Okayama Igakkai Zasshi (Journal of Okayama Medical Association)
Online ISSN : 1882-4528
Print ISSN : 0030-1558
Volume 89, Issue 7-8
Displaying 1-20 of 20 articles from this issue
  • Part I. Inhibitory effects of cepharanthine on the K+-release after the injury to cell membrane
    Masanobu MIYAHARA, Kozo UTSUMI, Katsumi SUGIYAMA, Kaname AONO
    1977Volume 89Issue 7-8 Pages 749-756
    Published: August 30, 1977
    Released on J-STAGE: March 30, 2009
    JOURNAL FREE ACCESS
    Concerining the physiological properties possessed by the cell membrane, recently an attention has been called on various problems such as the cell recognition mechanism to begin with, and the mutual relationships among the intercellular communication mechanism, as well as the metabolism adjustment of membrane binding enzymes, aside from the compartmentation of substances. Essentially the physico-chemical properties of the membrane of cancer cell and proliferating cells are important in relation to the treatment of cancer. And attempts are being made to change the cell metabolism by artificially altering the physicochemical properties of the cell membrane.
    Cepharanthine, one of the biscoclaurin alkaloids, is known from olden days to possess a thanatophidia hemolytic property, but this property seems to bring about the change in the physico-chemical properties of the cell membrane, and it is interesting in the point that this substance may have a membrane modifying property. For these reasons, during the investigation on the actions of alkaloids on the cell membrane many interesting phenomena have been elucidated. This report presents the results recently obtained about the changes in physico-chemical properties of biological membrane by the treatment with cepharanthine. The obtained results were as follows.
    1. Cepharanthine inhibits K+-release from red blood cells when these cells are treated with snake venom, phospholipase A, lysolecithine, lead acetate and hexane. But the alkaloid does not inhibit K+-release induced by ionophores, surface active agents and HVJ.
    2. Cepharanthine inhibits the hyperpolalization of membrane potential induced by lead acetate or hexane.
    3. Similar inhibitory effect of cholesterol on K+-release from the cells by the treatment with lead acetate is observed.
    4. From these results it is suggested that the inhibitory action of cepharanthine on K+-release from cells is due to the action of decrease in membrane fluidity.
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  • Masayuki MIZOBUCHI
    1977Volume 89Issue 7-8 Pages 757-780
    Published: August 30, 1977
    Released on J-STAGE: March 30, 2009
    JOURNAL FREE ACCESS
    The experiment was carried out to investigate the effects of total parenteral nutrition (TPN) on body weight, nitrogen balance and the histological or cytological changes of main organs in rats. TPN was introduced to 59 female Wistar rats of seven weeks after birth, for ten days. The animals were devided into six groups as follows:
    Group I Control, fed orally by pellets
    Group II TPN of amino acid-fat-glucose
    Group III TPN of amino acid-glucose
    Group IV Fat and glucose
    Group V Amino acid exclusively
    Group VI Starvation
    1) The body weight increases in Group II and III were 6.7% and 3.6% of the initial value, respectively. However, the increase in body weight of each group was less than that of control. Nitrogen balance study showed that cumulative nitrogens of Group II and III for seven days were 1.10 and 0.93 grams, respectively.
    2) Fatty infiltration in the liver was not due to whether TPN consisted of fat or not. When a Cal/N ratio was increased in TPN, liver showed fatty infiltration at the peripheral regions of the lobules. The fatty infiltration seemed to become more diffuse as caloric excess was greater. Electron microscopy of hepatocytes revealed multiple giant lipid droplets in cytoplasm and glycogen cluster localizing around lipid droplets. This suggests that glycogen may change to fat following TPN.
    3) TPN caused an atrophy of exocrine pancreas. The atrophy was much greater in Group IV, which was excluded amino acid. Ultrastructurally, pancreatic acinar cells revealed increased FCD (focal cytoplasmic degradation) of various size, augumented flat mitochondria, increased immature zymogen granules and dilatation of rough surfaced endoplasmic reticulum. These results show that deprivation of amino acid from TPN causes impairment to pancreatic acinar cells, even in short period.
    4) Infusion of hypertonic glucose or amino acid caused cloudy swelling and vacuolization of renal tubule epithelial cells, which ultrastructurally coincided with the swelling, deformity and vacuolization of mitochondria, low electron density of cytoplasm and increased lysosomes.
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  • Yoshihumi YOKOYAMA
    1977Volume 89Issue 7-8 Pages 781-791
    Published: August 30, 1977
    Released on J-STAGE: March 30, 2009
    JOURNAL FREE ACCESS
    The distribution of catecholamine of substantia nigra was observed under fluoromicroscopy by Falck-Hillarp method for catecholamine detection. Several mammals (monkey, goat, dog, cat, rabbit, guinea pig, rat and mouse) were used.
    Substantia nigra of all mammals used consisted of two regions, zona compacta and zona reticularis. Nerve cells and fibers with strong catecholamine-specific fluorescence were observed in zona compacta of all mammals used, but in zona reticularis nerve cells did not contain catecholamine and nerve fibers contained a little catecholamine. Nerve cells which consisted zona compacta of monkey, goat, dog and cat were large and sparsely distributed, while those of rabbit, guinea pig, rat and mouse were small and densely distributed. The strength of fluorescence of the latter animals were stronger than that of the former animals.
    Thus, the absence of significant differences among animals of distribution and cell shape of nerve cells of subatantia nigra suggest that this region has central role in the function of catecholamine, especially dopaminergic neurons of the brain.
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  • Yoshihiko ONO
    1977Volume 89Issue 7-8 Pages 793-817
    Published: August 30, 1977
    Released on J-STAGE: March 30, 2009
    JOURNAL FREE ACCESS
    The α1-antitrypsin (α1AT), which comprises more than 90% of the protein content of the α1-globulin, inhibits a variety of enzyme. α1 AT is the major trypsin inhibitor in human serum and has an important role as antiplasmin in fibrinolytic system.
    The experiments were carried out to investigate the effect of α1 AT on fibrinolytic system, measuring serum α1 AT levels and coagulative and fibrinolytic activities in man. The serum α1AT levels were measured in 30 controls, 93 cancer and 107 nonmalignant patients. They were also measured in the postoperative patients who were 7 cases of gastric cancer, 5 of lung cancer, 5 of breast cancer and 5 of cholelithiasis. The serum levels were measured by a single radial immunodiffusion plate (M-partigen) obtained from the Behring Institute.
    The results obtained were as follows:
    1) The levels of α1AT were significantly increased in patients with lung cancer, gastric cancer and colon cancer, while those with breast cancer and thyroid cancer remained in normal range. There was a significant correlation between α1 AT and the clinical stage of lung cancer and gastric cancer. Among nonmalignant patients, AT was significantly elevated in the inflammatory diseases such as lung abscess and hepatitis.
    2) α1AT had the lowerest level at the end of operation and it exceeded the preoperative value and returned to preoperative level at the 14th day. These changes of α1 AT were observed both cancer and nonmalignant patients. The plasma fibrinogen levels showed a similar pattern to α1 AT during postoperative period, whereas α2-macroglobulin had no significant changes. The concentration of plasma plasminogen and antithrombin III had been fallen below the preoperative levels by the 7th day and returned at the 14th day. Antitrypsin activities were increased to some extent by the 7th postoperative day.
    These results suggest that the elevation of α1AT is closely related to two factors as follows:
    a) α1AT may increase to neutralize a certain leucocytic protease when leucocytes increase in body.
    b) The increase in α1 AT might be due to regulating the plasmin which auguments in cancer, inflammation or postoperative patients, may induce release of antiplasmin whose activity is mainly dependent on α1 AT.
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  • Kensaku ONO
    1977Volume 89Issue 7-8 Pages 819-849
    Published: August 30, 1977
    Released on J-STAGE: March 30, 2009
    JOURNAL FREE ACCESS
    1) T cell population and blastogenic activity against both phytohemagglutinin (PHA) and pokeweek mitogen (PWM) were assayed in twenty patients, whose malignant neoplasma had been eradicated by surgery before more than six months, when the first clinical evidence of the recurrence was observed during the period of postoperative control.
    A deterioration of the blastogenic activity of the lymphocyte to PHA was highly significant, compaired with that in healthy subjects and patients who have had radical surgery for their neoplasm but showed no evidence of the recurrence. It was noted that this deterioration occurred regardlessly of the interval between the surgery and the recurrence and also of the mode of recurrent lesion.
    No remarkable differences were observed in the T cell population and the blastogenic activity to PWM among these three groups.
    2) Swiss mice were undergone subcutaneous autotransplantation of isolated tumor cells from their own methylcholanthrene-induced leg sarcoma after surgical removal of the tumor. The establishment of the autotransplantation and the growth of the tumor was daily observed and the blastogenic activity of the splenic lymphocyte to PHA was assayed 14 days after the tumor implantation.
    Neither a period before the tumor take nor cumulative incidence of the take were correlated with the blastogenic activity of the splenic cell. However, the individual size of the tumors 14 days after the transplantation was markedly smaller in mice having the blastogenic activity higher than the mean value in the group than in those having the activity lower than the mean. Thus the reverse correlation between the tumor size and the blastogenic activity ratio were statistically significant (p<0.01).
    An additional skin homograft performed simultaneously with the tumor inoculation resulted in the apparent increase in the number of the mice showing the blastogenic ratio higher than the mean in the non-grafted control, and this caused further inhibition of the tumor growth, showing also the significant reverse correlatio between the tumor size and the blastogenic ratio (p<0.01).
    B. C. G. inoculation, before, at and after the tumor transplantation, caused marked decrease in the mitogenic activity of the splenic cell, but the tumor growth was significantly detarded and the reverse correlation between the tumor size and the blastogenic ratio among the individual mice was preserved as in the above two experiments.
    In mice administered with Cephranthin, daily for 14 successive days starting with the tumor transplantation, the tumor growth inhibition of slight grade was observed as late as 14 days following the tumor inoculation and no significant effect on the blastogenic activity of the splenic cell was observed, and though the reverse correlation above was affirmed it was not highly significant (p<0.05).
    3) From the results mentioned above, the author intended to conclude that the reverse correlation between tumor growth and blastogenic activity of the lymphoid cell does exist and even the nonspecific immunopotentiation contribute to bestow the ability for the lymphoid cell to suppress the tumor growth by potentiation of the blastogenic activity, even in mechanism of establishment of autotransplantation of malignant tumor in the subject whose malignant neoplasma was eradicated, as in cancer bearing hosts.
    Additionally, the possibility that deterioration of the blastogenic activity of the lymphoid cell may play a role of triggering effect for dormant cancer cell to obtain a growing ability was presumed and a possibility of significant contribution to this mechanism of cell-mediated cancer immunity other than that can be assessed by mitogenic activity of the lymphoid cell was also discussed.
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  • I. Analyses of lipid component and fatty acid composition in rat incisor pulps before and after whole-body x-ray irradiation
    Gonosuke HASHIMOTO
    1977Volume 89Issue 7-8 Pages 851-858
    Published: August 30, 1977
    Released on J-STAGE: March 30, 2009
    JOURNAL FREE ACCESS
    Lipids and fatty acids extracted from rat incisor pulps before and after whole-body x-ray irradiation at 1000R were qualitatively and quantitatively analysed by means of thin-layer and gas-liquid chromatography, and the results are as follows.
    1) Ratio of the total lipid weight to protein weight was obtained 0.26 in normal rat dental pulps. The ratio in the irradiated group (3 days after irradiation) did not change.
    2) In fatty acid composition of lipid of normal rat incisor pulps, palmitic, oleic, stearic, arachidonic, and linoleic acids were much in this order, and palmitic, stearic and oleic acids were major component accounted for over 78% of the total fatty acids. In that of the irradiated group, the decrease of oleic acid and the increase of arachidonic acid were observed comparing those of normal pulp lipid.
    3) The major lipid of normal dental pulps composed of phospholipid, choresterol, triglyceride and choresterol ester, and phospholipid and choresterol accounted for about 45% and 30% of the total lipids, respectively. In the irradiated group, the decrease of choresterol and the increase of choresterol ester were represented.
    4) In results of tentative identification by various color reaction on the thin-layer chromatograph, the major phospholipids of dental pulps were lecithin, phosphatidyl ethanolamine and phosphatidyl serine, which accounted for over 90% of the total phospholipids. It is significant that phosphatidyl serine in dental pulp is greater than in any other mammalian soft tissue so far studied. In the irradiated group, it was slightly observed the increment of lecithin and the decrement of phosphatidyl serine, but they are not predominant.
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  • II. Lipid peroxidation of rat incisor pulp
    Gonosuke HASHIMOTO
    1977Volume 89Issue 7-8 Pages 859-863
    Published: August 30, 1977
    Released on J-STAGE: March 30, 2009
    JOURNAL FREE ACCESS
    Lipid peroxidation of dental pulp homogenate isolated from rat incisors was induced by ferrous ion. Lipd peroxidation of dental pulp homogenate isolated from rat 3 days after wholebody irradiation at 1000R x-ray was also induced by ferrous ion, and the activity was higher than that of control homogenate.
    In the extracted lipid components of dental pulps, the peroxidation of phospholipid were marked, but those of choresterol, choresterol ester, mono-, di-, tri-glycerides, and fatty acid (non-esterified) were slightly or negligible. In the phospholipid components, the peroxidation was highly observed in lecithin, phosphatidyl ethanolamine and phosphatidyl serine, and the peroxidation of phosphatidyl ethanolamine showed the most value in them.
    By the analysis of the fatty acid composition, arachidonic acid was included in the phospholipid but not in the triglyceride, suggesting arachidonic acid is substrate of the peroxidation of dental pulps.
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  • Yoshio UCHIDA
    1977Volume 89Issue 7-8 Pages 865-875
    Published: August 30, 1977
    Released on J-STAGE: March 30, 2009
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    As a link in the studies on biological activity of lymphocytes in the mice transplanted with Ehrlich ascites cancer and in cancer patients the correlation between machrophage migration inhibitory activity (MIF-activity) and the progress of cancer was investigated, and the results are presented briefly as follows.
    1. MIF activity in the mouse transplanted s. c. with 5×106 Ehrlich cancer cells first appears markedly in the regional lymph nodes and the MIF activity disappears about 10 days after the cancer transplantation.
    2. In three cases (8.6%) out of 36 having benign diseases MIF activity is positive.
    3. In measuring the MIF activity of peripheral blood lymphocytes of 71 gastric cancer patients before operation, those showing positive or pseudo-positive activity of MIF are 24 cases, being 33.9% of the total. MIF activity of peripheral blood lymphocytes before operation in the cases of Stage II is 41.7%, in those of Stage III it is 29.0% and in those of Stage IV it is 15.4%, indicating that the positive rate of MIF declines as the cancer progresses. Lymphocytes of the lymph nodes extirpated during operation show a similar tendency.
    4. In comparing MIF activity of peripheral blood lymphocytes of gastric cancer patients before operation with that after operation, the activity in Stage II before operation is 41.7% as against 0% after operation, that in Stage III it is 29.0% before operation and 10.5% after operation, showing a marked decrease, while that in Stage IV, in contrast, rises from 15.4% to 23.1%. Usually the MIF activity disappears when tumor removal is completely performed, but in the cases of progressive cancer the MIF activity seems to be sustained.
    5. Looking at the cross reaction, in gastric cancer cases with positive MIF activity for autochthonous antigen they show 42.9% positivity against heterogenous antigen. Besides we have observed the cross reaction between gastric cancer and colon cancer as well as between colon cancer and gastric cancer.
    The foregoing findings indicate that in human cancer just as in animal cancer MIF activity first appears in the regional lymph nodes, which tends to decrease and disappears as the cancer advances. Since there can be observed a marked cross reaction between gastric cancer patients, MIF would serve as a useful criterion for the discovery of gastric cancer and for the determination of prognosis.
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  • the evaluation for normal adults by echocardiography
    Mamoru TAGO
    1977Volume 89Issue 7-8 Pages 877-882
    Published: August 30, 1977
    Released on J-STAGE: March 30, 2009
    JOURNAL FREE ACCESS
    Isometric handgrip exercise were performed to evaluate the left ventricular performances in fifteen normal adults by echocardiography.
    The following parameters increased significantly during isometric handgrip exercise. blood pressure, heart rate, cardiac index, stroke work index, contractile index.
    The normal hearts responded to the increase of afterload during isometric exercise by increased heart rate but the promotion of contractile state was not seen, so it was thought that the wall stress or the wall thickness increased.
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  • Toshitake FUJII
    1977Volume 89Issue 7-8 Pages 883-894
    Published: August 30, 1977
    Released on J-STAGE: March 30, 2009
    JOURNAL FREE ACCESS
    Responding mechanism of regenerating hepatocytes induced by partial hepatectomy was studied.
    1) Fibroblast inhibitory and stimulatory factors on fibroblast proliferation co-exist in the same rat liver supernatant, and they were simply isolated each other by ethanol fractionation.
    2) These factors have protein components and lose the biological activities by heat-treatment, and they show the competitive activity each other, respectively.
    3) In regenerating liver, stimulatory factor(s) was not remarkably changed, but inhibitory one almost lost the inhibitory activity. These phenomena are very rational and suitable for hepatocyte regeneration.
    4) The inhibitory activity was separatedly eluted by DEAE-cellulose chromatography of inactive regenerating liver fraction, and new peak that did not exist in normal inhibitory fraction was detected in polyacrylamid gel electrophoresis of regenerating liver same fraction.
    5) These phenomena suggest that a possible new anti-inhibitory factor(s) would be formed by partial hepatectomy.
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  • Tadaatsu AKAGI
    1977Volume 89Issue 7-8 Pages 895-902
    Published: August 30, 1977
    Released on J-STAGE: March 30, 2009
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    The methods for cultivation of amniotic fluid cells were described.
    Transabdominal amniocenteses were performed for prenatal diagnosis of chromosome abnormalities. Successful cultures for karyotyping were accomplished in 12 of 13 samples obtained from 12 patients.
    In one case karyotypic analysis of cultivated amniotic fluid cells revealed a karyotype of 45, X and then therapeutic abortion was performed. The karyotype of cultivated fetal skin and umbilical cord blood cells was 45, X/45, XX mosaicism. The necropsy of the aborted fetus disclosed pure gonadal dysgenesis.
    Prenatal genetic diagnosis with cultivated amniotic fluid cells is useful for monitoring of high-risk pregnancies.
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  • Toshiyuki HAYABARA, Seiso YABUKI, Hideaki FUKUI, Masaki CHUDA, Reiko N ...
    1977Volume 89Issue 7-8 Pages 903-911
    Published: August 30, 1977
    Released on J-STAGE: March 30, 2009
    JOURNAL FREE ACCESS
    A follow-up study on 60 previously healthy patients with nomosymptomatic optic neuritis, retrobulbar neuritis and idiopathic optic atrophy is reported.
    73 per cent of the cases with insidious onset and slow progression were male, and 70 per cent of the cases who was troubled repeatedly on vision were female. In many cases, fever, common cold and overwork had precipitated the optic neuritis.
    During the following period, 6 patients (15.4% of cases with acute onset) suffered from the other neurological manifestations, and a half of them (7.7%) were diagnosed as multiple sclerosis. Patients, who was diagnosed as retrobulbar neuritis by ophthalmoscopic examination and, who was disturbed on vision bilaterally but unsimultaneously, and repeatedly have an affinity to multiple sclerosis.
    At the beginning of the disease, a mononuclear pleocytosis was noted in 20.5 per cent of the patients, and elevated total protein level in 35.9 per cent. Polyacrylamide gel disc electrophoresis of the lumbar CSF proteins from 39 patients and the measurement of IgG by single radial immunodiffusion from 10 patients, were carried out. It was characteristic that the electropherogram showed the increase of pre-albumin, fast α2-globulin+transferrin and γ-globulin, and the decrease of albumin. These results are similar to those in multiple sclerosis.
    IgG and IgG% is increased in cases of retrobulbar neuritis, although total protein content is high level in optic neuritis, and then, cases who was disturbed on vision unilaterally or bilaterally but unsimultaneously showed the high level of γ-globulin.
    Two of 3 patients who subsequently developed to multiple sclerosis, had the CSF findings indistinguishable from those in multiple sclerosis. In one patient, CSF protein level was normal but γ-globulin was increased selectively. Another patient showed the CSF findings with slightly elevated protein level, pleocytosis and increase of IgG and IgG%.
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  • Toshiyuki HAYABARA
    1977Volume 89Issue 7-8 Pages 913-927
    Published: August 30, 1977
    Released on J-STAGE: March 30, 2009
    JOURNAL FREE ACCESS
    Immunoglobulin E was measured in concentrated cerebrospinal fluid (CSF) of controls and patients with various neurological disorders, including multiple sclerosis (MS). The radioimmunoaasay method was used for IgE and the single radial immunodiffusion method was used for other immunoglobulins.
    IgE in CSF was able to quantify in 86.4% of 118 cases, and then, it is thought that there is quantitative IgE component in normal CSF. In controls (N=23) IgE contents ranged from 0.03 to 0.71 U/ml (M: 0.28 U/ml). IgE/100mg of total protein ratio in CSF ranged from 0.06 to 1.52 (M: 0.61). Then, it is suspected that normal value of IgE and IgE% is below 0.8U/ml and 1.6.
    The CSF/serum ratio of IgE was about 0.1% (0.08-0.12%). This value is larger than that of IgM, and smaller than that of IgA and suggests immunoglobulin permeability through the blood-brain-CSF barrier in reverse proportion to molecular weight.
    In pathological CSF, the increase of IgE was related to the increase of total protein, IgG, IgA and IgM levels but IgD. IgE level was increased in 19 of 84 cases (22.6%), especially high IgE level was seen in acute inflammatory diseases of central nervous system but in chronic disorders (for example dementia paralytica). Otherwise, polyradiculoneuritis and diseases with abnormality of CSF dynamics or brain atrophy, revealed the tendency of high IgE levels. In these disorders, the increase of IgE was parallel with total protein level, IgG and other immunoglobulins in most cases.
    The incidence of high IgE level in MS was 3 of 15 cases, but these increases of IgE were not corelated with abnormalities of other components in CSF and it was revealed the possibility of another pathophysiological process of IgE from IgG and other immunoglobulins in central nervous system. No relationship was present between the incidence of high CSF IgE level and various factors such as clinical stage, suspected lesion and severity of disturbance in MS.
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  • Toshiaki KAGEYAMA
    1977Volume 89Issue 7-8 Pages 929-948
    Published: August 30, 1977
    Released on J-STAGE: March 30, 2009
    JOURNAL FREE ACCESS
    Recent developments in microsurgical techniques have brought about the possibility of restoration of the main branches of the cerebral arterial blood flow resulting from obstructive conditions. These arteries usually indicate pathological alterations.
    However, there are only a few reports on the healing processes of the anastomotic sites on the pathological arterial wall.
    In the present study, the histological and histochemical healing processes were investigated in an anastomotic site of normal and pathological arterial walls.
    Ninety-two normal rats and 96 Goldblatt rats were used.
    End-to-end anastomosis was performed on the left common carotid artery by microsurgical technique.
    The patency rates of anastomosis were 71.6% in normal rats and 73.0% in Goldblatt rats. Patent anastomosed arteries were observed at various post-operative periods, ranging from 24 hours to 12 weeks. For histological studies, hematoxylin-eosin and Van Giesen stainings were used. For histochemical studies, lactic dehydrogenase, glucose-6-phosphate dehydrogenase, succinic dehydrogenase, cytochrome oxydase, acid phosphatase and alkaline phosphatase were examined.
    Goldblatt rats were divided into two groups histologically. One group had a slight degeneration of the medial muscle, and the other group had a marked degeneration. This histological difference between the two groups was probably due to blood pressure. A histological examination revealed a healing stage at four weeks in the former group and the histochemical study showed healing at eight weeks in the former group. There was no significant difference on the healing process between the former group and normal rats.
    Goldblatt rats which showed marked degenerative changes of the medial muscle indicated a poor healing process with granulation in the outer regions of medial necrotic areas. At 12 weeks after anastomosis, histological examination still showed the prominent and widespread degenerative changes on the medial wall. The histochemical examinations of this group revealed few enzymatic activities in the media in areas adjacent to the anastomosis, although rather higher activities were observed in the adventitia in granulation areas adjacent to the anastomotic site. But alkaline phosphatase in the adventitia around the anastomotic site markedly decreased in activity. These decreased activities of the alkaline phosphatase were probably due to destruction of the vasa vasorum.
    These findings suggested that the cause of these prominent and widespread necrotic regions were due to the absence of circulating blood which was stopped by temporarily clipping and destruction of the vasa vasorum during the operation.
    But, normal and slight degenerative arterial walls did not indicate such prominent changes.
    It was suggested from these experimental study that the pathological arterial walls were easily injured in the ischemic condition during the operation.
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  • Tadaatsu AKAGI, Yuzo MATSUDA
    1977Volume 89Issue 7-8 Pages 949-953
    Published: August 30, 1977
    Released on J-STAGE: March 30, 2009
    JOURNAL FREE ACCESS
    Membrane-associated (M-Ig) and cytoplasmic immunoglobulins (C-Ig) of 4 human lymphoblastoid cell lines and 3 clones isolated from one of them were examined by immunofluorescence test. It was found that individual cells produced only a single type of light chain components and one or more types of heavy chain components of Ig. There was no specific correlation between the Ig types of M-Ig and C-Ig.
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  • I) Effect of combination with shallow differential hypothermia and antimetabolites
    Takeo OHASHI
    1977Volume 89Issue 7-8 Pages 955-977
    Published: August 30, 1977
    Released on J-STAGE: March 30, 2009
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    How to treat the malignant brain tumors has been one of the biggest problems in neurosurgery. It has been in the past, and it is still now. In addition to surgical removal of tumors, various non-surgical method have been tried for the treatment of malignant gliomas. The use of heating or of cooling are some of these methods. They have been tried for a long time.
    Already in 1866 Busch, W. observed disappearance of sarcoma in patients suffering from erysipelas. Westermark, N. in 1927 observed that rat transplanted tumors were caused to disappear by exposing to heating, while the adjacent normal tissues were not damaged under conditions lethal to the tumors. In 1960 Woodhall, B. heated tumors locally that were perfused with chemotherapy. Shingleton, W. W., in 1962, heated localized tumor tissue on the patients with cancer, combined with regional chemotherapy under generalized hypothermia.
    According to Popovic, V. P. et'al. in 1965, they were first to report that differential hypothermia, keeping tumors normothermic under total body hypothermia at a temperature of 4°C during a period of 10 hours in experimental animals, induced disappearance of tumor without resuming their growth afterward. Popovic, V. P., et al. in 1966 carried out further experiments and observed that tumors of the animals disappeared also subsequent to cooling whole body to 4°C for 1 hours with anti-tumor agent, as well as to cooling whole body to 30°C for 24 hours without chemotherapy while the tumor kept at 37°C.
    However, in order to induce tumor disappearance the differential hypothermia has to last at least 4°C for 110 hours, or 30°C for 24 hours. Since deep hypothermia lasting several hours or shallow hypothermia for long time is not well tolerated in non-hibernators, namely human beings, present experiment has been performed in an attempt to simulate the conditions of clinical work as close as possible. For this purpose the bodies of 22 hamsters were mildly cooled to a temperature of 30°C, while cheek pouch transplanted tumors induced originally by adenovirus type 12 remained uncooled at 37°C for 10 hours (Fig. 1, 2). The dose of 50 mg/kg of 5-fluorouracil (5-FU) was administered intraperitoneally in single injection at the beginning of treatment of shallow differential hypothermia (Table 1, Fig. 2). This resulted in that within 7 days later 4 out of 22 tumors (20%) disappeared completely without resuming their growth afterward, and 12 out of 22 tumors (55%) regressed temporarily for a period of 10 days after treatment (Table 2, 3, Fig. 6-b, 8). When the same amount of 5-FU was administered into normothermic tumor-bearing animals or into hypothermic animals with hypothermic tumors, tumor size of the animals was not affected (Table 1, Fig. 5-a, 6-a). When shallow differential hypothermia was treated without any anti-tumor agent, neither normothermic tumor size nor hypothermic one of the animals was also affected (Table 1, Fig. 7-a).
    Histological findings were degeneration of tumor cells. Stainability of nucleolar RNA and nuclear DNA by means of methyl green-pyronine and Feulgen's methods was decreased already immediately after the shallow differential hypothermia treatment for 10 hours with 5-FU administration (Fig. 10-a). This was followed by, at 24 hours after the treatment, marked decreasing and loss of the stainability of nucleolar RNA and nuclear DNA as well as dismixture of chromatin and karyolysis of nuclei (Fig. 11-a, b). Scattered necrotic foci in the tumor tissue were obviously revealed 48 hours after the treatment. Tumor cells showed selectively these degenerative finding after treatment, while adjacent mucosal tissue of cheek pouch preserved their normal appearance almost wholly even 12 hours after the treatment (Fig. 10-c).
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  • II) A study of adenovirus type 12-induced T-antigen by immunofluorescence technique
    Takeo OHASHI
    1977Volume 89Issue 7-8 Pages 979-994
    Published: August 30, 1977
    Released on J-STAGE: March 30, 2009
    JOURNAL FREE ACCESS
    T-antigens of transplanted hamster tumors induced by human adenovirus type 12 were studied by fluorescent antibody technique within 24 hours after differential hypothermia (DH) treatment, in which one side cheek pouch tumor was warmed at 37±1°C under generalized hypothermia of the body and the other side tumor at 20±1°C for 10 hours.
    A high proportion of the tumor cells in the viable portions of the control tumors demonstrated specific fluorescent staining in three type. The most striking and consistent pattern was the presence of numerous fluorescent particles in the cytoplasm, which were granular and fleck-like shapes. A second type of fluorescence was fluorescent particles in the nucleus in addition to the cytoplasmic staining, and a third type was homogenous staining of nucleus.
    Immediately after DH treatment, the treated warmed tumor cells presented diminution of fluorescent staining, especially large numbers of granular fluorescences in the cytoplasms. Fleck-like and homogenous nuclear staining of the treated tumor cells scattered even at 10 and 15 hours after DH treatment, when treated tumor cells revealed necrobiotic findings in sections of HE and nucleic acid stain. But no fluorescence was observed at 20 and 24 hours after DH treatment, while not-treated hypothermic tumor cells demonstrated about same fluorescent staining as control tumor cells.
    It was suggested that cytoplasmic granular fluorescence diminished because DH treatment had an effect on thermo-sensitivity of T-antigen because DH treatment had an effect on thermo-sensitivity of T-antigen and/or inhibited T-antigen production, as well as loss of fleck-like and hemogenous nuclear fluorescence was due to tumor cell necrosis by DH treatment.
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  • Susumu OHARA
    1977Volume 89Issue 7-8 Pages 995-1019
    Published: August 30, 1977
    Released on J-STAGE: March 30, 2009
    JOURNAL FREE ACCESS
    198 male albino rats were used in this experiment. First, 42 rats were divided into 13 different dietary groups to test the effects of high fat and cholesterol in the diet on the level of serum cholesterol and the degree of atherogenesis. Then the effects of four different types of diet upon the healing of small vessel anastomoses were studied. Group one received a regular control diet (MF diet, Oriental KOBO Co. Ltd.), group two, A diet (regular diet with cholesterol 5%, peanut oil 5% and sodium cholate 2%), group three, AP diet (A diet with pyridinolcarbamate 0.05%), group four, AT diet (A diet with thiouracil 0.3%). All percentages were calculated on a weight basis. Each group received the above diets for 13 weeks before anastomosis. Then, end to end anastomosis of the common carotid artery was performed using the operating microscope. Rats were sacrificed in groups of 2 to 6 from 24 hours to 20 weeks postoperatively. Vessel patency was determined and histological and histochemical changes were noted.
    Average values of the level of s-cholesterol at the time of sacrifice were 80 mg/dl in the normal group, 484 mg/dl in the A group, 342 mg/dl in the AP group and 661 mg/dl in the AT group and patency rates were 23/30 (77%), 16/36 (44%), 17/32 (53%) and 3/8 (38%) respectively.
    Histologically, diffuse degeneration of smooth muscle cells in the media and marked deposition of fat granules in the intima and media were observed in the hypercholesteremic rats of groups A and AT. In the AT group mural thrombi were often observed around the site of anastomosis with narrowing of the lumen. In rats fed atherogenic diets, healing of the anastomosis was significantly delayed requiring 6 to 8 weeks as determined by histological evidence and about 12 weeks by histochemical. Some degree of degeneration with fat granules was noted in the walls of the vessels at the sites where the vascular clamps had been applied.
    Histochemical examination of anastomosed vessels from rats fed atherogenic diets A and AT showed markedly lower activities of LDH, G-6-PDH, SDH and Cy-O in the media and rather higher activities in the adventitia than in vessels from rats fed control diets. In all groups Ac-P activity increased in the adventitia following surgery then decreased gradually with the healing process and Al-P activity increased temporarily in granulation tissue. But the activity of this latter enzyme was found to be markedly decreased in the adventitia following surgery in the vessels of rats fed atherogenic diets and seemed to be correlated with the degree of medial necrosis.
    Patency rates were very low in rats fed the A, AP and AT diets as compared to those fed the control diet, there being a clear tendency for higher cholesterol levels to result in lower patency rates. It would appear that persistent hypercholesterolemia increases the risk of delayed occlusion at the anastomotic site caused by mural thrombi or atheromatous change. The rats in the AP group showed less prominent fat infiltration of the vessel wall and fewer mural thrombi than those of the A or AT groups. This work suggests that it would be advantageous to administer anti-coagulant and/or anti-atherogenic medication after vascular anastomosis and to maintain that therapy throughout the duration of the healing process.
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  • Part 1. Fundamental studies on identification of lymphocyte subpopulations and their application to lymphoid organs
    Kenji KORIYAMA
    1977Volume 89Issue 7-8 Pages 1021-1033
    Published: August 30, 1977
    Released on J-STAGE: March 30, 2009
    JOURNAL FREE ACCESS
    Thymus-derived lymphocytes (T-cells) and bone marrow-derived lymphocytes (B-cells) in the cryosection of human tissues were identified either by rosette formation with sheep erythrocytes (E) or with erythrocytes sensitized with antibody and complement (EAC), respectively. Invariable and stable reaction was obtained in EAC rosettes of the tissue lymphocytes while E binding of them was rather unstable. E could be adhered in the thymus sections which were obtained from normal infants during open heart surgery, but EAC did not bind to them at all.
    It was confirmed that B-cells located mostly in the lymph follicles of the lymph nodes as well as the palatine tonsils and T-cells were present predominantly in the paracortical area and interfollicular region. B-cells in the germinal centers would generally have more activated C3 receptors than those of the mantle zones. In addition, B-cells of the lymph follicles both in the lymph nodes and in the palatine tonsils could be also detected utilizing the membrane immunofluorescence technique. The proportion of T- and B-cells distribution in the lymphoid tissue was well correlated with that obtained on free cell suspension technique.
    Therefore, it could be concluded that T and B lymphocytes were able to identify respectively even in tissue level.
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  • Part 2. Identification of lymphocyte subpopulations infiltrating in autoimmune diseases
    Kenji KORIYAMA
    1977Volume 89Issue 7-8 Pages 1035-1047
    Published: August 30, 1977
    Released on J-STAGE: March 30, 2009
    JOURNAL FREE ACCESS
    The lymphocyte subpopulations were detected in the target organs obtained from various autoimmune disorders on the tissue cryosections. Thymus-derived lymphocytes (T cells) and bone marrow-derived lymphocytes (B cells) were detected by rosette formation with sheep erythrocytes (E) or with sheep erythrocytes coated with antibody and complement (EAC) respectively. Almost all infiltrating lymphocytes in the salivery glands in patients with Sjogren's syndrome were identified as B cells and T cells were detected only along the ducts. In Hashimoto's thyroiditis, infiltrating lymphocytes with the lymph follicles in the thyroid gland were mostly B cells whereas T cells localized as surrounded the thyroid follicles. T cells would not be detected in the spleen sections obtained from the patients with hypoplastic anemia at all. The white pulp was constituted by B cells, and the mononuclear cells in the red pulp might be macrophages of which these cell surfaces had IgG-Fc receptors.
    The lymph follicles of the thymus in a patient with myasthenia gravis complicated with thymoma was demonstrated to be formed with B cells.
    Therefore, it could be concluded that B cells were predominant lymphocytes in the target organs, while T cells localized in situ developing the autoimmune reactions in various autoimmune diseases.
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