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PubMed2001

The Finnmark Study. A prospective population study of the social support buffer hypothesis, specific stressors and mental distress.

BACKGROUND: According to the buffer hypothesis, social support or social network may affect mental health by buffering the negative effect of stress on mental health. Previous studies have mostly been cross-sectional or have been done on selected populations, and the results have been conflicting. METHODS: The buffer hypothesis was tested in three population-based health surveys which took place in 1987, 1990 and 1993 in five coastal municipalities in Finnmark, Norway. All persons aged 40-62 years and a random sample of those aged 20-39 years were invited, and 77%, 74% and 70% attended the three health surveys, respectively. Those who had attended the health surveys more than once and had answered the key questions about mental health and social network, social support (measured both as instrumental support as well as emotional support) and stress were included in the analyses. Stress was divided into acute stress, (somatic stress, civil stress and work stress) and chronic stress (having a chronic disease, disability pension, being a single parent or providing long term nursing care for someone in the family). Growth curve analyses using mental distress as the dependent variable were used, analysing the interactional effects of various types of stressors and social network or social support. RESULTS: When all possible stressors and the sum of social network/social support were taken into consideration, total social support/network buffered the deteriorating effect of total stressor score upon mental health. The effect was weak but significant, and stronger for women than men. When each stressor was analysed separately, only a significant buffer effect of social network could be detected for work stress. For one of the chronic stressors, receiving a disability pension, a buffer effect could be demonstrated for both social network and instrumental support. CONCLUSION: The results provide some support for the buffer hypothesis, and indication of specificity in the interactions between stressor and social network/social support was found.Women, in general, had a larger buffering effect from their social network than men.

Adult↗
PubMed1979

The buffering capacity of the internal phase of thylakoids and the magnitude of the pH changes inside under flashing light.

The buffering capacity inside thylakoids is determined and the magnitude of flash-induced pH changes inside is calibrated in the pH range from 6.4 to 8.1. The work is based on flash-induced absorption changes of neutral red in a chloroplast suspension in which the outer phase is strongly buffered by bovine serum albumin. It is shown that neutral red is bound inside thylakoids. The binding can be described by a simple isotherm with an apparent Km = 4 microM and satruation at 1 neutral red per 17 chlorophylls. The apparent pK of neutral red is shifted from 6.6 in solution to 7.25 when bound inside. It is demonstrated that neutral red is a clean indicator of pH changes inside, i.e. when properly used it shows no response to other events. Although bound it reports pH changes which occur in the internal osmolar (aqueous) volume of thylakoids. This is obvious from the influence of chemically very different buffers on the magnitude of the absorption changes of neutral red. These act in a manner proportional to their calculated buffering capacity in aqueous solution. The intrinsic buffering capacity of the internal phase is determined with the aid of these buffers, at pH 7.2 it is between 0.8 and 1 mM (at 60 mosM). The absence of large variations in the buffering capacity in the range from pH 6.4 to 8.1 suggests that proteinaceous groups are involved in addition to the lipids which may dominate the buffering capacity at lower pH. The magnitude of the internal pH change is arrpox. 0.6 (at pH 7.3) under stimulation of both photosystems with a short xenon flash of light.

Chloroplasts↗
PubMed2004

Quantitative review of riparian buffer width guidelines from Canada and the United States.

This paper reviewed the provincial, territorial, and state guidelines for the retention of treed riparian buffers after timber harvest in Canada and the United States. Comparisons amongst jurisdictions were facilitated through the use of a standardized template for the classification of waterbodies. Mean buffer widths varied from 15.1 to 29.0 m for different waterbody types when both countries were combined. However, Canadian jurisdictions had wider buffers (except for intermittent streams). In part, this was due to the high percentage of Boreal jurisdictions in Canada and Southeast jurisdictions in the United States. The Boreal region had the widest buffers while Southeastern jurisdictions had the narrowest buffers. Just under half (approximately 44%) of the jurisdictions investigated had three or more modifying factors in the guidelines. Of these, waterbody type, shoreline slope, waterbody size, and presence of fish were the most common. Boreal and Pacific jurisdictions tended to have a more diverse set of waterbody size classes, waterbody types, and other modifying factors. Jurisdictions from the Midwest, Northeast, and Southeast maintained relatively simple 'one-size-fits-all' guidelines. Jurisdictions without modifying factors for slope or presence of fish applied wider baseline buffers than jurisdictions with these factors. A large percentage of jurisdictions (approximately 80%) allowed some selective harvest in buffers. However, these were often accompanied by relatively restrictive prescriptions. In comparison to the ecological recommendations, buffer widths for most jurisdictions were adequate to protect the aquatic biota and habitats but were, generally, less than recommended widths for terrestrial communities. In the future, two management trends are likely to continue, the shift towards more complicated guidelines and the expansion to larger-scale, watershed planning of riparian areas.

Animals↗
PubMed2000

An estimate of rapid cytoplasmic calcium buffering in a single smooth muscle cell.

Cytoplasmic calcium increments in the absence of sarco (endo) plasmic reticulum function were measured with a low-affinity fluorophore Indo-1FF in single isolated smooth muscle cells from guinea-pig urinary bladder. To evaluate the Ca(2+)-buffering properties of the myoplasm, Ca2+ influx, measured as time integral of the Ica (integral of Ica), was compared with corresponding free Ca2+ increments (delta [Ca2+]i) in the cytoplasm. The ratio between integral of ICa and delta [Ca2+]i (integral Ica/delta [Ca2+]i), reflecting the Ca2+ buffering properties of the cytosol, was in the range of 4.9-9.3 pC/microM (mean 6.2 +/- 1.2, n = 12). It remained approximately constant (6.4 +/- 1.4 pC/microM, n = 8) during recordings lasting up to 25 min, suggesting that cytoplasmic Ca2+ binding does not change markedly during cell dialysis and that the endogenous Ca2+ buffer is not significantly washed out of the cell through the patch pipette. Wash-in or wash-out of BAPTA, a mobile high-affinity Ca2+ buffer, into or from the cell markedly changed the relationship between Ca2+ influx through Ca2+ channels and delta [Ca2+]i within minutes. Changes in integral of ICa/delta [Ca2+]i during the sequence of depolarizing steps, which increased free [Ca2+]i up to 5 microM, suggested lower limits for the apparent affinity of a rapid Ca2+ buffer (16 microM) and for the total buffer concentration (530 microM). Introduction of 4 mM DPTA (Kd for Ca2+ = 81 microM) into the cell more than doubled the total cytoplasmic Ca2+ buffer capacity. These results suggest that cytoplasmic Ca2+ buffer in smooth muscle cells has a low affinity for free Ca2+. The Ca(2+)-binding ratio of the cytoplasm in most cells was estimated to be between 30 and 40. The Ca(2+)-binding ratio did not differ markedly between cells isolated from neonatal (< or = 5 days) and adult animals.

Animals↗
PubMed1997

In vitro biocompatibility evaluation of a novel bicarbonate-buffered amino-acid solution for peritoneal dialysis.

BACKGROUND: Conventional lactate-buffered peritoneal dialysis fluids containing glucose as the osmotic agent have been shown to compromise important peritoneal host defence functions. The current study employed an in vitro model using activated peripheral blood mononuclear leukocytes (PBMC) for the preclinical biocompatibility assessment of a novel bicarbonate-buffered peritoneal dialysis fluid containing 1.0% amino acids as the osmotic agent. METHODS: PBMC (5 x 10(6)/ml) were pre-exposed (10-30 mm, 37 degrees C) to bicarbonate-buffered 1% amino-acid solution, bicarbonate- or lactate-buffered 1.5% glucose fluid, or control medium (RPMI). The cells were then washed and stimulated for 2 h at 37 degrees C in RPMI containing 100 ng/ml E.coli endotoxin from strain O55:B5. The cytokines IL-6 and TNF alpha in cell supernatants were assessed using specific enzyme immunoassays, cytokine mRNA expression by reverse transcription polymerase chain reaction. RESULTS: Short, i.e. 10 min, exposure to conventional, lactate-buffered glucose fluid resulted in a significant and time-dependent inhibition of cytokine release and mRNA expression by activated PBMC, whereas the cytokine response was improved even following prolonged (up to 2 h) exposure to bicarbonate-buffered 1% amino-acid solution or bicarbonate-buffered 1.5% glucose fluid. CONCLUSIONS: Our results suggest that very short, i.e. potentially clinically relevant, exposure to conventional dialysis fluid impairs the cytokine response by activated leukocytes. In this respect, the use of bicarbonate-buffered solutions containing 1.0% amino acids or 1.5% glucose may result in improved biocompatibility properties.

Amino Acids↗
PubMed1991

Citrate decreases contraction and Ca current in cardiac muscle independent of its buffering action.

Extracellular Ca (Cao) depletions that occur during cardiac muscle contractions are indicative of net Ca entry. Buffering Cao concentration ([Ca]o) with citrate can limit the magnitude of these Cao depletions [e.g., Shattock and Bers. Am. J. Physiol. 256 (Cell Physiol. 25): C813-C822, 1989] which theoretically would allow more Ca entry and consequently greater force at the same free [Ca]o. However, Shimoni and Ginsburg [Am. J. Physiol. 252 (Cell Physiol. 21): C248-C252, 1987] have shown that citrate inhibits cardiac contractions and suggested that this was due to its Ca-buffering action (i.e., dissipating a local elevation of [Ca] at the outer sarcolemmal surface and thereby decreasing Ca influx). To examine the effects of Ca buffering per se, we compared the effects of four low-affinity Ca buffers [citrate, nitrilotriacetic acid (NTA), dipicolinic acid (DPA), and acetamidoiminodiacetic acid (ADA)] on several cardiac preparations. In Mg-free medium with 2 mM free Ca (measured using murexide), citrate, DPA, and ADA (10 mM) decreased the force of twitch contractions in rabbit ventricle to 76 +/- 2, 60 +/- 2, and 85 +/- 2%, respectively, but 10 mM NTA increased force slightly to 105 +/- 2%. No simple correlation was observed between the Ca affinity of the buffer and its effect on tension. These effects were not due to changes in sarcoplasmic reticulum (SR) Ca loading because rapid cooling contractures were not affected and similar results were observed in the presence of caffeine or ryanodine. The depressant effects of citrate and ADA on tension were greater at pH 5.5-6 and ADA had no effect at pH 8.5. Thus the depressant effect is stronger with more protonated forms of citrate and ADA, which are also poorer Ca buffers. Citrate (but not NTA) decreased Ca current in whole cell voltage clamp and shifted the current-voltage relationship and reversal potential to more negative potentials. Citrate decreased Ca current more effectively at higher citrate and lower Ca concentrations. We conclude that citrate (and some other weak Ca buffers) may directly decrease Ca current and contraction in a manner independent of Ca buffering ability.

Action Potentials↗
PubMed2006

Baroreflex sensitivity, blood pressure buffering, and resonance: what are the links? Computer simulation of healthy subjects and heart failure patients.

The arterial baroreflex buffers slow (<0.05 Hz) blood pressure (BP) fluctuations, mainly by controlling peripheral resistance. Baroreflex sensitivity (BRS), an important characteristic of baroreflex control, is often noninvasively assessed by relating heart rate (HR) fluctuations to BP fluctuations; more specifically, spectral BRS assessment techniques focus on the BP-to-HR transfer function around 0.1 Hz. Skepticism about the relevance of BRS to characterize baroreflex-mediated BP buffering is based on two considerations: 1) baroreflex-modulated peripheral vasomotor function is not necessarily related to baroreflex-HR transfer; and 2) although BP fluctuations around 0.1 Hz (Mayer waves) might be related to baroreflex BP buffering, they are merely a not-intended side effect of a closed-loop control system. To further investigate the relationship between BRS and baroreflex-mediated BP buffering, we set up a computer model of baroreflex BP control to simulate normal subjects and heart failure patients. Output variables for various randomly chosen combinations of feedback gains in the baroreflex arms were BP resonance, BP-buffering capacity, and BRS. Our results show that BP buffering and BP resonance are related expressions of baroreflex BP control and depend strongly on the sympathetic gain to the peripheral resistance. BRS is almost uniquely determined by the vagal baroreflex gain to the sinus node. In conclusion, BP buffering and BRS are unrelated unless coupled gains in all baroreflex limbs are assumed. Hence, the clinical benefit of a high BRS is most likely to be attributed to vagal effects on the heart instead of to effective BP buffering.

Algorithms↗
PubMed2000

Buffering hypercapnic acidosis worsens acute lung injury.

Hypoventilation, associated with hypercapnic acidosis (HCA), may improve outcome in acute lung injury (ALI). We have recently reported that HCA per se protects against ALI. The current study explored whether the mechanisms of protection with HCA were related to acidosis versus hypercapnia. Because CO(2) equilibrates rapidly across cell membranes, we hypothesized that (1) HCA would afford greater protection than metabolic acidosis. We further hypothesized that (2) buffering HCA would attenuate its protection. Forty isolated perfused rabbit lung preparations were randomized to: control (normal pH, PCO(2)); HCA; metabolic acidosis; or buffered hypercapnia. After ischemia-reperfusion (IR) injury wet:dry ratio was greatest with control and buffered hypercapnia, and rank order of capillary filtration coefficient was: control approximately buffered hypercapnia > metabolic acidosis > HCA. Isogravimetric pressure reduction was greatest with buffered hypercapnia. Despite comparable injury, pulmonary artery pressure elevation was less with buffered hypercapnia versus control. In vitro xanthine oxidase (XO) activity depended on pH, not PCO(2). We conclude that: (1) HCA and metabolic acidosis are protective, but HCA is the most protective; (2) buffering HCA attenuates its protection; (3) buffering HCA causes pulmonary vasodilation; (4) because metabolic acidosis and HCA similarly inhibit in vitro XO activity, the differential effects cannot be explained solely on the basis of extracellular XO activity.

Acidosis, Respiratory↗
PubMed2005

Quantifying the impact of regular cutting on vegetative buffer efficacy for nitrogen-15 sequestration.

This study used the stable 15N isotope to quantitatively examine the effects of cutting on vegetative buffer uptake of NO3(-)-N based on the theory that regular cutting would increase N demand and sequestration by encouraging new plant growth. During the summer of 2002, 10 buffer plots were established within a flood-irrigated pasture. In 2003, 15N-labeled KNO3 was applied to the pasture area at a rate of 5 kg N ha(-1) and 99.7 atom % 15N. One-half of the buffer plots were trimmed monthly. In the buffers, the cutting effect was not significant in the first few weeks following 15N application, with both the cut and uncut buffers sequestering 15N. Over the irrigation season, however, cut buffers sequestered 2.3 times the 15N of uncut buffers, corresponding to an increase in aboveground biomass following cutting. Cutting and removing vegetation allowed the standing biomass to take advantage of soil 15N as it was released by microbial mineralization. In contrast, the uncut buffers showed very little change in 15N sequestration or biomass, suggesting senescence and a corresponding decrease in N demand. Overall, cutting significantly improved 15N attenuation from both surface and subsurface water. However, the effect was temporally related, and only became significant 21 to 42 d after 15N application. The dominant influence on runoff water quality from irrigated pasture remains irrigation rate, as reducing the rate by 75% relative to the typical rate resulted in a 50% decrease in total runoff losses and a sevenfold decrease in 15N concentration.

Biomass↗
PubMed2006

Significant Escherichia coli attenuation by vegetative buffers on annual grasslands.

A study was conducted to estimate the retention efficiency of vegetative buffers for Escherichia coli deposited on grasslands in cattle fecal deposits and subject to natural rainfall-runoff conditions. The study was conducted on annual grasslands in California's northern Sierra Nevada foothills, a region with a distinct wet-dry season Mediterranean climate. We used 48, 2.0- by 3.0-m runoff plots to examine the efficacy of 0.1-, 1.1-, and 2.1-m buffers at three land slopes (5, 20, and 35%) and four dry vegetation matter levels (225, 560, 900, and 4500 kg/ha) across 27 rainfall-runoff events during two rainfall seasons. Buffer width treatments were implemented by placement of cattle fecal material containing known loads of E. coli 0.1, 1.1, or 2.1 m upslope of the plot runoff collector. Mean total runoff to total rainfall ratio per plot ranged from 0.014:1 to 0.019:1 and reflected the high infiltration capacity of these soils. Approximately 94.8 to 99.995% of total E. coli load applied to each plot appears to be either retained in the fecal pat and/or attenuated within 0.1 m downslope of the fecal pat, irrespective of the presence of a wider vegetated buffer. Relative to a 0.1-m buffer, we found 0.3 to 3.1 log10 reduction in E. coli discharge per additional meter of vegetative buffer across the range of residual dry vegetation matter levels, land slope, and rainfall and runoff conditions experienced during this project. Buffer efficiency was significantly reduced as runoff increased. These results support the assertion that grassland buffers are an effective method for reducing animal agricultural inputs of waterborne E. coli into surface waters.

Animals↗
PubMed2004

Effectiveness of riparian buffers in controlling ground-water discharge of nitrate to streams in selected hydrogeologic settings of the North Carolina Coastal Plain.

Water-quality and hydrologic information were collected along ground-water flow paths from two well-drained and two poorly drained Coastal Plain settings in North Carolina to evaluate the relative effectiveness of riparian buffers in reducing discharge of nitrate to streams. At one well-drained site with a 100 m buffer, little or no effect was detected on surface-water quality by discharging ground water because extensive woody vegetation in the buffer was able to take up not only most nitrate, but also most ground water before discharging to the stream during the growing season (March-October). At the second well-drained site, ground water discharging to the stream from the side with a buffer contained about 2 mg/L of nitrate-nitrogen after passing through the bed of the stream compared to 6 mg/L in ground water discharging from the side with no buffer. In the poorly drained settings, nitrate in ground water decreased from about 6 mg/L in the recharge area to less than 0.02 mg/L downgradient from the riparian buffer. Ground water discharging from the side with no buffer contained 0.83 mg/L. Riparian buffers appear effective in reducing nitrate in ground water discharging to Coastal Plain streams.

Agriculture↗
PubMed1982

Metabolic and hormonal factors influencing extrarenal buffering of an acute acid load.

This study evaluates metabolic and hormonal factors influencing extrarenal buffering of an acute acid load. Phosphate deprivation of 2 weeks duration was associated with enhanced extrarenal acid buffering. The enhanced extrarenal buffering capacity of phosphate deprivation was not dependent on the presence of parathyroid glands. Parathyroid hormone administration to phosphate-deprived rats promoted a further enhancement of the buffering capacity of an acid load. Blood pH and HCO3 during acid loading were not significantly different between control and diphosphonate-treated rats and between phosphate-deprived rats and phosphate-deprived rats treated with diphosphonate. The mortality rate, however, was significantly higher in diphosphonate-treated rats than in rats not receiving the drug suggesting that diphosphonate blunts the buffering of an acid load in both control and phosphate-deprived rats. Chronic vitamin D administration and acute administration of arginine vasopressin in pharmacologic doses were associated with significant enhancement of buffering capacity as compared to control rats. Thyrocalcitonin administration to intact but not thyroparathyroidectomized rats was associated with diminished capacity to buffer an acid load. These data demonstrate that the buffering of an acute acid load is influenced by a number of dietary and hormonal factors probably acting at the level of the bone.

Acid-Base Equilibrium↗
PubMed1993

Antibodies bound to nitrocellulose in acidic buffers retain biological activity.

This report compares the binding of proteins to nitrocellulose membranes in acidic buffers (pH 2 and 3) with binding in neutral buffer (pH 7). Initially, similar amounts of antibodies and other proteins bound to the nitrocellulose membrane in both acidic and neutral buffers. However, the susceptibility of individual proteins to displacement (stripping) from the membrane by the milk blocking agent depended on the pH of the buffer used to bind the proteins to the membrane. Most proteins that bound to nitrocellulose in acidic buffers were relatively resistant to milk-stripping compared to proteins bound in pH 7 buffer. Acid-binding of proteins to nitrocellulose also decreased the amount of protein that was stripped from the nitrocellulose membrane when Tween 20 was included in the washing buffer. After correcting for the amount of antibody remaining on the membrane after the milk block, it was found that acid-bound antibodies were unchanged in biological activity when compared with the same antibodies bound at neutral pH. These results suggest that acid-binding of proteins could increase the sensitivity of nitrocellulose membrane assays that use milk and/or Tween 20.

Animals↗
PubMed1998

Faster capillary electrophoresis separation of wheat proteins through modifications to buffer composition and sample handling.

Studies were conducted to produce faster, simpler, more rugged protocols for separating wheat proteins by high performance capillary electrophoresis (HPCE). Three areas were targeted for improvement: initial capillary equilibration procedures, buffer composition, and post-separation rinsing procedures. For the initial equilibration of capillaries, a brief rinse with a hydroxypropylmethylcellulose (HPMC) solution was the most critical factor for successful separation of wheat proteins. To reduce separation time and maintain resolution, beta-alanine and glycine were each used in place of sodium phosphate as buffer ions. Two isoelectric buffers, aspartic acid and iminodiacetic acid (IDA) were also tested. Each of these four buffer systems generated substantially lower currents, and provided faster separations, than sodium phosphate-based buffers. Finally, post-separation rinsing procedures were re-examined with the goal of reducing the time necessary to rinse the capillary after each separation. A critical factor in achieving this goal was removal of albumins and globulins prior to separation. These proteins bind to the capillary wall and cause rising baselines and excessive peak tailing. Once these proteins were removed, capillaries could be rinsed with buffer for only 2 min between separations. Capillary equilibration procedures were shortened from 90 min to 30 min. Likewise, separation times were reduced by approximately 40% (25 min to 15 min) by using glycine in place of sodium phosphate in the separation buffer. Finally, post-separation times were reduced by 80% (10 min to 2 min). Overall, these factors resulted in a reduction in total separation time of 50% (35 to 17 min) and maintained high resolution separations and good run-to-run repeatability.

Albumins↗
PubMed2002

Effect of glycine on pH changes and protein stability during freeze-thawing in phosphate buffer systems.

Previous studies have established that the selective precipitation of a less soluble buffer component during freezing can induce a significant pH shift in the freeze concentrate. During freezing of sodium phosphate solutions, crystallization of the disodium salt can produce a pH decrease as great as 3 pH units which can dramatically affect protein stability. The objective of our study was to determine how the presence of glycine (0-500 mM), a commonly used bulking agent in pharmaceutical protein formulations, affects the pH changes normally observed during freezing in sodium phosphate buffer solutions and to determine whether these pH changes contribute to instability of model proteins in glycine/phosphate formulations. During freezing in sodium phosphate buffers, the presence of glycine significantly influenced the pH. Glycine at the lower concentrations (< or = 50 mM) suppressed the pH decrease normally observed during freezing in 10 and 100 mM sodium phosphate buffer, possibly by reducing the nucleation rate of salt and thereby decreasing the extent of buffer salt crystallization. The presence of glycine at higher concentration (> 100 mM) in the sodium phosphate buffer resulted in a more complete crystallization of the disodium salt as indicated by the frozen pH values closer to the equilibrium value (pH 3.6). Although high concentrations of glycine can facilitate more buffer salt crystallization and these pH shifts may prove to be potentially damaging to the protein, glycine, in its amorphous state, can also act to stabilize a protein via the preferential exclusion mechanism.

Ammonium Sulfate↗
PubMed2001

Lyophilization-induced protein denaturation in phosphate buffer systems: monomeric and tetrameric beta-galactosidase.

During freezing in phosphate buffers, selective precipitation of a less soluble buffer component and subsequent pH shifts may induce protein denaturation. Previous reports indicate significantly more inactivation and secondary structural perturbation of monomeric and tetrameric beta-galactosidase (beta-gal) during freeze-thawing in sodium phosphate (NaP) buffer as compared with potassium phosphate (KP) buffer. This observation was attributed to the significant pH shifts (from 7.0 to as low as 3.8) observed during freezing in the NaP buffer (1). In the current study, we investigated the impact of the additional stress of dehydration after freezing on the recovery of active protein on reconstitution and the retention of the native structure in the dried state. Freeze-drying monomeric and tetrameric beta-gal in either NaP or KP buffer resulted in significant secondary structural perturbations, which were greatest for the NaP samples. However, similar recoveries of active monomeric protein were observed after freeze-thawing and freeze-drying, indicating that most dehydration-induced unfolding was reversible on reconstitution of the freeze-dried protein. In contrast, the tetrameric protein was more susceptible to dehydration-induced denaturation as seen by the greater loss in activity after reconstitution of the freeze-dried samples relative to that measured after freeze-thawing. To ensure optimal protein stability during freeze-drying, the protein must be protected from both freezing and dehydration stresses. Although poly(ethylene glycol) and dextran are preferentially excluded solutes and should confer protection during freezing, they were unable to prevent lyophilization-induced denaturation. In addition, Tween did not foster maintenance of native protein during freeze-drying. However, sucrose, which hydrogen bonds to dried protein in the place of lost water, greatly reduced freezing- and drying-induced denaturation, as observed by the high retention of native protein in the dried state as well as the complete recovery of active beta-gal on reconstitution. These results indicate that addition of an effective stabilizer, such as sucrose, may minimize protein denaturation during freeze-drying in phosphate buffers, even if there are large-scale changes in solution pH during freezing.

Buffers↗
PubMed2001

Kinetics of chemical degradation of isoxaflutole: influence of the nature of aqueous buffers (alkanoic acid/sodium salt vs phosphate).

A kinetic study of the chemical degradation of isoxaflutole (5-cyclopropyl-1,2-oxazol-4-yl alpha alpha alpha-trifluoro-2-mesyl-p-tolyl ketone) into its diketonitrile derivative (DKN), which is its active herbicide principle, in organic buffers at different pH values was carried out using a HPLC/UV detection method. The values of the pseudo-first-order rate constants Kobs for the reaction were calculated and compared with those previously obtained in inorganic buffers. In both cases, Kobs was found to be dependent on pH and temperature, but at pH 5.2 the degradation of isoxaflutole in CH3COOH/CH3COONa buffers was considerably faster than in KH2PO4/Na2HPO4 buffers, indicating that the compound was sensitive to the nature of the reagents used to prepare buffered solutions. The influence of phosphate and acetate concentrations and the influence of the R-substituent in RCOOH/RCOONa buffers were investigated. For the HA/A- buffers studied, the values of Kobs were linearly dependent on HA and A- concentrations, which meant that the degradation of isoxaflutole was subject to general catalysis. The values of Kobs were also found to be dependent on the number and the position of the CH3 groups of the R-substituent. The known degradation product of DKN (a benzoic acid derivative) was not detected throughout this study.

Biodegradation, Environmental↗
PubMed1995

Stimulation of glycolysis by histidine buffers in mammalian liver during cold hypoxia.

This study was designed to address the reasons why glycolysis in mammalian liver is unable to function more efficiently during periods of cold hypoxia. Our hypothesis was that control of intracellular pH, by use of amino acid buffers with high pKa values, would allow prolonged flux through glycolysis and better maintenance of liver high-energy adenine nucleotide pool. The effects of two concentrations of histidine (90 and 180 mM) and one of carnosine (90 mM), a histidyl dipeptide, on energy metabolism and levels of glycolytic substrate (glucose) and anaerobic endproduct (lactate) were investigated during cold hypoxia using rat livers to model the mammalian system. The transition to anaerobic metabolism was apparent by an immediate rise in lactate levels upon entry into cold hypoxia. By 10-14 h hypoxia, contents of the endproduct had increased by 10, 13.5, and 14.5 mumol/g in buffers containing 90 and 180 mM histidine and 90 mM carnosine, respectively. As well, ATP, total adenylate contents, and "energy charge" ratios exhibited a rapid decline from initial values of 2.3-3.3 mumol/g, 4.3-5.5 mumol/g, and 0.64-0.75, respectively, over the first 2-4 h of cold hypoxia. With respect to efficacy, the 180 mM histidine buffer exhibited the most positive maintenance of adenylate levels, followed closely by 90 mM carnosine, and finally 90 mM histidine as the least effective of the three buffers. Nevertheless, all three buffers examined in this study showed positive effects compared to similarly treated livers stored in a solution of minor buffering capacity (a citrate-based solution) over the same time period. The data support the hypothesis that glycolytic flux and cellular energetics can be maintained by the inclusion of efficient buffering agents during periods of cold hypoxia.

Adenine Nucleotides↗