonomously replicate in yeast have also been mapped. The sequence and expression analyses indicate both prokaryotic and eukaryotic features of the chloroplast genes.

Journal Article↗