生物物理化学
Online ISSN : 1349-9785
Print ISSN : 0031-9082
ISSN-L : 0031-9082
9 巻, 3-4 号
選択された号の論文の7件中1~7を表示しています
  • 〔I〕実験的研究(Goldsol反応を中心として)
    石原 幸夫
    1963 年 9 巻 3-4 号 p. 175-193
    発行日: 1963/07/15
    公開日: 2009/03/31
    ジャーナル フリー
    Es scheint mir ein interessantes Problem, was für einen Zusammenhang zwischen einer neuen Untersuchungsmethode, d. h. bisher üblichen Methoden der Liquordiagnostik gefunden werden. Ich habe darauf von der “starch-column-zone elektrophoresis” das Serumeiweiss in die Eiweissfraktionen, wie Albumin, α1-, α2-, β- und γ-Globulin u. s. w., rein aufzutrennen probiert, so dass ich die Beziehung zwischen diesen elektrophoretisch auseinander aufgetrennten Eiweissfraktionen und der Goldsolreaktion experimentär analysiert.
    Die Resultate der Untersuchung sind unten kurzgefasst dargestellt.
    1. Die reine Albuminlösung gibt dem Goldsol keine Veränderung, d. h. es tritt keine Goldsolausfällung ein. Jede einzelne reine Globulinfraktionslösung wirkt flockend auf das Goldsol, d. h. sie zeigt verschiedene Wirkung, wie das Goldsol einmal schwach, andersmal stark zu fällen. Vor allem zeigt die γ-Globulinfraktion dieselbe Wirkung am stärksten. Nämlich findet man in der γ- Fraktion eine starke Fällungskraft, die bei der Goldsolreaktion eine typische Goldoslkurve des Paralysetyps hervorrufen kann. Und zwar zeigt uns die Zunahme der Fallungskräft in der Golreaktion eine parallele Neigung mit dem Aufsteigen jedes Globulinfraktionsgehalt, insbesondere ist dieses Verhältnis bei der γ-Fraktion am deutlihtsen.
    2. Ich untersuchte die Goldsolreaktion mit denjenigen Liquoren, die im normalen Liquorr einzelne Eiweissfraktion gelöst worden sind, was damit jede künstlich vermehrte Eiweissfraktion hat. Der Albuminfraktionsvermehrte Normalliquor zeigt dabei auf dem Goldsolreaktion keine Veränderung. d. h. seine Goldsolkurve ist immer von Normaltyp. Der α2- und der β-fraktionsvermehrte Normalliquor bewirken auf das Goldsol jeder für sich wie bei der reinen α2- und β-Fraktionslösung, um es zu verfärben und ausfällen zu lassen. Der γ-Fraktionsvermehrte Normalliquor reagiert mit dem Goldsol sehr empfindlich, so dass er den Normaltyp von Goldsolkurve in die typisch, so dass er den Normaltyp von Goldsolkurve in die typische Paralysekurve wechselt.
    3. Ich wendete dann die oben erwähnten Methoden auf den Liquor von Paralytikern an und unteruchte die Wirkungen der Eiweissfraktionen in diesem pathologischen Liquors. Der Goldsolfällungstyp des Paralyseliquors, der anfangs einen typischen Paralysetyp gezeigt hat, verändert sich mit der Vermehrung der Albuminfraktion sehr auffallend. D. h. die Ausfällung des Goldsols wird stark gehemmt, und sein Kurventyp bei Goldsolreaktion geht im Normaltyp über. Die Ursache dieser Erscheinung liegt in der kolloidalen Schutzwirkung des Albumins. Beim α2- und β2-Fraktionsvermehrten Paralyseliquor findet man, anders als die Vermehrung im Normalliquor, vielmehr eine kolloidale Schutzwirkung von leichtem Grad. Die Vermehrung der γ-Fraktion bewirkt noch kraftig über Intensität und Ausmass der goldsolfallung, so zeigt sie eine starke Kolloidfällungswirkung.
    4. Betreffs der Globulinfraktion-Albumin-Verhältnis und Goldsolreaktion, wirkt die Wechselung des Eiweissquotients, d, h. die Zunahme jeder Globuminfraktion überwiegt. Vor allem hemmt immer die Vermehrung der γ-Globulinfraktion eine Schutzwirkung des Albumins und zeigt eine starke Kolloidausfällungswirkung.
  • 〔II〕臨床的研究(進行麻痺髓液を中心として)
    石原 幸夫
    1963 年 9 巻 3-4 号 p. 195-219
    発行日: 1963/07/15
    公開日: 2009/03/31
    ジャーナル フリー
    Ich babe vorher über den Zusammenhang der elektrophoretisch aufgetrennten Eiweissfraktion und Kolloidreaktion, nämlich die Beziehung mit ihren Goldsolreaktion experimentär analysiert und berichtet. In dieser Abhandlung werde ich von den Totalzahlen der 109 paralytikern ihre Liquoreiweiss papierelektrophoretisch fraktionieren, und die aus der klinischen Praxis sich ergehenen Papierelektropherogramme mit den Goldsolreaktionen miteinander vergleichen, dann der Zusammenhang der Liquoreiweissfraktionen mit den Goldsolreaktionen klinisch analysieren.
    Die kurzgefassten Ergebnisse sind wie folgend.
    1. Man findet in demjenigen Liquor des progressiven Paralytikers, der durch antiluetische Behandlung noch nicht behandelt wurde, trotzdem der Befund einer luetischen Entzündung darin deutlich erkennbar ist, in beträchtlicher Anzahl die hohe γ-Globulin Vermehrung, und die Goldsolreaktion solcher Liquor zeigt die typische Paralysekurve. Bei diesen Fallen findet man nach der Therapie eine erhebliche Absinken der vermehrten Liquor-γ-Globulin sowie ein Abnehmen der Goldsolfällungsknaft bei Goldsolreaktion, deren typische Paralyseform der Goldsolkurve verändert sich ziemlich deutlich und verschob sich normalwärts. Anders zu sagen, gibt es einen deutlichen Zusammenhang zwischen dem Lipuor-γ-Globulinwert und der Goldsolreaktion.
  • I 濾紙面上の緩衝液流およびイオン強度の泳動像に及ぼす影響
    山口 延男
    1963 年 9 巻 3-4 号 p. 221-231
    発行日: 1963/07/15
    公開日: 2009/03/31
    ジャーナル フリー
    A systematic studies of the electrophoresis technique of human serum proteins (in sodium barbiturate buffer at pH 8.6) using the open horizontal method ware carried out.
    1. During electrophoresis, two kinds of the buffer flow occur on paper surface, one of which heads for the cathode on account of the electroendosmosis. The other one goes to the central part of paper from both of the anodic and cathodic buffer vessels in order to compensate for the relative dryness which occurs owing to the evaporation during electrophoresis. The apparent buffer flow on paper surface is the algebraic sums of these two kinds of buffer flow. The equlibrum points (zero points) of buffer flow are generally located on the cathodic part of the horizontal portion of paper.
    2. These equlibrium points, however, displace to the central part of paper as voltage, ionic strength and electrophoresis hour incerease. It is because, in these cases, the buffer flow due to the evaporation augments more rapidly than the one due to the electroendosmosis.
    3. The buffer flow on the more anodic part than the equlibrium points heads for the cathod and inversely on the more cathodic part. Their dimensions are proportional to the distances from the equlibrium points of buffer flow.
    4. On account of the buffer flow on paper, 1) the electrophoretic patterns are blurred, especially on the anodic part of paper, and displaced, as a whole, to the anodic or cathodic parts, leaving some extent of tailing on paper. This is disadvantageous for the precise determination of serum fractions because one fraction may be superimposed on the other's talings.
    5. As the buffer solution displaces under the continuous evaporation during electrophoresis, the ionic strength varies throughout on paper. To detect the variations of ionic strength, accumulated amounts of electrolytes on the dried papers, were examined by the high-frequency papyrography The ionic strength is proved to be small where the buffer flow is large. So, the ionic strength on paper decreases in proportion to the increases of distance from the equlibrium points of buffer flow.
    6. The time-migration relationship of B. P. B, stained Alb, is not linear but of inversely S-shaped curve, the formation of which is expounded to be due to the variations of ionic strength on paper.
    7. The most well separated electrophoretic patterns are obtained when serum is applied on the equlibrium points of buffer flow. It is because the applied serum migrates on the paper with relatively high ionic strength and relatively small buffer flow.
    8. In drying papers, the electrophoretically separated fractions are displaced owing to the water displacement due to the evaporation.
    9. The paper electrophoretic mobilities are only approximate in both of the closed and open method electrophoresis because the changes of buffer flow and ionic strength during electrophoresis, and the water displacement in drying papers do not take place uniformly on paper.
  • II 電気泳動像の分離と実験条件との関係について,ことに泳動像のコントロールの問題
    山口 延男
    1963 年 9 巻 3-4 号 p. 232-242
    発行日: 1963/07/15
    公開日: 2009/03/31
    ジャーナル フリー
    1. To make the voltage between both ends of the horizontal portion of paper as equal as possible in each experiment, it is necessary to keep the tensions of paper constant and to maintain the distances from the surface of buffer solution to the horizontal portion of paper constant, too.
    2. After electrophoresis, pH-values of barbiturate buffer solution deviate from the original pH, to some extent, in both of the anode and cathode containing vessels. They deviate in the paper ends containing vessels only when the applied voltage is relatively high or/and electrophoresis hour is extremely long. pH-Values of total buffer solutions, however, have little change if they are mixed and dissolved well after electrophoresis. Therefore, the same buffer solution can be used three or four times repeatedly, although some changes of ionic strength and some contaminations with paper fibres and bacilli may occur.
    3. At higher voltages, 1) the migration velocities are large, but the electrophoretic patterns are worse separated, especiaily when electrophoresis hour is short. 2) only in long electrophoresis hours, they are well separated and less blurred in their zones because of the enhancement of ionic strength on paper. 3) the length of Alb developped from γ-gl is short because both of the migration velocities and the buffer flow augment.
    4. When the ionic strength of buffer solution is stronger, 1) the migration velocities are small, 2) the electrophoretic patterns are well separated and their zones are less blurred because the zonal diffusion is restricted by the augmented ion transportation. 3) α1-gl is well separated from Alb. 4) globulins, especially, γ-gl displace to the more acidic part on paper if the migration distances of BPB stained Alb is fixed to a definite from the application point.
    5. The migration distance of BPB stained Alb from the application point is a good index for checking the electrophoretic separation of serum protein, because the electrophoresis hour required for the good separation is not definite but dependent on the experimental temperature and so on.
    6. In the author's laboratory, serum protein is submitted to the electrophoretic run usually at the constant voltage of 200V, using sodium barbiturate buffer of pH 8.6 and ionic strength of 0.075. When serum volumes of 0.01ml/2cm (width of a paper strip) or less are applied, BPB stained Alb is allowed to migrate 8cm from the application points. When serum volumes of 0.02ml or more, BPB stained Alb is allowed to 12cm, likewise. If the electrophoretic patterns are poorly separated and blurred, for example, owing to therelative low temperature, voltage and electrophpresis hour (expressed as the migration distance of BPB stained Al), for the first choice, and nextly the ionic strength are adjusted so as to ameliorate the results described in 3 and 4.
  • III 濾紙緩衝液系の変化と電気泳動像との関係,ならびに若干の技術的考案について
    山口 延男
    1963 年 9 巻 3-4 号 p. 243-251
    発行日: 1963/07/15
    公開日: 2009/03/31
    ジャーナル フリー
    1. For the precision of densitometry, each zone of serum fractions should be geometrically so figured as to be perpendicular to the direction of electromigration on its cathodic and anodic edges and to spread bilaterally just to the both edges of a paper strip. The electrophoretic patterns suiting this request will be got when the serum is applied as a rectangularly diffused zone at the starting line. The repeated streaking method proposed by Grassmann and Hannig, and the droplets depositing method are proved to be preferable in applying the serum on paper so far as the thorough attention to avoid the disfiguring effect of the first and last droplet running out from micropipett is paid. The author, in addition, devised a special micropipett which is fitted to push out a minute amout of serum by the pressure differences of fingers and to “handwrite” a uniform streak of serum on wet paper.
    2. On the paper, the migration velocities of serum fractions enhance as the water content of paper (g/cm2) increase. The increases of the tension lessen the water content of paper, subsequently leading to the slow migration velocities.
    3. The distortion and disfigure of a electrophoretic pattern on a paper strip are mainly due to the differences in tension.
    4. When several isolated paper strips are mounted in the same electrophoresis apparatus, the migration distance of each serum fraction on them are not generally equal, owing to the minute differences of tension among these paper strips.
    5. A sheet paper with slits as shown in Fig. 6 is newly designed for paper electrophoretic use.
    6. A new apparatus was devised for the facilitation of mounting a paper with slits and making the tensions of paper strips as equal as possible.
    7. As each fraction on paper strips on a paper with slits has almost the same migration distance, one of these strips can be used as a guide strip for checking the eletrophoretic separation as well as for cutting off the fractions from the strips for preparative experiments. The electrophoresis using a paper with slits is especially suitable for the preparation of serum fractions. This method is named the improved preparative electrophoresis by paper srips (Yamaguchi).
  • 山口 延男
    1963 年 9 巻 3-4 号 p. 253-265
    発行日: 1963/07/15
    公開日: 2009/03/31
    ジャーナル フリー
    Paper electrophoretic studies of serum protein iu the acidic Mc Ilvaine's buffer (pH 2.4 to 6.0) were carried out, using the open horizontal method. Further, the reproducible paper electrophoresis technique at pH 4.4 and the subsequent quantitative methods were reported.
    The following results were obtained.
    1. The acidic protein in serum, named M, is separated only between pH 4.8 and 4.0. At pH 4.8 and above, serum protein is separated into three fractions, namely, A, B1 and B2; no acidic protein is observed. At pH 3.6 and beneath, , serum protein is not separated in any fraction, and distributed only as a long and diffuse tailing on the cathodic part of paper.
    2. Serum fractions at pH 4.4 are named M1, M2, A and B from the anodic components. B, having no electric charge at pH 4.4, is displaced only by the buffer flow. M1, M2 and A migrate to the anode with the apparent mobilities in this order, but the migration of A is very small. B alone migrates to the cathode.
    3. Under the constant voltage of 200V, the electrophoretic separation at pH 4.4 is markedly influenced by the ionic strength of buffer solution and electrophoresis hour, namely, 1) with lower ionic strength, M1, M2, A and B are so far well resolved but their zones are diffuse and blurred, and 2) with much higher ionic strength, and also in too longer electrophoresis hour, they come together on the central part of paper because the buffer flow and the ionic strength on paper are exceedingly large.
    4. Serum should be applied at the places where A remains immobile after electrophoresis. It is because B fuses with A, when the applied volumes of serum are relatively large, and because the fast moving M1 and M2 are flowed back owing to the buffer displacement and superimposed on the other fractions, if the application points are on the more anodic part than the above mentioned places.
    5. In the author's laboratory, electrophoretic runs are routinely carried out at 200V for 14-16 hours, using Mc Ilvaine's buffer of pH 4.4 and 0.2 which contains 3.29g of citric acid (1 hydrate) and 10.53g of sodium diphosphate (12 hydrates) in a litre of destilled water.
    6. By amido black staining, M1 is stained only faintly, M2 weakly, A and B exceedingly densely. By PAS-staining, M1 and M2 are densely stained only in cases with increased mucoproteins. A and B, in general, are faint and diffuse. Therefore. these staining methods are inadequate for the quantitative densitometry of these fractions.
    7. Sera from some representative dysproteinemias including liver cirrhosis, gastric cancer, acute myelocytic leucemia, nephrotic syndrome and exudative pleuritis were run at pH 4.4 and fractionated. Subsequently, serum fractions and their protein-bound hexoses were chemically determined and compar ed.
    8. The improved preparative electrophoresis by paper strips (Yamaguchi) is proved to be valuable for the clinical observation of serum pH 4.4 as well as for the further investigation of M1 and M2 fractions.
  • 1963 年 9 巻 3-4 号 p. 268-289
    発行日: 1963/07/15
    公開日: 2009/03/31
    ジャーナル フリー
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