Okayama University Medical School

Insulin binding to erythrocytes was studied in diabetic patients. Insulin binding was lower in untreated diabetics and diabetic patients treated with diet or insulin than in normal subjects. Binding variation was mainly due to decreased binding site concentration in untreated and insulin-treated patients, and to lowered insulin binding site affinity in diet-treated patients. Several patients treated with hypoglycemic agents showed higher insulin binding due to increased binding site concentration. Insulin binding to erythrocytes may not always reflect the insulin binding status of insulin sensitive tissues.

The effect of L-glutamate of cyclic AMP levels in different areas of rat cerebral cortex was examined during the development of an iron-induced focus in the left sensorimotor cortex. The addition of L-glutamate resulted in increased cyclic AMP levels in slices from the quarter of cortex with the iron-induced focus. The levels lowered in slices from the quarter of cortex most remote from the focus. Thus L-glutamate showed stimulatory and inhibitory effects on the accumulation of cyclic AMP.

In order to get precise information about the movement of plasma membrane proteins in cap formation, cyto- and bio-chemical analyses were made of the plasma membranes from non-capped areas of Ehrlich ascites tumor cells (EATCs) exposed to concanavalin A (Con A). Blebs formed by treatment with cytochalasin B (CB) of the non-capped areas of cells having a cap were isolated and used as the plasma membranes from non-capped areas (ConA-CB-bleb fraction). This bleb fraction was compared with a bleb fraction prepared from cells without ConA-treatment (CB-bleb fraction). Cytochemical analysis of ConA-CB-bleb fraction revealed a decreased in conA binding sites (ConA-BS) compared to the CB-bleb fraction. SDS polyacrylamide slab gel electrophoresis also revealed a decrease in the major components of ConA-BS of the ConA-CB-bleb fraction. The minor components of ConA-BS showed no distinct quantitative difference between the ConA-CB-bleb and CB-bleb fractions. NA+ K+-adenosine triphosphatase (ATPase), 5' nucleotidase (5'ND) and gamma-glutamyl transpeptidase (gamma-GTP) did not show any decrease in activity in the ConA-CB-bleb fraction, but the activity of D+-stimulated phosphatase (K-Pase) was decreased. The findings indicate that there are two types of plasma membrane glycoproteins in EATCs; one includes those participating in cap formation due to ConA, e.g. the major components of ConA-BS and K-Pase, and the other, those not participating in such cap formation, e.g. some minor components of ConA-BS, ATPase, 5'ND and gamma-GTP, which keep their places without moving.

A 63-year-old man developed generalized lymphadenopathy with skin rashes, fever, hepatomegaly and polyclonal hypergammaglobulinemia, twice, in February 1972 and in June 1979, after taking allopurinol for gout. Cervical lymph node biopsy, performed each time, showed the presence of immunoblasts and plasma cells, effaced nodal structure with involvement of the pericapsular tissue, rich vascularity and numerous mitoses, indicative of angio-immunoblastic lymphadenopathy with dysproteinemia (Frizzera, Moran and Rappaport). The existence of hypersensitivity to drugs, in particular, allopurinol in certain patients was emphasized, and induction of immunoblastic lymphadenopathy with various other therapeutic agents was briefly discussed.

In order to investigate the immunological responsiveness of tumor-bearing hosts to tumor cells, splenic suppressor cells from Ehrlich tumor-bearing mice that inhibited anti-tumor effector cell activity were characterized. In vitro cell-mediated cytoxicity and cytostasis assays were performed to test for the existence of anti-tumor immunity. suppressive activity assayed by cell mixture experiments became apparent with decline of anti-tumor immunity and progressive tumor growth. The cells mediating the suppression were found to be nylon wool column adherent T cells and inhibited T cell dependent cytotoxicity rather than non-T cell dependent cytostasis. In vivo cell transfer experiments demonstrated that intravenous injection of suppressor cells to a host already inoculated with tumor cells mixed with antitumor effector cells resulted in significant enhancement of tumor growth. This inhibition of in vivo neutralization assay be suppressor cells was found in not only allogeneic but also syngeneic tumor system. Splenectomy at the time of tumor resection endowed the host with stronger resistance against subsequent reinoculated tumor than sham-splenectomy did, reflected by prolonged survival times. These results suggest that splenectomy combined with surgical removal of the tumor is a useful treatment of clinical malignancies.

The present status and the history of inbred strains of mice in the Experimental Animal Center, Okayama University Medical School, otherwise known as “The Colony of Okayama University Medical School” are described.

Marked IgE-mediated histamine release from rat mast cells sensitized in vitro with mouse antiserum occurs in the presence of added Ca++ and phosphatidylserine (PS), although a considerable degree of antigen-induced histamine release which may utilize intracellular or cell-bound calcium is also observed. The decay in the responsiveness to Ca++ of the sensitized cells stimulated by antigen in Ca++-free medium in the presence of PS is relatively slow, and maximum release is produced by Ca++ added 1 min after antigen. Histamine release also occurs when Ca++ is added after PS in the absence of antigen to the sensitized cells suspended in Ca++-free medium. Unlike the antigen-induced release, the intensity of this non-antigen-induced release varies depending on both mast-cell and antiserum pools. A heat-labile factor(s), which is different from antigen-specific IgE antibody and is also contained in normal mouse serum, is involved in this reaction. In the antigen-nondependent (PS + Ca++)-induced release, no decay in the responsiveness to Ca++ is observed after PS addition. Both the antigen-induced and non-antigen-induced release are completed fairly rapidly and are dependent of temperature, pH and energy.

Chromosome analysis was performed on cells from a patient of null cell lymphoma, well-differentiated type. A 14q12 translocation was observed in all the banded cells. In addition, there were multiple chromosome abnormalities. This case will be useful in considering the significance of the 14q1(1-3) translocation in malignant lymphoma disease.

Some critical experiments have been carried out on the microspectrophotometry using the lymphocytes of a mouse, stained with Feulgen reaction, revealing that most reliable value can be attained by illuminating the material with a small spot-light and integrating the area surrounded by the extinction curve drawn by tracing along the diameter of the smeared and fixed cell.

Der Verfasser macht die HARTERT sche Thrombelastographie bekannt; das Verfahren ist noch nicht in Ungarn und vielen anderen Laendern im Gebrauch. Auf Grunde seine Untersuchungen geht er auf die detaillierte Auswertung des Thrombelastograme ein und referiert ueber die Anwendung und Brauchbarkeit der Thrombelastographes und gibt im Tabelle 1. den Angaben verschiedenen Patienten bekannt.

After grinding the tubercle bacilli cells, both human virulent strain, H37Rv, and avirulent strain, H37Ra, cultured in 5auton's medium, and obtaining three fractions of R1, S1 and R2 (R1, the first sediment; S1, the second supernatant; and R2, the second sediment) by the ultracentrifugation, the authors studied the enzymatic activities and the antigenic capacity against infection of these fractions; and obtained the following results: 1) Although the R1-fraction confers the defensive forte to mice in some degree, because of the presence of living bacilli in the fraction, it is difficult to decide definitely whether the defensive force owes its capability to this fraction or to living bacilli at the present stage of our experiment. 2) The S1-fraction possesses enzymatic activity on various substrates, but it does not confer animal any defensive force against infection. 3) The R2-fraction specifically oxidizes lactate and succinate" and it can markedly impart animal the defensive ability against infection.

1. Histochemical and cytochemical studies with respect to the sites of reaction were made on the succinic dehydrogenase system activity of human and animal tissues using ditetrazolium salts, namely, neotetrazolium chloride, nitro-neotetrazolium chloride, and nitra-blue tetrazolium chloride. 2. The advantages and disadvantages of each ditetrazolium salt for histochemical and cytochemical purposes and the reaction taking place in frozen tissue sections and that in fresh tissue blocks were compared, and the method of procedure suitable for each condition was established with some modification. 3. Selecting conditions suitable for cytochemical purpose, it was shown that the reaction took place at the sites coinciding with mitochondria, and the distribution of the enzyme reaction was also examined. In addition, several new findings in the brains and other tissues cytochemically made clear were pointed out.

The present paper describes each pattern of the free amino acids in different parts of the dog brain determined by ion-exchange chromatography. The parts examined have been the cerebral cortex, cerebral white matter, cerebellar hemisphere, cerebellar vermis, caudate nucleus, thalamus, hypothalamus, and medulla oblongata. Gamma-aminobutyric acid concentration was the highest in the hypothalamus. Glutamic acid showed lower values in the white matter, hypothalamus, and medulla oblongata. Aspartic acid showed lower values in the white matter and caudate nucleus and higher values in the medulla oblongata. Glutathione and cystathionine showed higher values in the thalamus. N-Acetylaspartic acid showed lower values in the white matter and medulla oblongata. Glycine and alanine showed higher values in the medulla oblongata.

By physically destroying typhoid bacilli and centrifuging at a high speed, an insoluble granular fraction (P1) and soluble fraction (S1) were obtained. Chemical and enzymologic properties of these substances as well as their influences on the protective ability against infection were studied; and the following results were attained: 1. P1 contains an extremely small amount of proteins when compared with S1. 2. The enzymologic activity of P1 is entirely different from that of S1. In P1 the respiratory enzyme system of only lactate and succinate is localized. 3. Although both P1 and S1 possess the antibody-producing ability in serum of rabbit to the same high degree, P1 imparts to mice a markedly high protective ability against infection. 4. By the heat-treatment of P1 its antigenicity is lost at the same time.

From these results it is but natural to assume that the antigen-antibody reaction is involved in the phenomenon, eosinophilia. The antigen in this instance is the filtrate of hookworm emulsion, and the serum of hookworm disease as well as the bone marrow can be thought to contain the antibody. In any case, so long as the medium contains the serum or bone marrow or both of them obtained from the patient of hookworm disease, eosinophilia and the acceleration in the motility of eosinophils are brought about in the growth zone by addition of the filtrate of hookworm emulsion. Therfore, as for the mechanism inducing hookworm eosinophilia, it may by interpreted that the patient of hookworm disese is repeatedly sensitized by the antigen arising all probability from the metabolic products of hookworms or from the dead bodies of the worms; and producing the antibody in tissues and blood, thus the antigen-antibody reaction is elicited in vivo as long as hookworms live in the human body so that the increase in the mitosis and the acceration in the motility of eosinophils in the bone marrow are brought about with the resultant continuous discharge of a large quantity of eosinophils from the bone marrow parenchma into the sinusoids, there by inducing eosinophilia in the peripheral blood.

Of eosinophilias that we often encounter clinically, we selected two of the most representative ones, namely, hookworm diseae and bronchial astma, for our present sternal bone-marrow tissue culture, and studied the movement patterns and wandering capacity of eosinophils. As the results, even in those eosinophils that show no significant change other than the increase in number in ordinary stained-smear specimens of peripheral blood or bone marrow, it has been clarified that, when observed under living condition, they reveal a picture specific to individualistic behaviors according to diseases. Therefore, it can be assumed that in the pathologic condition what is known as eosinopilia not only eosinophils increase in number but also qualitative changes of eosinophlils specific to each disease are brought about, and consequently these specific changes are reflected on the movement patterns of the eosinophil.

1. With a view to grasp more simply and clearly the characteristics of this disease and in order to find a clue for prompt discovery of cases when encountered in future, the authors undertook a statistical study of the cases already reported by various authors. 2. The cases reported so far amount to 17 familial groups which consisted of 38 acatalasemic cases. These groups were distributed widely throughout Japan. The disease seemed to be prevalent in the rural communities where adherence to the custom of consanguineous marriage occurs. As yet, we have not heard of the occurrence of this disease in other countries. 3. The disease has equal distribution in both sexes. About one half of patients showed a peculiar oral gangrene (Takahara's disease). The great majority of these were noted in those less than 10 years of age. 4. The great majority of them were children whose parents were united in consanguineous marriage and have siblings with acatalasemia. 5. As for the treatment of oral lesions in this disease, extraction of tooth at the site of the lesions, removal of the diseased tissues en masse by resection, and penicillin treatment given concomitantly are effective. The course and the length of time required in healing of the wound due to the operation are about the same as in the case of normal persons. 6. The authors wish to call special attention to the phenomenon peculiar to the acatalasemic blood. The blood of acatalasemic individuals changes to brownish-black color in the absence of foaming or bubble formation upon the application of hydrogen peroxide to blood.

1. Methods of retrograde coronary perfusion and direct coronary artery perfusion in combination with a bubble oxygenator were investigated in dogs. 2. Ventricular fibrillation occurred more frequently during the operation in hypothermia than in the operation performed in combination with the extracorporeal circulation. 3. The optimal pressure of perfusion is considered to be 30 to 35 mm Hg in retroperfusion, whereas, 100 mm Hg in direct coronary artery perfusion. 4. Perfusion by the pressure bottle method is preferable to the gravity method because the fall of blood temperature in the irrigation tubing might cause ventricular fibrillation. 5. From the metabolic study of the methods is clear that there is a tendency to myocardial anoxia after 15 to 20 minutes of perfusion in both methods.

An improved chromatographic method for the analysis of sugar phosphates and nucleotides presented in the previous paper was applied on the analysis of the metabolites of fructose produced by the action of hexokinase in some tissues in vitro, and the excellency of this method was demonstrated. The results showed that by this method the metabolites of the sugar in its early stage of catabolism can be analysed very easy and exactly and estimated quantitatively as well, giving no inconsistency with the results presented by the routine method.