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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.

The usefulness of liposomes (in neutral, positively and negatively charged forms) as a carrier for adriamycin (ADM) was studied by examining the distribution of ADM and related fluorescent compounds in Ehrlich solid tumor-bearing mice. The mice were given free or liposome-entrapped ADM intraperitoneally. The distribution of ADM and related fluorescent compounds between the administration of the free form and liposome-entrapped form was measured by high performance liquid chromatography : The distribution was dependent on the form of the liposomes. The amounts of ADM and its metabolites in the mouse serum 20 min after administration of neutral-liposome-entrapped ADM were 10 times those after the administration of free ADM, 6 times those after the administration of a negatively charged form, and 3.5 times those in the administration of positively charged form. There was no marked difference in the concentrations of these compounds 5 h after administration. The concentration of these compounds in the liver 60 min after administration of each liposome-entrapped form of ADM were in inverse correlation with the concentrations in the serum obtained at 20 min after administration. Total concentrations of ADM and its metabolites in the tumors 20 min after administration of each entrapped form of ADM were 4-5 times that in administration of free ADM after 20 min. There were no marked differences in the concentration of ADM for administration of the various liposome forms. Statistically significant decreases in mean tumor weight were seen in the groups given neutral, positively and negatively charged liposome-entrapped forms compared to corresponding control groups given with free ADM.

To investigate the action of motilin on the sphincter of Oddi, the flow rate of the perfusate (FRP) discharged into the duodenal lumen through the orifice of the common bile duct was measured by means of an electric drop counter in decerebrated dogs. Motilin in doses above 0.5 micrograms/kg i.v. reduced or stopped the FRP. The fifty percent recovery time of FRP was 20 min and full recovery time was 30 min. The reduction of FRP induced by motilin was unaffected by denervation and atropinization. These results suggest that motilin caused an increase in tone of the sphincter of Oddi by acting on the sphincter muscle.

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.

In our study on the influences of various hormones and various endocrines on the megakaryocyte function by means of bone-marrow tissue culture, we obtained the following. 1. In the repeated administration of ACTH, cortisone, testosterone, progesterone, or thyroxin to guinea pigs, these hormones accelerated the megakaryocyte function, whereas estradiol on the contrary diminished the function. 2. The removal of such endocrines as the pituitary, adrenal, thyroid or testicles will diminish the megakaryocyte function, while removal of the ovaries accelerates it. 3. For the megakaryocytes in the hypophysectomized rats, ACTH acts most effectively to restore their function, followed by cortisone and pulverized thyroid, while testosterone has hardly any effect on the function. 4. ACTH, cortisone, prednisolone, testosterone, progesterone, and pulverized thyroid act directly on megakaryocytes so as to accelerate their function, while estradiol diminishes the megakaryocyte function. 5. For the megakaryocytes in idiopathic thrombocytopenic purpura ACTH is most effective in restoring the function; for the megakaryocytes in hypoplastic anemia cortisone is most effective; and for the megakaryocytes in Banti's disease prednisolone is most effective in restoring the megakaryocyte function.

1. In the stage later than the middle stage of pregnancy, morphological differences appear between the amniotic epithelial cells of placenta and those of the free part and the majority of cases the amniotic epithelial cells of placenta present more marked columnar shape than those in the surrounding area of ruptured orifice or those in the vicinity of placenta. However, there still remains a question whether or not such a phenomena is directly related to the secretory function of the placenta amniotic epithelium. 2. It seems that amniotic epithelial cells divide and multiply themselves by mitosis at least in the early and middle stages when their functions are at height. 3. Even in the stage later than the middle stage generally the amniotic epithelium of placenta is consisted of a single layer of columnar epithelial cells, and therefore, the author cannot agree to Forssell's theory. 4. In glycogen and lipid stainings, the amniotic epithelial cell layer shows more striking changes with the progress of gestational month when compared with those cells in other layers. 5. Glycogen in the amniotic epithelial cell layer is abundant in the early and middle stages of pregnancy, and it rapidly decreases near the late stage. Lipid granules on the contrary are less in the early stage, and start to appear in the middle stage, increasing rapidly towards the late stage. In general, the regressive degeneration picture of the late stage is not distinct histologically, but assuming glycogen to represent the cell activity and the lipid deposit to mean just the reverse, the amniotic epithelium functionally seems to fall into regressive degeneration from the middle stage. Other layers of fetal membranes likewise undergo fatty degeneration as the pregnancy progresses from the middle stage to the late stage. 6. There still remain problems to be solved on the question what role this regressive degeneration of the amniotic epithelial cell layer plays in de Watteville's theory, "Labor originates from the fetal membranes". However, granular PAS-positive substances in the amniotic epithelium are glycogen, and it seems difficult to connect simply the existence or non-existence of PAS-positive granules or Sudan-positive granules directly with the continuation or interruption of pregnancy.

According to the data obtained in this experiment by means of the geldiffusion technique, the specific antigen was not detected in MH134 ascitic tumor, comparing the anti-C3H liver sera with anti-MH 134 tumor sera, though a loss of organ specific antigen and weak antigenicity were found in MH134 tumor extract. In order to detect some qualitative alteration, supposedly a gain in antigenic components, the transplant rejection test was carried out. The result of this test indicates that the relative not absolute resistance could be induced to C3H mice by this prior sensitization with cell free active extract eluted from MH134 tumor tissue by Fluorocarbon treatment. During these experiments, it became clear that MH134 ascitic tumor cell has weak immunizing properties so that prolonged lapse of time and large dose of antigen are inevitably necessary. Moreover, through Fluorocarbon treatment of the tumor homogenate, the cellfree, serologically active antigen could be obtained, which will serve well for the induction of the isologous immunization.

In the experiments conducted with cells from the spleen and regional lymphoid cells derived from the axillary and regional lymph nodes of the mice receiving subcutaneous transplantation of Ehrlich tumor cells, it has been demonstrated that these lymphoid cells possess an ability to inhibit the proliferation of the subtrain of Ehrlich tumor cells (JTC-11), and that there is a parallel relationship between the rate of appearance of immunologicallly competent cells among these sensitized lymphoid cells and the inhibitory effect of these cells on JTC-11. In the experiment to reinforce the potency of sensitized cells conducted by means of the diffusion chamber technique, it has been proven that, while it is not an absolute prerogative, it is necessary to have the sensitized lymphoid cells come in contact with the target cells is order to wield the power of the former to the fullest extent. In the tissue cultures of lymphoid cells, it has been shown that sensitized lymphoid cells has a shorter life span than non-sensitized ones.

There is as yet no plausible and convincing explanation for the etiology of rheumatoid arthritis. The authors investigated anti-synovial tissue antibody in the body fluid of rheumatoid arthritis by means of indirect immunofluorescent technic using non-affected synovial tissues as antigen. As the result the anti-synovial tissue antibody was detected in 7 cases of the 15 synovial fluid samples of rheumatoid arthritis and in two out of the six serum samples. The site of the localization of this antibody was demonstrated to be in the synovial membrane, especially in synovial cells and in the small blood vessel walls situated immediately adjacent to the synovial surface, but it was found in no connective tissues other than synovial membrane. It seems that this anti-synovial tissue antibody should be considered as an independent factor from rheumatoid factor, and that rather than the rheumatoid factor it is more actively associated with the localization and progression of chronic inflammation within the rheumatoid arthritis joint.

From the histochemical study on the vascular distribution of limb muscles of cats and rats, the following results were obtained. 1. The red, white and intermediate fibers of the soleus and gastrocnemius individually present dissimilar patterns of the vascular distribution. In the transverse section of the muscle, most of the capillaries are found preferentially assembled around the small red fiber. Usually the red fibers are supplied with 4-6 capillaries, while the intermediate fibers 3-4 capillaries and the white 1-2 capillaries. 2. In the longitudinal section, the capillaries run parallel with the muscle fiber. Occasionally, these capillaries branching out for anastomoses cross the fiber almost at right angle. These anastomoses are more frequently observed on the red fibers. 3. The soleus muscle is better supplied with capillaries than is the gastrocnemius, because this muscle is composed only of two types of capillary rich fibers, namely, the red and intermediate fibers. 4. The mean capillary-fiber ratio of the cat gastrocnemius proves to be 0.6 and that of the soleus 1.4. 5. From these observations, it is considered that there is an intimate relationship between the capillary distribution pattern and the type of individual muscle fibers. 6. The red fibers being with a high oxidative enzyme activity and rich in mitochondria may require an abundant supply of oxygen and have a greater number of capillaries. In contrast to this, the white fibers with a lower enzyme activity and a fewer mitochondria need not to consume so much oxygen and are supplied with a fewer capillaries, because they participate only in phasic contraction. The intermediate fibers, with a moderate enzyme activity, may be considered to have a moderate number of capillaries.

Mononuclear cells from rabbit joint fluid were studied after synovitis was induced by various means, including the intra-articular injection of bacterial endotoxin or of aggregated human gamma globulin in normal rabbits, or of HGG in rabbits previously sensitized to this material. The large majority of mononuclear cells in all groups were monocytoid rather lymphocytoid, and these cells were most readily labeled with tritiated thymidine on the first day after injection. On day 2 and 3, the numbers of labeled cells decreased, except for the animals previously sensitized with HGG, in which there was an upswing of labeling on day 3. This upswing was associated with a considerable increase in numbers of cells resembing synovial cells, and may possibly be a reflection of synovial proliferation. Macrophages loaded with engulfed polymorphonuclear cells were observed in all experimental groups, a finding which emphasizes the lack of specificity of this reaction.

A high incidence of subnormal serum folic acid levels was observed in 48 patients receiving anticonvulsants (75 %). In peripheral blood, macrocytosis was detected in 46 % and an increase of hypersegmented neutrophils was also seen in 24 % of the patients. Correlation existed between these signs and low serum folate levels. The growth response of Lactobacillus casei and L. leichmannii was not suppressed by the addition of various anticonvulsants to the medium of the bioassay systems. Administration of 5 mg of folic acid for a month corrected macrocytosis and an increase of hypersegmented neutrophils significantly. Folic acid supply also decreased mean diameters of the nuclei of oral epithelial cells significantly. It is concluded that subclinical folic acid deficiency is common among the patients receiving anticonvulsants. Absorption of 3H.folic acid from the small intestine of rats was inhibited by large dose of diphenylhydantoin (20 mg) not by 5 mg. This fact suggests that in patients on diphenylhydantoin, the quantity balance of folic acid and diphenylhydantoin in the intestine regulates the absorption of folic acid.

I) On an identified giant neurone of the right parietal ganglion in a snail's subesophageal ganglion-complex, the synaptic contribution to the production of the plateau formation of biopotential or grouped spike discharges of the soma has been studied in the presence of a convulsant. 2) The orthodromic stimulation of a peripheral nerve (the intestinal nerve) can elicit the plateau formation of biopotential, instead of normal spike discharges, in the identified neurone treated with a convulsant. 3) With the application of a convulsant, for example bemegride which was in a concentration less than that necessary to produce the plateau formation, an EPSP accompanied a spike with a constant delay. This EPSP is a product of a proprioceptive reflex arc consisting of two excitatory synapses with a certain subordinate neurone. 4) Later, in the presence of a convulsant, spontaneously conveyed multiple EPSP's were observed on the biopotential of the identified neurone. These multiple EPSP's produced grouped spike discharges or the plateau formation of biopotential of the neurone. 5) The multiple EPSP's may be produced by the grouped spike discharges of the subordinate neurone, the membrane property of which would be changed by a convulsant. It is presumed that the grouped spike discharges or the plateau formation of biopotential often occurs synchron. ously in many neighboring neurones by means of synaptic triggering in the presence of a convulsant.

Circular DNA isolated from human kidney mitochondria was studied by electron microscopy. I. Mean contour length of monomers of the mitochondrial DNA was 4.96 ± SE 0.28 /μ 2. The complex molecules (oligomers) of mitochondrial DNA were observed in frequency of 6.2 per cent. Among them circular dimers accounted for two per cent of all circular DNA molecules. 3. Circular DNA fibers with an intermediate perimeter between the　monomer and dimer, and with a contour length shorter than 3 μ were occasionally observed. 4. Some discussions were made on the emergence of the circular dimer.

A non-invasive method for measuring portal blood flow by magnetic resonance (MR) phase contrast was evaluated in a flow phantom and 20 healthy volunteers. In a flow phantom study, the flow volumes and mean flow velocities measured by MR phase contrast showed close correlations with those measured by electromagnetic flowmetry. In 20 healthy volunteers, the cross-sectional areas, flow volumes and mean flow velocities measured by MR phase contrast correlated well with those measured by the Doppler ultrasound method. Portal blood flow averaged during the imaging time could be measured under natural breathing conditions by using a large number of acquisitions without the limitations imposed on the Doppler ultrasound method. MR phase contrast is considered to be useful for the non-invasive measurement of portal blood flow.

To clarify the influence of the introduction of metal bats on the physique and batting records of the players in the National Summer High School Baseball Tournaments, a comparative analysis was conducted between height, body weight, body-weight ratio (body weight/height), batting average and home run average of the best four teams' players (n = 493) and those of the other teams' players (n = 4,590) in three periods: the period of the use of wooden bats, that of the use of both wooden and metal bats and that of the use of metal bats. In the period of metal bat use, the mean values of physique of the best four teams' players were significantly larger (P < 0.05) and their average number of home runs was significantly higher than those of the other teams' players (P < 0.01). The only significant differences between the two groups in those indices for each time period were for height in the wood and metal/wood periods. This study demonstrated that the differences between the home run average and physique, including height, body weight and body-weight ratio of the best four teams and the rest of the teams were the greatest after the use of metal bats. These findings suggest that the importance of the home run average increased and was associated with large-size of physique after the use of metal bats in the National Summer High School Baseball Tournaments.

With the parameters of a flow-volume and a volume-time curve, the discriminant analysis of bronchial asthma is described. The subjects were classified into three groups (healthy adults, mild asthmatic patients and moderates ones). The difference of the mean vectors of the parameters of the three groups was made clear by the selection methods of the discriminant analysis between any two of the groups both with 6 parameters (%FVC, FEV1.0%, peak flow rate (PF), flow rate at 50% of FVC (V50), flow rate at 25% of FVC (V25), and V50/V25) and with 8 (6 parameters mentioned above and V75, V10). Forced expiratory volume in 1 second percent (FEV1.0%) or V50 was selected at the first step with 6 parameters, and V75 was selected at the first step with 8 parameters. Probabilities of misclassification with 8 parameters were lower than those with 6 ones and the probability of misclassification at the discriminant analysis between healthy adults and mild asthmatic patients with 8 parameters was 15.75% at the final step.

In urine of diabetics, a significantly great inhibitory activity of glyceraldehyde 3-phosphate dehydrogenase (GAP-Dehyd) was observed compared with that of normal subjects. The 0.2 ml of urine from 41 patients with diabetes mellitus inhibited 0.5 units of GAP-Dehyd by 27.2 +/- 3.0% (mean +/- S.E.M.), while that from 17 normal volunteers inhibited only by 9.0 +/- 1.0% (P less than 0.05). This inhibitory substance was extracted by 90% ethanol from diabetic urine and partially purified by anion exchange chromatography using Dowex-1 (HCOO type). The molecular weight of this substance was confirmed to be 100--300 daltons by an analysis on Biogel P-4 gel filtration chromatography. And analysis by thin layer chromatography using silicagel plate showed that this inhibitor was a ninhydrin reactive substance which has not been reported previously. From the above facts, it was assumed that the inhibitory substance of GAP-Dehyd in urine of diabetics was a new acidic compound of low molecular weight containing an amino residue in the molecule.