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Biomedical subjects

N Ishida

Publications and source records attributed to N Ishida.

At least 163 records · Page 9Linked to original sources
PubMed1996

PCR-RFLP analysis of the cytochrome b gene in horse mitochondrial DNA.

The mitochondrial DNA sequence of cytochrome b gene in a Thoroughbred horse was determined. By comparing DNA sequences between the Thoroughbred and published sequence data (two horses and one Grevyi zebra), polymerase chain reaction (PCR) primers were designed for amplification of a 590 bp DNA fragment in the cytochrome b gene, and PCR-restriction fragment length polymorphism (RFLP) analysis was studied in 140 horses of six breeds using three restriction enzymes (AciI, BamHI, RsaI). Two morphs were found using each of the three enzymes. By combining three enzymes morphs, the 140 horses examined were classified into four types. Type 2 was most frequent in all breeds.

Animals↗
PubMed1996

Preparation of a monoclonal antibody specific for 5-methyl-2'-deoxycytidine and its application for the detection of DNA methylation levels in human peripheral blood cells.

A monoclonal antibody specific for a modified nucleoside, 5-methyl-2'-deoxycytidine (m5dCyd), was prepared using 5-methylcytidine (m5Cyd)-keyhole limpet haemocyanin (KLH) conjugate, and was characterized. Termed FMC9, the antibody reacts with m5dCyd and slightly with m5Cyd and 5-methylcytosine (m5Cyt) but not with other nucleosides tested in this investigation. FMC-9 was used in an enzyme-linked immunosorbent assay (ELISA) system for the quantitation of m5dCyd levels. Sensitivity was in the picomole range. Methylation levels in peripheral blood cells of healthy donors were determined by inhibition ELISA. The percentage of m5dCyd in peripheral blood cells of 10 healthy donors was 5.08 +/- 0.50%. These results suggest that the inhibition ELISA using FMC9 is useful to monitor m5dCyd levels in the peripheral blood cells.

Adult↗
PubMed1996

Recombinant tissue plasminogen activator prevents intimal hyperplasia after balloon angioplasty in hypercholesterolemic rabbits.

The purpose of this study was to clarify whether continuous infusion of recombinant tissue plasminogen activator (rtPA) prevents intimal hyperplasia at the site of arterial injury. Forty-three Kurosawa and Kusanagi hypercholesterolemic (KHC) rabbits which underwent balloon angioplasty of the right common iliac artery were randomly divided into 2 groups; rtPA for 7 days (n = 29) and untreated (n = 14). The former group was subdivided into a high-dose group (n = 15) (2 mg/kg per day continuously for 7 days after 3 mg/kg for 4h) and a low-dose group (n = 14) (0.6 mg/kg per day for 7 days after 0.6 mg/kg for 4 h). Balloon angioplasty was performed with a 2.5-mm balloon (3 x 60-sec, 6-atm inflations at 60-sec intervals) 1 h after the initiation of rtPA infusion via an ear vein. The iliac arteries of the rabbits were histologically studied at 28 days. The cross-sectional areas of the intima, media, and adventitia were calculated at the site of intimal hyperplasia. The intimal cross-sectional area was 0.07 +/- 0.11 (mean +/- SD) mm2 for the high-dose rtPA group and 0.11 +/- 0.07 mm2 for the low-dose rtPA group, and both of these values were significantly less than that for the control group (0.57 +/- 0.21 mm2 p < 0.01). The ratio of the intimal to medial cross-sectional area was significantly (p < 0.01) lower for the rtPA groups than for the control group (high-dose rtPA; 0.10 +/- 0.13, low-dose rtPA; 0.21 +/- 0.19, control; 1.25 +/- 0.55). In conclusion, continuous infusion of rtPA for 7 days prevented intimal hyperplasia after balloon injury.

Angioplasty, Balloon↗
PubMed1996

Genomic sequence analysis of the bovine male-enhanced antigen-1 (Mea-1) and differential localization of its transcripts and products during spermatogenesis.

The male-enhanced antigen-1 (Mea-1) gene was previously isolated from a bovine testicular cDNA library. In the present study, we cloned the full-length bovine genomic Mea-1 gene and compared this with the Mea-1 cDNA. The 1035-nucleotide bovine mRNA for Mea-1 (excluding the poly (A) tail) is encoded in three exons distributed over 3123 base pairs of the genome. Analysis of the 5' flanking sequence by primer extension mapping identified two main transcription start sites and several minor ones. The 5' region contained transcription-related sequences such as TATA/CAAT boxes, GC-rich regions, and several cis elements. When chloramphenicol acetyltransferase (CAT) activities of 5'-deleted clones were measured in CHO, TM4, and BALB/3T3 cells, a critical region for transcription was identified around -249 to -113 bp region from transcription start site. In situ hybridization and immunohistochemistry indicate that transcripts of the Mea-1 gene were localized in primary and secondary spermatocytes, and spermatids, but the protein products were detected only in spermatids. Intensive transcription of Mea-1 gene and specific localization of the gene product suggest that Mea-1 may play a important role in the late stage of spermatogenesis.

Amino Acid Sequence↗
PubMed1996

[Trial of bladder sparing for patients with invasive bladder cancer by transurethral resection and intra-arterial chemotherapy].

BACKGROUND: We reviewed our experience with bladder conservative treatment for invasive bladder cancer in 20 patients to investigate which type of tumor was suitable for organ conservation. METHODS: We performed radical transurethral resection with dissection to deep layer of the muscle plus intra-arterial chemoinfusion of the cis-platinum at a dose of 80-100 mg/body twice at least, and the recurrent tumor was treated by the same procedure. There were 13 cases of stage T2 and 7 cases of stage T3a. All patients were pathologically diagnosed transitional cell carcinoma with 9 cases of grade 2 and 11 cases of grade 3. RESULTS: With a follow-up of 36 months (range 18 to 82), 15 (75%) of 20 patients were survived and 10 (50%) of 20 patients were free of tumor. Of 5 patients with residual or recurrent tumor, 2 patient underwent cystectomy and the others were required transurethral resection. Of 5 patients died, 4 patients died of cancer all with distant metastasis (lung meta: 2 cases, liver meta: 2 cases). In reference to tumor characteristics, all 4 patients had over 3 cm diameter tumors with non-papillary surface and 3 of 4 patients had invasive tumors extended to bladder neck and prostatic urethra. Of 15 patients alive, 7 (47%) patients had non-papillary tumors, 9 (60%) had grade 3 tumors, 5 (33%) had T3a tumors, but invasion of the prostate urethra was recognized in only 3 patients (20%). CONCLUSION: We had possibility of doing organ prevention therapy for the patients with invasive bladder cancer, even though it was non-papillary or grade 3 tumor. On the other hand, it was thought that a conservative approach should not be considered in patients with large non-papillary tumors that extended to bladder neck and prostatic urethra.

Adult↗
PubMed1996

[Prevention of left ventricular aneurysm formation and left ventricular remodeling caused by percutaneous transluminal coronary angioplasty performed 24-48 hours after onset of acute myocardial infarction].

The prevention of left ventricular aneurysm formation and left ventricular remodeling by percutaneous transluminal coronary angioplasty (PTCA) performed 24 to 48 hours after onset of acute myocardial infarction was investigated. Left ventriculography and coronary angiography were performed in 25 patients within 24 hours of onset of acute myocardial infarction. The patients were divided into two groups; reperfusion group (n = 17) and successful PTCA group (n = 8). The reperfusion group and the PTCA group included patients whose infarct-related coronary arteries were successfully reperfused to 99%, Thrombolysis in Myocardial Infarction (TIMI) grade II or III, immediately after coronary thrombolysis. However, the reperfusion group did not include the patients with spontaneous reperfusion or reperfusion after PTCA. The PTCA group consisted of patients who underwent successful PTCA performed within 24 to 48 hours after onset of infarction (mean 28.4 +/- 6.0 hours). Non-reperfusion in the acute phase was defined as TIMI grade 0-I. The size of the ventricular aneurysm in the PTCA group was significantly reduced compared with the reperfusion group (PTCA group 4.2 +/- 8.7%, reperfusion group 27.2 +/- 6.6%; p < 0.01). The increase of left ventricular end-diastolic volume (delta LVEDVI) from the acute to chronic phases was calculated to estimate left ventricular remodeling. There was a significant difference between the two groups in delta LVEDVI (PTCA group 4 +/- 19 ml/m2, reperfusion group 19 +/- 17 ml/m2; p < 0.05). PTCA performed within 24 to 48 hours after onset in patients with acute myocardial infarction and infarct-related coronary arteries, resulting in reperfusion to 99% TIMI grade II or III immediately after coronary thrombolysis, prevents left ventricular aneurysm formation and remodeling.

Aged↗
PubMed1995

Anisomycin induces phase shifts of circadian pacemaker in primary cultures of rat suprachiasmatic nucleus.

We have developed a cell culture system for the rat suprachiasmatic nucleus, in which a clear circadian oscillation of vasopressin release was observed. Using this culture system, the effect of anisomycin, an inhibitor of protein synthesis, on the circadian rhythm was studied. A phase-delay of more than 15 h could be produced by a 6-h anisomycin pulse. The magnitude of the phase-shift was dependent on the circadian time of the drug treatment and on its dose. The phase-response curve was similar to the response curves that have been measured for protein synthesis inhibitors in other organisms. These results strongly suggest that protein synthesis may be involved in the generation of circadian rhythms in mammals.

Animals↗
PubMed1995

Isolation and characterization of the HC8 subunit gene of the human proteasome.

For study of the molecular basis of regulation of proteasome gene expression, we isolated the gene encoding the alpha-type HC8 subunit of the human proteasome. About 2.3 kb of the 5' flanking region of this gene was tested for promoter function by chloramphenicol acetyltransferase assay. This analysis revealed that CAAT and TATA boxes, but not a GC box, are essential for its promoter activity. These results differed from previous findings that the genes for the alpha-type HC3 and beta-type HC5 subunits of the human proteasome have a TATA-less promoter and that two or three GC boxes function as the promoter sequences (Tamura, T. et al. (1994) J. Mol. Biol. 244, 1117-1124). We mapped the HC8 gene at q23 on human chromosome 14, which differs from the chromosomal locations of nine other proteasomal subunit genes mapped so far.

Animals↗
PubMed1995

Immunochemical detection of urinary 5-methyl-2'-deoxycytidine as a potential biologic marker for leukemia.

A monoclonal antibody against 5-methylcytidine was prepared and characterized. This antibody, termed AMC, was reactive with compounds that had 5-methylcytosine structure (i.e. 5-methyl-2'-deoxycytidine, 5-methylcytidine and 5-methylcytosine). AMC had the highest reactivity to 5-methyl-2'-deoxycytidine among reactive compounds and had no or very slight cross-reactivity to cytidine-related compounds and any other compounds. Analysis of immunoreactive materials in urine revealed that 5-methyl-2'-deoxycytidine rather than 5-methylcytidine was, contrary to our expectation, the major component. Then the inhibition ELISA system using AMC was established and urinary levels of 5-methyl-2'-deoxycytidine in healthy individuals and cancer patients were determined. The mean excretion levels of healthy individuals was 0.90 +/- 0.43 nmol/mumol creatinine and the cut-off value was set at the mean + 2 S.D. of healthy individuals (1.76 nmol/mumol creatinine). Among various types of cancer tested, elevated levels of 5-methyl-2'-deoxycytidine were detected in leukemia patients. From these results, urinary 5-methyl-2'-deoxycytidine might be applicable as a biologic marker for leukemia.

Animals↗
PubMed1995

Mitochondrial DNA sequences of various species of the genus Equus with special reference to the phylogenetic relationship between Przewalskii's wild horse and domestic horse.

The noncoding region between tRNAPro and the large conserved sequence block is the most variable region in the mammalian mitochondrial DNA D-loop region. This variable region (ca. 270 bp) of four species of Equus, including Mongolian and Japanese native domestic horses as well as Przewalskii's (or Mongolian) wild horse, were sequenced. These data were compared with our recently published Thoroughbred horse mitochondrial DNA sequences. The evolutionary rate of this region among the four species of Equus was estimated to be 2-4 x 10(-8) per site per year. Phylogenetic trees of Equus species demonstrate that Przewalskii's wild horse is within the genetic variation among the domestic horse. This suggests that the chromosome number change (probably increase) of the Przewalskii's wild horse occurred rather recently.

Animals↗
PubMed1995

Colonic and peritoneal tuberculosis associated with coloduodenal fistula.

We report a very rare case of tuberculous colitis that showed relatively long-segment involvement of the colon near the hepatic flexure with coloduodenal fistula that caused severe malnutrition. The formation of fistula in abdominal tuberculosis is very rare. This is the eighth reported case of abdominal tuberculosis with fistula and the first reported case with a coloduodenal fistula.

Adult↗
PubMed1995

Molecular biological approach to the circadian clock mechanism.

Many circadian phenomena have been described in a diverse range of species, from single cellular organisms to higher species of plants and animals. From several lines of evidence from Drosophila and Neurospora, the oscillation of the circadian clock seems to involve cycling gene expression. Although a great deal of information concerning the anatomy, neurophysiology and neurochemistry of circadian pacemakers has been obtained over the last decade, molecular and cellular approaches to this problem have only just begun. I will summarize recent progress of the molecular biological approach to the circadian clock mechanism. Finally, the importance of transcription factors to envision the common mechanism of circadian clock in the diverged species will be discussed considering with the existence of a hypothetical 'Time Box'.

Animals↗
PubMed1995

Diffusion of cell-associated water in ripening barley seeds.

Diffusion rate and restricted diffusion of cell-associated water in ripening barley seeds were examined by NMR microscopy using the pulse gradient spin-echo and the pulse gradient stimulated-echo methods. Changes in the mobility of cell-associated water and properties of cell membranes during ontogeny seed were assessed. Diffusion coefficients for bulk water transfer were high (greater than 0.9 x 10(-5) cm2/s) throughout the growth stages. The highest diffusion coefficient observed was comparable to the self-diffusion coefficient of pure water. Water compartment sizes and the permeability of the cell membranes in the seed were determined by Meerwall and Ferguson's modification of the model of Tanner or by the method of Callaghan et al. The endosperm consisted of large cells and vascular bundle small cells with permeable membranes.

Diffusion↗
PubMed1995

SYN987, SYN1193, and SYN1253, new quinolones highly active against gram-positive cocci.

SYN987, 1193 and 1253 are new fluorinated quinolones with enhanced activity against Gram-positive bacteria. The MICs of SYN 987, 1193 or 1253 were studied by an agar dilution method against 88 streptococci (including penicillin resistant Streptococcus pneumoniae), 30 enterococci and 62 Staphylococcus aureus. SYN 987, 1193 and 1253 were 2 to 4 times more active than sparfloxacin and 16-64 times more active than older fluoroquinolones against all streptococci and ciprofloxacin-resistant S. aureus. This study demonstrates that SYN 987, 1193 and 1253 are extremely potent antibacterial agents which deserve further evaluation.

Anti-Infective Agents↗
PubMed1995

SSCP analysis of pig mitochondrial DNA D-loop region polymorphism.

The sequence polymorphism that occurs in the mitochondrial DNA (mtDNA) displacement (D)-loop region is useful as a cytoplasmic DNA marker. We cloned the mtDNA D-loop regions of five breeds of pig by polymerase chain reaction (PCR) and determined their sequences. The sequence diversities in D-loop regions among five breeds of pig were located in the starting area of heavy-strand replication. From these sequences, we designed primers for PCR-mediated single-strand conformation polymorphism (PCR-SSCP) analysis that amplified the most polymorphic 227 bp fragment of the D-loop region. The results of PCR-SSCP analysis clearly showed that four types of polymorphism (A to D) are found in Landrace (A), Large White (A, B), Duroc (A), Göttingen miniature pig (B) and Meishan (C, D). The same polymorphisms were also detected from each porcine embryo by this method. Our results show that PCR-SSCP analysis is useful in detecting polymorphisms in the D-loop region of pigs and pig embryos.

Animals↗