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PubMed2002

Increased Ca2+ buffering function of sarcoplasmic reticulum in small mesenteric arteries from spontaneously hypertensive rats.

We compared the Ca2+ buffering function of the superficial sarcoplasmic reticulum (SR) during rest and during contraction in endothelium-denuded strips of small mesenteric arteries from 13-week-old spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto rats (WKY). The addition of caffeine (1-20 mM) caused a transient contraction in both strains, and the contraction was significantly larger in SHR. When the SR Ca2+ buffering function was eliminated by cyclopiazonic acid (CPA; 10 microM) or thapsigargin (100 nM), both of which inhibit SR Ca2+-ATPase, or by ryanodine (10 microM), which depletes the SR Ca2+, there was a larger contraction in SHR than in WKY, suggesting that the Ca2+ buffering function of the SR during rest is more important in SHR than in WKY. Judging from the augmenting effects of these three agents on the contractile responses to Bay k 8644 (1-300 nM), an agonist of L-type Ca2+ channels, or norepinephrine (10(-9)-10(-4) M), an alpha-adrenoceptor agonist, the effects were significantly greater in SHR than in WKY. We conclude that 1) the Ca2+ influx during rest and during stimulation with Bay k 8644 or norepinephrine is strongly buffered by Ca2+ uptake into the superficial SR in the small mesenteric arteries from SHR and WKY; and 2) these Ca2+ buffering functions are increased in SHR because of the larger capacity of SR for Ca2+ storage.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗
PubMed2006

Application of near-infrared and Fourier transform infrared spectroscopy in the characterization of ligand-induced conformation changes in folate binding protein purified from bovine milk: influence of buffer type and pH.

Fourier transform infrared (FT-IR) and near-infrared (NIR) spectroscopy have been applied to detect structural alterations in folate binding protein (FBP) induced by ligation in different buffer types. The amide I region pointed to a beta-sheet to alpha-helix transition upon ligation in acetate and phosphate buffers, and the formation of intermolecular beta-sheet was indicated at pH 5.0, in agreement with a dimerization of FBP taking place at this pH. The ligand-induced changes in the 2100-2300 nm NIR region were significant for FBP in acetate and phosphate buffers of pH 5.0, and the variations were interpreted as secondary structure changes, based on previous assignments of secondary structures to the combination bands in the NIR region. In the case of acetate buffer, variations in the amide combination bands agreed with the amide I analysis, but for the other buffer types some discrepancies were found and explained by side-chain contributions to the NIR, which could reflect the tertiary and quaternary structure differences. NIR spectra of FBP at pH 7.4 and 5.0 revealed contradictory effects on the side chains, reflecting different polymerization events at the two pH values, whereas the amide I region indicated similar changes at the two pH values. Therefore, we suggest that FT-IR and NIR spectroscopy may complement each other, such that the two techniques in combination may give information on all three types of protein conformational changes. While the secondary structure changes are revealed by FT-IR, the tertiary and quaternary structure changes are reflected in the NIR spectra, although the general influence of the latter changes on the NIR spectra remains to be confirmed.

Animals↗
PubMed2005

The concentrations of calcium buffering proteins in mammalian cochlear hair cells.

Calcium buffers are important for shaping and localizing cytoplasmic Ca2+ transients in neurons. We measured the concentrations of the four main calcium-buffering proteins (calbindin-D28k, calretinin, parvalbumin-alpha, and parvalbumin-beta) in rat cochlear hair cells in which Ca2+ signaling is a central element of fast transduction and synaptic transmission. The proteins were quantified by calibrating immunogold tissue counts against gels containing known amounts of each protein, and the method was verified by application to Purkinje cells in which independent estimates exist for some of the protein concentrations. The results showed that, in animals with fully developed hearing, inner hair cells had 110 of the proteinaceous calcium buffer of outer hair cells in which the cell body contained parvalbumin-beta (oncomodulin) and calbindin-D28k at levels equivalent to 5 mm calcium-binding sites. Both proteins were partially excluded from the hair bundles, which may permit fast unbuffered Ca2+ regulation of the mechanotransducer channels. The sum of the calcium buffer concentrations decreased in inner hair cells and increased in outer hair cells as the cells developed their adult properties during cochlear maturation. The results suggest that Ca2+ has distinct roles in the two types of hair cell, reflecting their different functions in auditory transduction. Ca2+ is used in inner hair cells primarily for fast phase-locked synaptic transmission, whereas Ca2+ may be involved in regulating the motor capability underlying cochlear amplification of the outer hair cell. The high concentration of calcium buffer in outer hair cells, similar only to skeletal muscle, may protect against deleterious consequences of Ca2+ loading after acoustic overstimulation.

Age Factors↗
PubMed1994

Brain buffering is restored in hyponatremic rats by correcting their plasma sodium concentration.

Previous studies from this laboratory showed that both acute and chronic hyponatremia impaired active brain buffering. These studies were performed to determine whether correcting the plasma sodium restored normal buffering in hyponatremic rats. Acute (1- and 2-day) and chronic (7- and 14-day) hyponatremia was induced in male Sprague-Dawley rats by constant desmopressin administration combined with a liquid diet. Plasma sodium was corrected by stopping desmopressin for 6 h, substituting solid chow, and allowing free access to water. Studies were performed 24 h later. Uncorrected hyponatremic rats who continued to receive desmopressin and liquid diet served as controls. Brain pH was determined by [31P]NMR in rats anesthetized with N2O and paralyzed with pancuronium. Brain buffering was determined by the response to CO2 loading. Resting brain pH was the same in corrected and uncorrected rats, but the two groups responded differently to CO2 loading. Thus, 55 min after ventilation with 20% CO2, corrected rat brain pH was 0.13 pH units higher than in uncorrected rats despite statistically similar changes in CO2 tension and arterial pH in both groups. Moreover, 15 min into recovery from CO2 exposure, brain pH in corrected rats overshot resting pH by 0.07, whereas no overshoot occurred in uncorrected rats. Buffering in corrected rats was identical to that shown previously in normonatremic rats. The complete restoration of late-phase buffering achieved by normalizing the plasma sodium of hyponatremic rats indicates that at least some portion of active hydrogen ion transport is sodium dependent in the brain.

Acute Disease↗
PubMed2005

Theoretical analysis concerning the characteristics of a dye-binding method for determining serum protein based on protein error of pH indicator: effect of buffer concentration of the color reagent on the color development.

In the dye-binding method based on protein error of a pH indicator, the color development has been reported to be markedly affected by the buffer concentration of the color reagent. In this study, the author analyzed this phenomenon by a theoretical calculation based on the chemical equilibrium of protein error. The calculation was performed on the assumption that both the dissociated dye anion and the anion contained in the buffer solution react with protein, forming a dye-protein complex and an anion-protein complex, respectively. The calculated results were compared with those obtained by the experiments using bromophenol blue, bromocresol green and bromocresol purple that are employed widely for determining the human serum albumin concentration clinically. The calculated results of this method are summarized as follows: (1) the color development decreases with the increase in the concentration of the anion contained in the buffer solution; (2) the calibration curve is more linear in the higher concentration of the anion than in the lower one. These calculated results agreed well with the experimental ones. From these results, it was concluded that the change in the color development by the buffer concentration of the color reagent is due to the change in the concentration of the buffer anion.

Blood Proteins↗
PubMed2006

Analysis of the difference in color development in the dye-binding method due to the kind of buffer solution.

In a dye-binding method using a pH indicator, color development has reportedly been affected by the kind of buffer solution used in the color reagent. This phenomenon was analyzed by using a calculation based on the assumption that the anion of the buffer solution also reacts with protein. Color development decreases with increases in the anion concentration of the buffer solution and in the equilibrium constant of the reaction between the anion and protein. The differences in color development due to the kind of buffer solution can be attributed to differences in the equilibrium constant of the reaction forming the anion-protein complex and to the concentration of the anion between the buffer solutions.

Anions↗
PubMed1980

Physicochemical characteristics of antimony microelectrode with special reference to selection of standard buffers.

1) Antimony (Sb) microelectrodes with tips of 2 to 5 micrometers in outside diameter were constructed, and their electromotive forces (EMF) were tested in response to the pH of several calibration buffers. The data were compared with those of glass pH electrodes. 2) Increasing ionic strength (I) caused a significant deviation in pH readings of the Sb-microelectrode (delta pHSb-Glass). The above salt effect was empirically given by delta pHSb-Glass = 0.017--0.125 I (I = 0.09--0.22) 3) Even with the correction for ionic strength, the values of delta pHSb-Glass were still dependent on the nature of calibration buffers employed. Among various buffers, Tris buffer [Tris(hydroxymethyl)-aminomethane, (THAM)] had the least disturbance on the Sb-microelectrode, and bicarbonate had also relatively minor effect. 4) The pH value according to the Sb-electrode was caused to deviate considerably by phosphate. The effect of phosphate within the range of 1 to 67 mM can be predicted by 1/delta pHSb-Glass = 1 + (0.174/[PO4]) 5) Like phosphates, BES [N, N-Bis (2-hydroxyethyl)-2-aminoethane sulfonic acid] also had a significant effect on the EMF of the Sb-microelectrode. 6) Upon consideration of the above facts, measurements of several biological fluids, such as those of the in vivo pH of intratubular fluid of bullfrog kidney were carried out with the Sb-microelectrode calibrated by 90 mM Tris buffer. 7) Thus, it was concluded that the Sb-microelectrode when properly calibrated could be used for the pH measurement of ultraminute amounts of biological samples.

Animals↗
PubMed1998

Calculation of the buffering capacity of bicarbonate in the rumen and in vitro.

We describe a model to calculate the buffering capacity of bicarbonate in the rumen. The addition of NaHCO3 results in the release of CO2 from solution and eventually from the rumen via eructation. This process directly neutralizes ruminal acidity. The degree to which the process continues depends on the partial pressure of CO2 in the gas phase, the pH, and a constant (7.74), according to the Henderson-Hasselbalch equation: pH = 7.74 + log([HCO3-]/pressure of CO2 in atmospheres). The addition of NaHCO3 to buffer solutions and ruminal fluid under high pressure of CO2 increased pH as predicted. The buffering capacity of ruminal fluid under CO2 was greater at low pH than was previously determined by titration in air. In contrast, in vitro systems in which CO2 is not permitted to escape may result in reduced buffering capacity. In vitro systems in which excess CO2 may escape (under N2 gas pressure) may result in uncontrolled pH elevation. Dilution of ruminal fluid under constant pressure of CO2 decreased ruminal pH as predicted by the model. The pH under different pressures at equilibrium and the buffering capacity are easily calculated for in vitro and in vivo systems.

Animals↗
PubMed1983

Effects of buffers on ruminal rate of passage and degradation of soybean meal in steers.

Eight rumen-fistulated steers were randomly assigned to medium- and high-concentrate diets supplemented with 0, .75, 2.5 and 5% of either sodium bicarbonate or an artificial saliva salts mixture. Each animal was fed for 21 d at 75 g dry matter/kg body weight . 75. Rates of soybean meal (SBM) degradation were predicted by adjusting digestion rates in nylon bags with respective passage rates of chromium-mordanted SBM particles. Measures of rumen fermentation were made during the incubation period. Liquid dilution rate was determined with 51Cr-EDTA. The high-concentrate diet produced higher (P less than .05) liquid dilution rates than the medium-concentrate diet, but there were no differences in response to the two buffers (P greater than .05). The liquid dilution rates averaged across diets for 0, .75, 2.5 and 5% levels of buffer were 6.2, 6.3, 8.5 and 8.7%/h (SE = .03) and passage rates for SBM were 5.2, 5.2, 6.5 and 6.7%/h (SE = .025). The 2.5 and 5% levels of buffer increased the rate of disappearance of SBM from the nylon bags and buffers fed at these levels also increased rumen pH and NH3-N concentration. Rumen pH was correlated with N disappearance from the nylon bag (r = .903, P less than .05). Buffer levels did not affect degradation rates of SBM.

Animals↗
PubMed1991

Effect of a buffering sugar-free lozenge on intraoral pH and electrochemical action.

Two double-blind crossover studies were performed to test a sugar-free lozenge containing bicarbonate and phosphate buffers (Profylin). The studies were performed in groups of 20 and 13 individuals. In Study I active buffering or placebo lozenge (not buffered) was given, and the pH of plaque and saliva was measured after 2, 5, 10, 20, and 30 min. In study II the lozenges were given 10 min after a sucrose rinse, and both the pH and the potential and polarization of amalgam restorations that made contact in the oral cavity were measured. In study I both lozenges increased the pH of plaque and saliva, but the values after sucking on the active lozenge were significantly higher than after placebo. In study II a pH recovery of plaque and saliva after the sucrose rinse was recorded for both types of lozenge, but it was most pronounced for the active, buffering lozenge. A statistically significant difference was, however, found only 5 min after sucking on the lozenge. No influence on the current magnitude was observed. The results thus indicate that the buffering sugar-free lozenge raises the pH of plaque and saliva and accelerates the pH recovery after a sucrose rinse but seems to have no influence on the galvanic current magnitude of amalgam restorations in contact.

Buffers↗
PubMed1990

Electron dense artefactual deposits in tissue sections: the role of ethanol, uranyl acetate and phosphate buffer.

The occurrence of electron dense deposits in sections of aldehyde-fixed tissue prepared for transmission electron microscopy has been attributed to a number of conflicting factors. In an attempt to clarify this, the precipitating effect of different combinations of phosphate or cacodylate buffer, glutaraldehyde, ethanol and uranyl acetate was investigated in test tubes. As a preliminary investigation the combination of phosphate buffer, ethanol and uranyl acetate was investigated in heart and kidney tissue fixed in glutaraldehyde with or without postosmication. The essential factors in the formation of electron dense deposits in these tissues appear to be phosphate buffer, ethanol, and uranyl acetate, although glutaraldehyde may contribute in some way. The nature and intensity of the deposits seem to vary with the sequence of combination of these factors. Osmium did not appear to be an essential factor in the reaction since deposits were observed in both osmicated and unosmicated tissue. To avoid such deposits, a postosmication distilled water wash for 20 to 30 min followed by en bloc staining with aqueous uranyl acetate is advised if phosphate buffer is used as a fixative vehicle or buffer wash after the primary fixative.

Animals↗
PubMed1987

An oxalate-sodium hydroxide buffer to study the role of milk replacer coagulation in preruminant calves.

An oxalate-NaOH buffer, which prevents coagulation in the abomasum of young calves, was added to a nonclotting milk replacer based on high heat skim milk powder, to investigate effects of this buffer on digestibility and metabolism of dietary components. Twelve Holstein male calves were allotted at random in two groups of six animals each and fed the experimental milk replacers from 3 to 17 d of age. Digestibility of dry matter, protein, and fat was similar for both control and buffered milk replacers. Concentrations of plasma glucose, triglycerides, alpha-amino nitrogen, and urea were the same in animals receiving buffer treated milk replacer and control diets. The data suggests that this oxalate-NaOH buffer would be suitable to prevent milk clotting in studies dealing with the importance of coagulation in digestion and metabolism in preruminant calves.

Abomasum↗
PubMed1988

Effects of a new multielement buffer on production, ruminal environment, and blood minerals of lactating dairy cows.

Twelve multiparous Holstein cows were used to evaluate the capacity of a multielement compound consisting mainly of northupite and sylvite to alleviate low milk fat percent. Possible mechanisms of action were assessed. Cows were arranged in a 4 X 4 Latin square design replicated three times. Basal diet was 55% concentrate:45% forage fed ad libitum. Dietary treatments were control, NaHCO3 at 1% of diet DM, and multielement buffer at 1 and 3% of diet DM. Feed intake and milk production were similar for all treatments. Milk fat percentages for the four treatments were 2.97, 3.21, 3.43, and 3.67%, respectively. A shift toward a higher molar percent of ruminal acetate and a lower molar percent of valerate appeared to coincide with changes in milk fat percentage. Milk protein percentage also was increased by supplemental multielement buffer. Ruminal fluid acidity was reduced by NaHCO3 and multielement buffer. Extent of in situ digestion of forage DM and cellulose was improved when cows consumed a buffering agent. Rate of corn silage digestion tended toward improvement. As mineral buffer was consumed, concentrations of Mg and K increased in ruminal fluid and blood. As dietary Na intake increased, ruminal and plasma Cl concentrations were depressed and plasma S and Ca were elevated.

Animals↗
PubMed1991

Relative buffering capacity of goat milk, cow milk, soy-based infant formulas and commercial nonprescription antacid drugs.

Buffering capacities of goat milk (Alpine, Nubian), cow milk (Holstein, Jersey), soy-based infant formulas, and non-prescription antacid drugs were estimated. Total N, protein, NPN, and P2O5 as major buffering entities were quantified for each milk category. Nubian goat milk had the highest levels of the three major buffering chemical entities, and the infant formulas contained less total N and NPN compared with natural goat and cow milks. Buffering capacities of the formulas also were lower than those of natural milks. Combinations of milk and antacid drugs had higher buffering capacities than either the milk or drug alone. Drug plus goat milk combinations upon addition of more than 2 ml of acid titrant exhibited fewer changes in pH than the respective drug plus cow milk combinations.

Acid-Base Equilibrium↗
PubMed1994

Effect of buffer addition to high grain total mixed rations on rumen pH, feed intake, milk production, and milk composition.

Lactating Holstein cows (n = 56) were fed high grain TMR in three feeding experiments to determine the effects of two rumen buffers on feed intake, milk production, and milk composition. The basal diet in Experiment 1 was 40% grass silage, 12% whole cottonseed, and 48% concentrate with treatments of 1) control (no added buffer), 2) 1.5% of dietary DM as Alkaten, and 3) 1.5% of dietary DM as Rumen 8. In Experiment 2, diets consisted of 12% corn and sunflower silage, 20% whole cottonseed, and 68% concentrate with treatments as in Experiment 1. The diets in Experiment 3 were identical to those in Experiment 2, except that buffers were increased to 2.2% of dietary DM. The DMI was greater for cows fed buffered diets than control diets in Experiments 2 and 3. Milk production was unaffected by treatment. Milk fat percentage was unaffected by diet in Experiment 1 and averaged 3.8%. Milk fat depression was alleviated for cows fed buffered diets in Experiments 2 and 3. No differences in rumen fluid pH or molar percentage of VFA were significant among treatments.

Animal Feed↗
PubMed1998

Effects of buffers on milk fatty acids and mammary arteriovenous differences in dairy cows fed Ca salts of fatty acids.

Ten Holstein cows in early lactation were used in a replicated 5 x 5 Latin square design to study the effects of MgO and three buffers added to diets containing Ca salts of canola oil fatty acids. Treatments were 1) control (basal diet; no buffer). 2) 1.1% NaHCO3 plus 1.1% KHCO3, 3) 1.9% NaHCO3, 4) 0.5% MgO, and 5) 2.0% Na sesquicarbonate (percentage of dry matter). The control diet contained 53% grass silage, 43% concentrate, and 4% Ca salts. Body weight, intake, milk yield, and percentages of milk fat, protein, and lactose were unaffected by treatments. Buffers and MgO tended to increase triacylglycerol extraction by the mammary gland and changed the proportions of some fatty acids in milk. Arterial concentrations of acetate and triacylglycerol were correlated with their respective arteriovenous differences. Extraction by the mammary gland was high for acetate (approximately equal to 58.2%), triacylglycerol (approximately equal to 47.3%) propionate (approximately equal to 34.6%), and glucose (approximately equal to 24.3%). Extraction of free fatty acids, phospholipids, or cholesterol was negligible. Mammary triacylglycerol arteriovenous difference tended to be higher than when MgO was fed than when NaHCO3 was fed. Sodium sesquicarbonate, NaHCO3, and the blend of bicarbonate buffers increased C18:2 in milk fat when compared with the control treatment. The concentration of C18:2 in milk fat decreased when MgO was fed, but the ratio of cis-C18:1 to trans-C18:1 increased compared with effects of dietary NaHCO3. Medium-chain fatty acids in milk fat tended to be higher than Na sesquicarbonate than with NaHCO3. Buffers and MgO modified the profiles of fatty acids in milk.

Acetates↗
PubMed1999

Influence of carbohydrate source and buffer on rumen fermentation characteristics, milk yield, and milk composition in early-lactation Holstein cows.

The effects of concentrate to forage ratio and sodium bicarbonate (buffer) supplementation on intake, ruminal fermentation characteristics, digestibility coefficients, milk yield, and milk composition were examined in 4 cannulated Holstein cows (100 +/- 20 d in milk). A 4 x 4 Latin square design with 2 x 2 factorial arrangement of treatments was implemented for 3-wk experimental periods. The 4 treatments were a 50:50 concentrate to forage ratio with 1.2% of dry matter (DM) and without added buffer and a 75:25 concentrate to forage ratio with (1.2% of DM) and without (0% of DM) buffer. The forage component of the ration was a 50:50 mixture of alfalfa and barley and triticale silage, and diets were fed ad libitum as a total mixed ration. Although feed intake was not influenced by treatments, substantial treatment differences were observed for milk yield and milk composition. Cows fed high-concentrate diet had lower ruminal pH, ruminal acetate, and butyrate concentrations, whereas propionate concentrations were significantly elevated. The addition of buffer, at both levels of concentrate inclusion, resulted in elevated total volatile fatty acids and acetate concentrations. We concluded that altering the forage concentrate ratio in the diet of lactation cows influenced milk yield and milk composition, but the addition of buffer to the diet prevented the elevation in trans-C18:1 fatty acids in milk fat, and related milk fat depression, associated with feeding high-concentrate diets.

Acetates↗
PubMed1999

A simple, two-component buffer enhances use of chromatofocusing for processing of therapeutic proteins.

To extend the feasibility of chromatofocusing to industrial use, we have developed a simple chromatofocusing buffer system capable of generating a smooth pH gradient without the use of an external gradient maker. Using two cationic buffering components, an internal pH gradient is produced on appropriate chromatography media over a broad pH range (9.5 to 5.0). The utility of this buffer system is demonstrated with PBE94 and DEAE Sepharose fast flow ion-exchangers, as well as with experimental fast flow chromatofocusing gels. Using a rapid flow rate, we evaluated this buffer system for recovery of a therapeutic protein from a bacterial cell extract. The simplicity of the buffer system requiring no external gradient maker, coupled with the use of fast flow chromatographic media to produce broad-range pH gradients, improves the scalability of chromatofocusing for processing of therapeutic proteins.

Biotechnology↗