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Polyamines are polycationic substances which are widely distributed in living organisms and have a close relation to rapid growth phenomena. We examined ornithine decarboxylase (ODC), which is the rate limiting enzyme of polyamine biosynthesis, and polyamine induction in primary cultured rat hepatocytes by various hormones which increase during pregnancy, and revealed differences in hormonal responses between adult and fetal rat hepatocytes. Thirteen hormones, including estrone, estradiol, progesterone, testosterone, human chorionic gonadotropin (HCG), cortisol, dexamethasone, insulin, glucagon, epinephrine and epidermal growth factor (EGF), were tested. Among these hormones, only insulin, dexamethasone and EGF induced ODC activity and polyamine biosynthesis, especially that of putrescine, in both adult and fetal hepatocytes. The effects of EGF were the most significant. The combined effect of insulin and dexamethasone was additive, while that of insulin and EGF was synergistic. The rate of ODC induction was higher in adult hepatocytes than in fetal hepatocytes, however, the reaction was earlier in fetal hepatocytes. These observations suggest that ODC and polyamines in the fetal stage of development are regulated by a mechanism different from that in the adult liver.

Thalamic neurons projecting to both the head of the caudate nucleus and the premotor cortex in the cat were studied by the retrograde fluorescent double labeling technique. After injections of Evans blue into the caudate nucleus, and diamidino-phenylindol into the premotor cortex, a small number of double labeled neurons appeared in the ventral anterior, ventral lateral, anteromedial, rhomboid, central dorsal, central lateral, central medial, paracentral and parafascicular nuclei, in addition to numerous single-labeled neurons. This indicates that some neurons in the thalamic nuclei send bifurcating axons to both the head of the caudate nucleus and the premotor cortex. The caudatal projections of these thalamic neurons are organized in a topical manner.

Catecholamines were measured in the amniotic fluid and in the first voided newborn urine obtained from appropriate-for-date infants of term deliveries. Catecholamine values in the amniotic fluid and urine were nearly equal when expressed in terms of creatinine. Significant positive correlations were observed between the amniotic fluid and urine of norepinephrine and epinephrine. In normal cases (n = 32) that underwent uneventful vaginal delivery, the 95% confidence limits for norepinephrine and epinephrine in the amniotic fluid were 1.53 to 2.33 ng/ml and 0.16 to 0.30 ng/ml, respectively. In cases of moderate stress (n = 12), only norepinephrine showed significantly higher values than the normal cases, while in cases of severe stress (n = 12), norepinephrine became more significantly high, and epinephrine was found to be elevated significantly. A significant difference was noted in the incidence of fetal stress between the infants with more than and those with less than 2.30 ng/ml of norepinephrine, the upper limits of the normal 95% confidence limits. However, for epinephrine such a significant difference was not noted. It was concluded that amniotic fluid catecholamines are of fetal origin and reflect fetal sympathoadrenal activity directly, even during labor, and that their level may be a good indicator of fetal condition and stress.

Age-associated changes in methionine-enkephalin (ENK) and thyrotropin releasing hormone (TRH) concentrations, and their receptors were examined in discrete regions of the rat brain. The ENK and TRH concentrations in aged rats were nearly identical to those in young adult rats, except for a slightly lower TRH value in the hypothalamus of the aged rats. On the other hand, the ENK and TRH receptor levels in the cerebral cortex of aged rats was markedly lower than that of young adults rats. The results suggest that determinations of both neuropeptide and receptor levels are indispensable for evaluation of peptide-mediated neural systems in the central nervous system.

1) The gaseous metabolism of guinea pigs (male and female) decreases to a certain extent after the removal of sexual glands, but it increases temporarily through the administration of the extract (of testis or ovary), up to the normal value as before the operation. 2) The testis extract has a little influence on increasing the gaseous metabolism of the ovariectomised guinea pigs and it is also temporary. 3) The oxidizable substances cause the gaseous metabolism of ovariectomised guinea pigs to increase markedly. 4) The gaseous metabolism of the castrated male is increased under the administration of the oxidizable substance of certain plant juices and the ratio of increase is proportional to the quantity of the oxidizable substance given, and the more, if the testis extract be added together. 5) The gaseous metabolism of the castrated animals fed with the basal diet free from vitamin C tends to diminish again. This diminished gaseous metabolism is only restored to the former level or more by the administration of vitamin C.

The results obtained in this investigation may be summarized as follows : 1. The CO2-output of the male muscle and other tissues is greater than that of the female. 2. The female muscle contains larger amount of water than the female muscle. 3. The muscle immersed 1/2 Ringer solution (or 1/2.5 R.) gave out smaller amount of CO2 per minute than the muscle in 2-Ringer's solution (or 2.5 R.). In spite of the difference in the water content of tissue between different sex, the salt content of the tissue liquid seems to be the same. In other words, larger the water content means larger content of tissue liquid in the tissue. Artificial introduction of water in the tissue or reduction of water content by immersing the tissue in 1/2 or 2-Ringer's solution is quite different from the natural condition occurring between different sex. However both of these conditions influence the gaseous metabolism in the same manner. On an assumption that the gas diffusion in liquid is proportional to the solubility of that gas, the above mentioned difference of CO2-output should be just reversed. Therefore it is not possible to interpret how the water content influences the gaseous metabolism. It may only be stated that the muscle which has a small amount of water to an extent which does not abolish excitability, gives out much CO2 and vice versa.

It was found, from the curves of mechanical registration that the precise determination of atrio-ventricular interval and the time interval from the beginning to the summit of ventricular contractions cannot be accurately measured. But it was clearly shown that the height of auricular contractions is obviously reduced by vagus stimulation as well as by the application of acethylcholin, which is generally known as "Vagusstoff" while on the other hand the ventricular contractions are not affected. It is considered, therefore, from the results obtained, that the acethylcholin like substance is produced from the endings of vagus nerve as a conseqence of its activity through stimulation, and that this substance causes a decrease in the height of auricular contractions. Examining the electrocardiogramm, it clearly showed a marked prolongation of the atrio-ventricular interval, and in the last three curves it failed to record the ventricular contraction after each P-spike. Hence it is considered that vagus stimulation affects the conductivity of atrio-ventricular bundle and causes diminution of its conductivity. The results above cited, agree with those of the former investigators. But with regard to the duration of the electrical response of the ventricle, it was found contrary for the vagus stimulation has no effect upon the function of ventricular muscle. From the results obtained, the following conclusions may be made. 1. The vagus stimulation affects the negative inotropic only on auricular contraction. 2. The vagus stimulation affects the negative dromotropic on the conductivity of atrio-ventricular bundle. 3. The vagus stimulation has no effect upon the function of ventricular muscle.

A. Um die Resultate obiger Untersuchungen ubersichtlich zu machen, habe ich in Tabelle I I die Antikorpermengenverhaltnisse zum Serumwert angegen. Danach muβ man einen Ubergang des Antikorpers zu beiden Korperflussigkeiten bei hohem Serumwert annehmen. Wegen der geringen Menge und der groβen Operations-schwierigkeit ist im allgemeinen der Antikorpernachweis bei der Ohrlymphe viel schwerer als beim Augenkammerwasser. B. Die Bindungszone des Prazipitins im Serum, Augenkammerwasser und in der inneren Ohrlymphe bei aktiv hoch immunisierten Kaninchen durch die wiederholte intravenose Antigeninjektion, ist uberall dieselbe. Ich bin noch keinem Fall begegnet, bei dem die Bindungszone des Prazipitins der 3 Versuchsflussigkeiten verschieden gewesen ware ; deshalb ist der Prazipitinnachweis etwas leichter als der andere Antikorpernachweis. C. Das Resultat stimmt mit den Angaben von Kimura11), daβ namlich die Bindungszone des Prazipitins im Serum, Augenkammerwasser und in der Zerebrospinalflussigkeit gleich ist, uberein. Es deckt sich auch mit den Angaben von Inoue32), daβ die Bindungszone des Prazipitins im Serum und in der Galle bei aktiv immunisierten Tieren gleich ist. Die Ergebnisse meiner obigen Untersuchungen durften sich folgendermaβen zusammenfassen lassen : Das Prazipitin, d.h. der Immunkorper, der im Augenkammerwasser und in der inneren Ohrlymphe auftritt, mag wohl teils lokal produziert werden, aber man muβ annehmen, daβ der groβere Teil dieses Immunkorpers aus der Blutbahn zum Augenkammerwasser und zur inneren Ohrlymphe ubergeht. Ich glaube, daβ dieses nicht nur fur Prazipitin gilt, sondern in gleicher Weise auch fur Agglutinin und Hamolysin.

A 76-year old farmer ingested 100 g of chlorphenamidine (Galectron), a plant acaricle, for the purpose of suicide. Gastric lavage was performed and the patient survived. Methemoglobinemia was noted after emergency treatment and was still present at 20 hours after ingestion of the compound. The patient was lethargic for at least 50 hours. Moderate neutrophilic leukocytosis and kidney injury were observed.

A simple method is described for determing thyroxine binding proteins in human serum by electrophoresis at pH 8.6, using cellulose acetate membrane as the supporting medium. The procedure had high reliability in sera of normal subjects, pregnant women and patients with decreased thyroxine binding capacity of thyroxine binding globulin.

Cells from methylcholanthrene-induced tumor (MC-tumor), Ehrlich ascites cancer or mouse ascites hepatoma (MH-134) were subcutaneously implanted in dorsal area of mice to examine the specific cell mediated immunity following implantation. The migration index (MI) of lymphocytes was determined at various time periods after cell transplantation. The MI-activity increased under all three implantations, reached maximum at a certain period, decreased gradually and disappeared. The maximum MI-activity coincided with the proliferation period of the implanted tumor cells. This peak occurred on the tenth postimplantation day with MC-tumors, on the fifth day with Ehrlich ascites cancer and on the sixth day with MH-134 cancer. In lymphoid tissues of animals with MC-tumor and Ehrlich ascites cancer, strong MI-activity appeared early in the regional axillary lymph nodes, while weak activity was observed consistently in the distant mesenterial lymph nodes. The MI-activity of the splenic lymphoid cells resembled the axillary lymph nodes cell activity. The MI-activity of venous blood lymphoid cells was parallel to the average value of lymphoid cells of the spleen and axillary and mesenterial lymph nodes.

The relationship between muscle activity at the terminal region of the common bile duct and the duodenal muscle was examined in rabbits. The rhythmic muscle activity in the terminal region was synchronous with duodenal muscle activity. The activity of the latter muscle preceded the former. The activity at the terminal region synchronous with the rhythmic activity of the duodenal muscle sometimes disappeared spontaneously. The muscle activity of the ampulla and the spincter at the terminal region was sometimes independently lost. The conduction of excitation from the duodenal muscle to the terminal region appeared to be performed at several sites. The existence of a "conduction-shunt path" between the terminal region and the duodenum, as well as between the ampulla and the sphincter appeared probably. Some quantitative differences were found between the spincter, ampulla and duodenum in inhibitory effects to stimulation of splanchnic nerves and reflex effects and to excitatory effects of cholecystokinin-pancreoxymin and caerulein. These results seem to indicate that the sympathetic nerves and the intramural cholinergic neurones controlling these region carry out activities quantitatively different from each other.

Hepatitis B core antigen (HBc Ag) and hepatitis B surface antigen (HBs Ag) were detected in the liver tissue of a patient with chronic aggressive hepatitis by the immunofluorescent complement technique. The presence of anti-HBc was examined by the same method in 67 human sera previously tested for HBs Ag, anti-HBs and s-GPT levels. HBc Ag was localized mainly in the nucleus and sometimes in the cytoplasm of the hepatic cells. HBs Ag was found only in the cytoplasm. The focal area of HBc Ag positive hepatic cells seemed to correspond to the HBs Ag positive cells. Double staining demonstrated the simultaneous presence of HBs Ag and HBc Ag in individual cells. Anti-HBc positive serum was found in 46 (68.7%) cases. Forty-eight (71.6%) indicated a combination of HBs Ag and anti-HBc.

In order to approach human cancer immunotherapy, the author carried out the immunotherapy with BCG on mice having homotransplanted cancer, observed the posttransplantation results with lapse of time, conduced daily macrophage inhibition test (MI test) and found the immunotherapy to be effective. At the same time the MI test proved to be a useful criterion in determining the course of cancer progress and effectiveness of the immunotherapy.

Phosphate-binding protein(s) was found in the inner mitochondrial membrane of calf heart by Sephadex G-200 and G-25 gel filtration. The binding activity was inhibited by N-ethylmaleimide and competed by a large amount of cold phosphate. The amount of phosphate bound to the fraction was 29 nmoles per mg of protein. Affinity chromatography with phosphate-bound Sepharose 4B confirmed the presence of phosphate-binding protein(s) in the active fraction of mitochondrial membrane fractionated by gel filtration.

In order to formulate an early diagnostic method for acute rejection after kidney transplantation, macrophage migration inhibition test (MIT) was carried out with lapse of time after inbred rat kidney allotransplantation. The mean survival time of rat kidney allograft was found to be 7.07 +/- 1.34 days. On the other hand, in the group treated with rabbit anti-rat lymphocyte serum (ALS) the mean survival time was lengthened to 14.15 +/- 2.14 days (p less than 0.05). The corresponding antigen used for MIT was prepared with donor kidney by ultrasonication, and its protein concentration at 180 mug/ml was the most optimal as not to elicit non-specific inhibition of macrophages. In the control group, activity of macrophage inhibitory factor (MIF activity) turned positive 3 days after the transplantation, and it became strongly positive by 5 or 7 dyas at the period when rejection crisis appeared frequently. ALS-treated group showed a lower MIF activity than the control group (p less than 0.05) and on 7-12 dyas before rejection crisis appeared frequently, MIF activity became strongly positive. These findings suggest that this MIT is simple and will be proved to be useful in predicting the acute rejection as well as in controlling the immunosuppression.

Human adenovirus type 12 (Ad 12) was inoculated through subtentorial route into inbred newborn mice (C3H/BifB/Ki), and sequential changes of the brain and tumor induction were examined by histological and immunofluorescent methods. Two days after virus inoculation, Ad 12 specific tumor antigen (fluorescent T-antigen) appeared in the cells of ependymal and subventricular matrix layers, choroid plexuses and leptomeninges in the subtentorial as well as the supratentorial brains. After 10 days, these fluorescent positive cells decreased gradually in number but still remained focally beneath the ependyma. Sixty days later, early tumor nodules were detected in the same regions in which remained the fluorescent cells. After 107 days, neurological signs and well-developed tumors were noted in 25 of 63 (30.1%) mice examined. In the cerebellum, both of T-antigens and tumors were limited around the IVth ventricle, but not in the granular layers. Histomorphologically, the tumors were of primitive neuroectodermal origin and consisted of the cells resembling immature matrix cells in the subventricular zone. These findings strongly suggest that the virus has a selective affinity to the remaining matrix cells, but not to cerebellar granular cells, at least, in newborn mice.

A cell line (HGC-27) was established by culture of the metastatic lymph node from a gastric cancer patient diagnosed histologically as undifferentiated carcinoma. HGC-27 cells were polygonal or short spindle-shaped and adhered to glass surfaces as a monolayer. The cells were probably derived from gastric cancer cells, as their origin from mesenchymal tissues can be excluded morphologically and enzyme-histochemically. Enzyme activities were generally negative or low, except for adenosine triphosphatase, lactic dehydrogenase and leucine aminopeptidase. These scanty findings might reflect the undifferentiated character of the original tumor cells. The cloning efficiency was 5.3% in liquid medium and 1.0% in soft agar. The doubling time was about 17 hr. Chromosomal analysis revealed a mode of 109 and 110 chromosomes.

A case with prolonged bacterial infection accompanied by an abnormal serum protein which migrated in the post-gamma region on electrophoresis is presented. The abnormal protein was identified as IgG with gamma-type light chain moiety. The patient suffered from prolonged pneumonia and cholecystitis, Bone marrow aspiration and skeletal x-rays did not indicate multiple myeloma.