AMAKASU, H., SUZUKI, H., MIYAKAWA, T., OOTSUKI, M, and GOTO, Y. AEnzymatic and Fluorimetric Determination of Total Bile Acid Sulfates in HumanUrine. Tohoku J. exp. Med., 1986, 149 (4), 343-350- A simple, precise and sensitive method for separation and determination of total bile acid sulfates in human urine is described. The sulfate fraction of urinary bile acids was separated with lipophilic anion exchange gel, piperidinohydroxypropyl Sephadex LH-20 after sample clean-up with Sep-Pak C18 cartridge. The obtained sulfate fraction was submitted to solvolysis with a small volume of dimethoxypropane-HCl solution and subjected to enzymatic-fluorimetrical assay using 3α-hydroxysteroid dehydrogenase and resazurin. In this method, no influence of existing salts in the reaction mixture on fluorescence intensity was observed and solvolysis reaction was almost complete. Overall recoveries of glycine- and taurine-conjugated bile acid 3-sulfates from normal urine ranged from 90.5 to 93.7% and those of unconjugates from 48.7 to 78.0%. The sensitivity of the described method enabled to estimate total bile acid sulfates with 0.5ml of normal urine and precision tests showed the satisfactory accurasy. The level of total urinary bile acid sulfates was estimated on some patients with hepatobiliary diseases and healthy subjects.
TAKAHASHI, H., HANANO, M., HAYASHI, S., ARAI, Y., YOSHINO, N., TAKAKUWA, E., TATEWAKI, W., NAGAYAMA, R., TAKIZAWA, S. and SHIBATA, A. Plasma Levelsof Protein C and Vitamin K-Dependent Coagulation Factors in Patients onLong-Term Oral Anticoagulant Therapy. Tohoku J. exp. Med., 1986, 149 (4), 351-357-Plasma levels of protein C (PC) and vitamin K-dependent coagulation factors (factors II, VII, IX and X) were measured in 100 specimens from patients on long-term warfarin therapy. Both activities and antigens of these coagulation factors were decreased, depending on the thrombotest values. Factor II activity/ antigen ratio and factor X activity antigen ratio were correlated well with thrombotest values, indicating that the concentration of inactive molecules (PIVKAs) relative to normal proteins increases with increasing intensity of anticoagulation. Although PC antigen (PC: Ag) was also decreased, the ratios between PC: Ag and vitamin K-dependent coagulation factor antigens remained constant, being independent of the intensity of warfarin therapy. These findings indicate that long-term oral anticoagulant therapy results in the suppression of the synthesis of both vitamin K-dependent coagulation factors and PC, but the production of the coagulant and anticoagulant proteins is well-balanced.
FUNAKOSHI, N., OHNO, T., KANOH, T., UCHINO, H., MIYAKE, T., ODA, T., KIYOSHIGE, K., ISHIDA, T. and KOHRIYAMA., K. Agammaglobulinemia in aPregnant Woman. Tohoku J. exp. Med., 1986, 149 (4), 359-365 - A case of common variable immunodeficiency in a pregnant woman is presented. Lobar pneumonia developed in the sixth month of the pregnancy and her serum immunoglobulin levels were found to be extremely low. She was treated successfully with immune human serum globulin and antibiotics. She delivered a full-term baby without any troubles. Various immunologic studies were done in the peripheral blood of the patient and the neonate. Consequently, functional abnormalities of helper T cells were considered to be responsible for the hypogammaglobulinemia.
ARAKI, S., MURATA, K. and YOKOYAMA, K. Socioeconomic Risk Factors forCerebrovascular, Ischemic Heart and Hypertensive Mortality in Japan. Tohoku J. exp. Med., 1986, 149 (4), 367-377- The effects of a wide variety of socioeconomic factors on the mortality from cerebrovascular, ischemic heart and hypertensive diseases in 46 prefectures in Japan for the years 1970 and 1975 were analyzed by stepwise multiple regression analysis. The results indicated the following risk factors for the three major vascular causes of death in males and females in the two years: (1) rural residence for cerebrovascular disease (and employment and high income in females); (2) rural and urban residence for Ischemic heart disease (in 1970 only); and (3) low income for hypertensive disease. The mortality from cerebrovascular disease within 46 prefectures was significantly higher in rural than urban areas in males and females in the two years, whereas the mortality from ischemic heart disease was higher in urban areas. The results were compared with recent data in Western countries. It is indicated that socioeconomic factors play important roles in the mortality from vascular causes of death as well as biological risk factors.
ARAKI, S. and MURATA, K. Social Risk Factors for the Mortality from MotorVehicle Accident. Tohoku J. exp. Med., 1986, 149 (4), 379-387- To clarify major social risk factors affecting the mortality from motor vehicle accident (MVA), we analyzed the effects of a wide variety of socioeconomic and demographic indicators (20 variables) on age-specific and age-adjusted mortality rates from MVA in 46 prefectures in Japan. The analysis was conducted twice at a 5-year interval, i.e., in 1970 and 1975, by stepwise regression analysis after classification of the indicators by factor analysis. The major social risk factors identified in the two years were as follows: (1) rural residence for persons, especially males, of major driving age (15-54 years) and young male children (0-4 years); (2) population growth by social mobility for schoolboys aged 5-14 years; and (3) high income (and employment) for schoolgirls, and proportion of old population for middle-aged females. Age-adjusted male rate of mortality from MVA within 46 prefectures was significantly higher in rural than urban areas in both 1970 and 1975.
MORITA, T., KONDO, S., TSUCHIDA, S. and WEISS, R.M. Characterization ofFunctional Beta-Adrenoceptor Subtypes in Rabbit Urinary Bladder Smooth Muscle. Tohoku J. exp. Med., 1986, 149(4), 389-395 - Spontaneous contractile force of muscle strips isolated from male rabbit urinary bladder dome (detrusor) and base (trigonal muscle) was significantly inhibited by isoproterenol (10-7-10-5M), a non-specific beta-adrenoceptor agonist or by terbutaline (10-8-10-5M), a selective beta2-adrenoceptor agonist. The EC50 values for isoproterenol and terbutaline in detrusor were the same those in trigonal muscle but the maximum relaxant response to isoproterenol or terbutaline was significantly greater in detrusor than in trigonal muscle. Dobutamine (10-5-10-4M), a relatively specific beta1- adrenoceptor agonist caused a small but significant relaxant response in trigonal muscle but no change in detrusor. In trigonal muscle the relaxant response to dobutamine was less than that to terbutaline. The relaxant response to 10-6M isoproterenol in detrusor was completely blocked by butoxamine (10-4M), a selective beta2-antagonist or by propranolol. (10-6M), a non-specific beta-antagonist but not by metoprolol (10-6-10-4M), a selective beta1-antagonist. Relaxation of trigonal muscle induced by 10-6M isoproterenol was inhibited by 10-5M metoprolol by 30%, by 10-4M butoxamine by 70%, or completely by 10-6 M propranolol. These findings are consistent with the view that the density of beta-adrenoceptors is higher in the detrusor than in trigonal muscle, and that the relaxant response to beta-adrenoceptor stimulation is mediated by beta2-subtype in the detrusor and by both of beta1- and beta2-subtypes in trigonal muscle of the male rabbit.
HANEDA, K., HESSEL, E.A., II., THOMAS, R. and HUNTSMAN, L.L. CardiovascularResponse to Rapid Infusion of Lactated Ringer's. Tohoku J. exp. Med., 1986, 149 (4), 397-406-Cardiovascular response to rapid infusion of lactated Ringer's was investigated in 5 adult dogs (average body weight=21.1kg) under 1% halothane anesthesia. Following implantation of aortic flow probe and left atrial line, the chest was closed and splenectomy was performed prior to the experiment. Warmed lactated Ringer's was administered at five different infusion rates (2.5, 5, 10, 15 and 20ml/kg/min in random sequence) to each dog until left atrial pressure (LAP) reached 20mmHg or a maximum of 50ml/kg had been infused. Subsequent infusions were done after stroke volume (SV) spontaneously returned to the control level. Cardiac output (CO), SV, heart rate (HR), mean arterial pressure (MAP), LAP and central venous pressure (CVP) were monitored simultaneously during infusions. HR was stable during infusions, whereas MAP increased by 39% of control. Response of LAP to volume infused was nearly linear at fast infusion rates (10, 15 and 20ml/kg/min). Response of LAP to slow infusion rates (2.5 and 5ml/kg/min) was curvilinear (decelerating curve). The relationship between CVP and volume infused was similar to LAP vs. volume infused. Ventricular function curves (SV, CO and stroke work vs. LAP) were also influenced by the rate of infusion with steeper curves at slow infusion rates than curves derived from fast infusion rates. However, initial changes in SV and CO curves were not significantly affected by the rate of infusion. We conclude that the cardiovascular response to rapid infusion of lactated Ringer's is rate dependent but initial changes in SV and CO curves are not significantly affected at infusion rates of 2.5, 5, 10, 15 or 20ml/kg/min.
FUNAYAMA, M., AOKI, Y., KUDO, T. and SAGISAKA, K. Sex Determination ofthe Human Skull Based upon Line Drawing from Roentgen Cephalograms. Tohoku J. exp. Med., 1986, 149(4), 407-416-The shape of the human skulls was studied with the aid of a personal computer to establish sex differences in quantitative anatomical terms. On the roentgen cephalograms from 50 adult males and 50 adult females, the lateral profile was transcribed onto an acetate sheet, on which the contour from the nasal apex to the forehead was digitized on a tablet digitizer into a series of dots which were input into the computer system. After this chain of dots was simulated by a spline function, the places most typically reflecting sex difference were determined in the profile based on the radius of curvature computed at each dot. The eminence of glabella and the nasal root, shown to be the places of skull apparently most characteristic of sex, were approximated to circular arcs with the least squares method, the radii of which were expected to serve for sexing. In both places, Student's t-test revealed a significant difference between the male and female groups (p<0.01). It was thus demonstrated that in male the eminence of glabella and the nasal concavity develop much more markedly than in female, presenting as a clear skeletal difference between both sexes.
SOU, S., TAKABAYASHI, T., OZAWA, N., SHO, K., Sasamoto, K., YAJIMA, A, and SUZUKI, M. Spontaneous and MMC Induced SCE in Lymphocytes from Patientswith Cervical Cancer. Tohoku J. exp. Med., 1986, 149 (4), 417-423- The frequencies of spontaneous and mitomycin C (MMC) induced sister chromataid exchanges (SCEs) were investigated in lymphocytes of the peripheral blood of women in different stages (O-III) of cervical cancer. Average generation time (AGT) was also examined by the replicative index (R.I.) method. The following results were obtained: (1) The spontaneous SCE frequency was found to be significantly higher in the cancer group than in the control group; 8.21±1.42 (mean±S.D.) against 5.62±0.55. (2) The SCE frequency gradually increased with the progression of the cervical cancer. (3) The SCE frequency in patients with carcinoma in situ (CIS), the lowest stage of cervical cancer, was significantly higher than that of the controls. (4) The frequency of MMC induced SCE was higher than that of spontaneous SCE in all groups, and the difference in frequency between MMC induced and spontaneous SCEs of cervical cancer group of the stages I-III was significant by different from that of the control group. (5) The AGT of the cervical cancer group was shorter than that of the control group, but it was not statistically significant.
MORI, K., SUZUKI, S., SUGAI, K., AKUTSU, Y., ISHIKAWA, M. and SAKAI, H. Morphological Changes of Platelets during the Process of Platelet Aggregation inGray Platelet Syndrome. Tohoku J. exp. Med., 1986, 149 (4), 425-436 Morphological changes of platelets during the process of platelet aggregation in gray platelet syndrome were observed by means of light microscopy, and scanning and transmission electronmicroscopy. Light microscopic examinations showed normal platelets and slightly enlarged, gray colored ones due to a deficiency in α-granules by May-Giemsa stain. At the time of maximum aggregation, normal platelets aggregated completely to form a mass by addition of ADP. On the other hand, patient's platelets only aggregated loosely and each platelet could be recognized separately, resembling primary aggregation of normal platelets. When collagen was added, almost no aggregation was noted in the patient's platelets. Scanning electronmicroscopic examinations revealed the same results as seen by light microscopy. Centralization of platelet organelles and almost complete aggragation and degranulation of platelets were noted immediately after 30 sec and 3-4 min after addition of ADP and collagen on the transmission electronmicrographs in normal platelets, whereas most patient's platelets remained to be unchanged. From these results, we concluded that these morphological changes were consistent with the changes of “release abnormalities” as shown in the functional examinations. Although some relationships between α-granule deficiency and functional abnormalities are considered, there is no definitive opinion.
KITAOKA, S., SUZUKI, H., NUMAZAKI, Y., KONNO, T. and ISHIDA, N. TheEffect of Trypsin on the Growth and Infectivity of Human Rotavirus. Tohoku J. exp. Med., 1986, 149 (4), 437-447- The effect of trypsin on the infectivity of human rotavirus (HRV) was examined using HRV strains that are cultivable only when treated with trypsin. Their infectivity in cell culture systems was enhanced in relation to enzyme concentration. The primary effect of trypsin appeared to be on the virus. Trypsin untreated HRV virions were capable of being adsorbed on the cell (MA104) but not of passing into it without trypsin. For multiple cycles of replication the presence of trypsin in the maintenance medium was required.
SHIRAI, K., SAITO, Y., YOSHIDA, S. and MATSUOKA, N. Existence of LipoproteinLipase in Rat Brain Microvessels. Tohoku J. exp. Med., 1986, 149 (4), 449-450- To clarify the possibility that brain microvessels could utilize very low density lipoprotein (VLDL)-triglyceride, the hydrolysis of [14C] triolein in VLDL by rat brain microvessels was examined. The rat brain microvessels were prepared under microscopy. In the homogenate, triolein hydrolyzing activity was observed only in the presence of apolipoprotein C-II. That activity was diminished by the addition of 1M NaCl. [14C] triolein incorporated in VLDL was hydrolyzed time- dependently. These results suggested that there exists lipoprotein lipase in brain microvessels and the brain can utilize VLDL triglyceride as a source of fatty acids.
TOMITA, Y., FUKUSHIMA, M. and TAGAMI, H. Stimulation of Melanogenesisby Cholecalciferol in Cultured Human Melanocytes: A Possible MechanismUnderlying Pigmentation after Ultraviolet Irradiation. Tohoku J, exp. Med., 1986, 149(4), 451-452- An increase in the amount of tyrosinase was demonstrated in the cultured human melanocyte after 6-day culturing with cholecalciferol (vitamin D3) by increased intensity of the immunofluorescent staining using monoclonal antibody against tyrosinase. Furthermore, the melanocytes became more dendritic as noted in those in the skin after the irradiation of ultraviolet. However, 7-dehydrocholesterol (pro-vitamin D3) or 1α, 25-dihydroxy-vitamin D3 (activated vitamin D3) did not induce any such effect on the cultured human melanocytes. Since cholecalciferol is known to be photo-chemically converted by the ultraviolet irradiation from pro-vitamin D3 produced in the skin, the so-far-unknown mechanism of human skin pigmentation after the ultraviolet irradiation may be partly explained by this stimulating effect of vitamin D3 on the melanocytes.