IWASE, M., NUNOI, K., SADOSHIMA, S., KIKUCHI, M. and FUJISHIIMA, M. Liver, Kidney and Islet Cell Tumors in Spontaneously Hypertensive and NormotensiveRats Treated Neonatally with Streptozotocin. Tohoku J. Exp. Med., 1989, 159 (2), 83-90-We studied the oncogenic action of neonatal streptozotocin (STZ) treatment in spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto rats (WKY) for 12 months. Two-day-old male neonates were intraperitoneally injected with STZ of which doses were 37.5-75.0mg/kg for SHR and 100.0-150.0mg/kg for WKY. The 12-month survival rate was 16 of 22 (73%) in SHR and 10 of 14 (71%) in WKY, respectively. The incidence of tumors in STZ-treated SHR was 27% in liver, 14% in kidney and 5% in liver and kidney, being related to the dose of STZ given, namely, 25% in 37.5mg/kg, 50% in 50.0 or 62.5mg/kg and 75% in 75.0mg/kg. In STZ-treated WKY which survived 12 months, all had tumors, namely, 70% in liver, 20% in kidney and 10% in liver and kidney. Histological features of liver and kidney tumors were characteristic of hepatoma and nephroblastoma, respectively. Islet cell tumor was evident in 4 of 10 (40%) in SHR treated with lower doses of STZ (≤50mg/kg) but not in SHR and WKY treated with higher doses (62.5-150.0mg/kg). The present study indicates that neonatal STZ treatment has the oncogenic action on liver, kidney and pancreatic islet.
TANNO, N., OIKAWA, S., KOIZUMI, M., FUJII, Y., HORI, S., SUZUKI, N., SAKUMA, E., KOTAKE, H., NAMAI, K. and TOYOTA, T. Effect of Taurine Administration onSerum Lipid and Biliary Lipid Composition in Man. Tohoku J. Exp. Med., 1989, 159 (2), 91-99-The effect of oral administration of taurine (3.2g/day, 2 weeks) on the metabolism of lipids and bile acids was studied with healthy humans. Four male subjects were fed taurine. Another five male subjects were administered 1g of cholesterol daily for two weeks and, at intervals of two weeks, cholesterol and taurine simultaneously. Serum lipoprotein and duodenal bile were analyzed. Oral administration of taurine resulted in the increase of taurine-conjugated bile acids. However, neither serum lipid nor biliary lipid composition was altered. Addition of taurine with cholesterol administration showed elevation of both the serum low density lipoprotein cholesterol level and the lithogenic index in bile. The ratio of glycine-to taurine-conjugated bile acids was changed from 4.1 to 0.63. The ratio of cholic acid/chenodeoxycholic acid was augmented from 0.57 to 0.81. The percentage of taurocholic acid, taurochenodexycholic acid and taurodeoxy- cholic acid were increased about 4-fold, 2.5-fold and 3-fold, respectively. Our results suggested that taurine administration alone did not influence the serum lipid level although taurine-conjugated bile acids were increased. The taurine intake would increase serum low density lipoprotein cholesterol and biliary cholesterol levels when excessive cholesterol is administered simultaneously.
MINEGISHI, N., MINEGISHI, M., TSUCHIYA, S. and KONNO, T. Effects of InVitro Treatment with 4-Hydroperoxycyclophosphamide and Hyperthermia on LeukemicProgenitor Cells. Tohoku J. Exp. Med., 1989, 159 (2), 101-111-A key point of autologous bone marrow transplantation for leukemic patients is how to remove leukemic cells from their own bone marrow grafts. In this study a leukemic progenitor cell assay was used to evaluate the antileukemic efficacy of marrow-purging protocols that employed hyperthermia or 4-hydroperoxycyclophosphamide (4HC) against leukemic blasts obtained from patients. After the treatment of 2×107 nucleated bone marrow cells/ml with 100μg/ml of 4HC in the presence of 7% suspension of packed autologous erythrocytes, leukemic colonies were eradicated in 10 of 13 cases and reduced to less than 0.3% as compared with the colony count in untreated cultures in two cases. More than 10% of leukemic progenitor cells survived after hyperthermia treatment (42°C 60 min) in 7 of 9 cases. It is suggested that treatment of leukemic cells and 7% autologous erythrocytes with 100μg/ml of 4HC is effective to eliminate leukemic progenitor cells. Treatment with hyperthermia may not be effective enough to eliminate leukemic progenitor cells from autologous bone marrow.
MINEGISHI, N., MINEGISHI, M., TSUCHIYA, S. and KONNO, T. Preservation ofImmature Hematopoietic Progenitor Cells Responding to Interleukin 3 in MarrowTreated with 4-Hydroperoxycyclophosphamide. Tohoku J. Exp. Med., 1989, 159 (2), 113-122-The toxic effects of 4-hydroperoxycyclophosphamide (4HC) on different human hematopoietic progenitor cells were determined. Day 7 colonies supported by granulocyte colony stimulating factor (G-CSF), day 10 colonies supported by granulocyte-macrophage colony stimulating factor (GM-CSF) were counted. Approximately 2×102 granulocyte-macrophage colonies per 5×104 bone marrow (BM) mononuclear cells were formed in each assy system. A combination of interleukin 3 (IL3) and erythropoietin (EPO) induces 3 types of day 14 colonies; erythroid burst-forming units (BFU-E, 40±29/5×104 BM cells), granulocytes-macrophage colony forming units (CFU-GM, 143±29/5×104 cells) and mixed colonies (CFU-GEMM, 24±13/5×104 cells). After treatment with 4HC, all the colonies were reduced. However, the recovery rate of day 14 colonies supported by the combination of IL3 and EPO (mean 12.1%) was significantly higher than the recovery rate of both day 7 colonies supported by G-CSF (mean 1.0%) and day 10 colonies supported by GM-CSF (mean 4.0%). The recovery rate of colonies supported by IL3 and EPO was also higher than that of day 14 colonies supported by GM-CSF and EPO. Immature pluripotent hematopoietic progenitor cells supported by IL3 and EPO appeared less sensitive to 4HC treatment than those supported by G-CSF and GM-CSF. Colony forming assays using IL3 may be usefull in order to predict the hematological reconstitution after autologous bone marrow transplantation with 4HC treated graft.
TASAKA, Y., MATSUMOTO, H., INOUE, Y. and HIRATA, Y. Contents andSecretion of Glucagon and Insulin in Rat Pancreatic Islets from the Viewpoint ofTheir Localization in Pancreas. Tohoku J. Exp. Med., 1989, 159 (2), 123-130- Regional difference in secretion of glucagon and insulin from the rat pancreatic islets and their contents in pancreatic tissue and islets were studied. The glucagon content in the normal rat pancreas was the highest at the splenic part followed in order by the gastric, choledochal and duodenal parts. The effect of alloxan on the glucagon content was stronger in the dorsal lobe (combined gastric and splenic parts) than in the ventral lobe (combined duodenal and choledochal parts), and more increase of glucagon was found. The size of pancreatic islets was similar between the dorsal and the ventral lobe. The glucagon content in the islets was significantly higher in the dorsal lobe than in the ventral lobe, but the insulin content in the islets was similar in both lobes. The release of insulin from cultured pancreatic islets from the dorsal lobe was similar to that from the ventral lobe, but the release of glucagon was significantly high from the cultured islets of the former in the presence of glucose or arginine compared with that from the latter. Also in isolated pancreatic islets the release of glucagon was significantly more marked in the islets from the dorsal lobe by arginine administration. These findings show that the islets from the dorsal lobe secrete and contain more glucagon than those from the ventral lobe in contrast to there being a similar amount of release of insulin between them.
AMANO, M., IMATAKA, K., SUZUKI, K., NAKAOKA, H. and FUJII, J. Age-Related Reduction in the Number of Rabbit Erythrocyte Na, K-ATPase. Tohoku J. Exp. Med., 1989, 159 (2), 131-137-The number of Na, K-ATPase units, assessed by [3H]-ouabain binding assay, was significantly less in erythrocytes from 17 mature (30-week) rabbits than in those from 17 young (16-week) ones (0.179± 0.010 vs. 0.263±0.014 pmol/109 cells, p<0.01). Consistent with this finding, the erythrocyte sodium concentration was significantly higher in mature rabbits as compared with young animals (17.1±1.30 vs. 10.8±0.68 mmol/liter, p<0.01). When the assays were repeated in 8 of the young rabbits 8 weeks after the initial determination, both variables were found to be similar to the levels from mature animals. The data suggest that the activity of erythrocyte sodium pump declines with aging or growth of donor rabbits, as a result of a reduction in the number of Na, K-ATPase units.
WAKO, Y., SUZUKI, K., ISOBE, A. and KIMURA, S. Increased HexokinaseActivity in Fetuses of Rats Developed under Maternal Hyperglycemia. Tohoku J. Exp. Med., 1989, 159 (2), 139-145-Hepatic and diaphragmatic hexokinase (HK) activities increased in fetuses of rats developed under maternal hyperglycemic conditions, compared to those in normal controls. In those organs, Type I and Type II hexokinase isozymes were detected by electrophoresis and densitometry with a chromatoscanner. Densitometric peak that corresponded to hepatic and diaphragmatic Type II hexokinase was also increased in the fetuses developed under hyperglycemia, compared to that of normal fetuses. These findings suggest that in the fetal liver, where significant activity of Type IV was absent, activity of Type II hexokinase varied depending on physiological state such as maternal hyperglycemia.
SAKUMA, T., ASAI, K., ICHIKI, T., SUGIYAMA, N., KIDOUCHI, K. and WADA, Y. Identification of Benzoylcarnitine in the Urine of a Patient of Hyperammonemia. Tohoku J. Exp. Med., 1989, 159 (2), 147-151-Benzoylcarnitine was identified in the urine of a patient with a carbamoyl-phosphate synthase I deficiency for whom sodium benzoate and L-carnitine had been used to treat hyperammonemia. This is a newly identified metabolite of benzoate. Its excretion in the urine was increased day by day at the administration of both sodium benzoate and L- carnitine from 0.10 to 2.25mmol/g creatinine. Since there is the possibility of a secondary carnitine deficiency and an increase of benzoyl toxicity after long-term therapy with benzoate supplementation and protein restriction, it is important to monitor the urinary excretion of benzoylcarnitine.
NAKAMURA, H., TAKASAWA, M., KASAHARA, S., TSUDA, A., MOMOTSU, T., ITO, S. and SHIBATA, A. Effects of Acute Protein Loads of Different Sources on RenalFunction of Patients with Diabetic Nephropathy. Tohoku J. Exp. Med., 1989, 159 (2), 153-162-To evaluate effects of acute loading of different proteins on renal function, glomerular filtration rate (GFR), albumin excretion rate (AER), and concentrations of plasma amino acids, 11 normal volunteers and 20 diabetic patients were studied before and after eating 1.0g/kg body weight of either tuna fish meal or bean curd on separate days. In normal subjects, the mean baseline GFR was 115.8±9.5ml/min/1.73m2, and the mean GFRs after ingestion of tuna fish meal were 134.1±15.5 (1hr), 146.2±18.8 (2hr), and 157.8±21.2 (3hr), respectively. GFR did not significantly increase in normal subjects after ingestion of bean curd. GFR in diabetic patients with normoalbuminuria after ingestion of each protein was similar to the response in normal subjects. In diabetic patients with microalbuminuria, GFR did not significantly increase after ingestion of each protein. In diabetes with macroalbuminuria, GFR decreased after ingestion of tuna fish meal and did not significantly change after intake of bean curd. In both normal subjects and diabetic patients, urinary AER did not increase after each kind of protein loading. Plasma concentrations of alanine, glycine, and arginine, known to induce glomerular hyperfiltration, increased to a greater degree after ingestion of tuna fish meal than after administration of bean curd. These findings suggest that responses of GFR to acute protein loading may differ according to the amino acid composition of the protein ingested and to the stage of diabetic nephropathy.
NISHIZAWA, O., ISHIDA, H., SUGAYA, K., KOHAMA, T., HARADA, T. and TSUCHIDA, S. Application of Doppler Color Flow Imaging Method of the Detectionof Vesicoureteral Reflux. Tohoku J. Exp. Med., 1989, 159 (2), 163-164-The doppler color flow imaging method was employed to detect the vesicoureteral reflux. The ultrasound examination was performed by using the doppler color flow imaging system. Ultrasonic sector scan was carried out by the use of the 3.75 MHz phased array and/or conbex array transducer with a transabdominal approach. The flow in the bladder from the ureteral orifice to the bladder was detected with a red color indicating the direction towards the probe and the flow from the bladder towards the ureteral orifice during the vesicoureteral reflux was detected with a blue color indicating the direction away from the probe. The doppler color flow imaging method appears to be a useful diagnostic procedure for the detection of the vesicoureteral reflux.
NAGAKURA, T., SUZUKI, H., MIYAZAKI, Y., OTSUKI, M., GOTO, J., NAMBARA, T. and TOYOTA, T. Determination of 3β-Hydroxylated Bile Acids in Human Serumand Liver Tissue. Tohoku J. Exp. Med., 1989, 159 (2), 165-166-3β, 7α- Dihydroxy-5β-cholanoic acid (3β, 7α-diOH) was detected in serum of 3 patients with intrahepatic cholestasis. 3β, 7β-dihydroxy-5β-cholanoic acid (3β, 7β-diOH) appeared in serum of those patients after treatment with ursodeoxycholic acid (UDC). These bile acids were also detected in only unconjugated fractions of serum of another 7 patients with chronic liver diseases, but not in liver tissue of them. The liver does not seem to from these bile acids itself because they were absent in liver tissue.
KOIWA, Y., HOSHI, N., OHYAMA, T., TAKAGI, T., KIKUCHI, J., HONDA, H. and TAKISHIMA, T. Effect of Left Ventricular Volume on the Magnitude of FunctionalDepression by External Minute Vibration. Tohoku J. Exp. Med., 1989, 159 (2), 167-168-We examined the effect of the left ventricular (LV) volume on the magnitude of the vibration induced functional depression (VID) using four canine cross-circulated isovolumically beating LV preparations. A sinusoidal, 50Hz vibration (1.5mm in amplitude) was applied to the ventricular anterior epicardium at different values of LV volume which was stepwisely increased by 1-2ml from 0 to 30ml. VID was estimated as the difference of the peak LV pressure between control and during external vibration. VID increased as an increment in LV volume when the volume was below 7-8ml and when peak LV pressure was less than 70mmHg. However, VID remained constant when the volume increased further. We concluded that VID was independent of LV volume at physiological range.
KOIWA, Y., TAKAGI, T., KIKUCHI, J., HONDA, H., HOSHI, N. and TAKISHIMA, T. The Improvement of Systolic Function of Depressed Left Ventricle by ExternalVibration at Diastole. Tohoku J. Exp. Med., 1989, 159 (2), 169-170-The left ventricular (LV) incomplete relaxation (IR) has been reported to play an important role in the pathophysiology of the congestive heart failure such as causing higher diastolic pressure and/or impeding the coronary perfusion during diastole. Therefore, using open chest canine preparations (n=4), we examined 1) whether the minute external vibration during diastole could release IR and 2) what occurred to LV systolic function in this perturbation.. LV failure with IR was induced by propranolol administration and, if necessary, by rapid atrial pacing up to 180 beats/min. When we applied a 50Hz, sinusoidal vibration of 2.1mm magnitude during diastole to the epicardium of LV with complete relaxation, there was no significant change in the ventricular function. However, the systolic functional improvement (3.8±1.1mmHg elevation in LV systolic pressure) was observed when the vibration was applied to LV with IR. We concluded that external vibration at diastole could release IR and would be useful to improve the systolic function of the depressed heart with IR.