TAKAHASHI, N., TAKAHASHI, S., KAGAYAMA, M. and YADA, K. Three-Dimensional Visualization of Golgi-Stained Neurons by a Projection X-RayMicroscope Converted from a Scanning Electron Microscope. Tohoku J. exp. Med., 1983, 141 (3), 249-256 - Trying to obtain the three-dimensional images of neuronal elements, Golgi-stained thick sections were examined by a projection X-ray microscope converted from a scanning electron microscope. An attachment unit for the projection X-ray microscope consists of a target holder, specimen stage and film cassette. Two kinds of target material, titanium and gold, were tested. The system was operated at 10 to 30kV. The electron beam into the target was about 20nA with the use of a pointed cathode. The stereopair of X-ray micrographs were taken by tilting the specimens. Quality and contrast of the X-ray micrograph taken with a titanium target was better than that with a gold target. The spatial resolution was estimated about 1 to 2μm for 100 to 350 μm thick sections with a titanium target. Compared with the light microscope, strong penetrating ability of X-ray permitted to visualize the internal structures of optically opaque materials. Further, almost all objects were in focus in X-ray micrographs even for relatively thick sections (100 to 350μm). The three-dimensional morphology of neurons and glial cells was demonstrated by a stereopair of X-ray micrographs.
FUJIMURA, Y., MATSUZAWA, T., KAWAMURA, M., TADA, K. and MIZUNO, K. Mass Screening of Urea Cycle Diseases: A New Mass Screening Method ofHyperornithinemia by Using Two Coupling Enzymes. Tohoku J, exp. Med., 1983, 141 (3), 257-261 - We devised a new microfluorometric mass screening method for determining ornithine in only one blood disc of 3mm in diameter (blood of 2-3μl) by use of two coupling enzymes, ornithine aminotransferase and Δ'-pyrroline-5-carboxylate reductase. We could apply this method to some cases of gyrate atrophy of the choroid and retina. Our method can be applied in range of 0 to 12-16 nmoles or about 54.8mg% or about 4mM ornithine in blood. Ornithine values by our method were well correlated with those by an amino acid analyser.
FUJIMURA, Y., KAWAMURA, M. and NARUSE, H. A New Mass ScreeningMethod for Determining UDP-Galactose in Blood. Tohoku J. exp. Med., 1983, 141 (3), 263-268 - We devised a new microfluorometric mass screening method for determining UDP-galactose by using only one blood disc of 3mm in diameter. This method contains two coupling enzymes of UDP-galactose-4-epimerase and UDP-glucose dehydrogenase, and conversion of NAB to NADH. The assay ranges are 0 to 1mM or 0 to 0.2mM UDP-galactose in blood. UDP-galactose contents were determined in galactosemia of epimerase-, transferase-and kinase-deficiencies, and several cases with high or low Paigen's values.
KIMURA, I., AYYAR, D.R. and LIPPMANN, S.M. ElectrophysiologicalVerification of the Ulnar to Median Nerve Communications in the Hand and Forearm. Tohoku J. exp. Med., 1983, 141 (3), 269-274 - One hundred and fifty upper extremities of 85 unselected subjects were studied. Supramaximal nerve stimulation of the median and ulnar nerves at the wrist and elbow was used and the compound muscle action potentials (CMAPs) from the abductor pollicis brevis (APB) muscle were recorded. The ulnar to median nerve communication in the hand - Riche-Cannieu anastomosis - was detected in 125 out of 150 hands (83.3%). The ulnar to median nerve communication in the forearm was demonstrated in only two out of 150 extremities (1.3%). The mean ulnar nerve innervation ratio (UNIR) in the APB was 27.6±16.4% (mean±S.D.). In the American black population, this UNIR percentage was statistically smaller than in the American caucasian or hispanic population (p<0.05).
FUJITANI, Y. and HOSOGAT, M. Circadian Rhythm of Electrical Activity andMotility of the Stomach in Cats and Their Relation to Sleep-Wakefulness Stages. Tohoku J. exp. Med., 1983, 141 (3), 275-286 - Polysomnographic recordings were done over continuous 3 days together with those of electrical and mechanical activities of the stomach using chronically implanted electrodes and force-transducer in unanesthetized and unrestrained state of 4 adult cats. Circadian rhythm of the basic electric rhythm (BER) and mechanical activity in the stomach accustomed to feeding schedule of once a day was divided into the two; the fed state from the end of feeding to 8 to 11hr later and the succeeding fasting state. However, differentiation of the fasting state into the contractile and uncontractile periods, which is known in dogs, was not evident in cats. The mean BER interval was longer and the standard deviation was smaller in the fed state than in the fasting state. Among stages of wakefulness (W), drowsiness (D), light slow wave sleep (LS), deep slow wave sleep (DS) and REM sleep, the mean BER intervals in every total time during the fed state were compared with one another. A definite unidirectional finding obtained in all 4 cats was only that the mean BER intervals in the stage D were longer significantly than those in the stage W.
TANI, K., FUJII, H., MIWA, S., IMANAKA, F., KURAMOTO, A, and ISHIKAWA, H. Phosphofructokinase Deficiency Associated with Congenital NonspherocyticHemolytic Anemia and Mild Myopathy: Biochemical and Morphological Studieson the Muscle. Tohoku J. exp. Med., 1983, 141 (3), 287-293 - Enzymatic and electron microscopical studies were performed on the muscle of a proband with phosphofructokinase deficiency. Enzymatic studies showed that muscle phosphofructokinase activity of the proband was decreased to about a half of normal. This enzyme was quite thermolabile and had low affinity for fructose 6-phosphate. Electron microscopical studies showed the accumulation of glycogen granules beneath the sarcolemma and between the myofibrils in spite of a lack of accumulation of the intermediates before the step of phosphofructokinase. The proband's clinical symptoms, i.e., hemolytic anemia and myopathy, were considered to be due to the unstable, mutant, muscle-type phosphofructokinase in the red blood cells and muscle.
WAGA, S., TAKAHASHI, Y., FUJITA, M., NAGATA, K., KURONUMA, T. and AOYAMA, R. Monocyte Function in Idiopathic Nephrotic Syndrome in Childhood. Tohoku J. exp. Med., 1983, 141 (3), 295-303 - The monocyte function of 112 specimens from 42 children with idiopathic nephrotic syndrome (INS) aged from 2 to 17 years was studied by the methods of nitroblue tetrazolium (NBT) reduction, phagocytosis of immunobeads (TB) and yeast cells, chemotaxis and acid alpha-naphthyl acetate esterase (ANAE) staining. The dissociation between phagocytosis and chemotaxis was observed in the fresh cases of steroid sensitive INS and in the cases of steroid non-sensitive INS. In the fresh cases of steroid sensitive INS, NBT reduction and phagocytosis were increased, but chemotaxis was decreased. In the cases of steroid non-sensitive INS, the phagocytosis of IB was decreased, but chemotaxis was increased. These findings suggest a different pathogenesis between steroid sensitive and steroid non-sensitive INS. The dissociation between phagocytosis and chemotaxis may be explained by the alteration of the surface receptors of monocyte and by lymphokines.
NAKAE, S., TSUCHIYA, S., KATAMINE, S., KONNO, T. and TADA, K. Ecto-5'-Nucleotidase Activities in B Lymphoblastoid Cell Lines at Various Stages ofMaturation. Tohoku J. exp. Med., 1983, 141 (3), 305-310 - B lymphoblastoid cell lines (LCL) at various stages of maturation, including possible precursor B cell lines without immunoglobulin (Ig) expression, pre-B cell-like LCL, and Ig-secreting LCL, were established from peripheral and bone marrow mononuclear cells of four patients with congenital agammaglobulinemia (CAG) by infection with Epstein-Barr virus (EBV) and estimated for their ecto-5'-nucleotidase (ecto-5'-NT, E.C. 220.127.116.11) activity. Ecto-5'-NT activities in Ig-secreting LCL derived from CAG patients fell into the same range of those in LCL derived from control subjects (from 7.3 to 33.0nmoles/hr/l06cells). When ecto-5'-NT activities of precursor B cell lines derived from the CAG patients were also compared with Ig-secreting LCL derived from control subjects, no significant difference in ecto-5'-NT activities was found among LCL with various Ig expression which might indicate various stages of B cell maturation. These results may suggest that ecto-5'-NT does not appear to be a biochemical marker of B cell maturation.
HAYASHIDA, C.Y., SUZUKI, K., FUJIYA, H., KOIZUMI, M., TOYOTA, T. and GOTO, Y. Morphometrical Quantitation of Pancreatic Endocrine Cells in Patientswith Carcinoma of the Pancreas. Tohoku J. exp. Med., 1983, 141 (3), 311-322 -A morphometrical study of the tumor-free pancreatic regions has been performed in 18 patients with carcinoma of the pancreas, who were divided into three groups, i.e., diabetics, patients exhibiting impaired glucose tolerance (IGT) and normoglycemics. Tissues were stained for A, B, D and PP cells by immunocytochemistry and, for comparative purposes, the results were expressed as the number of cells/mm2. The diabetic and the IGT groups presented reduced A and B cell populations, with B cell counting lower in daibetics than in the IGT group. In addition, an inverse correlation was found between the number of B cells and the fasting plasma glucose (FPG) values, which suggests that the destruction of B cells may be the primordial cause of the glucose abnormality found in those patients. Possibly, a destructive process and fibrosis, similar to those occurring in chronic pancreatitis, are implicated in the reduction of such endocrine cells.
FUKAYA, T., FURUHASHI, N., KONO, H., SHINKAWA, O., TAKAHASHI, T. and SUZUKI, M. The Effect of Sulpiride Administration on Maternal and FetalPlasma Prolactin Levels, and Fetal Growth in Rats. Tohoku J. exp. Med., 1983, 141 (3), 323-326 - Pregnant Wistar rats were daily injected with 5mg sulpiride into the peritoneal cavity starting on day 14 of gestation. Blood samples were obtained from the maternal carotid artery and fetal axillary vessels on day 20 of gestation. Serum prolactin (PRL) levels were determined by radioimmunoassay using a rat PRL RIA kit. The maternal serum and fetal plasma PRL levels were significantly higher in the sulpiride-treated group than in the saline control group. There was a significant positive correlation between fetal body weight and fetal plasma PRL level. These results suggest that sulpiride which reached the fetus stimulates fetal PRL secretion and that PRL may exert a growth-promoting effect on the fetus.
YAMANE, N., YUKI, M. and KYONO, K. Isolation and Characterization ofGroup B Streptococci from Genito- Urinary Tracts in Japan. Tohoku J. exp. Med., 1983, 141(3), 327-335 -The prevalence of group B streptococci in urine specimens submitted from both inpatients and outpatients to the Clinical Microbiology Laboratory were investigated. Out of 3, 780 urine specimens, forty-seven strains were identified as group B streptococci, and clinical features revealed that the isolation ratio from outpatients was higher than that from inpatients, and that the majority of isolates were from female urine. Whereas the isolation ratios of group B streptococci from vaginal swabs were 2.9% in 377 pregnant women at 37-40 weeks of gestation, and 4.8% in 335 non-pregnant women. Also, the newborns delivered from mothers tested were investigated. Only two cases were positive in ear, pharyngeal and/or nasal swabs, but no clinical manifestation was observed. An alternative nomenclature system of group B streptococci based on the serological classification of both heat-labile protein and heat-stable poly-saccharide antigens was evaluated for the clinical isolates and compared with the original Lancefield's procedure. The results indicated that the method described will provide a simple and reasonable technique for serotyping of group B streptococci.
UEMURA, Y. Dissociation of Aggregated IgG and Denaturation of MonomericIgG by Acid Treatment. Tohoku J. exp. Med., 1983, 141 (3), 337-349-Conditions for obtaining a large quantity of monomeric IgG which can be used for intravenous injections in humans was successfully established with human serum IgG preparation obtained using the ethanol fractionation method. When dissociation of IgG aggregates and denaturation of IgG monomers contained in the starting material were examined at various pH values, the treatment for 60min at 28°C at pH 3.8-4.0 was found to be optimum for obtaining such monomeric IgG. At this pH range, the dissociation of IgG aggregates into monomeric IgG reached maximum, without producing polymeric IgG. When monomeric IgG was treated at this acidic pH condition, its biological and physicochemical properties such as acid difference spectrum, molecular weight, plasmic digestion ratio, and complement-, antigen- and protein A-binding activities remained unchanged. The fact that large scale production of monomeric IgG can be easily performed under such conditions is discussed.
KIMURA, I., AYYAR, D.R., SATO, G. and KOGURE, K. ElectromyographicAssessment of Fatiguability and Recovery of Orbicularis Oculi in Myasthenia Gravis. Tohoku J. exp. Med., 1983, 141 (3), 351-358 - Orbicularis oculi fatiguability following maximal voluntary contraction and its recovery after administration of Tensilon (edrophonium chloride) were studied in 13 patients with myasthenia gravis (MG) and 8 normal controls. The number of EMG pulses per sec (C) was calculated using an experimentally manufactured EMG pulse counting system and the mean amplitude of the motor unit potentials per sec (A) was measured using the action potential analysis module (Medelec APA-6 MK-1) during maximal voluntary contraction of the orbicularis oculi muscle. Fatiguability was estimated as percentage change of C, the pre-exercise value being taken as 100%. It was 74.4±18.2% (mean±S.D.) in patients and was significantly different from 110.8±8.4% found in the controls (p<0.01). Recovery following intravenous injection of Tensilon was estimated as percentage change in C and A, the values before Tensilon injection being taken as 100%. The change in EMG pulse per sec (C) was 360.0±90.0% in MG and 113.0+21.2% in controls (p<0.005). The mean amplitude of the motor unit potential per sec (A) increased by 180.0±95.0% in MG and 120.0±42.0% in controls (p<0.01). This technique is thought to be valuable to confirm the diagnosis of MG when routine electrophysiological tests fail to demonstrate abnormalities in neuromuscular transmission.
YAMAGUCHI, Y., ISEMURA, M., YOSIZAWA, Z., SATO, A. and SUZUKI, M. Differences in the Carbohydrate Moieties of Plasma and Amniotic Fluid Fibronectinsas Revealed by Crossed Immunoaf nity Electrophoresis with Concanavalin A. Tohoku J. exp. Med., 1984, 141 (3), 359-367 - Human fibronectins isolated from pooled human plasma and amniotic fluid were studied as to differences in their carbohydrate moieties. The chemical analyses showed that amniotic fluid fibronectin is different from adult plasma fibronectin in carbohydrate content and composition while there seems to be no significant differences in amino acid composition. Crossed immunoaffinity electrophoresis with free concanavalin A, as well as rocket immunoelectrophoresis with immobilized concanavalin A intermediate gel, indicated that amniotic fluid fibronectin has little or no reactivity with this lectin while adult plasma fibronectin is strongly reactive. Fetal cord plasma fibronectin apparently interacted with concanavalin A, but its reactivity was weaker than that of adult plasma fibronectin. Fibronectin isolated from ascitic fluid of an ovarian cancer patient which was examined in additional experiments showed much weaker Con A-reactivity than fetal cord plasma fibronectin. These results suggest that fibronectins from various body fluids differ in their carbohydrate structures.
NISHIZAWA, O., HARADA, T., MORIYA, I., OHYA, A., SATOH, S., NAKAMURA, H., FUKUDA, T., MATSUZAKI, A. and TSUCHIDA, S. Application of UltrasonicM-Mode Scanning and Computer Evaluation on Urodynamics. Tohoku J. exp. Med., 1983, 141 (3), 369-370 - The M-mode scanning images of urethra, detrusor pressure and urine flow rate during voiding were recorded together on the line scan recorder, and allowed for plotting and calculation of some valuable urodynamic parameters by computer evaluation. This procedure will permit first application of M-mode scanning and new direction on urodynamics.
NISHIZAWA, O., OHYA, A., KIN, N., NAKAMURA, H., SATOH, S., MORIYA, I., HARADA, T. and TSUCHIDA, S. Percutaneous Introduction of suprapubic DoubleLumen Catheter into Bladder for Urodynamic Examination. Tohoku J. exp. Med., 1983, 141 (3), 371-372 - A modified method of percutaneous introduction of a suprapubic double lumen catheter (8 Fr) into the bladder for urodynamic examination is described. This method permits to fill the bladder at the filling rate about 50-60ml/min from the standard drip set, and the catheter can be percutaneously introduced into the bladder with safety.