Homogenates of mouse liver with isotonic sucrose solution were separated by the cell fractionation with repeating centrifugation. The supernatants were used for the inhibition test with the reagents such as 3, 5-diiodosalicylic acid lithium salt (LIS), guanidine and azide, heat, acid and alkali. After various treatments, the remaining catalase activities were measured and showed as a relative enzyme activity. Stability of catalase in liver supernatants was compared normal (C3H/CasCas) and mutant mice which were designated acatalasemia (C3H/CbsCbs), hypocatalasemia (C3H/CcsCcs) and acatalasemic heterozygote (C3H/CasCbs). In both treatments of LIS and guanidine, catalase of CasCas was most stable, CbsCbs were unstable, and catalase of CcsCcs and CasCbs showed midle stability between the stability of CasCas and CbsCbs in this order. On the contrary in azide treatment within the range from O to 1 mM azide, CbsCbs was most stable, and CasCas, CcsCcs and CasCbs were unstable in this order, but more than 1 mM azide, CcsCcs, CasCas and CasCbs were unstable in this order. After incubation at various temperatures changing from 30°C to 60°C for 10 min, CasCas was most stable, and CasCbs, CcsCcs and CbsCbs were unstable in this order. After acid and alkali treatments in the range of pH 5.5 to 9.0, relative activities of CcsCcs and CasCbs were similar to that of CasCas, but CbsCbs was less stable than CasCas, CcsCcs and CasCbs in the same range. It is considered that the structure of catalase molecule in the liver is different between normal, mutant mice and heterozigote of normal and mutant mice with regard to the difference of the stability to LIS, guanidine, azide, heat, acid and alkali treatments.
S. aureus can grow in high salinity condition by a salt-tolerance mechanism. We have shown that S. aureus accumulated a large amount of proline in cytoplasm for the osmoregulation in high osmotic condition. In this study, we tried to elucidate the dynamics of proline by altering the salt concentration and supplementing proline in semisynthetic or synthetic media. S. aureus required only 20μM of proline as an essential amino acid under the normal condition, but 700μM to accumulate proline as an osmoregulator under the 10% NaCl condition. When the supplementary proline was not enough, S. aureus accumulated glutamine to compensate the osmoregulation by proline.
On the 10th day of pregnancy, the mother rats were injected with water miscible vitamin A (200, 000IU/kg) intraperitoneally to gain fetuses with malformed ear induced by maternal hypervitaminosis A. Their fetuses were removed on the 21st day for preparation of their skeletal specimens by using bone and cartilage staining. In order to examine pathological findings of the malformed ear, chiefly of the middle ear region, a stereoscopic microscope was used. The results were as follows: No abnormalities were found in the majority of the tympanic bone, Meckel's cartilage and malleus. Abnormalities such as malformation and absence were found in the majority of the styloid process from Reichert's cartilage origin, the incus and the stapes. In view of the above phenomena, these are highly probable that the malleus has its origin in the first branchial arch that is associated with the Meckel's cartilage and the stapes has its origin in the second branchial arch that is associated with the Reichert's cartilage. It is also probable that the greater part of the incus also has its origin in the second branchial arch.
Vibratory mode of the infant skull was observed and analysed by holography to elucidate the mechanism of bone-conduction hearing in the childhood. Three dried human infant skulls, one of six months, two of eight months postnatal age, were used. As the cranial suture was incomplete with the wide fonticulus remaining and the composition was not so dense or strong as that of the adult, very careful manipulation was needed in the experiment with the infant skull. Consequently, an acoustic loud-speaker was utilized to make the infant skull vibrate in the free sound field, instead of using direct vibrator conventionally attached to the adult skull for the measurement of vibration. An area around the foramen occipitale magnum was fixed by means of bolts, and the skull was stood on a table. Attempts were made to change the frequency and intensity of the test sounds during the experiment. The sound pressure level of the sound field was measured with a noise meter. The value measured at the site of the skull closest to the loud-speaker was regarded as the intensity of the test sound. The resulting pattern of vibration was recorded and reconstructed by holographic interferometry. As the light source for holography helium-neon laser was used. Interference fringes were observed at 110 phon or more of the intensities of the test sounds. When frequency of the test tone was low such that 500 Hz or less, interference fringes of parallel lines were observed in the hologram. This meant the skull was found to vibrate as a whole, and the sound signal seemed to be transmitted by “inertia bone-conduction” in the mechanism of hearing when the frequency of the test sound was 500 Hz or less. on the other hand, at 1 kHz and more of the test frequency, the vibratory pattern changed into concentric fringes with multiple maximum points of amplitude. Then, it was clarified that when the frequency was 1 KHz or more, “compression bone-conduction” was added up in the hearing mechanism. At about 3 KHz of test sound, the vibratory amplitude was larger than any other frequencies used, suggesting that 3 KHz was the closest to the resonance frequency of the infant skull.
The role of substantia innominata in secondary generalization of amygdaloid kindled seizure was studied in cats and rats. Effects of the intracerebral injection of muscimol, a direct GABA agonist, and gabaculine, a GABA aminotransferase inhibitor, on amygdloid kindled seizure were examined. In addition, changes of regional brain GABA content after gabaculine injection into the substantia innominata were determined in rats. The prolongation of the latency to the onset of generalized seizure or the suppression of generalized convulsion was observed after the intra-substantia innominata injection in cats. In contrast, afterdischarge triggering threshold was elevated after the intra-amygdaloid injection. These effects were also observed in rats, the former effects were parallel to the increase of GABA content in the substantia innominata and frontal cortex, the latter effects were parallel to the increase of GABA content in the amygdala. The GABA system in substantia innominata might play an inhibitory role for the secondary generalization of amygdaloid kindled seizure.
This study was undertaken to examine the removal rate of guanidino compounds (GS) as uremic toxin in the small animal models of acute renal failure (ARF) undergoing hemodialysis. For this purpose, the methodological trial of hemodialysis was done to determine the clearance and removal rate of solutes, ultrafiltration rate and pressure in vitro, and extracorporeal blood flow in vivo. The mean removal rate of methylguanidine (MG) and guanidinosuccinic acid (GSA) was 33.8% and 56.0% respectively. Correlation was only observed between GSA and creatinine, GSA and arginine. But MG was not correlated with other GC. These results indicate that MG and GSA are important indicators to start hemodialysis and to understand metabolic disorder in ARF.
There was no distinct differnce between the two regression lines of total E. coli-form and fecal E. coli-form investigated from 1978 to 1980 and in 1985. Target value of fecal E. coli-form decided in the invstigation from 1978 to 1980 was applied to the investigation in 1985, which was recognized to be appropriate. Sampling stations where the ratio (%) of fecal E. coli-form against total E. coli-form is high are located in the suburbs of small and middle cities without river basin sewerage system. At present, even in a case having attained the quality standard value of total E. coli-form, the same disposition as that for the unattained should be done if the case has not attained the quality value of fecal E. coli-form.
The author studied the immunohistochemical localization of alpha and beta subunits of S-100 protein in normal human cerebral and glioblastoma tissues by an indirect immunoperoxidase method. The glial cells including astrocytes and oligodendrocytes showed positive staining for both alpha and beta subunits of S-100 protein, however, the neurons revealed positive staining for alpha subunit only. The positive staining for both alpha and beta subunits of S-100 protein was revealed in one case of 6 glioblastomas examined and the positive staining for either alpha or beta subunit in three cases. The other 2 cases showed no positive staining. By immunoelectron microscopy, the glial cells revealed positive reaction product for both alpha and beta subunits of S-100 protein throughout the cytoplasm. Occasionally, the positive reaction products for both alpha and beta subunits were observed in the glial nucleoplasm and outer membrane of cell organelles including rough endoplasmic reticulum. The subcellular localization of alpha subunit of S-100 protein in the neurons and glioblastoma cells is basically simillar to that of the glial cells. The immunoelectron microscopic study of the subcellular localization of S-100 protein subunits seems to be one of the useful methods in assuming the biological role of S-100 protein.
Silicone rubber, Flexicast (Nisshiri Corp., Tokyo) colored with four different dyes (acrylic polymer emulsion) was injected into the bile duct, hepatic artery, portal vein and hepatic vein. The liver parenchyma was corroded with potassium hydroxide solution. Because the liver cast with Flexicast had adequate flexibility, it was possible to observe the inner structure of the liver by separating the hindering small branches with the fingers. The three vascular systems and duct system were clearly distinguished by their different colors. Peripheral branches were easily removed with a scissors, if necessary, to expose the main branches. The details of the method for the corrosion preparation of the liver with Flexicast were described and discussed.
Regional cerebral blood flow was measured with 133Xe injected into the internal carotid artery in 23 cases of brain tumor and 6 cases of the control. Based on Stochastic method and compartmental method, mean regional cerebral blood flow measured 46±4 ml/100g/min and a model curve was induced from control cases as yy=2294 exp(-0.1153t)±706 exp(-0.019t). Deviation value of each curve from the model curve was analysed as an index for rCBF of tumor and non-tumor area. Regional cerebral blood flow on the tumor consisted of not only one compartment but also two compartments that abnormal high blood flow of arterio-venous channel accompanied with abnormal low blood flow or ordinary cerebral blood flow, namely intratumoral steal flow existed. On the other hand, patterns of cerebral blood flow were simple on non-tumor areas; high or low blood flow, however, inter-regional inhomogeneity of rCBF was demonstrated as follows. According to rCBF on tumor, three groups were classified to high blood flow group (3 cases of glioblastoma, 2 cases of meningioma, 2 cases of astrocytoma and 2 cases of AVM), low blood flow group (2 cases of glioblastoma, 2 cases of meingioma, 2 cases of metastatic tumor and 3 cases of intracerebral hematoma), and normal blood flow group (each case of glioblastoma, meningioma and AVM). Mean rCBF was 36±8 ml/100g/min on non-tumor areas: Perifocal area measured 37±9ml/100g/min and remote area from the tumor measured 36±7 ml/100g/min. No difference of mean rCBF between perifocal and remote area presented. However, 10 cases showed relative perifocal hyperemia that rCBF on perifocal area was higher than on remote area. Eight cases showed relative perifocal ischemia that vis-à-vis. Five cases showed no difference between non-tumor areas. In comparison of rCBF on tumor and non-tumor areas, relative focal hyperemia was demonstrated in 16 cases, relative focal ischemia in 4 cases, and relative focal isoremia in 3 cases. While 10 cases out of the 16 cases associated with relative perifocal hyperemia, all out of the 4 cases associated with relative perifocal ischemia. On another point of view, 6 cases out of 10 cases of relative perifocal hyperemia presented high blood flow on the tumor area and 7 cases out of 8 cases of relative perifocal ischemia presented low blood flow on the tumor area. Cerebral blood flow dynamics of brain tumor was basically characterized into two patterns: 1) In cases of high blood flow on the tumor area, rCBF on the non-tumor areas centrofugally decreased according to the distance from the tumor, that may be called intracerebral steal flow. 2) In cases of low blood flow on the tumor areas, centrofugal increase in rCBF on non-tumor areas that may be functioned as pressure difference in the hemisphere due to brain edema.
Serum TSH, T4, T3 rT3 and resin T3 uptake (RT3U) were daily or monthly measured in 71 full-term neonates and 146 healthy infants, to study the changes of these thyroid hormones levels in relation to the development during infancy. Serum TSH levels at the first day of life were very high and the geometric mean was 18.8 μU/ml. Following an abrupt fall at the second day (a mean of 7.7 μU/ml), the level decreased gradually to a mean of 3.4 μU/ml at the third week, which was not significantly different from the values in children aged 1 to 5 years. Serum T4 and T7 (T4 x RT3U) values remained high during the first 5 days of life (16.1-18.9 μg/dl for T4 and 5.2-6.4 for T7) as compared with the levels in cord blood. The levels decreased to values similar to those seen in children aged 1 to 5 years by the third week of life (10.8 μg/dl for T4 and 2.9 for T7). Serum T3 levels in the first 2 days of life were high as compared with those of cord blood. Following a transient fall until the age of 4 days, the level again increased slowly and reached the maximum mean level of 208 ng/dl at 3 months. The mean T3 levels between 3 and 21 days of life (113-155 ng/dl) were lower than those in children aged 1 to 5 years (169 ng/dl), but the levels in infants aged 3 to 6 months (190-208 ng/dl) were higher than those in children aged 1 to 5 years. On the other hand, serum rT3 levels were remarkably high in cord blood (184 ng/dl) and remained high during the first 5 days of life (158-231 ng/dl). An abrupt fall occurred at the second week (a mean of 70 ng/dl), and thereafter the levels decreased slowly to a mean of 26 ng/dl by the age of 7 to 8 months, a value similar to that seen in children aged 1 to 5 years. The difference in the T3/T4 ratio from that in children aged 1 to 5 years had become insignificant by the age of 22 to 28 days, while the differences in the rT3/T4 and T3/rT3 ratios had become insignificant by the age of 7 to 8 months. These results suggested that the varying maturation of the conversion mechanism of T4 to T3 in the peripheral tissues in developing infants together with the neonatal surge of TSH secretion may be responsible for these extraordinary changes of serum thyroid hormones during infancy, especially in the neonatal period. The present study also demonstrated that the maturation of the conversion mechanism has completed by the age of 7 to 8 months.
Effects of various factors including perinatal factors, neonatal feeding method and maternal thyroid hormones levels on the pituitary-thyroid function in neonates were studied. Perinatal factors, such as cesarean section, breech delivery, twin, neonatal jaundice and neonatal asphyxia, had no effects on serum TSH levels between 10 and 28 days of life. There were also no significant differences in serum TSH or T4 during 14-28 days of life between breast-fed and artificially fed infants. Though serum TSH levels in low birth weight neonates (preterm neonates) at the first day of life were significantly lower than those in full-term neonates, no significant difference between the two had been observed thereafter. Serum T3 levels in preterm neonates were significantly lower and serum T4 levels tended to be lower than those levels in full-term neonates during the first 2 weeks of life. On the other hand, in preterm neonates rT3 levels were significantly higher and T3/rT3 ratios were significantly lower than those in full-term neonates during the first 2 weeks of life. These results suggested that the conversion activity of T4 to T3 in the peripheral tissues is more immature in preterm neonates. No significant correlations of serum TSH, T4, T3 and rT3 levels were present between the maternal blood and cord blood, indicating that these hormones can hardly cross the placenta.