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Biomedical subjects

S Kanoh

Publications and source records attributed to S Kanoh.

At least 37 records · Page 2Linked to original sources

Pharmacokinetics of glutathione-dextran macromolecular conjugate in mice.

A fluorescein-labeled dextran-glutathione conjugate (FD-GSH) was synthesized in order to examine its disposition in the body. GSH was covalently attached to the FITC-labeled dextran by the cyanogen bromide activation method. Mice were injected with FD-GSH through the tail vein, and the levels of FD-GSH in the blood and various organs were measured fluorometrically. A substantial level of FD-GSH was found in the liver and this reached a maximum at 6-8 h after the injection. The hepatic uptake clearance was estimated to be 0.541 +/- 0.014 ml/h/g tissue or 42.4 +/- 9.8 ml/h/kg body weight. FD--GSH accumulated in the liver for a long period, while the half-life of the conjugate in the blood circulation was 1.45h. The cumulative urinary and fecal excretions of FD-GSH were 14% and 4% of dose at 72h after the injection, respectively. A molecular design of the conjugate was discussed on the basis of the results.

Animals↗

Actin-binding peptide from smooth muscle myosin light chain kinase.

The objective of this study was to localize the actin-binding site in the smooth muscle myosin light chain kinase. Limited proteolysis by thermolysin indicated that hydrolysis of the kinase at the N-terminal end of the molecule resulted in loss of actin-binding ability. Various methods of cleavage were investigated for the generation of a discrete actin-binding peptide. The method chosen was cleavage at the cysteine residues by the 5,5'-dithiobis(2-nitrobenzoic acid)-cyanide complex. This procedure yielded an actin-binding peptide of approximate M(r) 17,000. The peptide was purified and shown to possess the actin-binding properties of the native myosin light chain kinase. The binding constant of the isolated peptide and parent enzyme to actin was estimated as 7.5 x 10(4) M-1. From the amino acid composition of the peptide and comparison with the sequence of gizzard myosin light chain kinase, it was suggested that the actin-binding site is located within the N-terminal sequence 1-114. Comparison with other actin-binding proteins shows some similarities to gizzard alpha-actinin and caldesmon.

Actins↗

Tetrodonic acid-like substance; a possible precursor of tetrodotoxin.

A tetrodonic acid-like substance which was hardly distinguishable from authentic tetrodonic acid in thin-layer chromatography, high performance liquid chromatography, etc., was successfully purified from the ribbon worm and flatworm by a method consisting mainly of Bio-Gel P-2 column chromatography. The tetrodonic acid-like substance showed a specific toxicity of approximately 700 mouse units/mg as tetrodotoxin, unlike tetrodonic acid which is a completely non-toxic substance. Instrumental analyses including gas chromatography-mass spectrometry, secondary ion mass spectrometry and thin-layer chromatography/fast atom bombardment mass spectrometry disclosed that the tetrodonic acid-like substance had a structure similar to, but not the same as, that of tetrodotoxin. This, along with its high convertibility into tetrodotoxin during storage and other experimental operations, suggested that the tetrodonic acid-like substance is a precursor of tetrodotoxin.

Animals↗

Intra-arterial chemotherapy for bladder cancer by insertion of catheter from inferior gluteal artery.

Thirty-four patients with locally advanced bladder cancer have been treated with selective intra-arterial infusion of CDDP and/or ADM (IA therapy) prior to planned surgical resection. Follow-up ranged from 25 to 108 months (median 61). Initial tumor stage was cT2 in 10 patients, cT3 in 19 and cT4a in 5. Catheterization technique: gluteal muscles were dissected gently along the muscle fiber to expose the inferior gluteal artery with the patients in prone position, then the catheter was inserted. The tip was wedged in the internal iliac artery below the bifurcation of the superior gluteal artery. ADM 10-20 mg and/or CDDP 10-20 mg were infused once or twice a week. Total dose of ADM and CDDP were 40-580 and 60-240 mg. Thirteen patients received IA therapy + hyperthermia and 8 IA therapy + irradiation. Surgical resection included total cystectomy (22 patients), partial cystectomy (3 patients) and transurethral resection of the prostate (5 patients). Survival rate at 5 years is 57.9% (T2 = 90.0, T3 = 52.1, T4 = -). Eighteen patients are alive with no evidence of recurrences, and 11 patients were free of disease for more than 5 years. Side effects were bone marrow suppression (5 patients), vomiting (4), erosion of gluteal skin (7), and neurotoxicity, such as sensory disturbance in lower extremities or ischialgia (2); treatment was well tolerated in others. In conclusion, our results suggest that intra-arterial infusion of ADM and/or CDDP by insertion of catheter from inferior gluteal artery is safe with minimal systemic side effects, and prolongs survival for invasive bladder cancer.

Antineoplastic Agents↗

Signal peptide of the colicin E2 lysis protein causes host cell death.

Colicin E2 is encoded by the SOS-inducible colicin operon of plasmid ColE2-P9. Highly induced colicin E2 is excreted from the E. coli host cell, accompanied by quasi-lysis and cell death. These three events are all directed by the colicin lysis protein that comprises the signal peptide and a small mature lipoprotein. We constructed a mutant lysis gene encoding its complete signal peptide with addition of two external amino acids at the C-terminus. By using this mutant peptide as a reference, the wild type lysis signal peptide was identified and shown to remain quite stable after the cleavage. This mutant peptide also mimics the cleaved-off signal peptide avoiding the influence of the mature lysis lipoprotein, and suggested that cell death by the lysis protein, but neither colicin excretion nor quasi-lysis, is decided by the nature of its signal region.

Base Sequence↗

[Combination therapy of intra-arterial chemotherapy and local hyperthermia for bladder cancer].

Fourteen patients (age range 50-79 mean 64.2) with locally advanced bladder cancer or CIS have been treated with combination of selective intra-arterial infusion of adriamycin (ADM) and/or cisplatin (CDDP) and local hyperthermia, prior to planed surgical resection. The follow-up periods ranged from 33 to 70 months (median 52). Clinical staging was based on biopsy, CT, Echo, chest X-ray, bone scintigraphy, and IVP. The initial tumor stage was Tis in 1 patients, cT2 in 4, and cT3 in 9. Catheterization technique: with the patient in the prone position, the gluteal muscles were dissected to expose the inferior gluteal artery, then catheter was inserted. The tip was wedged in the internal iliac artery below the bifurcation of the superior gluteal artery. ADM 10-20 mg and/or CDDP 10-20 mg were infused once or twice a week, more than eight times during hyperthermia. The total dose of ADM and CDDP were 40-360 mg and 50-360 mg. Local bladder hyperthermia was achieved with a radiofrequency generator. Novatherm IH-500 (Inter-Nova Co. Ltd, Japan). Intravesical temperature was maintained over 40 degrees C for an hour. Surgical resection included total cystectomy (10 patients), partial cystectomy (3) and TUR (1). Of evaluable 8 patients, CR was obtained in 2 patients, PR in 4, NC in 1 and PD in 1. The survival rate (Kaplan Meier method) at 3 years was 71.4% (n = 9) and at 5 years, 62.5% (n = 4). Five patients died, but two of them died of other causes (traffic accident and spinal cord injury). Severe toxicity, consisting of bone marrow suppression (3) perineal erosion (3) and ischialgia (1), was well tolerated. In conclusion, our result suggests combination therapy of intra-arterial chemotherapy and local hyperthermia is safe with minimal systemic side effects, and may prolong survival with invasive bladder cancer.

Aged↗

Evaluation of the teratogenic potential of the rubber accelerator N-cyclohexyl-2-benzothiazylsulfenamide in rats.

The teratogenicity of N-cyclohexyl-2-benzothiazylsulfenamide (CBS) was studied in Wistar rats. Pregnant rats were given CBS at a dosage of 0.001, 0.01, 0.1 or 0.5% in the diet from Day 0 to Day 20 of pregnancy. Daily intakes of CBS were 0.7 mg kg-1 for the 0.001% group, 7.1 mg kg-1 for the 0.01% group, 69.6 mg kg-1 for the 0.1% group and 288.8 mg kg-1 for the 0.5% group. Maternal body weight gain during pregnancy in the 0.1 and 0.5% groups was significantly lowered. Food consumption during pregnancy in the CBS-treated groups, except for the 0.5% group, did not differ from that in the control group. Neither death nor clinical signs of toxicity were noted in the pregnant females of any group. Lowered weight in fetuses and the placentae were observed in the 0.5% group. There were no significant compound-related effects on the incidences of pre- and post-implantation losses and the number and ratio of live fetuses. Morphological examinations of the fetuses revealed no evidence of teratogenesis. It could be concluded that CBS possesses no adverse effects on the prenatal development of the offspring in rats at doses employed in the present study.

Animals↗

Evaluation of teratogenic potential of sodium sulfite in rats.

The teratogenicity of sodium sulfite was examined in Wistar rats. The pregnant rats were fed diets containing 5, 2.5, 1.25, 0.63 or 0.32% of sodium sulfite heptahydrate(Na2SO3.7H2O) ad libitum from day 8 to 20 of pregnancy. Maternal toxicity, as evidenced by decreased body weight gain and decreased food consumption was observed at the 5% group, but no clinical signs of toxicity were observed. A significant reduction in the fetal body weight of both sexes was observed in all dose groups except 2.5% group. No significant differences in the numbers of live fetuses and intrauterine death (dead fetuses and resorptions) or sex ratios of fetuses were found between the sodium sulfite-treated and control groups. Fetal external, skeletal and internal malformations were not observed at any dose level. However, several types of skeletal and internal variations as well as delayed ossifications were observed in some groups treated with sodium sulfite, but the incidences were not significantly different from controls. Also, some fetuses with dilatation of the renal pelvis and the lateral ventricle were found in all groups except 1.25% group, but there was no dose-response. The live birth index and survival rate of offspring within 4 weeks and their body weight gain at 3 weeks after birth were not affected by sodium sulfite-treatment. In conclusion, sodium sulfite (0, 0.32, 0.63, 1.25, 2.5 or 5.0% as Na2SO3.7H2O) administered in the diet to Wistar rats during days 8-20 of pregnancy produced related signs of fetal toxicity but no evidence of teratogenicity.

Animals↗

[Studies on the urophonography (1)--Basic investigation with the model of lower urinary tract].

The purpose of this study is to ascertain whether the urinary (turbulent) flow really causes the sounds during micturition in humans. For this purpose the artificial bladder and urethra, which were made of silicon rubber (3M Co. Minnesota, U.S.A.), were used. The manometric device was attached to the bladder to measure the intravesical pressure. Outside the bladder neck a prostate like protrusion surrounding the urethra was made. A microphone was attached to this protrusion to detect the sound. A recording system was also adopted. The bladder was filled with 200 ml of sterile eater. By pressurising the artificial bladder at 100 cmH2O, the water passed through the urethra. During this artificial voiding the sound detection was undertaken. At the same time uroflowmetry was also performed. It was demonstrated that sound spikes were recorded whenever water in the artificial bladder passed through the artificial urethra.

Auscultation↗

[Studies on the urophonography (2)--Clinical investigation on normal human subjects and patients with benign prostatic hypertrophy].

Recently we have developed an entirely new method of sonic detection of lower urinary tract disorders during micturition. The detection of these sounds was performed by a data catching and recording system, consisting of a detector, amplifier, and recorder. This procedure was designated as "urophonography" and its recording as "urophonogram". Twenty-five patients with BPH before and three months after suprapubic prostatectomy and 10 healthy male volunteers underwent urophonography. The urophonograms were classified into four types. Type 1 was characterized by its diamond shape, while Type 2 was characterized by its random sound spikes. Type 3 was a mixture of Types 1 and 2. Type 4 had only few sound spikes. Analysis of these urophonograms also revealed that the frequencies of these urethral sounds belonged to the range from 0.4 to 1.5 KHz. The amplitude (power) of the sounds was in the range of 40-60 dB. Healthy volunteers belonged to Type 4. BPH patients belonged to Types 1, 2 and 3 at equal frequency. Comparison of uroflowmetric parameters with urophonograms revealed that Type 3 patients with BPH showed a lesser degree of micturition disturbances, which was consistent with the weight of prostatic tumors surgically resected. Urophonography was undertaken before and after the treatment on these patients. It was clearly shown that the pattern of urophonograms shifted to normal (Type 4). It was considered that urophonograms were useful for urodynamic investigation.

Aged↗

[Studies on the urophonography (3)--Clinical investigation on prostatic cancer].

Recently we have developed an entirely new method of sonic detection of lower urinary tract disorders during micturition. This procedure was designated as "urophonography" and its recording as "urophonogram". Sixteen patients with prostatic cancer before and three months after anti-androgenic therapy and ten healthy male volunteers underwent urophonography. The urophonograms were classified into four types. Type 1 was characterized by its diamond shape, while Type 2 was characterized by its random sound spikes. Type 3 was a mixture of Types 1 and 2. Type 4 had few sound spikes. Analysis of these urophonograms also revealed that the frequencies of these urethral sounds belonged to the range from 0.4 to 1.5 KHz. The amplitude (power) of the sounds was in the range of 40-60 dB. Healthy volunteers belonged to Type 4. Prostatic cancer patients belonged to Types 1, 2 and 3 at 18.8, 68.8, nad 12.4%, respectively. After the treatment the percentages of Types 1, 2, 3, and 4 changed to 18.8, 50.0, 0, and 31.2%, respectively. Comparison of uroflowmetric parameters with urophonograms showed that Type 2 patients showed a lesser degree of micturition disturbances, evidenced by these parameters. It was considered that urophonograms were useful as urodynamic investigation.

Aged↗

[The long-term prognosis of intra-arterial chemotherapy in invasive bladder cancer].

During the period from August 1980 to April 1989, preoperative intra-arterial infusion chemotherapy was performed in 22 patients with resectable bladder cancer. An oblique incision approximately 12 cm long was made in the gluteal lesion to expose the inferior gluteal artery, into which a Teflon catheter was inserted and fixed. Via this catheter, a single dose of 10-20 mg CDDP and/or 10 mg ADM was injected 1-2/week. The majority of patients were treated with radiation and/or hyperthermia as a combined therapy. The 5-year survival rates (Kaplan-Meier method) were 81.6% in patients treated with intra-arterial infusion chemotherapy as an adjuvant to total cystectomy, compared with 37.5% in patients treated with total cystectomy only. There was a statistically significant difference between the two groups (p less than 0.01). Therefore, inferior-gluteal-artery infusion chemotherapy is effective as a preoperative adjuvant therapy with no serious side effects.

Aged↗

Detection of tetrodotoxin by thin-layer chromatography/fast atom bombardment mass spectrometry.

A new method for detection of tetrodotoxin (TTX) by thin-layer chromatography/fast atom bombardment (FAB) mass spectrometry was developed. TTX and/or related substances were separated by TLC on LHP-K high-performance precoated plates, with a solvent system of pyridine:ethyl acetate:acetic acid:water (15:5:3:4). The plates were subjected to positive FAB mass spectrometry, under scanning within a mass range from m/z 100 to 500. TTX was identified by selected ion-monitored chromatograms at m/z 320 (M + H)+ and 302 (M + H - H2O)+, along with full scan positive ion FAB mass spectrometry. The limit of detection for TTX was about 0.1 micrograms. TTX was also detected by cellulose acetate membrane electrophoresis/FAB mass spectrometry.

Animals↗

Tumor-bone interaction induced by transplantable human tumors in nude mice.

Tumor-bone interactions were experimentally studied using four transplantable human urogenital tumors in nude mice. The method consisted of subcutaneously (SC) inoculating tumor cells over the calvaria in nude mice after the periosteum has been disrupted. This resulted in a local tumor causing fragmentation of the bone. The degree of tumor-bone interaction also varied with the type of implanted tumors as shown in radiographic and histologic examinations. All tumors were associated with histologic patterns of classical bone remodeling, including bone destruction with osteoclast proliferation and reactive new bone formation. The evidence presented here suggests that the majority of tumor-bone interaction showed a combination of both features, bone destruction and new bone formation, and the mechanisms whereby tumors interact with bone may vary with the biological properties of the tumor. Our new system would be suitable for studying the biology of local interaction between bone and tumor cells and searching out a method to protect the bone from cancer cells.

Animals↗

Establishment of a model to evaluate inhibition of bone resorption induced by human prostate cancer cells in nude mice.

A model system of human prostate carcinoma in nude mice for searching out a method to protect the bone from cancer cells is described, in which the transplanted human prostate cancer cells were inoculated subcutaneously over the calvaria in nude mice after the periosteum wa disrupted. The tumor induced osteolysis associated with osteoclast proliferation accompanied with reactive new bone formation. This osteolysis was evaluated by measuring the increased area of bone resorption by its reduced opacity to X-ray, and histology. Etidronate disodium, a diphosphonate derivative, at a dose of three mg./kg. and 10 mg./kg. s.c. protected the bone by decreasing the extent of osteolysis as judged by the above criteria. This inhibition was obtained with no apparent effect on the growth of the tumor. These results are discussed in light of recent clinical work, showing that this animal model is a useful tool to test the effect of new drugs against osteolysis of cancer as well as to study the biology of local interaction between bone and cancer cell.

Adenocarcinoma↗

Teratology study of Tween 60 in rats.

The teratogenicity of Tween 60 was studied in Wistar rats. Pregnant rats were given Tween 60 at a dose of 0, 0.1, 1.0 or 10% in the diet from day 7 to day 14 of pregnancy. Daily intakes of Tween 60 were 99 mg/kg for the 0.1% group, 960 mg/kg for the 1.0% group and 7693 mg/kg for the 10% group. No change induced by Tween 60 was detected in the number, sex ratio and body weight of live fetuses. External, skeletal and internal examinations of the fetuses revealed no evidence of teratogenesis. It could be concluded that Tween 60 has no harmful effects on the prenatal development of the rat offspring at doses employed in the present study.

Animals↗

[Studies on the clinical effectiveness of combination therapy with irradiation and chemotherapy for advanced bladder cancer].

The effectiveness of a preoperative combination of chemotherapy and irradiation on advanced bladder cancer was evaluated. The combination therapy group (Arm I) included 21 patients, the preoperative chemotherapy group (Arm II) 14 patients, and a group without preoperative treatment, 35 patients. There were no distinguishing background factors among the three groups. Chemotherapy included vincristine, adriamycin, mitomycin, bleomycin/and 5-FU. A total of 3000 rads was irradiated locally, and the results were promising. The Arm I group was superior to the other two groups in terms of response rate, histological effectiveness and downstaging effects. Also, the 5-year-survival rate for Arm I was much better than in the other groups. There were no great differences in adverse effects between Arm I and II.

Aged↗