生物物理化学
Online ISSN : 1349-9785
Print ISSN : 0031-9082
ISSN-L : 0031-9082
17 巻, 2 号
選択された号の論文の6件中1~6を表示しています
  • 城石 和子, 田中 英子, 石本 美穂子, 久保田 憲太郎
    1973 年 17 巻 2 号 p. 49-54
    発行日: 1973/01/30
    公開日: 2009/03/31
    ジャーナル フリー
    A characteristic kidney disorder is commonly seen in Itai-itai disease in addition to the severe bone damage. Hence, urinary proteins of Itai-itai disease patients were analyzed by disc electrophoresis in order to diagnose this disease early and to distinguish it from other kidney diseases.
    The results are summarized as follows.
    1) Five different bands, designated as B-1 to 5 according to the decreasing order of the anodic mobility, were commonly detected in urinary proteins of Itai-itai disease. This pattern was specific for the disease and was provisionally named as I (Itai-itai disease) type.
    2) Three different bands, one corresponding in mobility to serum albumin and two other slower migrating bands, were seen in urinary proteins in other kidney diseases, while the urinary protein from healthy adults was composed solely of albumin. These patterns were provisionally named as K (kidney disease) and N (normal) type respectively.
    3) Among the five bands seen in the I-type pattern, B-3 and 4 specifically appeared in Itai-itai disease and seemed to be important in distinguishing I-type from K or N-type.
    4) Six protein fractions, prealbumin, albumin, α1, α2, β, and γ-globulins were prepared by cellulose acetate electrophoresis from a Itai-itai disease patient's urine and were utilized in disc electrophoresis to identify the above described five bands. It was revealed that B-1 was composed of α1-globulin, B-2 of albumin, B-3 of α1 and α2-globulins, and B-4 and B-5 of α2 and β-globulins, respectively.
    These results indicate that the disc electrophoretic analysis of urinary proteins is a useful screening test in the early diagnosis of Itai-itai disease. It also provides a clear distinction of this disease from other kidney diseases.
  • 高柳 尹立, 岩城 護
    1973 年 17 巻 2 号 p. 55-62
    発行日: 1973/01/30
    公開日: 2009/03/31
    ジャーナル フリー
    This investigation has been performed to detect the individual variances in protein components of the ascitic or pleural fluids obtained from 151 cases of various disorders.
    Quantitative determination of fibrinogen, α1-acid glycoprotein and IgG in the fluids was made by the single radial immunodiffusion method. Fibrinogen and its degradation product increased in parallel with the proliferation of cancer cells in the serous cavities, on the other hand, the increase of α1-acid glycoprotein reflected the histopathological classification of cancers.
    The immunoelectrophoretic analysis performed with the specific anti-human fibrinogen antiserum demonstrated the increase of fibrinogen degradation products in the ascitic and pleural fluids. The reaction patterns of fibrinogen and its derivatives were classified into five types and their relation to the morphological findings of cellular components was investigated. The majority of cancerous fluids was involved in Type 4 and 5 which revealed two distinct lines closer to the cathode. The cancer cell-negative transudates belonged to Type 1 or 2, forming very weak patterns and most of inflammatory effusions to Type 3.
    Fibrinolytic activity determined by the fibrin plate method showed a marked variety in each fluid. Based on the comparison of cytological pictures in active and in negative cases of fibrinolysis, it was suggested that the proliferation of macrophages correlated with the increase of fibrinolytic activity.
    It may be assumed that the immunological analysis and estimation of several protein components in the ascitic and pleural fluids are of help to the differentiation of cancerous and other disorders.
  • 荻田 善一, 佐子山 豈彦, 千葉 泰人, 益沢 学
    1973 年 17 巻 2 号 p. 63-70
    発行日: 1973/01/30
    公開日: 2009/03/31
    ジャーナル フリー
    薄層ポリアクリルアミドゲル電気泳動法を用いた血清蛋白成分の泳動分離における解像力はセルロースアセテート電気泳動法に比べはるかに優れている.しかしながら操作法の簡便さという点ではセルロースアセテート電清気泳動法に劣る.
    著者らはポリアクリルアミドゲルを支持媒質とする簡易薄層電気泳動法を考案し,血清酵素蛋白成分について泳動分離条件を検討してきた.操作法の簡便化という目的のために片面がスリガラス様の粗面をもった製図用のポリエステルフィルムの上に予じめ目的に応じた緩衝液を含んだアクリルアミドゲル薄層(1mm又は2mm)を形成させて湿室に保存しておく.このようにして,薄層アクリルアミドゲルの調製操作を省くことができる.
    支持シートとして,セロファン(124PDセロファン)又は片面がスリガラス様の粗面をもった製図用ポリエステルフィルムを用いた.
    試料として,血清は希釈することなくゲル薄層表面に予じめ作られた溝に添加した.1mm厚さのゲル薄層には3μl,2mm厚さのゲル薄層には15μlの血清を添加する.
    電極液槽用緩衝液を満した両液槽間に試料を添加したゲル薄層を置き,ゲル薄層の両端を緩衝液で湿らせた濾紙の1端を5mm程度重ねるようにのせ電極液槽との電気的連絡を行なう.
    温度の上昇を防ぐために電気泳動は5℃~10℃の保冷庫内で行なった.電気泳動はゲル幅1cm当り0.5~1.0mAで,120分行なった.
    泳動終了後,染色,脱色,固定したゲル薄層はガラス板上でセロファンシートの間にサンドイッチ状にはさんで室温で約2日間乾燥させ,乾燥ゲルフィルムとして保存する.
    また,この簡易薄層ポリアクリルアミドゲル電気泳動法はSDSを含んだゲル薄層を用いることによって,ペプチドの分子量を簡単に,しかも正確に推定できる有利な方法である.
  • 益沢 学, 鎌田 武信, 佐子山 豈彦, 千葉 泰人, 荻田 善一
    1973 年 17 巻 2 号 p. 71-76
    発行日: 1973/01/30
    公開日: 2009/03/31
    ジャーナル フリー
    Using a simple thin layer polyacrylamide gel electrophoresis, human serum alkaline phosphatase could be separated into up to 5 isozyme bands; 2 or 3 isozyme bands in normal adult's sera and 5 bands in patient's sera. They were numbered as Alp I, II, III, IV and 0 in order of decreasing anodic mobility. It was found that Alp I to IV corresponded in mobility to the major component of the respective tissue extracts, i, e, Alp I to hepatobiliary, Alp II to bone, Alp III to placental and Alp IV to intestinal alkaline phosphatase. Alp 0 was the activity remaining at the origin and was supposed to be polymers or complexes located on large particles, such as cell debris, etc.
    Alp III from pregnant sera could be devided into 3 subgroups, i. e., fast (Alp IIIF), intermediate (Alp IIII) and slow (Alp IIIS), according to its mobility.
    The activity of Alp I, sometimes with that of Alp IV, rose mainly in hepatobiliary disease, while activities of Alp II rose in bone and parathyroidal disease. Increase in activity of Alp III and 0 occurred mainly in pregnancy and malignant disease, respectively. The changes of alkaline phosphatase zymograms in pathological states gave us the key to differential diagnosis.
    It was shown that the appearance of Alp IV in normal adult's serum was related to the blood group B or O with salivary secretor type.
    Thus, the electrophoretic separation of alkaline phosphatase of human serum by using the simple thin layer polyacrylamide gel plate as the supporting medium was shown to give us many useful informations about clinical diagnosis. Treatments of serum before electrophoresis, such as diluting, freezing and thawing, heating and adding surface active agents, enzymes, lectins, etc., gave much more informations for the identification of alkaline phosphatase isozymes.
    It was indicated from these data that the migration of alkaline phosphatase isozyme bands was strongly affected by the structure of their end polysaccharide chains.
  • Bence-Jones蛋白尿の熱変性に関する超遠心分析による検討
    只野 寿太郎
    1973 年 17 巻 2 号 p. 77-80
    発行日: 1973/01/30
    公開日: 2009/03/31
    ジャーナル フリー
    Although Bence-Jones proteins are characterized by the property of precipitating on heating at 44-60°C and of redissolving on boiling, the factors governing this unusual solubility behavior have never been defined quantitatively, nor has the phenomenon been satisfactorily explained.
    Measurements of the property of precipitating phenomena at high temperatures have been performed by Putnam with various optical instruments.
    An ultracentrifugal study of the molecular size and shape changes occurring in Bence-Jones proteins in the temperatures ranging 30-100°C has become possible only with a reliable high temperature accessory to the analytical ultracentrifuge. This report describes the ultracentrifugal analysis of heat denaturations of Bence-Jones protein in solution at 30-100°C. The results obtained are as follows.
    1) The Bence-Jones protein forms unstable rapidly sedimenting boundaries above a critical temperature of 60°C, and the sedimentation coefficient could not be obtained.
    2) Molecular aggregation occurred when the solution of Bence-Jones protein was heated above 60°C and various types of aggregation products were suspected.
    3) The method was found to be not satisfactory for the study of heat denaturation of the urmary Bence-Jones protein in this temperature range.
    4) The technical problems associated with this study were discusssed.
  • 1973 年 17 巻 2 号 p. 81-131
    発行日: 1973/01/30
    公開日: 2009/03/31
    ジャーナル フリー
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