CHANGES OF APPARENT IONIC MOBILITIES IN PROTOPLASM : III. SOME EFFECTS OF GUAIACOL ON HALICYSTIS

Lowering the pH of sea water from 8.2 to 6.4 lowers the positive P.D. of Halicystis reversibly (this does not happen with Valonia). Exposure to sea water at pH 6.4 does not affect the apparent mobility of Na⁺ or of K⁺ (this agrees with Valonia). Guaiacol makes the P.D. of Halicystis less positive (in Valonia it has the opposite effect). Exposure to guaiacol does not reverse the effect of KCl in Halicystis which in this respect differs from Valonia. The P.D. can be changed from 66 mv. positive to 23 mv. negative by the combined action of KCl and guaiacol. Exposure to guaiacol affects Halicystis and Valonia similarly in respect to their behavior with dilute sea water. Normally the dilute sea water makes the P.D. more negative but after sufficient exposure to guaiacol dilute sea water either produces no change in P.D. or makes it more positive. In the latter case we may assume that the apparent mobility of Na⁺ has become greater than that of Cl^- as the result of the action of guaiacol. (Normally the apparent mobility of Cl^- is greater than that of Na⁺.) In Halicystis, as in Valonia and in Nitella, an organic substance can greatly change the apparent mobilities of certain inorganic ions (K⁺ or Na⁺).     

CHANGES OF APPARENT IONIC MOBILITIES IN PROTOPLASM : IV. INFLUENCE OF GUAIACOL ON THE EFFECTS OF SODIUM AND POTASSIUM IN NITELLA

In Nitella, as in Halicystis, guaiacol increases the mobility of Na⁺ in the outer protoplasmic surface but leaves the mobility of K⁺ unaffected. This differs from the situation in Valonia where the mobility of Na⁺ is increased and that of K⁺ is decreased. The partition coefficient of Na⁺ in the outer protoplasmic surface is increased and that of K⁺ left unchanged. Recovery after the action current is delayed in the presence of guaiacol and the action curves are "square topped."     

CHANGES OF APPARENT IONIC MOBILITIES IN PROTOPLASM : II. THE ACTION OF GUAIACOL AS AFFECTED BY pH

The normal P.D. across the protoplasm of Valonia macrophysa is about 10 mv. negative (inwardly directed). On adding 0.01 M guaiacol to the sea water the P.D. becomes positive and then slowly returns approximately to the normal value. In many cases this behavior is not much affected by raising the pH and so increasing the concentration of the guaiacol ion but in other cases such an increase makes the P.D. somewhat more negative. But if we wait until the exposure to guaiacol has lasted 5 minutes (and the P.D. has returned to its normal value) before we raise the pH, the result is very different. The cell then behaves as though it had been sensitized to the action of the guaiacol ion which appears to be far more effective than undissociated guaiacol in making the P.D. more positive. This may be due in part to the high apparent mobility of the guaiacol ion and in part to alterations which it produces in the protoplasm (such alterations increase the P.D. across the protoplasm whereas ordinary injury would be expected to lower it and the cells live on after this treatment and show no signs of injury). This action of the guaiacol ion is in marked contrast to the behavior of other anions whose effect resembles that of Cl^-.     

ON GUAIACOL SOLUTIONS : I. THE ELECTRICAL CONDUCTIVITY OF SODIUM AND POTASSIUM GUAIACOLATES IN GUAIACOL

1. Measurements on the densities, viscosities, dielectric constants, and specific conductances of pure anhydrous and water-saturated guaiacol at 25°C. are reported. 2. The solubility of water in guaiacol at 25°C., and its effect on the electrical conductivity of a sodium guaiacolate solution is given. 3. Electrical conductivity measurements are reported on solutions of sodium and potassium guaiacolates in water-saturated guaiacol at 25°C. 4. The decrease of electrical conductivity with increasing concentration for these salts is explained on the basis of an ionic equilibrium combined with the interionic attraction theory of Debye and Hückel. 5. The limiting equivalent conductances of sodium and potassium guaiacolates in water-saturated guaiacol at 25°C., the corresponding limiting ionic mobilities, and the dissociation constants are computed from the conductivity measurements. The salts are found to be weak electrolytes with dissociation constants of the order of 5 x 10^–6.     

THE EFFECTS OF CURRENT FLOW ON BIOELECTRIC POTENTIAL : I. VALONIA

The effect of direct current flow upon the potential difference across the protoplasm of impaled Valonia cells was studied. Current density and direction were controlled in a bridge which balanced the ohmic resistances, leaving the changes (increase, decrease, or reversal) of the small, normally negative, bioelectric potential to be recorded continuously, before, during, and after current flow, with a string galvanometer connected into a vacuum tube detector circuit. Two chief states of response were distinguished: State A.—Regular polarization, which begins to build up the instant current starts to flow, the counter E.M.F. increasing most rapidly at that moment, then more and more slowly, and finally reaching a constant value within 1 second or less. The magnitude of counter E.M.F. is proportional to the current density with small currents flowing in either direction across the protoplasm, but falls off at higher density, giving a cusp with recession to lower values; this recession occurs with slightly lower currents outward than inward. Otherwise the curves are much the same for inward and outward currents, for different densities, for charge and discharge, and for successive current flows. There is a slight tendency for the bioelectric potential to become temporarily positive following these current flows. Records in the regular state (State A) show very little effect of increased series resistance on the time constant of counter E.M.F. This seems to indicate that a polarization rather than a static capacity is involved. State B.—Delayed and non-proportional polarization, in which there is no counter E.M.F. developed with small currents in either direction across the protoplasm, nor with very large outward currents. But with inward currents a threshold density is reached at which a counter E.M.F. rather suddenly develops, with a sigmoid curve rising to high positive values (200 mv. or more). There is sometimes a cusp, after which the P.D. remains strongly positive as long as the current flows. It falls off again to negative values on cessation of current flow, more rapidly after short flows, more slowly after longer ones. The curves of charge are usually quite different in shape from those of discharge. Successive current flows of threshold density in rapid succession produce quicker and quicker polarizations, the inflection of the curve often becoming smoothed away. After long interruptions, however, the sigmoid curve reappears. Larger inward currents produce relatively little additional positive P.D.; smaller ones on the other hand, if following soon after, have a greatly increased effectiveness, the threshold for polarization falling considerably. The effect dies away, however, with very small inward currents, even as they continue to flow. Over a medium range of densities, small increments or decrements of continuing inward current produce almost as regular polarizations as in State A. Temporary polarization occurs with outward currents following soon after the threshold inward currents, but the very flow of outward current tends to destroy this, and to decondition the protoplasm, again raising the threshold, for succeeding inward flows. State A is characteristic of a few freshly gathered cells and of most of those which have recovered from injuries of collecting, cleaning, and separating. It persists a short time after such cells are impaled, but usually changes over to State B for a considerable period thereafter. Eventually there is a reappearance of regular polarization; in the transition there is a marked tendency for positive P.D. to be produced after current flow, and during this the polarizations to outward currents may become much larger than those to inward currents. In this it resembles the effects of acidified sea water, and of certain phenolic compounds, e.g. p-cresol, which produce State A in cells previously in State B. Ammonia on the other hand counteracts these effects, producing delayed polarization to an exaggerated extent. Large polarizations persist when the cells are exposed to potassium-rich solutions, showing it is not the motion of potassium ions (e.g. from the sap) which accounts for the loss or restoration of polarization. It is suggested that inward currents restore a protoplasmic surface responsible for polarization by increasing acidity, while outward currents alter it by increasing alkalinity. Possibly this is by esterification or saponification respectively of a fatty film. For comparison, records of delayed polarization in silver-silver chloride electrodes are included.     

EFFECTS OF HEXYLRESORCINOL ON VALONIA

The effects on Valonia of guaiacol and hexylresorcinol are similar but the latter is more effective. Both substances lower or abolish the potassium effect; i.e., the ability of the cell to distinguish electrically between Na⁺ and K⁺. Both substances change the order of mobilities so that v _Cl > u _Na becomes u _Na > v _Cl or u _Na = v _Cl.     

EFFECTS OF NITROBENZENE AND BENZENE ON VALONIA

The effects of nitrobenzene and of benzene resemble those of guaiacol and of hexylresorcinol. The P.D. changes in a positive direction and then in a negative direction. The latter change may bring the P.D. back to the starting point with guaiacol and hexylresorcinol but with nitrobenzene and benzene this is not always the case. The positive potential change produced by nitrobenzene and benzene may be antagonized to some extent by ammonia. Nitrobenzene and benzene raise the electrical resistance and this is antagonized to some extent by ammonia. The results afford a further illustration of the important fact that the behavior of inorganic ions can be changed by organic substances. The apparent mobility of Na⁺ is increased and that of K⁺ decreased by nitrobenzene and benzene (as is also the case with guaiacol and hexylresorcinol).     

CALCULATIONS OF BIOELECTRIC POTENTIALS : VI. SOME EFFECTS OF GUAIACOL ON NITELLA

Values have been calculated for apparent mobilities and partition coefficients in the outer non-aqueous layer of the protoplasm of Nitella. Among the alkali metals (with the exception of cesium) the order of mobilities resembles that in water and the partition coefficients (except for cesium) follow the rule of Shedlovsky and Uhlig, according to which the partition coefficient increases with the ionic radius. Taking the mobility of the chloride ion as unity, we obtain the following: lithium 2.04, sodium 2.33, potassium 8.76, rubidium 8.76, cesium 1.72, ammonium 4.05, ½ magnesium 20.7, and ½ calcium 7.52. After exposure to guaiacol these values become: lithium 5.83, sodium 7.30, potassium 8.76, rubidium 8,76, cesium 3.38, ammonium 4.91, ½ magnesium 20.7, and ½ calcium 14.46. The partition coefficients of the chlorides are as follows, when that of potassium chloride is taken as unity: lithium 0.0133, sodium 0.0263, rubidium 1.0, cesium 0.0152, ammonium 0.0182, magnesium 0.0017, and calcium 0.02. These are raised by guaiacol to the following: lithium 0.149, sodium 0.426, rubidium 1.0, cesium 0.82, ammonium 0.935, magnesium 0.0263, and calcium 0.323 (that of potassium is not changed). The effect of guaiacol on the mobilities of the sodium and potassium ions resembles that seen in Halicystis but differs from that found in Valonia where guaiacol increases the mobility of the sodium ion but decreases that of the potassium ion.     

EFFECTS OF CYANIDE ON THE PROTOPLASM OF AMEBA

The experiments seem to indicate that the toxicity of HCN and KCN for amebæ is due to their effect on the cell membrane and not on the internal protoplasm. Concentrated solutions (N/10–N/300) of HCN or KCN produce an initial increase in viscosity of the protoplasm of amebæ (immersed) which is followed by liquefaction and disintegration of the cell. Dilute solutions of HCN or KCN decrease the viscosity of the protoplasm of amebæ. Injections of HCN or KCN into amebæ produce a reversible decrease in viscosity of the protoplasm.     

APPARENT VIOLATIONS OF THE ALL-OR-NONE LAW IN RELATION TO POTASSIUM IN THE PROTOPLASM

In Vol. 37, No. 6, July 20, 1954, page 814, in the third paragraph, delete the first sentence and insert the following: This equation gives excellent results with Nitella for example with 0.001 M KCl outside and 0.05 M KCl in the sap we may use concentrations in place of activities in the equation and put a_s = 0.05 and a_o = 0.001. No allowance is made for any change in the concentration of incoming ions due to their combination with carrier molecules since this change would be small and difficult to estimate. If we put U = 73 and V = 1 we obtain P = 97 mv. which is close to the usual observed value. The expression (73 –1) ÷ (73 + 1) = 0.973.     

DIFFERING RATES OF DEATH AT INNER AND OUTER SURFACES OF THE PROTOPLASM : I. EFFECTS OF FORMALDEHYDE ON NITELLA

When protoplasm dies it becomes completely and irreversibly permeable and this may be used as a criterion of death. On this basis we may say that when 0.2 M formaldehyde plus 0.001 M NaCl is applied to Nitella death arrives sooner at the inner protoplasmic surface than at the outer. If, however, we apply 0.17 M formaldehyde plus 0.01 M KCl death arrives sooner at the outer protoplasmic surface. The difference appears to be due largely to the conditions at the two surfaces. With 0.2 M formaldehyde plus 0.001 M NaCl the inner surface is subject to a greater electrical pressure than the outer and is in contact with a higher concentration of KCl. In the other case these conditions are more nearly equal so that the layer first reached by the reagent is the first to become permeable. The outer protoplasmic surface has the ability to distinguish electrically between K⁺ and Na⁺ (potassium effect). Under the influence of formaldehyde this ability is lost. This is chiefly due to a falling off in the partition coefficient of KCl in the outer protoplasmic surface. At about the same time the inner protoplasmic surface becomes completely permeable. But the outer protoplasmic surface retains its ability to distinguish electrically between different concentrations of the same salt, showing that it has not become completely permeable. After the potential has disappeared the turgidity (hydrostatic pressure inside the cell) persists for some time, probably because the outer protoplasmic surface has not become completely permeable.     

PATHOLOGICAL CHANGES IN ULTRASTRUCTURE: EFFECTS OF VICTORIN ON OAT ROOTS

Cells in the interior of susceptible oat roots treated with the disease-inducing agent victorin exhibit many of the ultrastructural features which characterize the epidermal or outermost root cap cells of untreated roots. An increase in electron density of cell walls fixed in permanganate is the first effect of victorin seen in the root interior. Other early victorin-induced changes are formation of enlarged, densely stained vesicles by the Golgi apparatus and organization of the endoplasmic reticulum into roughly parallel profiles. All of these features are characteristic of untreated epidermal cells. Victorin also induces the formation of large numbers of lomasome-like wall lesions and causes a marked increase in the number of nearly spherical, membrane-bounded structures tentatively identified as spherosomes. Similar lomasome- and spherosome-like structures are much more abundant in the outermost cells of the root cap than in other regions of untreated roots. This suggests that these structures may be characteristic of cells destined to undergo disintegration. Victorin-induced lesions appear to arise within the cell wall as the result of an activation of wall-degrading enzymes. An early change which makes the unit structure of the plasma membrane visible over extended areas may account for victorin-induced changes in permeability. Disrupted plasma membranes and swollen mitochondria are found only in cells heavily damaged by victorin. Many of the effects of victorin resemble those of calcium deficiency and calcium is known to suppress victorin-induced disease symptoms. This suggests that calcium nutrition may play a role in the pathological changes induced by victorin.     

EFFECTS OF GUAIACOL AND HEXYLRESORCINOL IN THE PRESENCE OF BARIUM AND CALCIUM

Guaiacol was applied at two spots on the same cell of Nitella. At one spot it was dissolved in 0.01 M NaCl, at the other in 0.01 M CaCl₂ or BaCl₂. The effect was practically the same in all cases, i.e. a similar change of P.D. in a negative direction, involving a more or less complete loss of P.D. (depolarization). When hexylresorcinol was used in place of guaiacol the result was similar. That Ca⁺⁺ and Ba⁺⁺ do not inhibit the effect of these organic depolarizing substances may be due to a lack of penetration of Ca⁺⁺ and Ba⁺⁺. The organic substances penetrate more rapidly and their effect is chiefly on the inner protoplasmic surface which is the principal seat of the P.D.     

猪精子体外获能前后离子成分变化

应用扫描电镜和镜射电镜能谱技术,为猪精子获能前后质膜表面和内部的离子成分进行了研究,结果表明,猪精子获能后质膜表面的Na~+和Al~(3+)升高,而Cl~-和Ca~(2+)降低;精子顶体内Na~+和Cl~-降低,Ca~(2+)、K~+和Fe~(2+)升高;中段线粒体内的Na~+、Ca~(2+)和Fe~(2+)升高,而K~+和Cl~-降低。文章分析了精子获能后顶体内Na~+、Cl~-、K~+、Ca~(2+)和Fe~(2+)变化的浓度比和摩尔比。     

IONIC EFFECTS ON LIGNIFICATION AND PEROXIDASE IN TISSUE CULTURES

Crown-gall tumor tissue cultures release peroxidase into the medium in response to the concentration of specific ions in the medium. This release is not due to diffusion from cut surfaces or injured cells. Calcium, magnesium, and ammonium were, in that order, most effective in increasing peroxidase release. The enzyme was demonstrated cytochemically on the cell walls and in the cytoplasm. Cell wall fractions, exhaustively washed in buffer, still contained bound peroxidase. This bound peroxidase could be released by treating the wall fractions with certain divalent cations or ammonium. The order of effectiveness for removing the enzyme from the washed cell walls is: Ca⁺⁺ ≈ Sr⁺⁺ > Ba⁺⁺ > Mg⁺⁺ > NH₄⁺. These data support the thesis presented that specific ions can control the deposition of lignin on cell walls by affecting the peroxidase levels on these walls.     

CHANGES IN THE APPARENT CYTOPLASMIC HYDROGEN ION CONCENTRATION OF AMEBA DUBIA ON INJECTION OF EGG ALBUMIN

1. Egg albumin when injected into an ameba or discharged into the solution about it raises the apparent pH of the cytoplasm of the ameba. 2. With time the cytoplasm returns to the original pH 6.9 if the nucleus is present. Amebae that have received repeated injections of albumin in some cases extrude their nuclei. In these cells the cytoplasm remains at the more alkaline pH induced by the albumin for at least 12 hours. 3. When a 2 per cent solution of albumin is introduced into a suspension of amebae there is a temporary marked rise in the rate at which CO₂ is given off with no corresponding rise in O₂ uptake. 4. The results observed can be explained if the albumin discharged onto the surface of the ameba rapidly enters the cell and there becomes distributed in a phase of the cytoplasm other than the one which contains the phenol red.     

气候变化对我国红松林地理分布影响的研究

在红松林地理分布规律研究的基础上,应用地理信息系统ＩＤＲＩＳＩ和专门计算机软件——生态信息系统ＧＲＥＥＮ,找出适宜红松林分布的地理气候参数区间,并以此确定了红松林适宜分布区,在此基础上,根据全球气候预测模型ＧＣＭｓ预测的２０３０年的气候变化结果,就气候变化对我国红松林地理分布的可能影响进行了预测。结果表明：到２０３０年,因气候变化的影响,我国适宜红松分布的面积将有所增加,但增加幅度不大,仅占当前气候条件下适宜红松分布面积的３．４％,局部地区变化的情况是：在黑龙江省的西北部适宜红松分布的面积将有所增加;在辽宁省的西南部适宜红松分布的面积将有所减少。红松林现实分布区的南界将向北移动０．１～０．６个纬度,北界将向北扩展０．３～０．５个纬度,黑龙江省境内的红松林分布区的西界将向西扩展０．１～０．５个经度。我国适宜红松分布的面积将由当前气候条件下的２．９×１０７ｈｍ２,增加到３．０×１０７ｈｍ２。就当前气候变化影响预测中存在的问题及未来研究的方向进行了讨论。