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PubMed1994

Adventitious redox-active metals in Krebs-Henseleit buffer can contribute to Langendorff heart experimental results.

Adventitious redox-active metals in Krebs-Henseleit buffer exhibit a significant enhancement of damage to isolated rat hearts. Using atomic absorption spectroscopy, it was determined that Krebs-Henseleit buffer contains substantial amounts of contaminating iron and copper. Significant copper contamination was found in ACS Reagent grade sodium chloride and sodium bicarbonate; iron contamination in sodium chloride, potassium chloride, sodium bicarbonate, and calcium chloride. Chelating resin treatment of individual reagents was found to decrease copper content of Krebs-Henseleit buffer from 0.32 to 0.17 microM. Using salicylate as a probe for .OH formation, it was determined that considerable amounts of this radical are formed when 0.25 mM ascorbate is added to the buffer indicating significant metal-catalysed autoxidation. Isolated rat hearts, perfused with non-chelexed Krebs-Henseleit buffer plus 0.25 mM ascorbate for 60 min, sustained moderate injury with developed systolic pressure, +dP/dtmax and -dP/dtmax decreased by 30 to 35% by the end of experiment. Hearts perfused with chelating resin-treated Krebs-Henseleit buffer sustained no significant injury within the same time frame. Furthermore, it was observed that hearts perfused with non-chelexed Krebs-Henseleit buffer accumulate significant amounts of copper depending on the amount of contamination and length of perfusion. Significant effects on post-ischemic end diastolic pressure were observed in hearts perfused with a Krebs-Henseleit buffer subsequently found to be contaminated with high levels of copper. These results clearly demonstrate that adventitious redox-active transition metals may be a confounding factor in experimental results. Further, it is recommended that all perfusion media be routinely examined for adventitious metals and treated if deemed necessary.

Animals↗
PubMed1994

Frog striated muscle is permeable to hydroxide and buffer anions.

Hydroxide, bicarbonate and buffer anion permeabilities in semitendinosus muscle fibers of Rana pipiens were measured. In all experiments, the fibers were initially equilibrated in isotonic, high K2SO4 solutions at pHo = 7.2 buffered with phosphate. Two different methods were used to estimate permeabilities: (i) membrane potential changes were recorded in response to changes in external ion concentrations, and (ii) intracellular pH changes were recorded in response to changes in external concentrations of ions that alter intracellular pH. Constant field equations were used to calculate relative or absolute permeabilities. In the first method, to increase the size of the membrane potential change produced by a sudden change in anion entry, external K+ was replaced by Cs+ prior to changes of the anion under study. At constant external Cs+ activity, a hyperpolarization results from increasing external pH from 7.2 to 10.0 or higher, using either CAPS (3-[cyclohexylamino]-1-propanesulfonic acid) or CHES (2-[N-cyclohexylamino]-ethanesulfonic acid) as buffer. For each buffer, the protonated form is a zwitterion of zero net charge and the nonprotonated form is an anion. Using reported values of H+ permeability, calculations show that the reduction in [H+]o cannot account for the hyperpolarizations produced by alkaline solutions. Membrane hyperpolarization increases with increasing total external buffer concentration at constant external pH, and with increasing external pH at constant external buffer anion concentration. Taken together, these observations indicate that both OH- and buffer anions permeate the surface membrane. The following relative permeabilities were obtained at pHo = 10.0 +/- 0.3: (POH/PK) = 890 +/- 150, (PCAPS/PK) = 12 +/- 2, (PCHES/PK) = 5.3 +/- 0.9, and (PNO3/PK) = 4.7 +/- 0.5. PNO3/PK was independent of pHo up to 10.75. At pHo = 9.6, (PHCO3/PK) = 0.49 +/- 0.03; at pHo = 8.9, (PCl/PK) = 18 +/- 2 and at pHo = 7.1, (PHEPES/PK) = 20 +/- 2. In the second method, on increasing external pH from 7.2 to 10.0, using 2.5 mM CAPS (total buffer concentration), the internal pH increases linearly with time over the next 10 min. This alkalinization is due to the entry of OH- and the absorption of internal H+ by entering CAPS- anion. The rate of CAPS- entry was determined in experiments in which the external CAPS concentration was increased at constant external pH. Such increases invariably produced an increase in the rate of internal alkalinization, which was reversed when the CAPS concentration was reduced to its initial value.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1977

The influence of carbon dioxide, bicarbonate and other buffers on the potential of antimony microelectrodes.

Antimony microelectrodes were calibrated at 37 degrees C in phosphate buffers, in different bicarbonate solutions at various CO2-partial pressures and in buffers like TRIS1, TES2, MES3 and malonic acid. By use of the latter buffers (with exception of malonic acid) the most reliable calibration curves were obtained ("normal values"). The usual calibration in 67 mmol/l standard phosphate buffers turned out to be unacceptable because the obtained mV-values were too high (negative) in comparison to all other buffers. Different calibration curves resulted from the use of pure bicarbonate solution whether the pH-values were changed by variation of pCO2 or of the bicarbonate concentration. Low bicarbonate concentrations in combination with low pCO2 gave mV-values which were too low relative to the other buffers. Both the increase of pCO2 as well as of the bicarbonate concentration caused a shift of the potential of the antimony electrodes toward "normal values". In solutions containing other buffers the influence of bicarbonate and pCO2 became negligible with increasing buffer concentration. Decreasing oxygen partial pressure was found to cause an increase of the potential of the antimony electrodes. The influence of liquid junction potentials at the reference electrode is discussed.

Antimony↗
PubMed1982

The adverse effects of HEPES, TES, and BES zwitterion buffers on the ultrastructure of cultured chick embryo epiphyseal chondrocytes.

Chick embryo epiphyseal chondrocytes cultured in media containing HEPES, TES, and BES zwitterion buffers, used in combination or independently, consistently developed cytoplasmic vacuoles. This cytoplasmic vacuolation was resolved when the zwitterion buffered media was replaced by media containing bicarbonate:CO2 enriched air buffer. Vacuoles were infrequent or absent in cultures grown in bicarbonate:CO2 enriched air. Chondrocytes with an established extracellular matrix showed less vacuolation than fibroblastlike and polygonal shaped cells that lacked such a matrix. The granular endoplasmic reticulum and Golgi dictyosomes of zwitterion buffered chondrocytes were distended and contained a flocculent amorphous material. Cytoplasmic vacuoles (0.5 to 3.0 micron diam) formed by the fusion and intracellular accumulation of Golgi vesicles and vacuoles also contained a flocculent material enhanced by ruthenium red. Membrane bound extracellular vacuoles containing ruthenium red stained proteoglycan aggregates were common in the extracellular matrix of zwitterion buffered cultures but were generally absent from bicarbonate treated cultures. Electron dense calcium deposits seemed much larger and more numerous in the presence of zwitterion buffers. It is suggested that HEPES, TES, and BES buffers, used alone or in combination, may adversely affect cell membrane systems, and thus the transport or secretory mechanisms operative in cultured chondrocytes, or both, resulting in vacuole formation and the intracellular accumulation of synthesized export material. Although the mechanism by which HEPES, TES, and BES induce these changes remains unclear, the use of zwitterion buffers in biological preparations should be treated with caution.

Alkanesulfonates↗
PubMed1998

Maintenance of acidic lateral intercellular spaces by endogenous fixed buffers in MDCK cell epithelium.

The lateral intercellular spaces (LIS) of MDCK cell epithelia grown on permeable supports are about 0.4 pH units acidic to the bathing solutions, presumably because of buffering by the fixed negative charges on the surface of the lateral cell membranes. To test the hypothesis that fixed buffers are responsible for the acidity, a theoretical and experimental approach was developed for the determination of the concentration and pK of the fixed buffer constituted by the glycocalyx. The pH of the solution in the LIS was measured by ratiometric fluorescence microscopy while the buffer concentration or composition of the bathing solutions was altered. In addition, the divalent cation Sr2+ was added to the perfusion solutions to displace protons from the fixed buffer sites for the determination of the fixed buffer properties. We conclude that the LIS contain 3.7 mm of pK 6.2 fixed buffer and that this buffer is responsible for the acidic microenvironment in the LIS.

Animals↗
PubMed2004

Nuclear telomerase is less accessible to antibody probing than known nuclear antigens: retrieval with new immunostaining buffer.

Telomerase is an important tumor marker but few antibodies to the enzyme have been described or used without difficulty in histochemical detection. Here we report specific detection of the enzyme in cell and tissue preparations using a new monoclonal antibody (mAb 476) and a new antigen-retrieval buffer (Enhancing buffer). When used to detect telomerase under normal immunostaining conditions in HL-60 cells or tissue sections of hepatocellular carcinoma or metastatic choriocarcinoma, unexpectedly, the antibody stained the cytoplasm rather than the nucleus. Nuclear staining, however, was revealed using the Enhancing buffer. Since other nuclear antigens in the HL-60 cell could be stained both ordinarily and in the Enhancing buffer, nuclear telomerase appears to be shrouded by the nuclear matrix or blocked by accessory proteins. The cytoplasmic activity seen in normal buffer but absent largely from the Enhancing buffer may be an artifact or the nascent, "naked" enzyme. With a known cytoplasmic antigen (proteinase-3) chosen arbitrarily for comparison, the antigenicity was found enhanced, instead, by the Enhancing buffer. The mode of action of the Enhancing buffer differs from that of microwave irradiation or the signal amplification (CSA) used by some investigators. The latter was found to enhance the cytoplasmic reactivity rather than the nuclear reactivity of mAb 476.

Animals↗
PubMed1987

The role of fixed and mobile buffers in the kinetics of proton movement.

We derive a simple expression for the effective diffusion coefficient of protons in Fick's second law, Deff, when both spatially fixed, HF, and mobile, HM, buffers are present. These buffers are present at moderately high concentrations ([Ftot], [Mtot] greater than 1 mM) in most biological systems. We consider only the case where the protonation reactions remain at equilibrium during the diffusion process. When the pH is to the alkaline side of the pK values of the fixed and mobile buffers ([H+] less than KF, KM), the effective diffusion coefficient of protons in Ficks second law is: (Formula: see text) where DH is the diffusion coefficient of the protons free in the aqueous phase and DHM is the diffusion coefficient of the mobile buffer. The equation illustrates three features of diffusion in a buffered system. Firstly, the effective diffusion coefficient of protons is always lower than the diffusion coefficient of free protons. Secondly, increasing the concentration of fixed buffers always decreases Deff. Thirdly, increasing the concentration of mobile buffer can increase Deff when fixed buffers are present.

Biological Transport↗
PubMed1979

Formation of inside-out vesicles of Bacillus licheniformis. Dependence on buffer composition and lysis procedure.

1. The extent to which the cytoplasmic membrane of the Gram-positive bacterium Bacillus licheniformis formed inside-out vesicles was studied with the freeze-fracture technique. The membrane orientation appeared to be dependent on the buffer compositon as well as on the lysis procedure used. 2. By manipulating these conditions, membrane preparations were obtained with the percentage of inside-out vesicles varying from 15 to 80%. 3. More vesicles had the opposite orientation when the cells were lysed in potassium phosphate buffer than when they were lysed in sodium phosphate buffer. Tris-HCl buffer favoured the formation of inside-out vesicles more than phosphate buffer. 4. Lysis of protoplasts in hypotonic buffers resulted in more inside-out vesicles than did direct lysis of cells in hypotonic media. 5. In an attempt to explain the observed differences, experiments were performed in which the morphology of thin-sectioned lysing cells in sodium phosphate buffer was compared with that in potassium phosphate buffer. The results from these experiments indicate that the formation of inside-out vesicles is brought about by an effect on the membrane itself rather than on the cell wall, on the cell wall membrane association, or on the cytoplasm.

Bacillus↗
PubMed1989

Assay sensitivity and differentiation of monoclonal antibody specificity in ELISA with different coating buffers.

Buffers of different pH and ionic strength were employed as coating buffers for antigen adsorption to microtitre plates. Their efficiency for coating plates with rinderpest virus (RPV) and foot-and-mouth disease virus (FMDV) antigens was studied by ELISA with polyclonal and monoclonal antibody preparations. While the adsorption and detection of RPV antigen with polyclonal antiserum was highly dependent on the ionic strength and pH of coating buffer, adsorption of antigenically active FMDV antigen was relatively unaffected by the buffering conditions. Both antigens were adsorbed optimally in 0.01 M phosphate buffer, pH 8.0. When monoclonal antibodies were used to detect antigen, there was a greater degree of dependence on the coating buffer than that found with polyclonal antisera. Moreover, when they were used to detect antigen adsorbed under several buffering conditions, monoclonal antibodies showed a variety of preferred buffers. The usefulness of this differential reactivity in distinguishing epitope specificity is demonstrated.

Antibodies, Monoclonal↗
PubMed1986

Isoelectric focusing using non-amphoteric buffers in free solution: I. Determination of stable concentration profiles.

For large-scale separations of proteins, the use of simple non-amphoteric buffers in free solution and in multicompartment electrolyzers seems promising for industrial applications. The stabilization of a pH profile with this type of buffer requires the strict observation of two conditions: choice of an adequate buffer; stationary profiles of concentrations. During electrolysis in free solution, the ions of the buffer are displaced across the compartments by migration and by diffusion. To keep a stationary composition, the inflow and outflow of all individual ionic species through each compartment must be identical. At high current, diffusion may be neglected against migration and the ionic flows will be identical if the transport number of each ion is constant at each location within the cell. In these conditions, stationary compositions will be independent of the electric current. This condition of constant transport numbers implies the use of profiles of buffer concentrations different from those published up to now. The new equations for these profiles of concentrations are given in the present paper. The constant migration of the ions must be compensated in the end compartments of the isoelectric focusing cell to provide a stable steady state. Two methods are proposed in the literature: the buffer renewal method and the external recycling method (rheoelectrolysis). Here modified buffer renewal method is proposed. Using stationary mass balances, analytical equations are given to calculate the flows and the composition of the solutions to be recycled or added. Using these equations and the profiles of concentrations to keep constant transport numbers, it is demonstrated that only a renewal of the buffers in the end compartments may lead to stable pH profiles and thus to valid conditions of separation.

Buffers↗
PubMed1992

Kinetics of diffusion in a spherical cell. II. Solute buffering included.

This paper on diffusion kinetics in neurons presents an analysis of diffusion in the presence of solute buffering. Computational rather than theoretical methods are usually necessary since buffering generally precludes an analytical solution to the diffusion equations. As in the companion paper, our methods are illustrated in the context of calcium diffusion in a spherical cell. However, the same methods can be applied to the spread of any second messenger and other geometries. Analytical or computational predictions of the time course of diffusion and buffering may help guide further experiments and simulations. For example, simulations of calcium diffusion in a model of the bullfrog sympathetic ganglion cell show that buffering at depths greater than 5-6 microns is almost instantaneous compared to diffusion from sources at the cell membrane. Since buffering complicates the design of multicompartmental models, we demonstrate that a few compartments designed on the basis of diffusion alone (Carnevale and Rosenthal, 1992) may be a satisfactory framework for a model that includes bimolecular buffering. An analytical solution may be possible if the buffering reaction can be linearized. We describe a method for linearizing bimolecular saturating buffering, i.e., approximating it by a unimolecular non-saturating process that immobilizes solute. The analytical solution for the linearized reactive diffusion problem fits a non-linear model of calcium movement in the bullfrog sympathetic ganglion cell quite well after a few milliseconds.

Animals↗
PubMed1991

Alkali cation effect on carbonyl-hemoglobin's and -myoglobin's conformer populations when exposed to freeze-concentration of their phosphate-buffered aqueous solutions.

Freeze-concentrated aqueous phosphate-buffered (pH 6.8) solutions of carbonyl-hemoglobin (HbCO) and -myoglobin (MbCO) were investigated by Fourier-transform infrared spectroscopy for the effect of alkali cation on the population of conformers. When using sodium phosphates as buffer components, HbCO was transformed from conformer III (at approximately 1951 cm-1) which is the dominant form at ambient temperatures, into conformer IV (at buffer concentration at a given temperature. The conformational changes started slightly below the temperature where ice began to crystallize and the remaining solution became freeze-concentrated, and they were reversible for HbCO. For MbCO in 0.5 M sodium phosphate buffer solution, however, they were irreversible and MbCO denatured completely. When potassium phosphate salts were used for preparing the buffer at the same pH of 6.8, little or no transformation of conformer III into conformer IV was observed. The conformational changes induced by sodium salts are attributed to a decrease in pH and it is shown by infrared spectroscopy that during freeze concentration drastic changes in composition of the two buffer components H2PO4-/HPO(2)4- occur, the acid component increasing strongly relative to the base component. Supersaturation is also important because change from conformer III to IV requires a minimum concentration of sodium salts: whereas 0.1 M sodium phosphate buffer concentration shows a strong effect, 0.03 M concentration does not and therefore behaves like a potassium phosphate buffer.

Animals↗
PubMed1996

Influence of alkaline buffers on cytoplasmic pH in myocardial cells exposed to metabolic acidosis.

The influence of different clinically used alkaline buffers on cytoplasmic pH in normal as well as acidotic rat myocardial cells was investigated in this study by means of the fluorescent intracellular probe 2',7'-bis-(carboxyethyl)-5,6-carboxyfluorescein acetoxymethyl ester (BCECF-AM). It was shown that both sodium bicarbonate and Tris buffer mixture (Tribonat) caused a significant and dose-dependent acidification of the cytoplasm of suspended myocardial cells with normal initial intracellular pH. This decrease was followed by a slow increase during the observation period. The initial cytoplasmic pH value was more easily reached when Tris buffer mixture was used. Ringer's acetate also caused a decrease of intracellular pH, but this change persisted and was further amplified during the experiment. Carbicarb in larger dosages as well as pure trometamol (Tris) caused a pronounced dose-dependent and lasting intracellular alkalinization. Intracellular acidosis was achieved by preincubating the cells in sodium acetate. Addition of sodium bicarbonate caused an initial and dose-dependent acidification of the cytoplasm followed by a slow increase to values slightly above the induced acidosis. In contrast, Tris buffer mixture showed a tendency towards an initial acidification only when larger dosages were used, and correction of the induced acidosis was possible by use of moderate to large volumes. Ringer's acetate produced a lasting and dose-dependent decrease of cytoplasmic pH, while Carbicarb and pure trometamol caused an immediate, pronounced and persistent alkalinization. Myocardial cells with low initial cytoplasmic pH due to preincubation in an acid buffer also showed an early decrease of intracellular pH after addition of sodium bicarbonate and Tris buffer mixture. In the case of sodium bicarbonate correction of the acid-base disturbance was not achieved during the observation period, while this was accomplished by use of larger volumes of Tris buffer mixture. Carbicarb in larger volumes caused an increase in intracellular pH. The most significant and persistent increases of cytoplasmic pH was achieved by use of pure trometamol. In conclusion, the present in vitro study implies that Tris buffer mixture (Tribonat) is well-suited for correction of intracellular acidosis since it acts without causing a pronounced initial intracellular acidosis or a later potentially hazardous huge cytoplasmic alkalinization.

Animals↗
PubMed2005

Oxidation of Good's buffers by hydrogen peroxide.

Good's zwitterionic buffers are widely used in biological and biochemical research in which hydrogen peroxide is a solution component. This study was undertaken to determine whether Good's buffers exhibit reactivity toward H(2)O(2). It is found that H(2)O(2) oxidizes both morpholine ring-containing buffers (e.g., Mops, Mes) and piperazine ring-containing zwitterionic buffers (e.g., Pipes, Hepes, and Epps) to produce their corresponding N-oxide forms. The percentage of oxidized buffer increases as the concentration of H(2)O(2) increases. However, the rate of oxidation is relatively slow. For example, no oxidized Mops was detected 2h after adding 0.1M H(2)O(2) to 0.1M Mops (pH 7.0), and only 5.7% was oxidized after 24h exposure to H(2)O(2). Thus, although all of these buffers can be oxidized by H(2)O(2), their slow reaction does not significantly perturb levels of H(2)O(2) in the time frame and at the concentrations of most biochemical studies. Therefore, the previously reported rapid loss of H(2)O(2) produced from the ferroxidase reaction of ferritin is unlikely due to reaction of H(2)O(2) with buffer, a conclusion supported by the fact that H(2)O(2) is also lost rapidly when the solution pH of the ferroxidase reaction is controlled by a pH stat apparatus in the absence of buffer.

Buffers↗
PubMed2004

Influence of buffering effects of dentifrices and fluoride gels on abrasion on eroded dentine.

OBJECTIVE: Evaluation of abrasion resistance of eroded and non-eroded dentine brushed with experimental dentifrices and gels of varying buffering capacities. DESIGN: One hundred and twenty human dentine specimens were distributed among six groups (A-F) according to the different buffering capacities of experimental toothpastes and gels. The dentifrices (pH: 4.5) and gels (pH: 4.5) differed in fluoride content, RDA-value and buffer capacity (given as mg KOH needed for neutralisation): (A) 0.125%F, RDA: 77, 6mg KOH; (B) 0.125%F, RDA: 125, 24mg KOH; (C) 0.125%F, RDA: 129, 12mg KOH; (D) 0%F, RDA: 81, 1.55mg KOH; (E) 1.25%F, RDA: 21, 8.5mg KOH; (F) 0.125%F, RDA: 7, 3.11mg KOH. Ten specimens were cycled through three alternating demineralisations (1% citric acid, 5min) and remineralisations (artificial saliva, 1min) including brushing abrasion in an automatic brushing machine (2 x 2000 strokes). Ten samples of each group were not eroded, but only brushed. RESULTS: After three cycles, loss of dentine was determined by profilometry (mean +/- S.D. [microm]). Eroded samples: (A) 37.12+/-4.95; (B) 48.67+/-4.77; (C) 39.88+/-3.76; (D) 32.92+/-2.67; (E) 3.65+/-1.35; (F) 6.47+/-1.31. Uneroded samples: (A) 30.78+/-6.47; (B) 37.84+/-7.75; (C) 30.07+/-3.05; (D) 26.93+/-2.30; (E) 1.76+/-0.94; (F) 0.51+/-0.18. Analysis of variance revealed significantly higher abrasion values for the eroded compared to the non-eroded samples (P<or=0.05). Dentifrices with same pH-value, fluoride content and similar RDA-values but a higher buffering capacity exhibited significantly more abrasion than toothpastes with lower buffering capacity. Highly-concentrated fluoride gel with higher buffering capacity lead to less abrasion values compared to the gel with low fluoride concentration and low buffering capacity. CONCLUSION: Toothbrushing abrasion behaviour of eroded dentine may be influenced by the fluoride content, RDA-value and buffering capacity of the applied dentifrice or gel.

Buffers↗
PubMed2006

Corneal calcification after chemical eye burns caused by eye drops containing phosphate buffer.

PURPOSE: Chemical burns with calcium containing corrosives as well as irrigation with phosphate buffer solutions after eye burns bear the risk of corneal calcification. The aim of this study was to evaluate the correlation between the occurrence of corneal calcification after chemical injuries and the usage of phosphate buffer containing local therapeutics. METHODS: We reviewed the data of 179 patients who have been treated in the University Eye Clinic Aachen, Germany, between 1941 and 2000. Only when the corrosive did not contain calcium and when the initial irrigating solution did not contain phosphate buffer, respectively, were patients included in the study. The cases were analyse, if the patient was treated with phosphate buffer containing eye drops/ointment during the first 7 days of hospitalization or as an out-patient, and if corneal calcification was visible by slit-lamp examination during the follow-up. Statistical analysis was performed using Fischer's exact test. RESULTS: 152 eyes were included. From 63 eyes treated with phosphate buffer containing eye drops, 31 eyes (49%) developed corneal calcification. From 89 eyes treated without phosphate buffer containing eye drops, only 23 eyes (26%) developed corneal calcification. The two-sided p-value of Fischer's exact test is 0.0036. CONCLUSION: During follow-up after chemical eye burns, eye drops containing phosphate buffer double the risk of corneal calcification. We recommend avoiding these agents in order to prevent the burned cornea from additional opacity. Substances containing phosphate buffer are listed in this article.

Buffers↗
PubMed2004

Role of the buffer in retention and adsorption mechanism of ionic species in reversed-phase liquid chromatography. I. Analytical and overloaded band profiles on Kromasil-C18.

The influence of the pH, the concentration, and the nature of the buffer on the retention and overloading behavior of propranolol (pKa = 9.25) on Kromasil-C18 was studied at 2.75 < pH < 6.75, using four buffers (phosphate, acetate, phthalate, and succinate), at three concentrations, 6, 20, and 60 mM. The propranolol band profiles were recorded for three sample sizes, less than 1 microg and 375 microg (sample less concentrated than the buffer), and 7500 microg (band more concentrated than the buffer). Results showed that the buffer concentration, not its pH, controls the retention time of propranolol, in agreement with the chaotropic model. The retention factor depends also on the nature of the buffer, particularly the valence of the basic anion. At moderate loading, the band profiles are well accounted for by a simple bilangmuir model (no adsorbate-adsorbate interactions) with the monovalent anions H2PO4- (pH 2.75), HOOC-Ph-COO- (pH 2.75), HOOC-CH2-CH2-COO- (pH 4.16) and CH3COO- (pH 4.75), and by a bimoreau model (significant adsorbate-adsorbate interactions) with the bivalent anions OOC-Ph-COO- (pH 4.75), OOC-CH2-CH2-COO- (pH 5.61) and HPO4(2-) (pH 6.75). The isotherm were determined using the inverse method. The results show that both the saturation capacity and the equilibrium constant on the low-energy sites increase with increasing buffer concentration, a result similar to that observed with neutral salts. For bivalent anions, the adsorbate-adsorbate interactions are much stronger on the low than on the high energy sites. The density of high energy sites is lower and the equilibrium constant on the low energy sites are higher with bivalent than with univalent anions. These results are consistent with the formation of a propranolol-buffer (2:1) complex with bivalent anions.

Adsorption↗
PubMed2003

Buffers and oscillations in intracellular Ca2+ dynamics.

I model the behavior of intracellular Ca(2+) release with high buffer concentrations. The model uses a spatially discrete array of channel clusters. The channel subunit dynamics is a stochastic representation of the DeYoung-Keizer model. The calculations show that the concentration profile of fast buffer around an open channel is more localized than that of slow buffers. Slow buffers allow for release of larger amounts of Ca(2+) from the endoplasmic reticulum and hence bind more Ca(2+) than fast buffers with the same dissociation constant and concentration. I find oscillation-like behavior for high slow buffer concentration and low Ca(2+) content of the endoplasmic reticulum. High concentration of slow buffer leads to oscillation-like behavior by repetitive wave nucleation for high Ca(2+) content of the endoplasmic reticulum. Localization of Ca(2+) release by slow buffer, as used in experiments, can be reproduced by the modeling approach.

Buffers↗