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PubMed1985

Exposure concentration versus environmental concentration: a field survey in organic solvent workplaces.

The relationship was investigated between the environmental concentrations in workplaces and the concentrations in the breath zones of the workers therein. The former was measured in 47 organic solvent workplaces of various types taking both the grid-sampling strategy (Measurement A) and a spot-air sampling to detect possible maximum exposure (Measurement B), while the latter was determined by the application of personal samplers to 328 workers. Although a low yet significant correlation was observed between the results of Measurement A (i.e., the combination of the geometric mean and the geometric standard deviation of the environmental concentration) and the exposure concentration, excess exposure (i.e., over occupational exposure limit exposure) would occur in some workplaces when the occupational exposure limit itself be used as the limit value for the setting of criterion for environmental concentration. The analyses of the experiences suggest no excess exposure would be expected if half the occupational exposure limit be employed as the limit value.

Acetates↗

PubMed1992

Performance of feedlot steers fed diets containing laidlomycin propionate or monensin plus tylosin, and effects of laidlomycin propionate concentration on intake patterns and ruminal fermentation in beef steers during adaptation to a high-concentrate diet.

Two hundred eighty-eight beef steers (British x Continental x Brahman) were fed a 90% concentrate diet containing either no ionophore (control), laidlomycin propionate at either 6 or 12 mg/kg of dietary DM, or monensin plus tylosin (31 and 12 mg/kg of DM, respectively). Neither of the two levels of laidlomycin propionate nor monensin plus tylosin affected (P greater than .10) ADG or feed:gain ratio. Monensin plus tylosin reduced (P less than .01) daily DMI for the 161-d trial period compared with the other three treatments. Laidlomycin propionate at 6 mg/kg increased (P less than .05) DMI relative to the control, laidlomycin propionate at 12 mg/kg, and monensin plus tylosin diets during the 2nd wk of the trial and from d 57 to 84. Treatments did not affect carcass measurements. In a second experiment, 12 ruminally cannulated steers were fed diets containing no ionophore or laidlomycin propionate at either 6 or 12 mg/kg of DM. Samples were obtained for two consecutive days while the dietary concentrate level was 75%, after which the diet was switched abruptly to 90% concentrate, and samples were collected on several days during a 21-d period. The rate at which steers consumed their daily allotment of feed was not altered markedly by laidlomycin propionate. Likewise, laidlomycin propionate did not affect total ruminal VFA concentrations or proportions. Ruminal concentrations of D-lactate were reduced (P less than .10) by 6 but not by 12 mg/kg of laidlomycin propionate.(ABSTRACT TRUNCATED AT 250 WORDS)

Ammonia↗

PubMed1996

[The effect of the albumin concentration on the relation between the concentration of ionized calcium and total calcium in the blood of dogs].

Based on the results of 367 healthy dogs of different age, it could be demonstrated that the concentration of ionized calcium corrected to the pH-value of 7.4 ([Cai (7.4)]) as well as the concentration of total calcium ([Catot]) clearly decreased with increasing age. The most obvious changes were found during the first four months. The [Cai (7.4)] was not influenced distinctly by sex or by breed. The reference range (2.5-97.5% quantil) for [Cai (7.4)] in heparinized plasma was 1.32-1.51 mmol/l in 4-months- to 1-year-old dogs and 1.22-1.46 mmol/l in dogs older than one year, corresponding to a proportion of Cai to Catot of 44.9-54.9%. A moderately close correlation existed between [Cai (7.4)] and the [Catot] (r = 0.754) (n = 393 adult dogs: 180 healthy animals and 213 unselected patients). A similar correlation coefficient was found between the concentrations of Catot and albumin (r = 0.718) or total protein (r = 0.617), respectively. The proportion of Cai to Catot decreased with an increasing concentration of albumin, whereas [Cai (7.4)] tended to increase. The correction of the [Catot] for albumin did not lead to an increased correlation coefficient for the relation with [Cai (7.4)] (r = 0.676). In addition to albumin concentration, the relation between [Cai (7.4)] and the Catot is primarily influenced by complex-forming ions. This became clear by the transient citrate-induced decrease of [Cai (7.4)] whereas [Catot] increased after infusion of fresh frozen plasma in dogs suffering from diarrhea. This investigation shows the limits of the estimation of calcium homoeostasis on the basis of the [Catot].

Aging↗

PubMed1984

The development of pseudohypoparathyroidism. Involvement of progressively increasing serum parathyroid hormone concentrations, increased 1,25-dihydroxyvitamin D concentrations, and 'migratory' subcutaneous calcifications.

The hormonal changes in the development of pseudohypoparathyroidism ( PSH ) have not, to our knowledge, been previously reported. The male sibling of a child with PSH was studied for 2 1/2 years. At 1 year of age he had generalized subcutaneous calcifications that subsequently migrated over his body. At 3 years of age and over a six-month period, serum calcium levels fell; serum phosphorus, parathyroid hormone (PTH), and 1,25-dihydroxyvitamin D (1,25-[OH]2D) concentrations increased. There was no calcemic, phosphaturic, or urinary cyclic adenosine monophosphate response to PTH. The concentration of serum PTH was suppressed by infusion of calcium and doubled with edetic acid infusion, indicating that the parathyroids were sensitive to changes in calcium levels. Thus, increasing PTH and increased 1,25-(OH)2D concentrations occur in the development of PSH . Migratory skin calcifications may occur. We speculate that increasing the serum PTH level reflects increasing compensatory parathyroid production to overcome a progressive PTH receptor defect and serves, with increased 1,25-(OH)2D concentrations, to prevent severe falls in serum calcium concentrations in the early stage of the disease.

Calcinosis↗

PubMed1996

Cell cycle-dependent regulation of cellular ATP concentration, and depolymerization of the interphase microtubular network induced by elevated cellular ATP concentration in whole fibroblasts.

In the present work, evidence is presented indicating that an increased cellular ATP concentration during mitosis may, in conjunction with other factors [Verde et al., 1990: Nature 343:233-238; Andersen et al., 1994: J Cell Biol. 127:1289-1299], induce depolymerization of the interphase microtubular network in cultured fibroblasts. It is shown here that the cellular ATP concentration varies through the cell cycle, reaching a peak at G2M- and minimum at late G1/early S-phase. Furthermore, we have found, using indirect immunofluorescent staining with an antitubulin antibody, that depolymerization of the interphase microtubular network may be induced by increasing the intracellular ATP concentration in cultured fibroblasts from 2.2 mM to 4.1 mM. This may be obtained through addition of adenosine and P1 to the growth medium. Our results indicate that this effect of adenosine and Pi is not mediated via adenosine receptors, but through an elevated cellular ATP concentration. ATP is suggested to act through a concentration-dependent effect on the exchangeable GTP site on tubulin, and not through the action of protein kinases or microtubule-associated proteins.

3T3 Cells↗

PubMed2002

Quantitative determination of cellulase concentration as distinct from cell concentration in studies of microbial cellulose utilization: analytical framework and methodological approach.

In analyzing microbial cellulose utilization, it would be useful to independently measure the mass concentration of cells and cellulase enzymes. Such measurements would allow investigation of the allocation of cellular resources between synthesis of cells and cellulase, in vivo cell- and cellulase-specific cellulose hydrolysis rates, and bioenergetics. Methodological protocols are not established for independent determination of cell and cellulase concentrations for the common case in which a substantial fraction of cellulase is attached to the cell surface. Alternative analytical approaches by which to develop such protocols are examined from the perspective of error minimization. For cell concentration measurement, acceptable accuracy is expected when the concentrations of a cell-specific component (e.g., DNA) is determined or when total protein is determined in conjunction with a measurement specific to cellulase. For cellulase concentration measurement, acceptable accuracy is expected when a measurement specific to cellulase such as ELISA is used. Several analytical approaches are rejected based on large expected errors.

Bacteria↗

PubMed1988

Nuclear magnetic resonance relaxation in experimental brain edema: effects of water concentration, protein concentration, and temperature.

Proton relaxation times T1 and T2 of macromolecular solutions, bovine brain tissues, and experimental cat brain edema tissues were studied as a function of water concentration, protein concentration, and temperature. A linear relation was found between the inverse of the weight fraction of tissue water and the spin-lattice relaxation rate, R1, based on a fast proton exchange model for relaxation. This correlation was also found for the spin-spin relaxation rate, R2, of gray matter samples and macromolecular solutions at low concentrations. Concentrated solutions of protein-water samples showed an enhanced relaxation due to viscosity effects. The T2 of white matter was considerably lengthened with elevated water concentration, but showed no straightforward relation with the total tissue water content. The relaxation times of all samples increased with temperature, supporting the assumption of fast proton exchange in the model for relaxation. This was not found for white matter, in which T2 decreased with increasing temperature, which indicated that intermediate or even slow exchange was present. The relation found between relaxation times and tissue water content can be used to predict the amount of and/or increase in tissue water due to water-elevating processes such as edema.

Animals↗

PubMed1992

Effect of polyethylene glycol-400 at low concentrations on long-term growth of muscle phosphoglucomutase crystals from concentrated salt solutions.

Although rabbit muscle phosphoglucomutase occasionally deposits tetragonal crystals from solutions of ammonium sulfate at about 47% of saturation, low concentrations of polyethylene glycol-400 (PEG), 1 to 4.5% w/v, must be included to sustain crystal growth. A comparison of long-term growth rates for macroscopic crystals in the presence and absence of added PEG suggests that at high salt concentration this cosolute exerts its primary effect on disordered protein aggregates, either in the external medium or at the surface of the crystal, and thereby allows the growth of much larger crystals. Since the observed effects may arise from a PEG-induced increase in the "solubility" of the aggregate that exceeds the induced increase in solubility of the crystalline phase under these conditions, the physical basis for a cosolute-induced increase in solubility in the presence of a precipitant is considered. The applicability of such a rationale to the present system is supported by an assessment of the relative effects of polyethylene glycol and beta-octylglucoside on amorphous, salt-induced precipitates of phosphoglucomutase. PEG also produces what appears to be a differential effect on nucleation efficiency and crystal growth rate. Thus, seed crystals cannot be enlarged at a significant rate at high salt concentration without producing showers of extraneous nucleation centers when the concentration of added PEG is 3% or less. But PEG concentrations of 4.5% essentially eliminate the showering problem, ostensibly by increasing the supersaturation required for nucleation to a greater extent than that required for crystal growth. The same type of effect is observed during de novo growth. Again a solubility-based mechanism is posed. Hysteretic effects related to properties of amorphous aggregates of the protein also are described.

Ammonium Sulfate↗

PubMed1998

Mouse spermatozoa in high concentrations of glycerol: chemical toxicity vs osmotic shock at normal and reduced oxygen concentrations.

The cryobiological preservation of mouse spermatozoa has presented difficulties in the form of poor motilities or irreproducibility. We have identified several likely underlying problems. One is that published studies have used concentrations of the cryoprotectant glycerol that are substantially lower (0.3 M) than the approximately 1 M concentrations that are optimal for most cells. Another may arise from the known high susceptibility of mouse sperm to free radical damage. We have identified two contributors to damage from higher concentrations of glycerol, namely, chemical toxicity proportional to concentration and exposure time and osmotic damage arising from too rapid an addition and removal of the glycerol. When toxicity is minimized by restricting the exposure time to 1 or 5 min and osmotic shock is minimized by adding and removing the glycerol stepwise, relatively high percentages of the sperm survive contact with 0.8 M glycerol. Free-radical damage in mouse sperm is known to be proportional to the oxygen concentration. We have determined the consequences of reducing the oxygen to <3% of atmospheric by the use of a bacterial membrane preparation, Oxyrase. Oxyrase reduced damage from centrifugation and substantially reduced damage from osmotic shock; however, it did not significantly reduce glycerol toxicity.

Animals↗

PubMed2002

Alkanediyl-alpha,omega-bis(dimethylalkylammonium bromide) surfactants 10. Behavior in aqueous solution at concentrations below the critical micellization concentration: an electrical conductivity study.

Ion pairing and premicellar association have been often invoked to explain results obtained in studies of aqueous solutions of ionic dimeric surfactants (gemini surfactants), mainly by means of surface tension and electrical conductivity, at concentrations below the critical micellization concentration (cmc). The present work was undertaken in an attempt to find out under which conditions these effects come into play. For this purpose the electrical conductivity of solutions of many dimeric surfactants of the type spacer-alpha,omega-bis(alkyldimethylammonium bromide) have been measured. The alkyl chain contained m=10-18 carbon atoms. The spacer group was either an alkanediyl with s carbon atoms (m-s-m surfactants) or a xylylene m-xylyl-m surfactants). The results show that ion pairing occurs in solutions of m-s-m dimers with m< or =10, mostly as a result of their high cmc values. The results for 12-s-12 dimers with s< or =10 and for 12-xylyl-12 showed no evidence of either ion pairing or premicellar association. Premicellar association was present for 12-s-12 dimers with s> or =12, for m-8-m dimers with m> or =14, and for 16-xylyl-16. It showed through a positive curvature of the specific conductivity versus concentration plot and the presence of a maximum in the equivalent conductivity vs (concentration)(0.5) plot at concentrations below the cmc. The free energy associated with the premicellar association of m-8-m dimers has been estimated from the available cmc and micelle ionization degree data.

Journal Article↗

PubMed1999

A general approach to modeling conduction and concentration dynamics in excitable cells of concentric cylindrical geometry.

This paper discusses mathematical approaches for modeling the propagation of the action potential and ion concentration dynamics in a general class of excitable cells and cell assemblies of concentric cylindrical geometry. Examples include myelinated and unmyelinated axons, single strands of interconnected cardiac cells and outer hair cells. A key feature in some of the cells is the presence of a small working volume such as the periaxonal space between the myelin sheath and the axon in the myelinated axon and the extracisternal space between the plasma membrane and the subsurface cisterna of the outer hair cell. Proper treatment of these cell types requires a modeling approach which can readily address these anatomical properties and the non-uniform biophysical properties of the concentric membranes and the ionic composition of the volumes between the membranes. An electrodiffusion approach is first developed in which the Nernst-Planck equation is used to characterize axial ion fluxes. It is then demonstrated that this "full" model can be stepwise reduced, eventually becoming equivalent to the standard cable equation formulation. This is done in a manner that permits direct comparisons between the full and simplified models by running simulations using a single parameter set. An intermediate approach where the contributions of the axial currents to ion concentration changes and the effect of varying ion concentrations on solution conductivities are ignored is derived and is found adequate in many cases. Two application examples are given: a "cardiac strand" model, for which the intermediate formulation is shown sufficient and a model of the myelinated axon, for which the full electrodiffusion formulation is clearly necessary. The latter finding is due to spatial inhomogeneities in the anatomy and distribution of ion channels and transporters in the myelinated axon and the restricted periaxonal space between the myelin sheath and the axon.

Action Potentials↗

PubMed1979

Steady-state plasma concentrations as a function of the absorption rate and dosing interval for drugs exhibiting concentration-dependent clearance: consequences for phenytoin therapy.

Model-dependent relationships describing the effects of absorption rate and dosing interval on steady-state phenytoin plasma concentrations are presented and discussed. Utilizing a range of operative Michaelis-Menten parameters that characterize phenytoin elimination via a single capacity-limited pathway, a situation assuming instantaneous absorption (case I) is compared with the situation in which continuous constant-rate absorption occurs (case II). The results of these comparisons demonstrate that average steady-state concentrations do not differ significantly between the two cases. Ratios (case I/case II) of areas under the curve during a steady-state dosing interval substantially deviating from unity are associated with high plasma clearances and clinically low phenytoin concentrations. When the same daily dose is maintained, but the dosing interval is altered (0.25, 0.33, 0.50, 1.0 day), little difference in the average steady-state levels is observed even when absorption is instantaneous. Differences between steady-state maximum and minimum concentrations increase with prolonged dosing intervals as well as faster absorbed phenytoin formulations, but for most patients these fluctuations are therapeutically insignificant. A dimensionless parameter, Q, which is a function of the individual patient's parameters and the dosing regimen, is introduced, and its relationship with steady-state phenytoin concentrations is discussed. Formulation-related differences in phenytoin dissolution rates that may result in significantly altered absorption rates should not affect average steady-state levels unless the extent of absorption is altered. More frequent dosing is not necessary to avoid increases in the average steady-state levels when rapidly absorbed phenytoin products are administered, but may be desirable if the required daily dose is high or the individual patient exhibits a narrow therapeutic range for this drug.

Drug Administration Schedule↗

PubMed1999

Serum concentrations of various environmental contaminants and their relationship to sex steroid concentrations and phallus size in juvenile American alligators.

Recent studies have reported a number of abnormalities in the hatchling and juvenile alligators of Lake Apopka, FL (USA). These abnormalities include modifications of plasma concentrations of sex steroids in males and females as well as abnormalities in gonadal morphology, gonadal enzyme activity, and steroidogenesis. Embryonic exposure to environmental contaminants in the eggs has been hypothesized to be the causal agent for these changes. However, posthatchling exposure can also contribute to changes in reproductive and endocrine functioning. We have detected serum concentrations of 16 of 18 organochlorine pesticides or metabolites (OCs) and 23 of 28 congener-specific polychlorinated biphenyls (PCBs) examined in juvenile alligators from Lake Apopka, Orange Lake, and Lake Woodruff National Wildlife Refuge. Lake Apopka juveniles had significantly elevated serum concentrations of p,p'-DDE, dieldrin, endrin, mirex, oxychlordane, SigmaDDTs, and SigmaPCBs compared to juveniles from the other lakes. Further, we observed no correlations between serum contaminant concentrations and sex steroid concentrations (estradiol-17beta and testosterone). However, serum testosterone was significantly lower in males from Lake Apopka and Orange Lake compared to Lake Woodruff NWR. We did not observe relationships between phallus size or other body parameters and serum contaminant levels. Phallus size was smaller in males from Lake Apopka even after adjustment for body size. We suggest that the observations previously reported for juvenile alligators-and observed again in this study-are apparently not associated with the current serum levels of the environmental contaminants we measured, but could be due to exposures during embryonic development to these or other pollutants. Future studies must determine if a causal relationship exists between the contaminants found in alligator eggs and abnormalities observed in the hatchlings and persisting in juveniles.

Alligators and Crocodiles↗

PubMed1994

[Transferrin concentration in seminal plasma in testicular varicocele with various sperm concentrations and various forms of azoospermia].

Seminal plasma transferrin concentrations were measured in semen samples from 424 men attending a fertility unit. There was a clear positive correlation between seminal plasma transferrin concentration and sperm density: transferrin concentrations decrease with decreasing sperm density and are lowest in patients with azoospermia and those who have undergone vasectomy. The differences between the various groups in sperm density are highly significant, but individual data counts also vary widely. Furthermore, it was demonstrated that in polyzoospermia the seminal plasma transferrin concentration is increased, suggesting primary tubular hyperactivity. No correlations with other seminal parameters or hormonal values were found. Seminal plasma transferrin concentrations in normozoospermia and in varicocele testis or in cases with increased numbers of immature germ cells in the ejaculate were not significantly different. This may be interpreted as indicative of intact secretory activity of Sertoli cells in both varicocele and increased desquamation of immature germ cells. The absence of correlation with any of several important spermatological parameters and our inability to differentiate between azoospermia caused by obstruction and by tubular impairment indicate that seminal plasma transferrin is not a useful marker for Sertoli cell function or for seminiferous tubular dysfunction.

Adolescent↗

PubMed2005

[Background concentrations of molds in house dust. Determination of mold concentrations in dwellings without known mold infestations in three parts of Germany].

Mold concentrations of house dust samples from dwellings reportedly free from mold infestations were analysed to obtain background values. Samples from carpet floors were taken from 80 dwellings in three parts of Germany in winter and summer. Samples were analysed with the cultivation method (using suspension). This resulted in the detection of 35 mold species or genera. Concentrations of the genera Alternaria, Cladosporium, Fusarium and yeast increased in summer. In contrast concentrations of Aspergillus and Penicillium in the summer samples were nearly equal to those in the winter samples. The majority of the various molds were only found in a number of samples too small for a reliable comparison of the winter and summer findings to be made. The results were compared with findings of other authors. The authors propose the concentrations of the 95th percentile of each species (when representing over 10,000 KBE/g of dust) as background values, while a threefold higher value is regarded as an indication of mold presence. An exception should be made for the concentrations of molds which increase sharply in the outdoor air in summer. Because of the difficulty in estimating the outdoor influence, no evaluation proposals can be given for these species for the summer season.

Air Pollution, Indoor↗

PubMed2005

[Background concentrations of molds in air. Determination of mold concentrations in dwellings without known mold infestations in three parts of Germany].

The mold concentration of indoor air and outdoor air in three parts of Germany was investigated in both winter and summer. In total, air samples from 80 dwellings, reportedly free from mold infestations, were analysed with both the cultivation method and the total spore count method. With the cultivation method, 40 mold species and genera were differentiated, while with the total spore count method, 11 spore types were distinguished. The concentration of the genera Alternaria, Cladosporium, Fusarium and yeast increased in summertime. In contrast, the concentrations of Aspergillus and Penicillium measured in summer were nearly equal to those measured in winter. The majority of the various molds were only found in a small number of samples, too small for a reliable comparison of the winter and summer findings. The 95th percentile of the indoor mold concentrations is suggested as the upper limit of the background concentration. The results are discussed comparing the assessment proposals of various authors, and a new assessment proposal is described.

Air Pollution, Indoor↗

PubMed1989

Inhibition of post-ischemic ventricular recovery by low concentrations of prostacyclin in isolated working rat hearts: dependency on concentration, ischemia duration, calcium and relationship to myocardial energy metabolism.

The objective of this study was to characterize the effect of prostacyclin (PGI2) on ventricular function following total global ischemia in isolated working rat hearts and to investigate the mechanism of its action. Ischemia was initiated for 10, 15, 20 or 25 min with or without treatment with PGI2. Increasing durations of ischemia resulted in a progressive decline in high energy phosphate (HEP) stores, an elevation in tissue lactate, and incomplete recovery of function with reperfusion. Prostacyclin at either 1 or 10 ng/ml had no effect on HEP levels or total adenine nucleotides, and tissue lactate was not significantly affected by PGI2 in hearts made ischemic for 10 to 20 min, but both PGI2 concentrations significantly elevated lactate levels after 25 min ischemia. Reperfusion recovery of left ventricular function was complete following 10 and 15 min ischemia, but incomplete recovery was evident following 20 min ischemia (77% of pre-ischemic function); and although PGI2 had no direct effect on the function of aerobically perfused hearts, recovery of aortic flow with 1 ng/ml PGI2 after 20 min of ischemia was reduced to approximately 20% (P less than 0.01). This depression in recovery was associated with significantly increased lactate levels during reperfusion. At a concentration of 10 ng/ml PGI2 did not depress ventricular recovery or elevate lactate content after 20 min ischemia. When hearts made ischemic for 20 min were analyzed, a significant negative correlation was found between ventricular recovery (aortic flow rate) and lactate concentration; however, no correlation existed between recovery and ATP levels. After 25 min of ischemia, five of eight (62.5%) untreated hearts demonstrated some degree of ventricular recovery, however, only two of ten hearts studied demonstrated any measurable functional recovery with either PGI2 concentration. This effect of PGI2 to reduce or prevent recovery of ventricular function following either 20 or 25 min of ischemia as well as the corresponding elevation in lactate levels was prevented by treatment with the calcium channel blocker verapamil. This study therefore shows that PGI2 at critical low concentrations can depress left ventricular recovery following total ischemia. This effect of PGI2 becomes more pronounced as ischemia duration is prolonged and is associated with elevated tissue lactate levels. The studies with verapamil suggest that PGI2 may be acting via the slow calcium channel to increase lactate levels and depress ventricular recovery following prolonged periods of ischemia.

Adenine Nucleotides↗

PubMed1986

Cytoplasmic concentrations of inorganic phosphate affect the critical concentration for assembly of actin in the presence of cytochalasin D or ADP.

Skeletal muscle actin labelled with pyrene was used to measure the critical concentration (Cc) for assembly in conditions designed to approximate the ionic environment in the cytoplasm. Under these conditions (0.1 M-KCl, 2 mM-MgCl2, 1.1 mM-ATP, 0.1 mM-CaCl2, 0.5 mM-ethyleneglycol-bis(beta-aminoethylether)N,N'-tetraacetic acid, 0.25 mM-2-mercaptoethanol, 20 mM-imidazol X HCl, pH 7.0), the steady-state Cc value was estimated to be 0.07 microM (3.0 micrograms/ml), and, consistent with previous observations, the Cc increased to 0.20 microM (8.7 micrograms/ml) in the presence of 10(-6) M-cytochalasin D, and to 1.10 microM (47 micrograms/ml) after conversion of ATP to ADP using hexokinase and glucose. Addition of inorganic phosphate (Pi) at concentrations up to 20 mM caused only a slight decrease in the steady-state Cc, but at 2 mM-Pi (a reasonable estimate of cytoplasmic concentrations) the increase in Cc due to cytochalasin D was abolished, and at higher Pi concentrations there was even a slight decrease. Increasing Pi concentrations also progressively reduced the steady-state Cc for ADP-actin close to that for ATP-actin. These results are consistent with an increased affinity of ADP-actin for the polymer in the presence of Pi. To determine whether these effects of Pi were simply mass action effects on hydrolysis of bound ATP by polymerized actin, the stoichiometry of ATP hydrolysis during actin assembly was estimated and found to be at unity within the limits of experimental error and to be unaffected by Pi up to 20 mM. In addition, actin depolymerized by removal of ATP using glucose and hexokinase rapidly reassembled after addition of 20 mM-Pi. These results are interpreted by a mechanism involving the formation of ADP-Pi-actin species and are discussed in relation to the phenomenon of treadmilling and the theory of dynamic instability, and the potential for their occurrence in cells.

Actin Cytoskeleton↗