Biomedical subjects

M Takanami

Publications and source records attributed to M Takanami.

At least 37 records · Page 2Linked to original sources

PubMed1990

[Treatment using CDDP and radiation therapy in treating bladder tumor].

Combination therapy consisting of cis-diamine-dichloroplatinum (CDDP) and radiotherapy was used to treat bladder cancer. The subjects were 20 patients with bladder cancer we experienced since September, 1986. They were between 33 and 83 years old and consisted of 13 males and 7 females. Among the histotypes and grades, there were 3 cases of G2, 7 of G2-G3 and 10 of G3 metastatic epithelial carcinoma. The classification of primary tumors was T2 in six cases, T3a in 12 cases and T4 in two cases. In 12 cases where the bladder was preserved, this therapy was performed after TUR biopsy or TUR, and the effects were evaluated after observation periods of 5-17 months. Side effects caused by CDDP were anorexia in 15 cases, nausea and vomiting in 10 cases, leukopenia in 10 cases and thrombocytopenia in four cases. Those caused by the radiotherapy were diarrhea in seven cases and skin disorders in six cases. All of these side effects were mild and the patients recovered spontaneously.

Adult↗

PubMed1989

Signal structure for transcriptional activation in the upstream regions of virulence genes on the hairy-root-inducing plasmid A4.

The inducibility of the vir genes (virA, -B, -C, -D, -E, and -G) on pRiA4 was examined at the transcriptional level, and the RNA-starting sites were determined by S1-nuclease mapping and primer-extension experiments. All of these genes were inducible, while virA, -E, and -G were transcribed even under noninducing conditions. Each transcription of virB, -C, -D, and -E was initiated at one particular site, but that of virA and -G occurred at two and three sites, respectively, depending on the conditions used. In the DNA region upstream from each inducible transcript, one or more blocks of six base-pairs, 5'TGATAACT3' (vir box), were found to be placed characteristically. These blocks were phasing with an interval of 11 base-pairs, and the most upstream one in each upstream region was preceded by an additional block in the inverted orientation. Although the distance between the block(s) and the promoter varied with the vir gene, every block was placed in a phase nearly opposite to the -35 and -10 regions of the promoter.

Bacterial Proteins↗

PubMed1989

Reconstitution of nucleosomes in vitro with a plasmid carrying the long terminal repeat of Moloney murine leukemia virus.

The potential of nucleosome assembly along the sequence of a plasmid carrying the long terminal repeat (LTR) and its flanking region of Moloney murine leukemia virus was analyzed by in vitro reconstitution experiments with histones from chicken erythrocytes. The results of electrophoretic mobility-shift and micrococcal nuclease-digestion assays indicated that the plasmid DNA contained four preferred sites for nucleosome formation. However, all of these sites were mapped on the vector moiety but not on the LTR moiety. Computer analysis of the sequences in the four preferred sites, each spanning about 150 bp, indicated that short runs of (dA,dT) containing two kinds of triplets, AAA/TTT and AAT/ATT, occurred frequently. Furthermore, many of these triplets tended to occur in the same side of the DNA helix, suggesting that DNA curvature was involved in the preferred sites for nucleosome assembly. Consistent was the observation that DNA fragments carrying these preferred sites showed anomalous electrophoretic mobilities at a low temperature.

Base Sequence↗

PubMed1989

The FokI restriction-modification system. II. Presence of two domains in FokI methylase responsible for modification of different DNA strands.

Based on the previous findings that the FokI methylase (MFokI) consists of 647 amino acid residues and contains two copies of the segment specific for adenine methylase, Asp-Pro-Pro-Tyr, at amino acid positions 218-221 and 548-551, the role of these copies in the methylation reaction was investigated by introduction of a mutation into each segment. The MFokI gene was inserted into M13 vectors, and the Asp residues in the two segments were converted to Gly and Ala by oligonucleotide-directed mutagenesis. The wild-type and mutant genes were recloned into an expression vector, from which gene products were purified. A short DNA fragment carrying the FokI recognition site was treated with each of these enzymes, and after separation of the two strands by duplex formation with M13 viral DNAs carrying the respective strands, the presence or absence of modification was judged from susceptibility to FokI endonuclease. The results of analysis showed that different strands were modified in an asymmetric way by the introduction of mutations into one of the two segments, and that the segments at the N-terminal and C-terminal moieties participated in modification of the strands carrying 5'-GGATG-3' and 3'-CCTAC-5', respectively. We concluded that MFokI contained two functional domains each of which was responsible for modification of different strands in the target DNA.

Deoxyribonucleases, Type II Site-Specific↗

PubMed1989

The fokI restriction-modification system. I. Organization and nucleotide sequences of the restriction and modification genes.

A DNA fragment that carried the genes coding for FokI endonuclease and methylase was cloned from the chromosomal DNA of Flavobacterium okeanokoites, and the coding regions were assigned to the nucleotide sequence by deletion analysis. The methylase gene was 1,941 base pairs (bp) long, corresponding to a protein of 647 amino acid residues (Mr = 75,622), and the endonuclease gene was 1,749 bp long, corresponding to a protein of 583 amino acid residues (Mr = 66,216). The assignment of the methylase gene was further confirmed by analysis of the N-terminal amino acid sequence. The endonuclease gene was downstream from the methylase gene in the same orientation, separated by 69 bp. The promoter site, which could be recognized by Escherichia coli RNA polymerase, was upstream from the methylase gene, and the sequences adhering to the ribosome-binding sequence were identified in front of the respective genes. Analysis of the gene products expressed in E. coli cells by gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that the molecular weights of both enzymes coincided well with the values estimated from the nucleotide sequences, and that the monomeric forms were catalytically active. No significant similarity was found between the sequences of the two enzymes. Sequence comparison with other related enzymes indicated that FokI methylase contained two copies of a segment of tetra-amino acids which is characteristic of adenine-specific methylase.

Amino Acid Sequence↗

PubMed1989

Methylation strongly enhances DNA bending in the replication origin region of the Escherichia coli chromosome.

Two-dimensional gel electrophoresis, at high and low temperatures, and gel mobilities of circularly permuted DNA segments showed a large bending locus about 50 bp downstream from the right border of the 245 bp oriC box, a minimal essential region of autonomous replication on the Escherichia coli chromosome. Bending was strongly enhanced by Dam methylation. In DNA from a Dam- strain, the mobility anomaly arising from altered conformation was much reduced, but was raised to the original level by methylation in vivo or in vitro. Enhancement of the mobility anomaly was also observed by hybrid formation of the Dam- strand with the Dam+ strand. Near the bending center, GATC, the target of Dam methylase, occurs seven times arranged essentially on the same face of the helix with 10.5 bp per turn. We concluded that small bends at each Dam site added up to the large bending detectable by gel electrophoresis.

Bacterial Proteins↗

PubMed1989

Construction of an ordered cosmid collection of the Escherichia coli K-12 W3110 chromosome.

A cosmid library of the Escherichia coli K-12 W3110 chromosome was constructed in which clones were assigned to locations on the chromosome map by hybridization and genetic marker complementation tests. Approximately 70% of the genome was represented by this library. The identified clones can be maintained in the homologous system and would facilitate genetic studies of E. coli.

Chromosome Mapping↗

PubMed1989

[The mechanism of blood outflow from the cavernous tissue of human penis by computer graphics].

There are two theories concerning the mechanisms of human penile erection and its maintenance. One theory goes that the artery responsible for blood inflow into the cavernous space relaxes, while the vein responsible for blood outflow actively contracts. The other theory asserts that no active closing mechanism in present in the penile venous system, but rather the erection is totally controlled by the amount of blood inflow. In order to corroborate the vascular construction of the penis, we prepared serial sections of the penis specimens obtained at autopsy and observed these sections by light microscopy, and we investigated the construction of veins which let blood flow out from the corpus cavernosum penis by computer graphics. As a result, we were unable to find any valves in the blood outflow system. Moreover, efferent vessels were seen to let blood flow from the cavernous spaces, after running immediately below the tunica albuginea for a small distance, to the tunica albuginea, running obliquely through inner veins toward the outside of the tunica albuginea in the direction of the long axis of the penis; then these veins are continuous to the veins on the tunica albuginea. From the above-mentioned structures we can infer the following mechanism of penile erection.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1989

Signal structure of the cis-acting element recognized by VirG protein, a positive regulator in Agrobacterium.

VirG protein is known to be the positive regulator for vir genes of Ri and Ti plasmids in Agrobacterium. To investigate the cis-acting element recognized by VirG, we have determined the transcriptional start points of the vir genes on Ri plasmid (pRiA4b). From the analysis of the upstream sequences, it has been found that the sequences generally similar to 5'TG(A/T)AA(C/T)3' appear in phase with an 11-base pair interval, and that the -35 and -10 regions of the promoters are located nearly in the reversal phase with those phasing sequences. These sequence characteristics are conserved in the vir genes on Ti plasmids (pTiA6 and pTiC58). Moreover, the similar sequence structures were found in the regulatory regions of the genes controlled by PhoB and OmpR, which are the E. coli positive regulators closely related to VirG in their amino-acid sequences.

Bacterial Proteins↗

PubMed1989

[A clinical evaluation of AMS 700 penile prosthesis in the treatment of impotence].

The AMS 700TM & 700 CXTM penile prosthesis was implanted in 34 organic and mixed-type impotent patients between November, 1984 and July, 1988. The patients' ages ranged between 32 and 70, with an average age of 57.2. Twelve weeks after implantation, the prosthesis was evaluated in terms of patient satisfaction with sexual intercourse and postoperative complications. Twenty three patients (67.7%) were highly satisfied, 7 (20.3%) satisfied, 3 (8.8%) dissatisfied and 1 patient worsened (2.9%). Severe complication was found in two cases, both of them being complicated by infection; as a result the prosthesis was removed. Utility of the prosthesis, e.g. effectiveness and safety was; high utility in 26 cases (76.5%), moderate utility in 7 cases (20.6%), and no favorable in one case (2.9%). Thus, overall effectiveness and safety of the AMS 700 penile prosthesis, with its excellent cosmetic appearance upon implantation, were demonstrated by the above findings.

Adult↗

PubMed1989

[Clinical results of endourologic technique for upper urinary calculi].

Between May, 1985 and September, 1987, percutaneous nephrolithotripsy (PNL) and transurethral ureterolithotripsy (TUL) have been performed at our hospital and five affiliated hospitals. We report on the therapeutic results of 216 cases treated during this period. PNL and TUL were performed 168 times on 120 patients (119 males and 41 females) and 57 times on 56 patients (35 males and 21 females), respectively. Rate of success with PNL was 80.7% (103/130) for renal calculus, 85.7% (18/21) for upper ureteral calculus, and 47.1% (8/17) for middle-lower ureteral calculus. Rate of success with TUL was 57.9% (33/57). Of these, cases of ureteral lower edge calculus showed the lowest rate of success, being 33.3% (2/6). Complications of the cases undergoing PNL and TUL were: ureteral damage in 13; hemorrhage in 6; renal pelvic damage in 4; ureteral lower edge stenosis in 3; pyrexia in 3; cardiac insufficiency in 2 and retention of perfusate in posterior peritoneal cavity, pneumonia and renal insufficiency in 1 case each.

Adolescent↗

PubMed1988

Supercoiling response of E. coli promoters with different spacer lengths.

The effect of negative supercoiling on a series of synthetic Escherichia coli promoters has been investigated. These promoters carry perfect consensus sequences at the -35 and -10 regions, but with different spacer lengths (Aoyama, T. et al. (1983) Nucleic Acids Res. 11, 5855-5864). Topoisomeric plasmids carrying these synthetic promoters were constructed, and their activities were compared by detecting in vitro transcripts with the probe-hybridization method. In the relaxed state, the one with 17 basepairs (bp) spacing showed the highest activity, and the activity steeply decreased both sides of the optimal spacing. Similar results have been observed by run-off transcription. By introducing negative superhelicity, the 17 bp spacing promoter showed a relatively little response to supercoiling. In contrast, the activities of those with 16 and 18 bp spacings were markedly stimulated by supercoiling, with the mean, negative superhelical density (-sigma) which gave the maximum activity being about the same for the 16-18 bp spacing promoters (-sigma = 0.03 to 0.04). The promoter with 19 bp spacing, which showed no activity in the relaxed state, exhibited a significant activity at higher superhelicities (-sigma = 0.06). Even the 20 bp spacing promoter showed some activity by increasing superhelicity, while the 15 bp spacing promoter did not. On the basis of these observations, possible mechanisms by which negative supercoiling of DNA stimulates the protomer activity are discussed.

DNA, Bacterial↗

PubMed1988

Primary lipogranuloma of male genitalia.

Lipogranulomas developing secondarily in the genitourinary system have been reported rather frequently, but primary lipogranulomas without any past history of etiologically related conditions are rare. We report on 2 cases recently encountered in which tumors were diagnosed as sclerosing lipogranuloma on histopathologic examination.

Adult↗

PubMed1986

Structure of viral DNA in a rat cell line, GY1, transformed by Ad12 HindIII fragment-G.

The cell line GY1, established by transformation of a rat cell line 3Y1 with the Ad12 HindIII fragment-G (leftmost 6.8%, nucleotide 1 to 2322), contains more than 100 viral copies per haploid genome. The viral DNAs in this cell line were cloned into a phage vector, lambda gtWES lambda B, and recloned into pBR322 with their flanking cellular DNAs. Independently isolated 39 clones were analyzed by restriction enzyme cleavage and Southern blot hybridization experiment and divided into 11 classes. Some of classes contained multiple identical clones, at maximum 16 clones. It may be interpreted that amplification of some of the recombined sequences had occurred after the multiple integrations of transfected DNAs within cells. Using five clones from different classes the sequences of recombination sites were determined. Viral DNAs deleted with varying degrees at both ends were flanked by quite different cellular sequences in different clones and no common sequences were revealed around viral-cellular junctions. Tandemly repeated viral DNAs were found in one of the clones to be integrated in a head to tail manner into cellular DNA. The linkage of these two viral DNAs had occurred at the site where parental viral DNAs shared 2 bp. Palindrome structures could be constructed around viral-cellular and viral-viral junction sites and around the regions of parental viral DNAs corresponding to the junction sites in all of the cases investigated.

Adenoviridae↗

PubMed1986

Evaluation of therapeutic efficacy in psychogenic impotence by means of logarithmic scoring.

In an attempt to develop as objective a method for the evaluation of impotence as possible, the authors assigned numerical values to four parameters considered essential to sexual function, namely, libido, erection, ejaculation, and orgasm, and used the sum totals of these scores, collected before and after initiation of therapy, as indicators of overall sexual function. Numerical values were assigned according to a logarithmic scale, in four stages from 0 to 10, i.e., 0, 1, 3, and 10; a score of 0 signified "normal" and a score of 10, "abnormal." By comparing sum totals of scores computed before and after initiation of therapy, the authors were able to evaluate therapeutic efficacy on the basis of changes in these sum totals. Using this method, the mean total score for a control group of 24 normal subjects was 1.67 +/- 0.26 (mean +/- standard error). For the test group, which consisted of 24 patients of psychogenic impotence, the mean total score prior to initiation of therapy was 16.46 +/- 3.55, an extremely high score in comparison with the control group. After four weeks of therapy, the mean total score dropped to 9.37 +/- 1.77, indicating a statistically significant (p less than 0.05) decrease over the pre-therapy mean total score.

Adult↗

PubMed1985

Sites of dnaA protein-binding in the replication origin of the Escherichia coli K-12 chromosome.

On the basis of the observation that dnaA protein binds preferentially to DNA fragments carrying the Escherichia coli chromosomal replication origin (oriC), the binding sites were investigated by DNase I footprinting. As a result, three strong binding sites were identified in the minimal oriC sequence. The respective binding sites were 16 to 17 base-pairs long, and contained a common sequence (5') T-G-T-G-(G/T)-A-T-A-A-C (3') in the middle, although their polarities were not the same. Since mutants defective in function for autonomous replication have been isolated in the corresponding positions of the common sequence at each binding site, dnaA protein-binding at these sites seems to be significant for replication initiation.

Bacterial Proteins↗

PubMed1985

Essential structure of E. coli promoter II. Effect of the sequences around the RNA start point on promoter function.

Starting from a synthetic E. coli promoter with the consensus sequences at -35 and -10 regions, a sequence CAT frequently occurred in the RNA start points of natural promoters was introduced in the downstream of the consensus sequences, and the sequences around the RNA start points as well as the relative positions of CAT from the consensus sequences were altered. Analysis of the RNA start points and strength of these synthetic promoters in an in vitro transcription system provided evidence that the RNA start point was principally fixed by distance from the -10 consensus sequence (TATAAT). Neither the promoter strength nor the RNA start point was significantly influenced by the CAT sequence. The sequences around the RNA start points rather seemed to exert influence on the response of promoter to temperature and salts.

Base Sequence↗

PubMed1985

Nucleotide sequence coding for the insecticidal fragment of the Bacillus thuringiensis crystal protein.

The insecticidal crystal protein (ICP) gene, icp, from a 68-kb plasmid derived from Bacillus thuringiensis subsp. sotto was cloned in Escherichia coli. The icp expression in E. coli cells was confirmed by both immunological and insect-toxicity assays of the cell extract. The entire icp gene resides in the 6.6-kb PstI fragment, which codes for a 144-kDal peptide identical to the intact ICP, as determined by its size and reaction with anti-ICP antibody. Deletion analysis further revealed that the 2.8-kb region within the 6.6-kb PstI fragment codes for ICP. Analysis of the nucleotide sequence indicated that a peptide of 934 amino acid residues truncated at the C-terminal end is encoded by this 2.8-kb fragment. A unique feature of this truncated ICP is the abundance of cysteine and lysine residues within its C-terminal region.

Bacillus thuringiensis↗