HUSSEIN, K.A. and MEDOUKH, N.T. A Simple Method for the Determinationof Plasma Extrinsic Coagulant Activity. Tohoku J. exp. Med., 1986, 150 (2), 103-116-A spectrophotometric study of plasma coagulation is presented with a method for the determination of plasma coagulant activity, via the extrinsic pathway. Dilute citrated plasma was coagulated by recalcification after the addition of a very small amount of thromboplastin. The maximum increase in absorbance per min during coagulation was used as a measure of coagulant activity. The method was applied to normal platelet-rich plasma samples, and the effect of changing the concentration of plasma, thromboplastin, fibrinogen, and heparin was studied.
SASANO, H., WATANABE N. and SASANO, N. Corticosteroid Binding in HumanArtery. Tohoku J. exp. Med., 1986, 150 (2), 117-125-Mineralo- and glucocorticoid binding in cytosol of surgically obtained human arteries and umbilical arteries were analyzed. Scatchard analysis of glucocorticoid binding in femoral and umbilical arteries revealed the binding parameters of Kd=4.3, 8.2×10-9M; Bmax=42, 90fmol/mg protein, respectively. Scatchard analysis of mineralocorticoid binding in inferior mesenteric artery showed a curvilinear pattern suggestive of the presence of two binding components, one with a higher affinity and a lower capacity for the ligand (Kd=3.3×10-10M, Bmax=0.8fmol/mg protein) and the other with lower affinity and higher capacity (Kd=10×10-7M, Bmax=36fmol/ mg protein). Polyacrylamide gel electrophoresis of mineralo- and glucocorticoid binding in the cytosol of popliteal artery also showed the presence of the corticosteroid binding components with limited binding capacity. Sucrose density gradientcentrifugation of corticosteroid binding in cytosol of femoral artery revealed the presence of dexamethasone binding molecule sedimenting in the 8-9 S region and deoxycorticosterone binding molecules sedimenting in the 7-8 and 8-9 S regions of the gradinent. Those results revealed the presence of mineralo- and glucocorticoid receptors in human artery and suggest the possible direct actions of corticosteroids upon human artery.
SHIMIZU, T. Serotonin Uptake at 22°C of Stored Platelets. Tohoku J. exp. Med., 1986, 150 (2), 127-133 In order to ascertain the possibility that platelet serotonin uptake may occur during storage of platelet concentrates (PC) at 22°C with agitation, the high-performance liquid chromatographic procedure was applied to determine serotonin uptake by platelets. Studies at 22°C showed that platelets stored for 4 days exhibited a significant serotonin uptake with a Vmax value of 2.4×10-19 mole/platelet/min and a Km value of 0.62×10-6M. Incubation of PC with 5×10-6M serotonin for 1 day at 22°C increased their serotonin contents from 2.2 to 4.2×10-7 mole/1011 platelets. Thrombin stimulation caused about 80% release of intracellular serotonin from fresh as well as stored platelets, which contained standard serotonin in the same amount as the original amount. These results suggest that a significant serotonin uptake of platelets might occur during in vitro storage at 22°C and stored platelets have retained abilities to sequester extracellular serotonin into dense granules.
ISHIZAWA, E., HORIUCHI, T., TADOKORO, M., SUZUKI, Y. and YAMAKI, S. Surgical Management of Pulmonary Atresia, and Critical Pulmonary Stenosis withIntact Ventricular Septum. Tohoku J. exp. Med., 1986, 150 (2), 135-144-Our experience with the surgical management of hypoplastic right ventricle with intact ventricular septum has included 20 patients with pulmonary atresia and 3 patients with critical pulmonary senosis. Group 1 consisted of 7 infants who were treated by transarterial pulmonary valvotomy. Four of them were lost shortly after surgery. Group 2 included 11 neonates and one infant who were treated by systemic-pulmonary shunt with 2 operative death. Group 3 included 4 infants who underwent primary prosthetic enlargement of the right ventricular outflow tract. Two of the 4 were lost in the early postoperative period. Eight of 13 patients in groups 1 and 2 who survived the initial palliative procedures underwent additional operations to enlarge the right ventricular outflow tract, and 5 patients survived. However, adequate right ventricular and tricuspid valve growth was obtained in only patients in whom the prosthetic enlargement of the right ventricle was performed in early infancy. These findings strongly suggest the necessity to decompress the right ventricle completely, as early as possible. In conclusion, 1) transarterial pulmonary valvotomy seldom provided satisfactory decompression of the right ventricle, and operative risk was high. 2) systemic-pulmonary shunt to re-establish pulmonary blood flow improved neonatal survival, although this procedure may not provide a satisfactory long term palliation. 3) primary or secondary operative enlargement of the right ventricle in early infancy provided decompression of the right ventricle and accelerated substantial right ventriculargrowth. For patients without an infundibulum or for those in whom attempts to induce tricuspid valve growth had failed, a Fontan type procedure seemed to be the procedure of choice.
NAKANE, T. and CHIBA, S. Regional Differences of Responses to AdrenoceptorAgonists in Isolated and Perfused Canine Coronary Arteries. Tohoku J. exp. Med., 1986, 150 (2), 145-154-Regional differences of the responses to adrenoceptor agonists were investigated in isolated canine coronary arteries by use of a cannula inserting method. Acetylcholine induced a dose-dependent vasodilation. Norepinephrine and epinephrine produced a vasoconstriction followed by a strong vasodilation in large coronary arteries and only a weak vasodilation in small coronary arteries. Phenylephrine (a selective alpha-1 agonist) induced a strong vasoconstriction in both arteries. The threshold dose and ED50 value for phenylephrine in small coronary arteries were much larger than those in large coronary arteries, although the vasoconstrictions by KCl and prostaglandin F2α were not different between in large and small coronary arteries. Clonidine and xylazine (selective alpha-2 agonists) produced a slight vasoconstriction but not dosedependently and a vasodilation with extremely large doses. ED50 value of vasodilation for salbutamol (a selective beta-2 agonist) was approximately 80 times greater than that for isoproterenol (a non-selective beta-agonist) in large coronary arteries, but was approximately 20 times in small coronary arteries. The maximal dilator response to salbutamol was about the same as that to isoproterenol in small coronary arteries, whereas it was much smaller than that to isoproterenol in large coronary arteries. These results suggest that adrenoceptors are heterogeneous according to the distance from the coronary orifice in canine epicardial coronary arteries.
FUNAKOSHI, A., SHINOZAKI, H., MIYAZAKI, K., NAKANO, I. and IBAYASHI, H. Effects of Cisapride on the Secretion of Pancreatic Polypeptide in Healthy Volunteers. Tohoku J. exp. Med., 1986, 150 (2), 155-160-Plasma human pancreatic polypeptide (hPP) responses to intravenous injections of a new gastric and intestinal motility promoting agent. R51 619 (cisapride), were studied in healthy volunteers. The mean fasting concentrations of plasma hPP (39±5.4pg/ml; mean± S.E.) were elevated significantly to the peak values of 118±25.1pg/ml 30min after the drug had been given in a dose of 4mg over 5min. However, plasma insulin (IRI), glucose and prolactin levels were not elevated. Atropine diminished the cisapride-induced hPP elevation, thereby suggesting that cisapride induced release of acetylcholine but had no antidopaminergic action.
SASAKI, I., MIYAKAWA, H., KAMEYAMA, J., KAMIYAMA, Y. and SATO, T. Influence of Obstructive Jaundice on Acute Gastric Ulcer, Intragastric pH andPotential Difference in Rats. Tohoku J. exp. Med., 1986, 150 (2), 161-168-To elucidate the influence of obstructive jaundice on the process of acute ulceration of the stomach under stress loading and to investigate the preventative effect of vagotomy, a series of experimental studies were carried out using SD rats. Animals were divided into four groups: Control, jaundice, vagotomy and jaundice with vagotomy. Each group was subjected to the water immersion-restraint stress and changes in the potential difference of gastric mucosa (PD), the intragastric pH (pH) and the score of ulcer index at sacrifice (UI) were measured. The process of acute ulceration under the stress loading was shown to be accelerated bythe weakening of defensive factors in the case of obstructive jaundice. To monitor changes of pH and PD was found to be useful to predict the manifestation of gastric leision. Vagotomy seemed to be partly effective to prevent the ulceration evidenced by lowering aggressive facter (an increase of pH), but it may also decrease the defensive factor because the vagotomy produced a significant lowering of PD.
SATO, Y., HANAI, J., TAKASUGI, N, and TAKEDA, T. Determination ofUrinary Vanillylmandelic Acid and Homovanillic Acid by High PerformanceLiquid Chromatography for Mass Screening of Neuroblastoma. Tohoku J. exp. Med., 1986, 150 (2), 169-174-In order to develop a mass screening method for the early detection of neuroblastoma, we measured urinary vanillylmandelic acid (VMA) and homovanillic acid (HVA) by a high performance liquid chromatography (HPLC) equipped with an electrochemical detector. Urinary samples were collected on a piece of filter paper and then pre-treated by a simple method. Measurement of VMA and HVA was performed within a short period of time, and by about 80 samples per a day could be analyzed by automatic system. The method described herein proved to be reliable, and we thus recommend it as a useful method for measurement of both VMA and HVA, leading to more accurate detection of neuroblastoma in mass screening test.
KOJIMA, M. and OHE, H. Experimental Study on the Regulation of TesticularFunction by the Cremaster Reflex in Rats. Tohoku J. exp. Med., 1986, 150 (2), 175-180-The present study was performed to clarify the role of the cremaster reflex in regulating the testicular function in rats. The impairment of cremaster reflex by bilateral dissection of the genitofemoral nerves resulted in a decrease of testicular weight and degenerative changes of the semeniferous tubules in the rats kept in a cold room (4°C), but not in the rats kept at room temperature (22°C). The results indicate that the cremaster reflex contributes much to the temperature regulation of the testicle located within the scrotum.
WASADA, T., KODAMA, Y., INOUE, K., HISATOMI, A., SAKO, Y. and IBAYASHI, H. Plasma Somatostatin Response in Normal and Gastrectomized Subjects. Tohoku J. exp. Med., 1986, 150 (2), 181-188-The present work was undertaken to investigate a possible contribution of the stomach to plasma concentrations of somatostatin-like immunoreactivity (SLI) in humans. For this purpose, plasma SLI responses were determined in previously gastrectomized subjects during lipid-heparin induced elevation of plasma free fatty acids (FFA), which gave a consistent rise of plasma SLI in normal controls. Fasting plasma SLI levels in both the subjects with antral and total gastrectomy did not differ from the values in normal controls. Acute elevation of plasma FFA resulted in about 5-fold increase of plasma SLI above basal level in the controls. In contrast, in both the gastrectomy groups there was no significant rise in plasma SLI, despite a comparable elevation of plasma FFA levels. In conclusion, these findings demonstrate that the stomach, probably the antral region of the stomach, accounts for the most part of increase of plasma SLI when hyperfatty-acidemia is induced and suggest that following the stimulation by nutrients the stomach makes a greater contribution to plasma SLI levels than does the pancreas or extragastric intestinal tract in humans.
SUZUKI, Y. Fluid Secretion and Electrical Properties of Pancreatic Acini ofSyrian Golden Hamster. Tohoku J. exp. Med., 1986, 150 (2), 189-208-Mechanisms of electrolyte and fluid secretion in the hamster pancreatic acini were investigated by measurements of fluid and amylase secretion in vivo and in vitro, and membrane potential and resistance in vitro. Unlike mouse and rat pancreas the hamster pancreas secreted only a small amount of juice in response to acinar stimulants such as acetylcholine (ACh) and caerulein. The mean resting potential was -62mV and the mean resting input resistance was 29MΩ. Ionophoretic application of ACh evoked membrane depolarization accompanied by a reduction in input resistance. The reversal potential for this ACh-evoked depolarization (EAch) was -11mV. The EAch was shifted by changing extracellular K, Na and Cl concentration in a similar way to that of mouse pancreatic acinar cells. The relative permeability and current for K, Na and Cl at the EAch calculated by using the Goldman-Hodgkin-Katz equations were PK/PNa/PCl=1/0.9/0.43 and IK/INa/ICl=1/1.4/0.4 respectively (In the mouse Pk/PNa/PCl=1/1.2/5 and IK/INa/ICl=1/ 2.3/1.3, Petersen et al. 1981). The results indicate that mechanisms of electrolyte and fluid secretion in hamster pancreatic acini are qualitatively similar to those in the mouse and rat pancreas. The less permeability of the acinar cell membrane to Cl ions in the hamster than in the mouse and rat appears to be associated with the fact that the acinar stimulant evokes a far smaller amount of fluid secretion in this species than in the mouse and rat.
TAKUSAGAWA, K., ASOO, N., SATO, T., NAGAI, H., MOTOMIYA, M. and KONNO, K. Immunoelectron Microscopic Observations on the Localization of Fibronectin inNormal Human Lung. Tohoku J. exp. Med., 1986, 150 (2), 209-223-The localization of fibronectin was examined in normal human lung using immunoelectron microscopy. Fibronectin staining was present not only in the endothelial, alveolar epithelial and bronchial epithelial basal laminae and associated with interstitial collagen fibrils and elastic fibers, but also in basal laminae of fibroblasts and smooth muscle cells. There was a periodicity in the staining of fibronectin on collagen fibrils. Reaction products against fibronectin were present in intracellular organelle including cisternae of smooth endoplasmic reticulum of endothelial cells and those of rough endoplasmic reticulum of fibroblasts. No other cells contained reaction products. The localization of fibronectin was compared with that of ruthenium red staining in normal human lung. The localization of fibronectin was consistent with that of proteoglycan.
TANAKA, M., NISHIZAWA, M., INUZUKA, T., BABA, H., SATO, S., MIYATAKE, T., AOKI, S. and SHINADA, S. Chronic T Cell Leukemia with a NK PhenotypeReacting with Anti-Myelin-Associated Glycoprotein (MAG) Mouse MonoclonalAntibody. Tohoku J. exp. Med., 1986, 150 (2), 225-226-We describe a patient demonstrating chronic T cell leukemia with a natural killer (NK) phenotype. The leukemic cells could be stained by OKT 3 (T cells), anti-Leu-7 and anti-myelinassociated glycoprotein (MAG) (NK cells) but not anti-Leu-11 monoclonal mouse antibody (activated NK cells). Fresh mononuclear cells showed a very low NK activity, although this activity returned to normal levels after 18 days incubation with interleukin-2 and some stimulants. It was not known why the NK activity of fresh mononuclear cells was low. This report is the first on leukemia characterized by anti-MAG monoclonal antibody.
YAMAGUCHI, S., KODA, N. and OHASHI, T. Diagnosis of Alkaptonuria byNMR Urinalysis. Rapid Qualitative and Quantitative Analysis of HomogentisicAcid. Tohoku J. exp. Med., 1986, 150 (2), 227-228-Urinalysis of alkaptonuria using NMR spectroscopy revealed the abnormal amount of homogentisic acid. Qualitative and quantitative determination was performed simultaneously with untreated urine sample and within 15min.
ITO, K., MIYAKAWA, H. and KATO, H. The Long-Term Facilitation ofNeuronal Activity Produced by Repeated Pairing of an Orthodromic Stimulus andAntidromic Stimuli in the Sliced Hippocampal Formation. Tohoku J. exp. Med., 1986, 150 (2), 229-230-Long-term facilitation in extracellulary recorded population spikes was demonstrated by repeated paired stimulation (RPS) in CAl neurons of the guinea pig hippocampal slices. Paired stimulation, consisting of a single orthodromic “test” stimulus and a train (5 pulses at 200Hz) of antidromic stimuli with an interstimulus interval (ISI) of 20, 500msec or 1sec was repeated every 5sec over a 250sec period. The amplitude of the population spike in response to the test stimulus increased gradually after RPS and reached a plateau level 30-50min after RPS. The plateau value was dependent on ISI; the shorter the ISI the greater the increase of responses. In contrast, no facilitation was observed when either the test stimulus or the antidromic stimuli were applied separately, without pairing.