Characterization of calcium, phosphate and peroxide interactions in activation of mitochondrial swelling using derivative of the swelling curves

We describe a new method for the analysis of mitochondrial swelling curves. Using classical swelling curves, only the maximum extent of the swelling can be calculated in a numerical form. However, taking the derivative of the classical swelling curves enables the evaluation of two additional parameters of the swelling process in a numerical form, namely, the maximum swelling rate after the addition of the swelling inducer (as dA???/10 s) and the time (in sec) at which the maximum swelling rate after the addition of the swelling inducer is obtained. The use of these three parameters enables the better characterization of the swelling process as demonstrated by the evaluation of calcium and phosphate interactions in the opening of the mitochondrial permeability transition pore and by the characterization of the peroxide potentiating action.     

Development of the sexual skin with pubertal maturation in female chimpanzees

We examined the adolescent development of the sexual skin of chimpanzees (Pan troglodytes) by daily observation, evaluation of swelling, and weekly photogrametry. Although the size of swelling differed with the individual, the development of sexual swelling followed four stages: (1) initial stage, the labial region began to show a slight swelling and recovery; (2) labial stage, swelling at the labial region became maximal; (3) anal stage, another swelling center appeared in the anal region and enlarged; and (4) full maturity stage, the labial and anal regions merged into a full swelling. Menarche occurred after the beginning of the anal stage, and the regular cycle was then established. All of the swelling stages and the peak swelling size are regarded as good indicators of reproductive maturation in chimpanzees.     

Structural data on the intra-crystalline swelling of beta-chitin

The intra-crystalline swelling of the highly crystalline beta-chitin from Tevnia jerichonana was investigated by X-ray crystallography and Fourier transform infrared (FTIR) spectroscopy, using hydrogenated and deuterated hydrochloric acids as swelling agents. Three levels of swelling were identified that could be defined as inter- and intra-sheet swelling. A moderate and reversible swelling in water and methanol gave crystalline beta-chitin cystallosolvates, namely dihydrate and methanolate, respectively. In these, an inter-sheet swelling was observed, corresponding to an expansion of only the b parameter of the unit cell of beta-chitin. Under these swelling conditions, the use of deuterated reagents had no effect on the amide N&z.sbnd;H⋯O&z.dbnd6;C hydrogen bonds that hold the structure of beta-chitin together, but only induced a partial and reversible deuteration of the chitin hydroxymethyl groups. A more severe swelling - but still reversible - occurred with 6 N HCl or DCl, which converted the crystals of beta-chitin into a paracrystalline gel-like product resulting from inter-sheet+intra-sheet swelling. With this acid strength, the deuteration pattern indicated that a fraction of the amide hydrogen bonds was broken and became susceptible to an irreversible deuteration. A very severe and irreversible swelling occurred with 8 N HCl or DCl. In that case, the inter- and intra-sheet swelling was extensive to the point where all memory of the parallel-chain beta-chitin was lost. In addition, this swelling was accompanied by a drastic and rapid depolymerization. The treatment with 8 N HCl led invariably to crystalline alpha-chitin when the samples were neutralized.     

On the mechanism of lactate dehydrogenase release from skeletal muscle in relation to the control of cell volume

The mechanism of enzyme release from isolated skeletal muscle was illustrated by the study of the release of lactate dehydrogenase (LDH). In hypotonic media of different composition but of same tonicity the increase of LDH permeability was triggered at the same range of relative osmolality R (0.45 less than R less than 0.55), although the swelling in the respective media showed appreciable differences. The kinetics of muscle swelling showed that a deviation from the theoretically computed swelling curve to lower values of swelling was connected with an increased LDH permeability. The reduction of swelling was ATP- and Ca2+ and/or Mg2-dependent. It is concluded that swelling of cells generally precedes the leakage of soluble enzymes, and the cross-linking of filaments at the sarcoplasmic side of the sarcolemma under appropriate conditions can counteract swelling, thereby blebbing off the cell membrane from the filament meshwork. In the course of this process, sufficiently large membrane lesions are produced through which macromolecules may escape into the extracellular space.     

Development of a Model for Analyzing the Swelling Rate of Ionic Gels on the Basis of the Diffusion of Mobile Ions: Application to the pH-Sensitive Swelling of a Polyelectrolyte Complex Gel Prepared from Xanthan and Chitosan

A model for analyzing the swelling rate of ionic gels was developed on the basis of the diffusion of a species of mobile ion. This model was applied to the analysis of pH-sensitive swelling of a xanthan/chitosan complex gel in NaOH solutions of pH 9–12, using the sodium ion as the reference mobile ion. The time–course for swelling of gel beads with a pH change from 11 to 10 was successfully described by the developed model. The values for the diffusion coefficient obtained by fitting the model to the data were of the same order as those for the diffusion coefficient of the sodium ion measured for a membrane of the complex gel. Thus, it was confirmed that the swelling rate of the gel due to pH change was mainly controlled by the diffusion of mobile ions. However, the time-course for swelling of the gel at pH values below 10 was not satisfactorily explained by the model developed, suggesting that the change in the degree of ionization during swelling also affected the swelling rate of the xanthan/chitosan complex gel.     

Glutamine in the mechanism of ammonia-induced astrocyte swelling

Brain edema and the subsequent increase in intracranial pressure are the major neurological complications in fulminant hepatic failure (FHF). Brain edema in FHF is predominantly "cytotoxic" due principally to astrocyte swelling. It is generally believed that ammonia plays a key role in this process, although the mechanism by which ammonia brings about such swelling is yet to be defined. It has been postulated that glutamine accumulation in astrocytes subsequent to ammonia detoxification results in increased osmotic forces leading to cell swelling. While the hypothesis is plausible and has gained support, it has never been critically tested. In this study, we examined whether a correlation exists between cellular glutamine levels and the degree of cell swelling in cultured astrocytes exposed to ammonia. Cultured astrocytes derived from rat brain cortices were exposed to ammonia (5 mM) for different time periods and cell swelling was measured. Cultures treated with ammonia for 1-3 days showed a progressive increase in astrocyte cell volume (59-127%). Parallel treatment of astrocyte cultures with ammonia showed a significant increase in cellular glutamine content (60-80%) only at 1-4 h, a time when swelling was absent, while glutamine levels were normal at 1-3 days, a time when peak cell swelling was observed. Thus no direct correlation between cell swelling and glutamine levels was detected. Additionally, acute increase in intracellular levels of glutamine by treatment with the glutaminase inhibitor 6-diazo-5-oxo-L-norleucine (DON) after ammonia exposure also did not result in swelling. On the contrary, DON treatment significantly blocked (66%) ammonia-induced astrocyte swelling at a later time point (24 h), suggesting that some process resulting from glutamine metabolism is responsible for astrocyte swelling. Additionally, ammonia-induced free radical production and induction of the mitochondrial permeability transition (MPT) were significantly blocked by treatment with DON, suggesting a key role of glutamine in the ammonia-induced free radical generation and the MPT. In summary, our findings indicate a lack of direct correlation between the extent of cell swelling and cellular levels of glutamine. While glutamine may not be acting as an osmolyte, we propose that glutamine-mediated oxidative stress and/or the MPT may be responsible for the astrocyte swelling by ammonia.     

Alterations of cell volume regulation in the development of hepatocyte necrosis

Intracellular Na+ accumulation has been shown to contribute to hepatocyte death caused by anoxia or oxidative stress. In this study we have investigated the mechanism by which Na+ overload can contribute to the development of cytotoxicity. ATP depletion in isolated hepatocytes exposed to menadione-induced oxidative stress or to KCN was followed by Na+ accumulation, loss of intracellular K+, and cell swelling. Hepatocyte swelling occurred in two phases: a small amplitude swelling (about 15% of the initial size) with preservation of plasma membrane integrity and a terminal large amplitude swelling associated with cell death. Inhibition of Na+ accumulation by the use of a Na+-free medium prevented K+ loss, cell swelling, and cytotoxicity. Conversely, blocking K+ efflux by the addition of BaCl2 did not influence Na+ increase and small amplitude swelling, but greatly stimulated large amplitude swelling and cytotoxicity. Menadione or KCN killing of hepatocytes was also enhanced by inducing cell swelling in an hypotonic medium. However, increasing the osmolarity of the incubation medium did not protect against large amplitude swelling and cytotoxicity, since stimulated Na+ accumulation and K+ efflux. Altogether these results indicate that the impairment of volume regulation in response to the osmotic load caused by Na+ accumulation is critical for the development of cell necrosis induced by mitochondrial inhibition or oxidative stress.     

Role of blood lipoproteins in adaptive reorganization of the rat liver mitochondria

It was shown in experiments in vitro that high density lipids (HDL), very low density lipids (VLDL) and cAMP taken separately did not affect the kinetics of mitochondrial swelling in the liver of control and 72-hr-fasting rats. ApoHDL and particularly ApoVLDL obtained by delipidation of appropriate lipoproteins had pronounced capacities to stimulate the process of swelling. Mitochondrial swelling increased to an ever greater degree under the action of cAMP coupled with apoproteins. The maximum stimulation of the swelling was attained as a result of administering apoVLDL in conjunction with cAMP. The effects of enhanced swelling were the most remarkable in experiments with mitochondria from the control animals.     

Structural theory of the swelling pressure of corneal stroma in saline

The swelling pressure of the corneal stroma is discussed in terms of a stromal model of collagen fibrils interconnected via protein-mucopolysaccharide molecular complexes. The analysis leads to an expression for the swelling pressure in terms of molecular parameters of the complexes regarded as polymeric chains. Assigning reasonable values to these parameters we find that the model is consistent with the swelling pressure experiments. These parameters indicate the electrostatic (Donnan) pressure component accounts for most of the swelling pressure at normal thickness in 0.174 M NaCl, while the pressure components deriving from a free energy of mixing account for most of the swelling pressure at normal thickness in 1.74 M NaCl.     

Relationship between the induction of swelling and the inhibition of elongation caused by oryzalin and colchicine in corn roots

In corn seedling roots, colchicine and the dinitroaniline herbicideoryzalin cause swelling and inhibition of elongation which seemto be interconnected responses, yet they can be separated. First,both colchicine (10^–3 M) and oryzalin (10^–6 M) affectelongation (2–4 hr) well before swelling (8–10 hr).Second, a short (4 hr) oryzalin treatment produces maximal inhibitionof elongation but not maximal swelling. Third, a combinationof colchicine and oryzalin (at concentrations which give maximalor saturating effects on swelling and elongation when appliedsingly) produces a greater effect on swelling (but not elongation)than either alone. Thus swelling and inhibition of elongationare not related in a simple reciprocal relationship and maybe, at least in part, separate responses, and colchicine andoryzalin may not act in completely identical ways. (Received June 14, 1977; )     

Influence of Antioxidants on the Peroxidative Swelling of Mitochondria in vitro

To clarify the role of prooxidative processes during in vitro swelling of freshly isolated rat liver mitochondria, the influence of different antioxidants and free-radical scavengers was tested. Ascorbate below 10 mmol/L without externally added Fe²⁺ acted as a prooxidant and enhanced swelling. Higher concentrations in the presence of Fe²⁺ showed antioxidant properties and a decrease in swelling and lipid peroxidation. Swelling was abolished by -tocopherol and reduced to 50% by butylated hydroxytoluene. Glutathione supplementation decreased both swelling and lipid peroxidation. Oxidized glutathione caused swelling without any effect on peroxidation. Hydrogen peroxide, cumene hydroperoxide and t-butyl hydroperoxide caused progressive decreases in glutathione and reduced niacinamide coenzyme levels, suggesting prooxidative changes. Dithiothreitol was found to abolish this effect. Thus, antioxidants reverse superoxide-induced mito chondrial swelling and lipid peroxidation in vitro.     

Simulation of the influences of bathing solution and crosslink density on the swelling equilibrium of ionic thermo-sensitive hydrogels

The influences of the bathing solution and crosslink density on the swelling equilibrium of ionic thermo-sensitive hydrogels due to temperature stimulus are studied by mathematical modeling. The model used is termed the multi-effect-coupling thermal-stimulus (MECtherm) model with consideration of multiphases and multiphysics. It is used for the steady-state numerical simulation of the hydrogels in swelling equilibrium after it is verified well by comparison with available experimental data concerning the variation of volume swelling ratio with temperature. The phenomenon of volume phase transition is simulated for the thermal-stimulus responsive hydrogel. Simulations predict well the influences of the bathing solution concentration and crosslink density of polymeric network on the swelling equilibrium of the hydrogels.     

Structural-mechanical properties and swelling of the protein complexes of hyaluronate and protein-chondroitin-4-sulfate]

It was shown that the mechano-structural strength and swelling of gels consisting of gelatine and potassium hyaluronate (PH) or potassium protein-chondroitin-4-sulphate (PPCS) depended on the correlation of the components. Low PH and PPCS concentrations showed a minimal mechano-structural strength and a maximal swelling of gels. In the zones of neutralization of the positive electrical charges of gelatine by macropolyanions a high mechano-structural strength coincided with a minimal swelling. With high PH concentrations the mechano-structural strength and swelling of gels became equal to those of pure gelatine; and with high PPCS concentrations there was noted an additional parallel increase in the strength and swelling of gels.     

Re-assessment of the potential impact of physiologically relevant pH changes on the hydration properties of the isolated mammalian corneal stroma

The pH sensitivity of the swelling of the mammalian corneal stroma was reinvestigated to assess whether or not there were detectable differences in the hydration properties of this collagen-keratocyte matrix within a physiologically relevant range (as opposed to extremes of acid or alkaline pH) and at a physiologically relevant temperature. From recent post-mortem eyes of adult cows, square (8 x 8 mm) samples of corneal stroma were prepared and incubated in an isotonic, buffered (HEPES etc.), mixed salts solution with added glucose at 37 degrees C. The time-dependent changes in wet mass were assessed over 24 h. The rate and magnitude of stromal swelling were different within the range of pH 6.5-8.5. The wet mass of stromal samples increased almost 2-fold within 1 h, and then at lesser rates to realise 3.25-3.75-fold and 4-5-fold increases in wet mass by 9 h and 24 h respectively. The maximum increases were observed at pH 7.25-7.5, with most of the effect being the result of differences in the initial rate of swelling. The discontinuous swelling and the pH effect on the rates of swelling were also evident when the data were fitted to a previous kinetic model (Elliott et al., J. Physiol. (Lond.) 298 (1980) 453-470). It is concluded that pH changes in the physiological range can have a small but reproducible impact on the swelling kinetics of the isolated mammalian corneal stroma ex vivo.     

“八风”、“八邪”穴切开减压治疗蛇伤肢体肿胀疗效观察

目的观察"八风"、"八邪"穴切开减压对蛇伤肢体肿胀的治疗效果。方法选取我院治疗的毒蛇咬伤肢体肿胀患者60例,随机分为对照组和治疗组各30例,对照组给予常规治疗,治疗组在常规治疗基础上加予"八风"、"八邪"穴切开减压治疗,观察比较两组患者肿胀最大值时周径差及肿胀消退时间。结果治疗组患者肿胀最大值时周径差明显小于对照组(P0.05),肿胀消退时间明显短于对照组(P0.05)。结论 "八风"、"八邪"穴切开减压对减轻蛇伤肢体肿胀程度、加快肿胀消退有确切效果。     

Assessment of the effects of cetylpyridium chloride on water content of the collagen-keratocyte matrix of the mammalian corneal stroma ex vivo

The effects of cationic surfactants on the time-dependent increases in hydration of the corneal stroma were investigated to assess if the contribution of the proteoglycans could be titrated and how it might relate to the maximum and minimum swelling properties of the corneal stroma. From recent post-mortem eyes from adult sheep, square (8 x 8 mm) samples of corneal stroma were prepared and incubated in isotonic neutral pH mixed salts solution with added glucose, or pure water, at 37 degrees C. The time-dependent changes in wet mass were assessed over 24 h in the absence or presence of 0. 001-2% w/v cetylpyridium chloride (CPC) or benzalkonium chloride (BAC). The rate and magnitude of stromal swelling was reduced in a concentration-dependent fashion by the surfactants. In mixed salts solution, 100% inhibition of swelling could be achieved at 2% CPC and BAC. In pure water, the relative swelling was much more substantial and could only be attenuated by CPC.     

Characterization of the swelling of a size-exclusion gel.

The swelling of a dextran gel, Sephadex G-75, was observed in an aqueous environment at room temperature by a noninvasive technique that uses light microscopy coupled to an image analysis system via a video camera. The rate of swelling was found to follow the Tanaka and Fillmore theory, from which the overall gel diffusion coefficient was estimated as 6.3 x 10(-7) cm2/s. In addition to giving a quantitative measure of gel swelling that could be useful in the mechanical design of liquid chromatography columns, this approach provides data on wet particle size and particle size range, which is needed for the modeling of diffusional and mass transfer effects in size-exclusion chromatography. In this context, key observations are that the gel particles are nearly spherical with an elliptical shape factor of 0.98 (perfect sphere = 1) and that there is little difference between sizes of particles obtained in water, 50 mM Tris-glycine buffer (pH 10.2), and buffer containing 1 mg/mL protein. The diameter of the dry material ranged from 20 to 100 microns, while the hydrated particles had diameters of 40-350 microns. The rate of swelling is rapid, with 50% swelling occurring in about 10 s and swelling to 99% of the final wet particle size being obtained in less than 90 s.     

Development of the Stem Tuber in Kohlrabi

Stem swelling in kohlrabi (KR) normally occurred between thethird and fifth node and followed initiation of meristematicactivity in pith parenchyma cells; this usually began about7 weeks after germination and was associated with the deathof parenchyma cells in the centre of the pith lower down thestem. Cell division began just above this region, at about thethird node and zones of dividing cells extended upwards andlaterally, giving rise to more centres of active division. Carbohydrateanalyses showed that fructose, glucose, starch, and an unidentifiedsugar accumulated in the stem. Melezitose appeared in the stemand raffinose in the leaves only after visible bulbing had occurred.Sucrose content fluctuated in both leaves and stems and in thelatter was positively correlated with hours of sunshine on theday before sampling. The contents of gibberellin-like substancesin the leaves of KR were compared at two stages of developmentwith those of marrowstem kale (MSK). Gibberellins were presentin both acidic and neutral fractions in KR tissues and the amountswere similar to those in MSK. Stem swelling in KR could notbe attributed to a gross deficiency of gibberellins. Inter- and intra-varietal approach-grafts were made with KRand MSK plants. With KR as ‘scion’ partner, stemelongation of the scion was not greatly affected by the natureof the stock, but stem swelling was much reduced on MSK stocks.Stem swelling in MSK scions was greater on KR stocks than onMSK. Retention of stock leaves and buds above the union tendedto inhibit elongation of the scion partner and also inhibitedstem swelling in KR scions on MSK stocks. Application of gibberellicacid to leaves of MSK stocks from which the terminal bud hadbeen removed, increased both stem elongation and swelling inKR scions. KR scions top-grafted on defoliated MSK stocks showedvigorous stem elongation without swelling. MSK scions top-graftedon KR stocks showed reduced stem elongation with abnormal stemthickening. It is suggested that high auxin levels may preventbulbing in KR and that gibberellin-like substances may facilitateboth stem elongation and bulbing when auxin levels are low. Applying gibberellic acid (GA₃) or indole-3-acetic acid (IAA)alone or in combination, to young intact plants of KR increasedstem height and checked stem swelling. Application of benzyladenine(BA) at 10^–4 M accelerated swelling and checked stem elongation.GA₃ applied with BA (10^–4 M) increased both stem heightand girth, both with or without added IAA. It was concludedthat the requisite conditions for stem swelling included thepresence of a kinin² and a low level of auxin. High levels ofauxin prevented stem swelling, whether applied as IAA to youngintact plants or as natural auxins from leaves and buds of MSKplants grafted to KR scions. Although applied gibberellins usuallytended to prevent swelling, native gibbereltins appeard to playa minor role in controlling its initiation which probably occurswhen the auxin/kinin ratio in the plant reaches a critical lowlevel.     

Red cell volume regulation: the pivotal role of ionic strength in controlling swelling-dependent transport systems.

A volume increase of trout erythrocytes can be induced either by beta-adrenergic stimulation of a Na+/H+ antiport in an isotonic medium (isotonic swelling) or by suspending red cells in an hypotonic medium (hypotonic swelling). In both cases cells regulate their volume by a loss of osmolytes via specific pathways. After hypotonic swelling several volume-dependent pathways were activated allowing K+, Na+, taurine and choline to diffuse. All these pathways were fully inhibited by furosemide and inhibitors of the anion exchanger (DIDS, niflumic acid), and the K+ loss was mediated essentially via a 'Cl(-)-independent' pathway. After isotonic swelling, the taurine, choline and Na+ pathways were practically not activated and the K+ loss was strictly 'Cl(-)-dependent'. Thus cellular swelling is a prerequisite for activation of these pathways but, for a given volume increase, the degree of activation and the degree of anion-dependence of the K+ pathway depend on the nature of the stimulus, whether hormonal or by reduction of osmolality. It appears that the pattern of the response induced by hormonal stimulation is not triggered by either cellular cAMP (since it can be reproduced in the absence of hormone by isotonic swelling in an ammonium-containing saline) or by the tonicity of the medium in which swelling occurs since after swelling in an isotonic medium containing urea, the cells adopt the regulatory pattern normally observed after hypotonic swelling. We demonstrated that the stimulus is the change in cellular ionic strength induced by swelling: when ionic strength drops, the cells adopt the hypotonic swelling pattern; when ionic strength increases, the isotonic swelling pattern is activated. To explain this modulating effect of ionic strength a speculative model is proposed, which also allows the integration of two further sets of experimental results: (i) all the volume-activated transport systems are blocked by inhibitors of the anion exchanger and (ii) a Cl(-)-dependent, DIDS-sensitive K+ pathway can be activated in static volume trout red cells (i.e., in the absence of volume increase) by the conformational change of hemoglobin induced by the binding of O2 or CO to the heme.     

Na-K-Cl Cotransporter-1 in the mechanism of ammonia-induced astrocyte swelling

Brain edema and the consequent increase in intracranial pressure and brain herniation are major complications of acute liver failure (fulminant hepatic failure) and a major cause of death in this condition. Ammonia has been strongly implicated as an important factor, and astrocyte swelling appears to be primarily responsible for the edema. Ammonia is known to cause cell swelling in cultured astrocytes, although the means by which this occurs has not been fully elucidated. A disturbance in one or more of these systems may result in loss of ion homeostasis and cell swelling. In particular, activation of the Na-K-Cl cotransporter (NKCC1) has been shown to be involved in cell swelling in several neurological disorders. We therefore examined the effect of ammonia on NKCC activity and its potential role in the swelling of astrocytes. Cultured astrocytes were exposed to ammonia (NH(4)Cl; 5 mm), and NKCC activity was measured. Ammonia increased NKCC activity at 24 h. Inhibition of this activity by bumetanide diminished ammonia-induced astrocyte swelling. Ammonia also increased total as well as phosphorylated NKCC1. Treatment with cyclohexamide, a potent inhibitor of protein synthesis, diminished NKCC1 protein expression and NKCC activity. Since ammonia is known to induce oxidative/nitrosative stress, and antioxidants and nitric-oxide synthase inhibition diminish astrocyte swelling, we also examined whether ammonia caused oxidation and/or nitration of NKCC1. Cultures exposed to ammonia increased the state of oxidation and nitration of NKCC1, whereas the antioxidants N-nitro-l-arginine methyl ester and uric acid all significantly diminished NKCC activity. These agents also reduced phosphorylated NKCC1 expression. These results suggest that activation of NKCC1 is an important factor in the mediation of astrocyte swelling by ammonia and that such activation appears to be mediated by NKCC1 abundance as well as by its oxidation/nitration and phosphorylation.