Search

Drug effects were studied on anaphylactic histamine release from rat mast cells sensitized in vitro with mouse IgE antibody. When histamine release was elicited by adding Ca-++ at various times after antigen-stimulation of sensitized cells in Ca++-free medium, the drugs to be tested were added shortly before each Ca++ addition. Quercetin was effective only when added before or immediately after antigen. Theophylline and disodium cromoglycate (DSCG) were active irrespective of the time interval between antigen and Ca++ addition. Verapamil was more effective when added before or simultaneously with antigen than when added later. When tested in the two-stage experiments, quercetin showed inhibition only in Stage 1 and verapamil was inhibitive primarily in Stage 1, while theophylline and DSCG wee only inhibitive in Stage 2. It seems that quercetin selectively and verapamil primarily act to block calcium-gate opening resulting from antigen-antibody interaction on the mast cell membrane, while theophylline and DSCG selectively inhibit the passage of calcium through open calcium channels.

The antimycotic action of 1, 4-bis-(m, m'-amidinophenoxymethyl)-cyclohexane dilactate (MAC), a synthetic diamidine compound, on Candida albicans was studied. The minimum inhibitory concentration (MIC) ranged from 3.31 to 6.25 micrograms/ml against both standard and clinically isolated strains. MAC was fungistatic at MIC and weakly fungicidal at the concentration of 100 micrograms/ml. MAC did not affect the cell wall or cause cell lysis. Intracellular constituents, such as 260 nm and 280 nm absorbing materials, were released from the cells by treatment with MAC indicating that MAC affected membrane permeability. The release of 260 nm absorbing material was inhibited by the presence of Ca2+ and Mg2+. Acidic phospholipids such as cardiolipin and phosphatidylglycerol inhibited the anti-Candida activity of MAC, but sterols and lecithin were not inhibitory, indicating that MAC interacted with acidic phospholipids of the cell membrane. Freeze-fracture electron microscopy showed that MAC caused aggregation of membrane particles and patch formation on the P face, which may indicate that MAC is a membrane disrupting agent. It appeared that MAC affected C. albicans at the cell membrane by interacting with acidic phospholipids and caused disorganization of the membrane structure resulting in the release of intracellular constituents without lysis.

To investigate cellular interactions between human T and B lymphocytes in various diseases, we established a technique to prove terminal differentiation of B lymphocytes into immunoglobulin synthesizing and secreting cells. We also established a double antibody radioimmunoassay to measure the amount of IgG, IgA and IgM synthesized and secreted in culture supernatants. Purified immunoglobulins were obtained from sera of patients with myeloma or macroglobulinemia. The peripheral blood lymphocytes from 25 normal individuals had the geometric mean synthetic rates of 1886 ng for IgG, 1607 ng for IgA and 1173 ng for IgM per 1 X 10(6) cells when cultured for nine days in the presence of pokeweed mitogen. The method is simple and sensitive, and is thought to be useful for examining human lymphocyte function in vitro.

Insulin and human erythrocyte cell membrane interactions were studied with respect to binding and dissociation. The per cent of specific binding of 125I-labeled insulin to erythrocytes was directly proportional to the cell concentration. The optimum pH for binding was 8.1. The initial binding rate was directly proportional to, and the steady state insulin binding was reversely proportional to, the incubation temperature. The per cent of specific binding of 125I-labeled insulin was 12.10 +/- 1.13 per cent (mean +/- SD)/4 X 10(9) cells (n = 10) at 0.8 ng/ml insulin. Native insulin competed with 125I-labeled insulin for binding and showed almost complete inhibition at 10(4) ng/ml. The Scatchard plots were upward concave. Maximum binding capacity was 230 binding sites per cell. The average affinity constant decreased as the per cent of fractional occupancy increased. Affinity constants for the empty and filled sites were 1.49 and 0.16 X 10(8) M-1 respectively. Bound insulin was displaced by native insulin. The dissociation rate by "dilution + native insulin" was higher than that by "dilution only". The dissociation rate was accelerated even by the physiological concentration of insulin and maximum at 100 ng/ml. It is concluded that human erythrocytes have insulin binding sites which are indistinguishable from insulin receptors on the target tissues for insulin.

Serum specimens from 12 patients with type A hepatitis were analyzed for immunoglobulin M-type antibody to hepatitis A virus (IgM anti-HA). A recently developed solid-phase radioimmunoassay kit for IgM anti-HA (HAVAB-M, Abbott Laboratories) and a competitive binding radioimmunoassay kit (HAVAB, Abbott Laboratories) with or without 2-mercaptoethanol treatment, as modified by Yano et al. (Acta Hepatol. Jpn. 21, 704-712, 1980) were used to obtain an M-index. All specimens obtained within 60 days of the onset of illness and specimens from 2 of 4 patients later than 60 days after the onset were positive with the HAVAB-M test. This test gave negative results to sera which were positive for anti-HA by a standard HAVAB test in the following: 3 patients with type B hepatitis; 5 with non-A, non-B hepatitis; 11 healthy adults; and 10 sera strongly positive for rheumatoid factor. The M-index for type A hepatitis in sera within 30 days of the onset (mean value of the M-index, m, = 1.52; standard deviation, SD, = 0.25) was significantly higher than that for non-A hepatitis (m = 1.05; SD = 0.15) and for healthy adults (m = 1.02; SD = 0.10). The simplicity and usefulness of the HAVAB-M test in diagnosis of acute type A hepatitis over those measuring the M-index by HAVAB tests were shown by direct comparison of the results.

Partial excess of chromosome 1 (q25-q32) was noted in malignant cells from all of 10 patients who had disorders such as non-African Burkitt's lymphoma, adult T-cell leukemia, myelofibrosis, malignant lymphoma, chronic lymphocytic leukemia or chronic myelocytic leukemia in blast crisis. The break points on chromosome 1 were at centromere, q12, q21, q23, q25 and q32. Variations in the specific region of the long arm of chromosome 1, q25-q32, were thought to be important in the evolution of malignant cell proliferation.

Most of the common organic solvents are excreted into urine as metabolites. A correlation exists for several organic solvents between the amount taken in and the amount of metabolites excreted. Many methods have been developed for the measurement of these urinary metabolites. The methods were classified into three group: colorimetry, gas chromatography and high performance liquid chromatography. The characteristics and availability of these methods in a laboratory for routine work were reviewed. The correction equation for the amounts of metabolites in spot urine is discussed.

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.

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.