Ornithine carbamoyltransferase (OCT) is almost exclusively located in the liver mitochondria'matrix of ureoteric animals. A trace activity of OCT in normal serum has been found and the enzyme activity increased markedly in patients with acute hepatic diseases. Therefore, those phenomena have been used as an index for diagnosing some hepatic diseases, although few studies have been made on the properties of the enzyme in serum, and there is no direct evidence whether the serum enzyme was released from the matrix of liver mitochondria. Here I report that OCT has been homogeneously purified by ammonium sulfate fractionation, heat treatment,2nd ammonium sulfate fractionation, Phenyl Sepharose CL-4B and CM-Sephadex C-50 column chromatographies. The purified enzyme exhibited the molecular weight of 39,500as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and of 115,000 by gel filtration. In addition, the characteristics of serum OCT in galactosaminetreated rats and variations in the amount and the activity of the serum enzyme after treatment with galactosamine were investigated. A single injection of D-galactosamine (90mg/100g of body weight) into rats caused acute liver cell injury, and the activity of OCT in the serum increased about 600 times as compared with that in normal serum. The serum and mitochondrial enzyme activities gave the similar pH profiles, showing an optimum of pH 8.5. Apparent Km values of the serum enzyme for ornithine under the assay conditions at pH 7.4 and 7.7 were 1.59mM and 0.94mM, respectively, and those of the mitochondria' enzyme for the corresponding substrate were 1.69 mM and 0.97 mM, respectively. The Km value of the serum enzyme for carbamyl phosphate was 0.34 mM, which was similar to that of the mitochondrial enzym e. The mitochondrial and serum enzymes had a molecular weight of 115,000 by Sephacryl S-300gel filtration. The antibody against the mitochondria' enzyme was raised, and the immunological properties of the serum enzyme were examined. The experiments of immunoinhibition showed that an increase in the antibody caused both the serum enzyme activity and the mitochondrial enzyme activity similarly to decrease. Analysis by SDS-PAGE showed that the molecular weights of the serum and mitochondrial enzymes were identical. The amounts of the enzyme varied in a manner similar to the variations in the activity. Those results indicated that OCT was released into the serum in an intact form from the mitochondrial matrix during hepatic damage.
It has been suggested that fentanyl, a synthesized narcotic analgesic, causes bronchoconstriction and elevates airway resistance. However little work has been done to investigate influences of fentanyl upon the lung airway in detail. The purpose of this study is to examine the changes in such parameters of the airway dynamics as lung compliance, airway resistance, and respiratory power in house rabbits. The animals underw ent artificial ventilation by means of Servo 900 B ventilator while total lung dynamic compliance (C), airway resistance (R) and respiratory power were calculated on the analogue tracings of pressure, flow, volume and multiplication of pressure by flow, respectively. The calculation of airway resistance was carried out after the method developed by O. P. Norlander who had made it possible to elicit the elastic component of the airway pressure independently from the nonelastic one, and thus attained more accurate value. There was also contained another similar investigation with benzonatate, a new antitussive, in this study. The results were as follows; Fentanyl did not increse R, but decreased it signific antly either in massive dose (20μg/kg), or in another arbitrary dose (8μg/kg) given after droperidol in a similar fashion to clinical NLA. In the propranolol pretreated animals, fentanyl (8μg/kg) produced significant decrease in R. However in the phentolamine pretreated animals, the same dose of fentanyl caused no change in R. These findings suggest that the decrease in R caused by fen tanyl is due to a blocking action of α-receptors in the airway smooth muscle. C was decreased in all of the investigated groups of animals which included the one serving as control and having artificial ventilation by ventilator alone without any investigated drug administration. However the decrease in C was smaller in the propranolol pretreated group. Power was increased at twenty minutes after the administration of fentanyl in a ll of the groups except the propranolol pretreated group. The increase in power was elicited by the concomitant rise in the airway pressure. High dose of benzonatate (2mg/kg) decreased R, while C was decreased in all of the groups with benzonatate and in the control group as well. It is concluded that benzonatate not only inhibits cough reflex in the alveoli and the small airway but relaxes the bronchial smooth muscle in the large airway.
To investigate the mechanisms of development and prevention of adrenal apoplexy induced by 7,12-dimethylbenz (a) anthracene (DMBA) in adult rats, two experiments were carried out. In the first experiment,31 JCL Sprague-Dawley female rats divided to the following 6 groups; (1) untreated control, (2) singly DMBA-intubated, (3) twice Heparin-combined, (4) twice Phenobarbital (PB) -pretreted, (5) quadruply PB-pretreated, (6) 3-Methylcholanthrene (MC) pretreated. All animals of groups (2) - (6) were intubated by DMBA,25mg, disolved in 2 ml of sesame oil on the age of 55 days. Heparin,240 units, was injected subcutaneously 15 and 24hr after the intubation of DMBA. PB,100mg/kg, was injected subcutaneously either 2 times (24 and 48 hr before the intubation) or 4 times (24,48,72 and 92 hr before the intubation). MC,80mg/ kg, was given orally with a stomach tube 48 hr before the DMBA administration. All rats were killed by decapitation 40 hr after the DMBA administration (on the age of 57days). Both adrenals were observed morphologically, and the induction rates of adrenal apoplex y in the experimental groups were obtained as follows; Control 0/8, DMBA solo 8/8, Heparincombined 10/10, twice PB-pretreated 10/10, quadruply PB-pretreated 9/10, MC-pretreated 0/16. Moreover, the histologic finding of adrenal apoplexy was more severe in the heparincombined group than the DMBA solo group. In the second experiment using adult female rats, both hepatic clearance and adrenal incorporation of tritiated DMBA were observed biochemically 1and 3 hr after the single intraperitoneal injection of 3H-DMBA,800μCi/ kg, emulsified in DMSOsaline. The radioactivities in the fresh hepatic tissues and in the organic solubent-extractable fractions from the adrenal glands were counted by a Packard liquid scintillation spectrometer, and they were calculated to dpm/mg of wet weight. In comparison of DMBA solo and PB-pretreated groups, the pretreatment of MC accerelated significantly the hepatic clearance of DMBA and decreased prominently the adrenal incorporation of DMBA and organic solubent extractable fraction of its metabolites 3 hr after the intraperitoneal administration. In conclusion, DMBAinduced adrenal apoplexy may initiate from multiple adrenocorticolytic phenomena evolked by a direct action of DMBA in inner zone of the adrenal cortex, and may develope, without an intravascular coagulation, under a special microcirculaion of the adrenal cortex. The ultimate adrenocorticolytic agent was known 7-hydroxymethyl-12-methylbenz (a) anthracene, which introduced by the hepatic metabolism of DMBA. But, the pretreatment of MC accerelates prominently the detoxication of DMBA in the liver, decreases the incorporation of DMBA in the adrenal gland, and finally prevents DMBA-induced adrenal apoplexy. PB had no effect on the accerelation of DMBA detoxication and the prevention of adrenal apoplexy by DMBA.
Abnormalities in ECG frequently occurring in patients associated with cerebrovascular lesions, especially subarachnoid hemorrhage, have been documented in many reports. Similar abnormalities in ECG have also been reported on such experiments as following electric stimulation of various sites of the brain, direct intracranial injection of space taking substances or induced ICP elevation. Drugs are currently administered commonly by injection into the cisterna magna o r via the transorbital approach, however approach to the circle of Willis is difficult by either method or hemorrhage due to puncture of adjacent vessels may frequently occur. The author produced in rabbits a method injecting small amounts of drugs via the craniopharyngeal duct into the basal cistern with reduced surgical damage and minimal influences on the systemic circulation. 1. Direct administration of 0.5ml or less of physiological saline into the basal cistern of the rabbit via craniopharyngeal duct produced no changes in the systemic circulation or ECG. 2. Withdrawal of 1.0ml of cerebrospinal fluid from the craniopharyngeal duct and subsequent injection of serotonin (10mg-1.0ml) induced no changes in ECG except bradycardia. 3. When prostaglandin F2α (500pg-0.5ml) was injected into the basal cistern, supraventricular and ventricular arrhythmia were observed in ECG after a mean interval of 301sec. ± 74sec., and S-T depression and lower T wave remained for more than 45 minutes. An increase i n blood pressure and a decrease in the heart rate were also noted. 4. Intravenous administration of the same dose of PGF2a caused no changes in ECG except bradycardia. 5. Injection of the same dose into the basal cistern after cervical spinal cord transection at the C2-C3 level induced no changes in ECG. 6. Thus the stimulation by the injection of PGF2a was shown to be conducted down to the heart via the cervical spinal cord. 7. A decrease in PaCO2 was observed following injections of serotonin or PGF2α into the basal cistern.
the quality of ceroid deposited in Kupffer cells of CCI4-inhalated mouse liver by the color intensity developed with the leucomalachite green stain or zinc leuco-dye stain for ceroid (zinc leucosafranin and leucofuchsin stain). Materials and Methods: For the purpose of measuring ceroid 3-4 serial paraffin sections were prepared from each localized area at random selected from CCI4-inhalated mouse livers. Measurements were made on 10 individual Kupffer cells filled with ceroid granules on each section. Results: The spectra of absorption curves obtained indicated that a gradual increase in the color intensity developed with both the leucomalachite green and zinc leucosafranin stain in the Kupffer cell ceroid has a tendency to run parallel with the length of the experimental period of time until 105 times of CCI4 inhalation carried out once on alternate days, the leucofuchsin stain being less effective. The above findings suggest a gradual increase in pigment maturity with the la pse of the experimental period. In order to obtain more morphological evidence, the present authors could compare with success the above absorption spectra with a diagram gained by means of dipicting “multilayer” absorption curves in which the quality of ceroid deposited in Kupffer cells is illustrated as an image of a hill, leaving an area of a nucleus blanks.
Here reported on the unexpected anti-erythrocyte antibodies found in our transfusion unit of Kansai Medical University Hospital during last ten years from Oct.,1975 to Sept.,1984. The unexpected antibodies could be detected in 2.11% of all materials examined. Among these antibodies, anti-Lea, anti-P1 and anti-E (alone) were detected more frequently than others, occupying 29.0%,19.4% and 8.2% of them respectively, and each one case having anti-K1 or anti-Fyb antibody scarcely detectable in Japanese, was found. The case with anti-K1 was 64 years old woman who had experi enced three pregnancies and had been suffering from chronic bronchitis concomitant with liver cirrhosis. Thrombocytopenia was found at the time of the blood examination. Her anti-K1 antibody was of IgG type, and was detectable only by means of Coombs' test. Although her red blood cell antigen was kk as to the Kell system, we could not examine that of her husband, because he had already died. As for the cause induced her anti-K1 antibody, her previous pregnancies should be considered, while its autologous occurrence due to chronic bronchitis could not be ruled out. The case with anti-Fyb was 39 years old man, who had been healthy so far, and had never received any blood transfusion. His red cell antigen was Fy(a+b-) as to Duffy system, his anti-Fyb antibody being of IgG type, too, and could be detected only by means of albumi n-Coombs' or Liss-Coombs' test. In this case, causes which might have induced his antibody could not be presumed.
The present study was made in an attempt to clarify the changes in the cephalic circulation and cardiovascular function occurring after stimulation or destruction of the basal ganglia of rabbits in reference to electrical activities of the central nervous system. The results were as follows. 1. Stimulation of th e globus pallidus with 3Hz resulted in a change in the spike and wavelike pattern in the EEG of the frontal cortex (FC) and hippocampus (HPC). This stimu lation had almost no effect on the blood pressure, heart rate and bilateral common carotid blood flow volume, but there were a few cases with a slight drop in blood pressure and slight decrease in heart rate. 2. Stimulation of the globus pallidus with 8Hz resulted in a change in the recruiting responselike pattern in the EEG of FC and HPC. With this stimulation, there was a drop in blo od pressure, decrease in heart rate and decrease in bilateral common carotid blood flow volume. 3. When the globus pallidus was stimulated with 100Hz, a pattern of desynchr onization became apparent in the EEG of FC and HPC. This stimulation induced a drop in blood pressur e, decrease in heart rate and decrease in bilateral common carotid blood flow volume. 4. The drop in blood pressure and decrease in heart rate induced by stim ulation of the globus pallidus took place even when the bilateral depressor and carotid sinus nerves were sev ered. But, when the vagus nerve was severed bilaterally and atropine sulfate injected intrave nously, the effect decreased. 5. The decrease in bilateral common carotid blood flow volume induced by stimulation of the globus pallidus became even more prominent when the effect of change of blood gases w as eliminated by artificial respiration. 6. Stimulation of th e putamen with 3Hz resulted in a spike and wave-like pattern in the EEG of FC and HPC. When stimulated with 8Hz, a change in the recruiting response-like pa ttern occured and with 100Hz, a pattern of desynchronization was observed in FC and a pattern of synchronization or desynchronization was observed in HPC. Any one of these stimuli had almost no effect on blood pressure and heart rate, but there was an increase in bilateral common carotid blood flow volume. 7. This increase in bilateral common carotid blood flow volume induced by stimulation of the putamen became less when the effect of blood gas variation was eliminated by artificial respiration, and this phenomenon was almost completely suppressed by intravenous injection of atropine sulfate. 8. When the caudate nucleus was stimulated, a change in the EEG was almost same pattern as the change induced by stimulation of the putamen but there was little effect on the blood pressure, heart rate and bilateral common carotid blood flow volume. 9. On destruction of the globus pallidus, putamen, caudate nucleus or internal capsule, slow wave became apparent in the EEG of FC but it was not observed apparently in the EEG of HPC. 10. Blood pressure and heart rate showed little change with any one of these destruction but the common carotid blood flow volume increased with destruction of the globus pallidus and decreased with destruction of the putamen, caudate nucleus or internal capsule. 11. The increase of blood flow volume in bilateral common carotid and bilateral internal carotid arteries, induced by destruction of the globus pallidus, was unaffected even when the influence of blood gas variation was eliminated by artificial respiration, but this trend tended to be lessened by the administration of atropine sulfate. 12. The decrease of blood flow volume i n bilateral common carotid and bilateral internal carotid arteries, induced by destruction of the putamen, was also observed under artificial respiration, but this trend was significantly lessened by the intravenous administration of phenoxybenzamine compared to the untreated control.
We analyzed 138 CVD patients during 5 year period to obtained the important factors contributing to prognosis. (1) High blood sugar and Ecg abnomalities were related to poor prognosis in the cerebral bleeding group. (2) Brain herniation, deep coma, internal bleeding in CT and ventricular bleeding were the major factors related to hospital mortality. Hepatorenal failure in the cerebral bleeding and SAH group and renal failure in cerebral infarction group had close relation with poor prognosis, indicating “below of neck” therapy is necessary for life support. (3) Similar condition exists in DIC, therefore prevention of brain edema and early therapy of DIC is necessary. (4) Traine d staff and well organized institution are necessary in order to evaluate the patients condition and to benefit from emergency operation.
As a part of the study to elucidate the influence of exteroceptive stimulation on the excitability of the brain stem reticular formation (RF) in the presence of flash stimuli, the effect of hippocampus stimulation on the recruiting response was investigated in the rabbit and the following observed. 1) The recruitin g response in the cerebral cortex EEG induced by 8 Hz stimulation of the nucleus ventralis anterior (VA), belonging to the diffuse thalamocortical projection system, was inhibited by simultaneous 100 Hz stimulation of the hippocampus. 2) The recruiting response by 8 Hz stimulation of th e VA was inhibited or facilitated following simultaneous 8 Hz stimulation of the hippocampus. 3) The recruiting response induced by 8 Hz stimulation of the VA was facilitated by 3 Hz stimulation of the hippocampus .4) The rate of change in the amplitude of this recruiting response following stimulation of the hippocampus was analyzed quantitatively by determination of the wave pattern, using the digital computor (ATAC-201). The findings suggest that the previously reported (I) increase in excitability of the RF by attachment of contact lens in the presence of the flash stimulus, corresponds to the effect of 100 Hz stimulation of the hippocampus and the decrease in excitability of the RF by contact lens in the absence of the flash stimulus corresponds to the effect of 3 Hz stimulation of the hippocampus, respectively.