The chromogenic method for amylase activity using blue starch is widely used in clinical laboratories. This method is simpler, quicker and more reproducible than the amyloclastic method using iodine staining or the saccharogenic method using dinitrosalicylic acid. Recently, Levitt et al, found that the chromogenic method often showed considerably lower value on urinary amylase than the amyloclastic and saccharogenic methods. This discrepancy of amylase values was clearly improved by the addition of albumin or other proteins to reaction mixture. Among four proteins tested, albumin, . γ-globulin, α-casein and submaxillary mucin, albumin was most effective for the improvement of assay value.
The four proteins tested did not affect the K_m value for a substrate, blue starch, appreciably, indicating that they did not affect the affinity of amylase for its substrate. But they did increase the V_max value, showing that they increased the effective amount of amylase during incubation. Thus they will increase amylase activity by protecting the enzyme from inactivation.
Amylase activity was higher in urine samples containing proteins than in those without any protein. Therefore, amylase activity in urine samples showing proteinuria may be evaluated as higher than that in urine samples without detectable protein. Thus albumin should be added to the reaction mixture for assay of urinary amylase at a final concentration of 1mg/ml. The results also indicate that albumin should be added at the step of dilution of enzyme. Routine addition of albumin at these steps should make the assay more reliable for clinical purpose.

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Previous works show that macrophages promote the in vitro proliferative response of T cells of the guinea pig. However, in most of the past experiments peritoneal exudate cells have been used as macrophages and macrophages separated from the other organ sources were rarely employed up to date.
In order to elucidate the differences in the function of macrophages from different organ sources in an individual non-treated guinea pig, the following experiments were undertaken using purified lymph node T cells passed through nylon wool column, T-cell mitogens (Con A, PHA-P) and Mitomycin C (MMC) -treated accessory cells from three different organ sources (lymph nodes, spleen and lung). The present studies were centered on the demonstration of accessory cell function of alveolar cells on the in vitro proliferative response of T cells because accessory cell functions are not yet fully clarified at present.
The optimal cell number of MMC-treated whole cells in the determination of ³H-TdR uptakes was 1×10⁶ cells and the optimal incubation time in this experimental system was 48 hours. Similar results were obtained in all experimental systems employing the different organ sources of MMC-treated whole cells and in Con A- or PHA-P-stimulated mitogenesis.
In case of MMC-treated alveolar whole cells, however, there was a difference in the levels of T cell ³H-TdR incorporation by mitogenic stimulation in comparison with that from other two organ sources. The levels of ³H-TdR uptake stimulated by Con A were rather lower, while the levels of ³H-TdR uptake by PHA-P were not so much different.
In order to separate the cell fraction having accessory function, adherent cell-rich fractions were separated from the whole cells in three organ sources. The levels of ³H-TdR uptakes in these cell fractions were elevated.
From these rusults, it was clarified that alveolar cells, especially adherent cell-rich fraction, have accessory function on the in vitro proliferative responses of lymph node T cells by mitogenic stimulation.

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There are many reports related to the delayed-type hypersensitivity (DTH) following to group A streptococcal infection. The roles of cell-mediated immunity to streptococci, however, isn't fully understood. Lymphocytes from normal guinea pigs and from guinea pigs displaying DTH to group A streptococcal cell wall antigen were cultured in the presence of the antigen. The supernatant fluid from the lymphocyte culture was assessed for their roles in the cell-mediated immunity to streptococcal cell wall antigen.
Guinea pigs were sensitized with a single, 4 or 12 times injections of a emulsion containing 500μg of antigen with Freund's complete adjuvant into the four foot-pads and subcutaneously. Lymphocytes from regional lymph nodes of normal or antigen sensitized guinea pigs were incubated with various concentrations of antigen. The culture supernatant fluid was then collected.
Using the culture supernatant fluid, macrophage migration inhibition test (MIF), ³H-TdR incorporation of normal guinea pig lymphocytes, ¹⁴C-glucosamine up take by normal guinea pig macrophages, the activity to accelerate phagocytosis against viable streptococci, and cytotoxic activity on ³H-labeled L-cells were investigated.
DTH to streptococcal cell wall antigen was observed in immunized guinea pigs before the production of serum antibody. The peritoneal macrophages of normal guinea pigs were activated and phagocytosis of the macrophages against viable streptococci were accelarated by lymphokines which obtained from culture supernatant fluid of lymphocytes of immune animals in the presence of antigen. This fact may play the very important role in the streptococcal immunity at the early stages of streptococcal infection before the production of serum antibody. The production of cytotoxic factor from immunized lymphocytes decreased following to the increase of sensitization times, but the production of macrophage migration inhibitory factor didn't change following to the increase of sensitization times. The activity was observed distinctly in the culture supernatant fluid from lymphocytes 12 times' sensitized animals, but couldn't observe in that from normal, a single and 4 times' sensitized animals.
Thus the production of macrophage migration inhibitory factor was not necessarily correlated with that of the mitogenic factor and that of the cytotoxic factor. These facts suggest that there may exist T-cells displaying different function in the process of immunization.

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Pyelorenal backflows which have been often clinically observed by retrograde pyelography are usually classified in the following four types; pyelotubular, pyelo-interstitial, pyelovenous and pyelolymphatic.
These types were found in our corrosion-casting specimen study of the fourteen human kidneys used with acrylic resin, “TECHNOVIT” (Kurzer Co., West Germany) : i. e. red, blue and yellow coloured resins were infused in the artery, vein and ureter, respectively. The fourteen cases in this communication were consisted of following categories.
1) Pyelotubular backflow (seven cases, photo 1). In these specimens the fine structures of the tubuli which were radially distributed on the center of the calyx were observed. These tubuli belong to the collecting ducts and the papillary ducts, and are estimated approximately at 50.400 microns in diameter.
2) Pyelo-interstitial backflow (thirteen cases, photo 2). In these cases, the rocky masses of the acrylic resin around the fornix of the renal calyx were observed.
3) Pyelovenous backflow (thirteen cases, photo 3). The injected acrylic resin teared or ruptured the fornical angle of the renal calm and flowed into the peripapillary veins was observed. Therefore, the arcuate, interlobares, interlobulares and renal veins are filled with resin.
4) Pyelolymphatic backflow (five cases, photo 4). In these specimens, the very fine streaks or threadlike tubuli passing outside the renal fornix were recognized. These countless tubuli are probably the lymphatic vessels, and are estimated at 20.40 microns in diameter.

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Recently, it was demonstrated that not only histamine and isoproterenol but also bradykinin elevate cyclic AMP (cAMP) levels of murine lymphocytes and also it was suggested that the agents increasing intracellular cAMP have in general an antiproliferative effect to lymphocytes.
In this study, the effects of bradykinin as well as histamine and isoproterenol on mitogen induced proliferation of murine splenic lymphocytes were examined. The results obtained were as follows:
1) Bradykinin as well as histamine and isoproterenol at the noncytotoxic concentrations suppressed in vitro proliferation of splenic lymphocytes induced by PHA-P or Con-A. The most remarkable inhibition was observed at the concentrations of 10^-5M bradykinin, 10^-4M histamine and 10^-7M isoproterenol. The inhibition percents against PHA-P induced proliferation were 44% at the presence of 10^-5M bradykinin, 27.6% at the presence of 10^-4M histamine and 27% at the presence of 10^-7M isoproterenol. The inhibition percents against Con-A induced lymphocyte proliferation were 50.6% at the presence of 10^-5M bradykinin, 28.4% at 10-4M histamine and 32.9% at 10^-7M isoproterenol.
2) Both bradykinin and histamine did not suppress LPS-induced lymphocyte proliferation, but isoproterenol suppressed it and the inhibition percent was 28.4% at the presence of 10^-7M isoproterenol.
3) Against LPS-induced proliferation of splenic lymphocytes isolated from nu/nu mice, bradykinin and histamine did not show suppressive effect, but isoproterenol showed inhibitory effect and the inhibition percent was 22.9% at the presence of 10^-7M isoproterenol.
4) From the results of this experiment it was suggested that isoproterenol may affect both T and B-lymphocytes, but bradykinin as well as histamine may affect rather T-lymphocytes among lymphocyte populations.

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In order to clarify the incidence mechanism of the disturbance of glucose metabolism recognized during the acute stage of cerebrovascular disease, a study was made by performing an intravenous glucose tolerance test, and the relationship between the clinical symptoms and glucose metabolism disturbance was investigated. Moreover, rabbits were used as experimental animals, and cerebral hemorrhage was produced experimentally by the cold injury method deviced by Klatzo. The levels of the blood glucose (B. G.), insulin, and cortisol were examined periodically. Furthermore, the influence of β-adrenergic blockade (propranolol) on the blood glucose level measured on the rabbits with experimental cerebral hemorrhage was investigated, and the following results were obtained.
Through the clinical observations on the cases of cerebrovascular diseases, the disturbance of glucose metabolism was severe in the cases of cerebral hemorrhage, particularly intraventricular hemorrhage, in comparison to that of cerebral infarction. Those with poor prognoses such as severe disturbance of consciousness, high C.S.F. -pressure, and intraventricular hemorrhage revealed severer than all the others. That is, the more intense stress on the brain caused the severer disturbance of glucose metabolism.
In the experimental cerebral hemorrhage, the group of severe injury revealed higher B.G. level, that is hyperglycemic response curve. In the same group, the cortisol level in blood was recognized higher. The hyperglycemic response curve was controlled by administering propranolol. Also, the B.G. level of the internal carotid vein on the damaged hemisphere during the earlier stage of the injury was lower than the other. This means that the glucose uptake in the brain was increasing on the damaged hemisphere.
The above results explain that the disturbance of glucose metabolism in cerebrovascular disease is due to the higher cortisol level resulted from the active function of pituitary adrenocortical system and also to the β-adrenergic receptors by epinephrine. This hyperglycemia is possibly related to biological reaction to repair cerebral injury and the lowered function of the brain.

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CNS complications of M. pneumoniae infections are not widely known yet. The relation between M. pneumoniae and CNS disease has been based mainly on clinical observations and serological data from serial serum samples. Isolations of this organism from CSF, blood and throat culture have been tried, but direct evidences of an etiologic relationship have been very few.
In this paper, we reported three patients complicated with meningitis (case 2) and meningoencephalitis (case 1 & 3) who had respiratory tract infections diagnosed on the basis of clinical symptomes and serological findings of M. pneumoniae infection.
Although nourologic symptomes have been improved, in two cases with meningoencephalitis, abnormal findings of EEG were respectively noticed about one and three months after onset. In one case (case 1), focal spike and wave were recognized in the left temporal area with a natural sleep activated EEG. In another case (case 3), 3-4c/s atypical spike and wave complex were recognized dominantly over the left hemisphere with a natural sleep activated EEG. These abnormal EEG findings have persisted for months or longer period. Accordingly, a long term follow-up by EEG should be required for these cases. R. J. Lerer stated that 21% of CNS complications of M. pneumoniae infections had no antecedent respiratory symptomes.
This suggests that culture of M. pneumoniae and the serological tests for those cases of aseptic meningitis and meningoencephalitis, the etiology of which is unknown, should be carried out.

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