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Biomedical subjects

Y Kaneko

Publications and source records attributed to Y Kaneko.

At least 91 records · Page 5Linked to original sources
PubMed2001

Use of human recombinant DNase I expressed in COS-7 cells as an immunogen to produce a specific anti-DNase I antibody.

To obtain human deoxyribonuclease I (DNase I) as an immunogen, we have developed a procedure that is more useful and effective than the conventional procedure, which uses human urine as a starting material. In the new procedure, we culture COS-7 cells transfected with expression vector carrying human DNase I cDNA, and then purify the enzyme from the culture medium. The enzyme can be easily isolated to apparent homogeneity by passage through only three chromatography columns. The rabbit antiserum that we used against the recombinant DNase I was not inferior to that used against DNase I from human urine, in terms of both its ability to discriminate DNase I phenotypes and its ability to neutralize enzyme activity. Therefore, our procedure may be useful for producing an antibody specific for human DNase I.

Animals↗
PubMed2001

New antifungal 1,2,4-triazoles with difluoro(substituted sulfonyl)methyl moiety.

New 1,2,4-triazoles (2) having a difluoro(substituted sulfonyl)methyl moiety were designed and synthesized via alpha,alpha-difluoro-alpha-(substituted thio)acetophenones (3). Compounds (2) showed potent antifungal activities against C. albicans, C. krusei, A. flavus and A. fumigatus in vitro and against C. albicans in vivo for oral and i.v. administrations. Especially, (-)-2a, (-)-2b and (-)-2d showed potent antifungal activities.

Antifungal Agents↗
PubMed2001

Application of the PHO5-gene-fusion technology to molecular genetics and biotechnology in yeast.

Modern biological scientists employ numerous approaches for solving their problems. Among these approaches, the gene fusion is surely one of the well-established valuable tools in various fields of biological sciences. A wide range of applications have been developed to analyze a variety of biological phenomena such as transcriptional regulation, pre-mRNA processing, mRNA decay, translation, protein localization and even protein transport in both prokaryotic and eukaryotic organisms. Gene fusions were also used for the study of protein purification, protein structure, protein folding, protein-protein interaction and protein-DNA interaction. Here, we describe applications of gene fusion technology using the Saccharomyces cerevisiae PHO5 gene encoding repressible acid phosphatase to molecular genetics and biotechnology in S. cerevisiae. Using the PHO5 gene fusion as a reporter, we have identified several cis- and trans-acting genes of S. cerevisiae which are involved in splicing of pre-mRNA, biosynthesis of amino acids, ubiquitin-dependent protein degradation, signal transduction of oxygen and unsaturated fatty acid, regulation of transcription by the nucleosome and chromatin. The PHO5 gene fusions exhibiting the mating-type specific expression were also generated to develop a breeding technique for industrial yeast. It is concluded that the PHO5 gene fusion is extremely useful and should be further exploited to investigate various cellular steps of the eukaryotic gene expression.

Journal Article↗
PubMed2001

Evaluation of signal-averaged electrocardiography for clinical diagnosis in arrhythmogenic right ventricular dysplasia.

Arrhythmogenic right ventricular dysplasia (ARVD) is a heart muscle disorder of unknown etiology that is characterized pathologically by fibrofatty replacement of the right ventricular myocardium. We investigated the relationship between the electrocardiogram (ECG) appearances and signal-averaged ECG (SAECG) in 7 cases with ARVD, and evaluated the usefulness of SAECG as a screening test to detect patients with ARVD. Compared with the conventional 12-lead ECG, the SAECG detects abnormalities at a higher rate in ARVD patients (57% versus 86%). SAECG was more sensitive as a screening test to detect patients with ARVD than 12-lead ECG.

Adolescent↗
PubMed2001

Intracranial meningeal malignant fibrous histiocytoma mimicking parasagittal meningioma--case report.

A 69-year-old female presented with a rare intracranial meningeal malignant fibrous histiocytoma (MFH). The neuroimaging appearance of this tumor was very similar to parasagittal meningioma. The tumor was grossly totally removed, and local irradiation of 50 Gy was performed. The histological diagnosis was MFH. The patient recovered from the preoperative deficits, and no recurrence was observed by 27 months after surgery.

Aged↗
PubMed2001

Measurement of pulpal blood flow in dogs with nonradioactive colored microspheres.

The viability of dental pulp depends largely on regional blood flow, as in other organs. Measurement of absolute pulpal blood flow (PBF) and comparisons with blood flow in other organs allow the prediction of microvascular regulation in dental pulp. In previous studies, PBF was measured in dogs mainly with radioactive microspheres. However, this established technique is inaccessible to many investigators due to concerns over radiation safety and radioactive waste. To overcome these limitations, a new method has been introduced that involves the use of nonradioactive colored microspheres for measuring regional blood flow in the myocardium and in other organs in animals. However, no previous studies have investigated the use of this method to measure PBF in dogs. We attempted to determine whether blood flow in dental pulp, which comprises a small amount of the total tissue in dogs, could be measured using this technique by comparing the measured values with those for regional myocardial blood flow. Mean blood flow values were between 0.148 and 0.182 ml/min/g for dental pulp at four different sites and about 1.0 ml/min/g in regional myocardium. These values are comparable to those previously reported using radioactive microspheres. As nonradioactive colored microspheres safety permitted measurement of absolute PBF in dogs, this technique appears to be useful for research into microvascular blood flow in dental pulp.

Analysis of Variance↗
PubMed2001

Effects of recombinant human endostatin on a human neuroblastoma xenograft.

New antitumor agents must be added to the current neuroblastoma treatment regimens to improve the clinical results. We investigated whether recombinant human endostatin (rhEndostatin), an antiangiogenic agent, is effective against human neuroblastoma in the human neuroblastoma xenograft model designated TNB9. When tumors on the back of nude mice grew to a weight of 90-95 mg, rhEndostatin 10 mg/kg/day was administered subcutaneously every day for 10 consecutive days. Mean relative tumor weight in mice administered rhEndostatin (n=5) was significantly less than that in controls (n=12) on days 2, 4, and 6 after the start of administration (p<0.01 on day 2, p<0.05 on days 4 and 6), and regression of tumor growth (TRW<1.0) was marked on day 2. The maximum inhibition rate (MIR) by rhEndostatin was 46.4%, indicating inefficacy, but it may not be appropriate to apply Battelle Columbus Laboratories criteria to this experimental model because rhEndostatin is a protein. After day 8, tumors in the experimental group increased in weight and were not statistically significantly different from those in controls. Recombinant human endostatin was used in tumors in the arterial system of the mouse in this experiment because eventually rhEndostatin, not recombinant mouse endostatin, may be used to treat advanced neuroblastoma in the clinical setting. The results show that there is little cross-reactivity of rhEndostatin with the human and mouse models and indicate that rhEndostatin could become an effective agent for the treatment of human neuroblastoma.

Animals↗
PubMed2001

Quantitative analyses of messenger RNA of human endogenous retrovirus in patients with systemic lupus erythematosus.

OBJECTIVE: Human endogenous retrovirus (HERV) has emerged as a possible causative agent of systemic lupus erythematosus (SLE). To investigate the role of HERV in the etiology of SLE, we performed quantitative analyses of messenger RNA (mRNA) of the HERV clone 4-1 in patients with SLE. METHODS: Reverse transcriptase (RT)-polymerase chain reaction (PCR) and real-time quantitative PCR (RQ-PCR; TaqMan methodology) were used in this experiment. RESULTS: The quantities of mRNA of the HERV clone 4-1 gag region in patients with SLE were significantly higher than in healthy controls, and the amounts of such mRNA in the patients were decreased by steroid treatment. CONCLUSION: These phenomena may be related to the production of viral components derived from HERV clone 4-1 and contribute to the pathogenesis of SLE; studies using a larger number of patients are required to confirm these points.

Adult↗
PubMed2001

[Changes of fatty acid composition of brain synaptosomal phospholipids in isoflurane exposure rat].

We examined effects of inhalational anesthetic drug, isoflurane, on phospholipid and fatty acid in brain synaptosome. Wistar strain male rat was treated by inhalation of isoflurane. Rats were divided into 3 groups each 6 rats, one was 1 minimum alveolar concentration (MAC) exposure group, 1MAC group, and another was 2 times of MAC exposure group, 2MAC group, and the other was non exposure group, control group. The animals were kept in to box (0.343 m3) and the gas flow rate was set in 4 L/min by anesthetic instrument. After 60 minutes of exposure, rats were decapitated. Immediately, cerebrums were removed and fraction of synaptosome was sampled. In 2MAC group, C14:0 of phosphatidylcholine (PC) increased significantly as compared to the control group, but C16:1, C18:0, C18:2 and C20:3 decreased significantly. And also, C18:2 and C20:3 decreased significantly in 1MAC group. In terms of phosphatidylethanolamine(PE), C18:1 in 1MAC group, C14:0 and C16:1 in 2MAC group increased, but C20:3, C20:4 and C22:5 in 2MAC group decreased significantly as compared to the control group. Regarding phosphatidylserine + phosphatidylinositol, C14:0 in 2MAC group increased, but C22:5 decreased. In lysophosphatidylcholine, C12:0 and C14:0 in 2MAC group and C18:0, C20:4 in 1MAC group increased significantly, but C18:1 in 2MAC decreased. The changes of phospholipids and fatty acid in synaptosome were due to the metabolism of phospholipids of basic matrix and this was caused by effects of isoflurane on neural cellular membrane. The results indicated the suppression of membrane activity. Isoflurane has physiological activity on metabolism of phospholipid of cellular membrane. Thus, it has effects on neural cellular functions in brain.

Anesthetics, Inhalation↗
PubMed2001

Comparative study on anesthetic potency depending on concentrations of lidocaine and epinephrine: assessment of dental local anesthetics using the jaw-opening reflex.

Anesthetic potency of a local anesthetic on the dental pulp was investigated by increasing or decreasing the concentration of lidocaine and that of epinephrine. An electromyogram of the digastric muscle in Japan White male rabbits was recorded during the jaw-opening reflex induced by electrical stimulation of the dental pulp. Probit analysis was used for the determination of the 50% effective volume (ED50) values of the anesthetic. The anesthetics used were plain 2% lidocaine solution (2Lid-0 group), 2% lidocaine solution with 12.5 microgram/mL of epinephrine (2Lid-1/8 group), 2% lidocaine solution with 6.25 microgram/mL of epinephrine (2Lid-1/16 group), and 4% lidocaine solution with 5 microgram/mL of epinephrine (4Lid-1/20 group). No anesthetic effect was shown in the 2Lid-0 group. The 2Lid-1/8 group indicated adequate anesthetic potency with the smallest dosage at all observation periods. The potency in the 2Lid-1/16 group was 0.3-0.5 times, and that in the 4Lid-1/20 group was 0.3-0.4 times as much as the 2Lid-1/8 group. The decrease in epinephrine concentration produced the decrease in the anesthetic potency on the dental pulp independent of lidocaine concentration. These results suggest that the increase in lidocaine concentration may not compensate the decrease in epinephrine concentration.

Anesthesia, Dental↗
PubMed2001

[Intermittent androgen deprivation treatment of prostate cancer restarted at low level of serum prostate specific antigen: a pilot study].

We report a pilot study on a novel protocol of intermittent androgen deprivation (IAD) treatment of prostate cancer (PC), in which androgen deprivation is restarted when serum prostatic specific antigen (PSA) level reached more than 2 ng/ml and is stopped when PSA level decreased below 0.3 ng/ml. Thirty-two patients (aged 60 to 86 years, median 74 years) with prostate cancer (Stage A in 4 patients, B in 20, C in 1, D in 5, and relapse after radical prostatectomy in 2) were treated with IAD. Median serum PSA prior to the start of endocrine therapy was 15.65 (range 2.67 to 306.3) ng/ml. Eleven patients were treated with lutenizing-hormone-releasing hormone (LHRH) agonist alone and 21 were treated with LHRH agonist plus an antiandrogen. Median duration of first endocrine therapy was 572 (range 100 to 1,543) days. Median serum PSA at the start of first off-phase was 0.038 (range 0.003 to 0.489) ng/ml. After a median of 207 days (range 140 to 843) of follow-up, 19 patients were in the first cycle, 9 in the second cycle, 3 in the third cycle, 1 in the fourth cycle. Two patients developed androgen-independent PC. The median duration of first off-phase of IAD was 287 days. There was a significant inverse relation between the duration of the first on-phase and testosterone level measured 4 months after the cessation of first on-phase therapy (R = -0.518). These results suggest that our protocol provides a reasonable length of off-phase duration and that the long term-androgen deprivation phase might delay the recovery of the testicular endocrine function which should be maintained during the off-phase of IAD.

Aged↗
PubMed2001

[Nationwide sensitivity surveillance of various antibiotic activities against bacteria isolated from patients with severe infections].

The susceptibility of 3,058 bacterial strains isolated between January and March, 1997 from patients with severe infections in Japan to ciprofloxacin and other injectable antimicrobial agents was measured using broth microdilution method. Methicillin-resistant Staphylococcus aureus (MRSA) strains were generally sensitive to vancomycin, teicoplanin and arbekacin, and resistant to CPFX and other antibacterial agents. MIC90 of CPFX against Streptococcus pneumoniae, to which MIC of ampicillin was more than 4 micrograms/mL, was below 2 micrograms/mL. PRSP (Penicillin resistant S. pneumoniae), which was also resistant to cephalosporins and carbapenems, showed no cross-resistance to CPFX. The susceptibility of Gram-negative bacteria to CPFX was as high as that to carbapenems. Especially, MIC90 against Pseudomonas aeruginosa was 2 micrograms/mL. 3 strains of isolated 446 P. aeruginosa strains had blaIMP gene. CPFX and pazufloxacin demonstrated good susceptibility with 0.25 microgram/mL of MIC to 2 strains of these 3 strains. The susceptibility rate of the most common isolates from patients suffering from lower respiratory tract infections excluding MRSA to CPFX was more than 80% (indication: % strains < pneumonia break point).

Anti-Bacterial Agents↗
PubMed2001

[The usefulness of autologous blood transfusion for urologic surgery].

To avoid potential risks associated with homologous blood transfusion including viral infection and graft versus host disease (GVHD), autologous blood donations have been promoted in urologic surgery. We assessed its necessity in the patients undergoing radical retropubic prostatectomy and total cystectomy. A total of 27 patients ranging from 54 to 78 years old donated 400 to 1,200 ml of blood prior to radical prostatectomy (17 patients) and total cystectomy (10 patients). Recombinant erythropoietin was administered in 26 out of 27 patients. The mean hemoglobin concentration was 14.1 g/dl before donation and 12.8 g/dl before operation. The mean volume of surgical blood loss was 1,659 ml ranging from 529 to 2,990 ml in total cystectomy, and 1,438 ml ranging from 553 to 2,841 ml in radical prostatectomy. Overall, 22 out of 27 patients (82%) did not require an additional homologous blood transfusion. In conclusion, autologous blood donation is a safe and useful method to avoid homologous transfusion in radical prostatectomy and total cystectomy. Eight hundred ml of blood donation is suggested to be appropriate prior to these surgeries.

Aged↗
PubMed2001

[Extracorporeal shock wave lithotripsy for upper urinary tract stones using piezoelectric shock wave lithotripsy].

Between August 1999 and July 2000, 123 cases of renal stones and 52 cases of ureteral stones in 116 males and 59 females were treated with the New Piezolith 2500. The average number of sessions required for renal and ureteral stones was 2.50 and 1.48, respectively. At 3 months postoperatively, stone-free rates for renal and ureteral stones were 64.2% and 72.7%, respectively. Assessing residual stones less than 4 mm in diameter as an effective treatment outcome, the efficacy rates for renal and ureteral stones were 94.3% and 86.4%, respectively. Side effects were encountered in 5 cases (2.9%) of high fever and one case (0.57%) of renal subcapsular hematoma. New Piezolith 2500 is effective and safe for the treatment of upper urinary stones.

Adult↗
PubMed2000

Positive regulation of transcription of homeoprotein-encoding YHP1 by the two-component regulator Sln1 in Saccharomyces cerevisiae.

The IME1 gene is essential for initiation of meiosis in Saccharomyces cerevisiae. Transcription of IME1 is induced under starvation for nitrogen and glucose and in the presence of the MATa1 and MATalpha2 gene products. We have shown in our previous work that homeoprotein Yhp1 binds to a 28-bp region between nt -702 and -675 of the IME1 promoter in vivo and in vitro. We also revealed that the 28-bp region fused with a reporter gene harbored Yhp1-dependent URS (upstream repression sequence) activity and that the transcription of YHP1 was repressed by nonfermentable carbon source. In this study, we found, using a 5'-deletion series of the YHP1 promoter fused with a reporter gene, that the URS responsible for repression of YHP1 transcription with a nonfermentable carbon source is located at a region from nt -696 to -466 of the YHP1 promoter. We also identified and delimited a UAS (upstream activation sequence), which confers activation in both fermentable and nonfermentable carbon source media, to be from nt -356 to -306 of the YHP1 promoter. The UAS of the YHP1 promoter contained an MCE (Mcm1 control element) that is a target of the general transcription factor Mcm1 and is known to be involved in positive regulation by the two-component regulator Sln1. Consistent with this fact, the YHP1 transcription level was reduced in the Deltasln1 mutant, indicating that the two-component regulator Sln1 is involved in activation of YHP1 transcription.

Base Sequence↗
PubMed2000

Characterization of S818L mutation in HERG C-terminus in LQT2. Modification of activation-deactivation gating properties.

We examined the mechanism(s) for HERG channel dysfunction in an S818L mutation in the HERG C-terminus using the heterologous expression system in Xenopus oocytes. Injection of S818L cRNA alone did not produce expressed currents. Coinjection of an equal amount of S818L cRNA with wild-type (WT) cRNA into oocytes did not exhibit apparent dominant-negative suppression. However, coinjection of excess amounts of S818L cRNAs with WT cRNA into oocytes decreased HERG current amplitudes and shifted the voltage dependence of activation to negative potentials, accelerated its activation and deactivation. The data suggest that S818L alone cannot form functional channels, whereas S818L subunits can, at least in part, coassemble with WT subunits to form heterotetrameric functional channels, and imply that the HERG C-terminus may contain a domain involving the activation-deactivation process of the channel. These findings may provide new insights into the structure-function relationships of the HERG C-terminus.

Amino Acid Substitution↗