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The role of cardioplegic solution buffering in myocardial protection. A biochemical and histopathological assessment.

The advantages of buffering cardioplegic solutions to improve adenosine triphosphate preservation and postarrest hemodynamic function have been previously promoted. We evaluated the benefit of histidine buffering (195 mmol/L) in a low sodium (27 mEq/L) cardioplegic solution (Roe's) in a canine model of multidose cardioplegic arrest. Four solutions, two unbuffered (K+ = 10 mEq/L and K+ = 30 mEq/L) and two buffered (K+ = 10 mEq/L and K+ = 30 mEq/L), were tested in four groups of dogs for a 4 1/2 hour arrest period followed by 1 hour of reperfusion. Use of the unbuffered solution resulted in a drop in myocardial adenosine triphosphate from 29 +/- 1 mmol/kg (mean +/- standard error of the mean) (K+ = 30 mEq/L) and 28 +/- 2 mmol/kg (K+ = 10 mEq/L) to 8 +/- 2 mmol/kg and 7 +/- 2 mmol/kg, respectively, during the arrest period. In both buffered groups, adenosine triphosphate remained at preischemic levels during the entire arrest period. Myocardial glycogen followed the same pattern as adenosine triphosphate in the buffered groups. Lactate production was markedly elevated in all groups during ischemia. Postarrest hemodynamic function, as assessed by intraventricular isovolumic developed pressure measurements, was better (p less than 0.05) in the buffered low-potassium group than in the other three groups. The extent of myocardial necrosis, measured by triphenyl tetrazolium staining and confirmed by electron microscopy, was minimal (2% +/- 1% of biventricular mass) in the buffered low-potassium group, significantly greater (7% +/- 2% and 10% +/- 2%) in the unbuffered high-potassium and low-potassium groups, respectively, and highest (35% +/- 9%) in the buffered high-potassium group. These findings indicate that significant buffering capacity (similar to that of blood) in a crystalloid cardioplegic solution can be effective in preserving myocardial adenosine triphosphate stores, improving postarrest contractile function, and minimizing myocardial necrosis, provided the combination of high extracellular potassium and high pH levels is avoided.

Adenosine Triphosphate↗

Imidazole-buffered cardioplegic solution. Improved myocardial preservation during global ischemia.

Progressive acidosis is a constant finding in global myocardial ischemia and is associated with reduced myocardial contractility after ischemia. The hypothesis tested in these experiments was that imidazole (pKa = 6.7 at 37 degrees C), a commonly used buffer in physiology and microbiology, would provide superior buffering capacity when used in lieu of bicarbonate (pKa = 6.1 at 37 degrees C) in a cardioplegic solution. Twenty-eight isolated, working rabbit hearts were perfused, and preischemic and postischemic determinants of performance were measured. The 30 minute interval of normothermic global ischemia was altered by the injection at 0 and 15 minutes of 2 ml/gm wet weight of a buffered cardioplegic solution. Control hearts received a bicarbonate-buffered cardioplegic solution and experimental hearts received a solution buffered with imidazole. In the imidazole-buffered group, there was a superior recovery of coronary flow, developed left ventricular pressure, peak rate of rise of left ventricular pressure, peak rate of relaxation, and stroke work indices (p less than 0.05). Recovery of mechanical parameters was coincident with an improved acid-base status of the coronary sinus effluent at the end of ischemia. Coronary sinus effluents in the imidazole group had significantly higher pH values and lower partial pressures of carbon dioxide than coronary sinus effluents in the bicarbonate-buffered group (p less than 0.001). The data suggest that improved buffering of the extracellular and possibly intracellular space during global ischemia with a nonbicarbonate buffer is beneficial and provides improved postischemic myocardial recovery.

Animals↗

The endogenous calcium buffer and the time course of transducer adaptation in auditory hair cells.

Mechanoelectrical transducer currents in turtle auditory hair cells adapt to maintained stimuli via a Ca2+-dependent mechanism that is sensitive to the level of internal calcium buffer. We have used the properties of transducer adaptation to compare the effects of exogenous calcium buffers in the patch electrode solution with those of the endogenous buffer assayed with perforated-patch recording. The endogenous buffer of the hair bundle was equivalent to 0.1-0.4 mM BAPTA and, in a majority of cells, supported adaptation in an external Ca2+ concentration of 70 microM similar to that in turtle endolymph. The endogenous buffer had a higher effective concentration, and the adaptation time constant was faster in cells at the high-frequency end than at the low-frequency end of the cochlea. Experiments using buffers with different Ca2+-binding rates or dissociation constants indicated that the speed of adaptation and the resting open probability of the transducer channels could be differentially regulated and imply that the endogenous buffer must be a fast, high-affinity buffer. In some hair cells, the transducer current did not decay exponentially during a sustained stimulus but displayed damped oscillations at a frequency (58-230 Hz) that depended on external Ca2+ concentration. The gradient in adaptation time constant and the tuned transducer current at physiological levels of calcium buffer and external Ca2+ suggest that transducer adaptation may contribute to hair cell frequency selectivity. The results are discussed in terms of feedback regulation of transducer channels mediated by Ca2+ binding at two intracellular sites.

Animals↗

The free solution mobility of DNA in Tris-acetate-EDTA buffers of different concentrations, with and without added NaCl.

The free solution mobility of a high-molecular-weight DNA, linear pUC19, and a 20-bp oligomer called dsA5 have been studied as a function of Tris-acetate-EDTA (TAE) buffer concentration, with and without added NaCl. The two DNAs migrate as separate peaks during capillary electrophoresis, because the mobility of linear pUC19 is higher than that of the 20-bp oligomer. In TAE buffers ranging from 10-400 mM in concentration, the migration times and peak areas of the two DNAs are independent of whether they are electrophoresed separately or in mixtures, indicating that DNA-DNA and DNA-buffer interactions are absent in these solutions. The migration times of the two DNAs vary and the peak areas are not additive when the TAE buffer concentration is reduced to 5 mM or below, indicating that DNA-DNA and DNA-buffer interactions are occurring at very low TAE buffer concentrations. The mobilities of linear pUC19 and dsA5 decrease slowly with increasing conductivity or ionic strength when the conductivity is increased by increasing the TAE buffer concentration. When the Tris buffer concentration is held constant and the conductivity is increased by adding various concentrations of NaCl to the solution, the mobilities of linear pUC19 and dsA5 first increase slightly, then become independent of solution conductivity (or ionic strength), and finally decrease when the NaCl concentration is increased above approximately 50 mM. The mobility variations observed in the various TAE and TAE-NaCl solutions are described qualitatively by Manning's theory, although quantitative agreement is not achieved. The free solution mobilities of single-stranded pUC19 and two 20-base oligonucleotides have also been measured. The free solution mobility of single-stranded pUC19 is approximately 15% lower than that of native pUC19, in agreement with other results in the literature. Somewhat surprisingly, the mobilities of the single- and double-stranded 20-mers are equal to each other in TAE buffers with and without added NaCl.

Acetates↗

Setting priorities for research on pollution reduction functions of agricultural buffers.

The success of buffer installation initiatives and programs to reduce nonpoint source pollution of streams on agricultural lands will depend the ability of local planners to locate and design buffers for specific circumstances with substantial and predictable results. Current predictive capabilities are inadequate, and major sources of uncertainty remain. An assessment of these uncertainties cautions that there is greater risk of overestimating buffer impact than underestimating it. Priorities for future research are proposed that will lead more quickly to major advances in predictive capabilities. Highest priority is given for work on the surface runoff filtration function, which is almost universally important to the amount of pollution reduction expected from buffer installation and for which there remain major sources of uncertainty for predicting level of impact. Foremost uncertainties surround the extent and consequences of runoff flow concentration and pollutant accumulation. Other buffer functions, including filtration of groundwater nitrate and stabilization of channel erosion sources of sediments, may be important in some regions. However, uncertainty surrounds our ability to identify and quantify the extent of site conditions where buffer installation can substantially reduce stream pollution in these ways. Deficiencies in predictive models reflect gaps in experimental information as well as technology to account for spatial heterogeneity of pollutant sources, pathways, and buffer capabilities across watersheds. Since completion of a comprehensive watershed-scale buffer model is probably far off, immediate needs call for simpler techniques to gage the probable impacts of buffer installation at local scales.

Agriculture↗

Hydrochemical buffer assessment in agricultural landscapes: from local to catchment scale.

Non-point-source pollution of surface and groundwater is a prominent environmental issue in rural catchments, with major consequences on water supply and aquatic ecosystem quality. Among surface-water protection measures, environmental or landscape management policies support the implementation and the management of buffer zones. Although a great number of studies have focused on buffer zones, quantification of the buffer effect is still a recurring question. The purpose of this article is a critical review of the assessment of buffer-zone functioning. Our objective is to provide land planners and managers with a set of variables to assess the limits and possibilities for quantifying buffer impact at the catchment scale. We first consider the scale of the local landscape feature. The most commonly used empirical method for assessing buffers is to calculate water/nutrient budgets from inflow-outflow monitoring at the level of landscape structures. We show that several other parameters apart from mean depletion of flux can be used to describe buffer functions. Such parameters include variability, with major implication for water management. We develop a theoretical framework to clarify the assessment of the buffer effect and propose a systematic analysis taking account of temporal variability. Second, we review the current assessment of buffer effects at the catchment scale according to the theoretical framework established at the local scale. Finally, we stress the limits of direct empirical assessment at the catchment scale and, in particular, we emphasize the hierarchy in hydrological processes involved at the catchment scale: The landscape feature function is constrained by other factors (climate and geology) that are of importance at a broader spatial and temporal scale.

Agriculture↗

In vitro buffering capacity of cardiac and skeletal muscle tissue in normotensive and hypertensive rats. Assessment using PCO2 equilibration of homogenates.

Experiments were performed to assess the in vitro buffering capacity (physicochemical buffering) of cardiac and skeletal muscle tissue in normotensive and hypertensive rats using the techniques of PCO2 equilibration of tissue homogenates. We have previously shown (Oldershaw PJ, Cameron IR. Int J Cardiol 1988;18:131-141 and Int J Cardiol 1988;18:143-149) that the ability of myocardial cells to regulate intracellular pH is improved in association with left ventricular hypertrophy (induced by experimental hypertension) and this study purports to investigate the role of physicochemical buffering in this improved control. Rats were made hypertensive using the technique of sub-diaphragmatic aortic constriction and left for 1 or 4 weeks at which time samples of left ventricular and skeletal muscle tissue were taken for analysis; sham-operated animals were used as controls. The tissue was homogenised and in vitro buffering capacity assessed using PCO2 equilibration. The slope of the log PCO2-pH plot was calculated for each tissue and taken as a measure of buffering capacity. In control animals, in vitro buffering was greater in skeletal than cardiac muscle (slopes 1.89 +/- 0.5 cf. 1.64 +/- 0.06 P less than 0.05). In association with hypertension of 1 or 4 weeks duration the buffering capacity of left ventricular tissue increased to 2.00 +/- 0.08 (P less than 0.01) at 1 week and to 2.05 +/- 0.06 (P less than 0.01) at 4 weeks. There was no comparable change in the buffering capacity of skeletal muscle tissue. At least part of the observed in vivo changes in intracellular pH control may therefore be explicable on the basis of improved intracellular physicochemical buffering.

Animals↗

Baroresistant buffer mixtures for biochemical analyses.

Hydrostatic pressure is a useful tool in the study of varied fields such as protein aggregation, association, folding, ligand binding, and allostery. Application of pressure can have a significant effect on the pK(a) values of buffers commonly used for biochemical analysis. Consequently, cationic buffers, rather than neutral ones, are generally used to minimize pH effects; however, even with these buffers, the change in pH over 3 kbar may be consequential in highly pH-sensitive biochemical systems. Using fluorescence-based assays, we have systematically examined the effects of pressure on various buffers in the neutral pH range. We show that many commonly used cationic and Good's buffers increase in pH with pressure on the order of 0.1 to 0.3 pH units/kbar, in agreement with other published values. Carboxylates and phosphate decrease in pH to a similar extent. Buffer mixtures, composed of both cationic and carboxylate or phosphate components, are shown to be an order of magnitude less pressure sensitive than the individual component buffers. Using various relative concentrations of Tris and either phosphate, tricarballylate (1,2,3-propanetricarboxylate), or CDA (1,1-cyclohexane diacetate) at pH values between 7 and 8 yields baroresistant buffer mixtures. Buffer mixtures can be optimized for a specific pH, and a list of mixtures is presented for general laboratory use.

Carboxylic Acids↗

Effects of different buffers on the thermostability and autolysis of a cold-adapted protease MCP-01.

A cold-adapted protease MCP-01 was obtained from deep-sea psychrotrophic bacterium Pseudoaltermonas sp. SM9913. The effects of four different buffers, all at 50 mmol/l concentration, on its thermostability and autolysis were studied. The autolysis process of MCP-01 was studied by capillary electrophoresis. The thermostability of MCP-01 increased successively in the following order: carbonate < Tris < phosphate < borate. The optimum temperature for casein hydrolysis also increased in the same order. This suggested that the conformation of MCP-01 was flexible and its autolytic susceptibility was affected by some factors in the buffers such as charge and ionic species. The results also showed that different buffers, in addition to affecting the autolysis speed, gave different patterns of autolysis products. In carbonate buffer, Tris buffer, phosphate buffer and borate buffer, the autolysis patterns of MCP-01 were different. These results suggested that protease MCP-01 probably have different conformations in different buffers, thus exposing different autolysis sites on the enzyme surface. In addition, the loss of activity correlated with the speed of autolysis in the four different buffers, showing that autolysis may be a reason for the low thermostability of the enzyme.

Borates↗

Randomized long-term evaluation of bicarbonate-buffered CAPD solution.

BACKGROUND: Over the past 15 years, lactate has been used successfully as a buffer in peritoneal dialysis solutions, although its effectiveness in the correction of uremic acidosis and its biocompatibility on peritoneal resident cells have been questioned. In addition, some investigators have suggested other potential adverse metabolic effects resulting from the unphysiologically high lactate flux into the body during CAPD. These potential problems associated with lactate-containing CAPD solution prompted the search for alternative buffer-containing solutions. Bicarbonate, the physiological buffer, was considered when the problem of calcium and magnesium carbonate solubility was solved by the use of a two-compartment bag system, allowing the mixing of bicarbonate and divalent cations immediately before infusion. The long-term tolerance, safety, efficacy and therapeutic value of a bicarbonate-buffered peritoneal dialysis solution were evaluated in this study. METHODS: This open, randomized, controlled, multicenter study comparing a 34 mmol/liter bicarbonate- with a 35 mmol/liter lactate-buffered peritoneal dialysis solution was performed in two consecutive 12-week-treatment phases. Fourteen Centers participated in this trial. RESULTS: A total of 69 out of initially 123 randomized patients completed the six-month study period (36 patients in the bicarbonate group and 33 in the lactate group). While the arterial acid base status of the total study population did not change during the study period and no significant difference was observed between the two treatment groups, the acid-base status of patients in the bicarbonate group entering the study with a metabolic acidosis significantly improved (mean +/- SD; blood pH: baseline = 7.361 +/- 0.05, week 12 = 7.380 +/- 0.04, P < 0.05; week 24 = 7.388 +/- 0.03 P < 0.05; plasma bicarbonate: baseline = 19.49 +/- 3.01 mmol/liter, week 12 = 21.16 +/- 2.63 mmol/liter, P < 0.01; week 24 = 21.51 +/- 2.42 mmol/liter, P < 0.01). No significant changes were recorded in acidotic patients treated with the conventional lactate-buffered solution. The changes in plasma bicarbonate from baseline during the study was significantly different between the groups (week 12: lactate = +0.11 +/- 2.21 mmol/liter, bicarbonate = +1.69 +/- 2.55 mmol/liter, P < 0.05, 95% confidence interval for the difference 0.21 to 2.94 mmol/liter; week 24: lactate = +0.03 +/- 2.48 mmol/liter, bicarbonate = +1.82 +/- 2. 96 mmol/liter, P < 0.05, 95% confidence interval for the difference 0.16 to 3.42 mmol/liter). The normalized protein catabolic rate (nPCR) slightly but significantly decreased in the lactate group (baseline -0.90 +/- 0.23 g/kg/day, week 24 -0.83 +/- 0.21 g/kg/day, P < 0.01) and increased in the bicarbonate group (baseline +0.89 +/- 0.28 g/kg/day, week 24 +0.92 +/- 0.26 g/kg/day, P < 0.05). Changes from baseline between groups were significant (week 24, lactate = -0. 099 +/- 0.15 g/kg/day, bicarbonate = 0.049 +/- 0.12 g/kg/day, P < 0. 01, 95% confidence interval for the difference 0.068 to 0.229 g/kg/day). Other evaluated parameters (biochemical profile, peritoneal function parameters, dialysate protein loss) did not differ significantly between the two groups. No adverse effects related to the study solution were recorded. CONCLUSIONS: These results support the efficacy and safety of bicarbonate-buffered peritoneal solutions in a controlled randomized comparison for up to six months. Peritoneal dialysis solutions containing the physiological buffer bicarbonate might effectively replace conventional lactate-buffered CAPD solutions.

Acid-Base Equilibrium↗

A change from HCO3(-)-CO2- to hepes-buffered medium modifies membrane properties of rat CA1 pyramidal neurones in vitro.

1. Intracellular recordings were obtained from CA1 pyramidal neurones in rat hippocampal slices. Perfusion with a HCO3(-)-CO2-free, HEPES-buffered medium at pH 7.4 produced a wide variety of reversible effects on neuronal excitability, compared to responses obtained under standard (21 mM-HCO3-, 5% CO2, pH 7.4) conditions. 2. Introduction of HCO3(-)-CO2-free medium most commonly elicited, within 5-20 min, a fall in resting membrane potential (Vm), a rise in threshold for Na(+)-dependent action potential generation, and a reduction in input resistance. Anomalous inward rectification in the hyperpolarizing direction and subthreshold inward rectification were commonly reduced in HEPES-buffered medium. More prolonged exposure (> or = 25 min) to HCO3(-)-CO2-free medium produced, on occasion, Na+ spike inactivation. 3. The amplitudes of the fast and medium after-hyperpolarizations (AHPs) following a single depolarizing current-evoked action potential were attenuated during perfusion with HEPES-buffered medium at pH 7.4, as was the composite AHP following a train of action potentials. 4. Perfusion with HEPES-buffered medium at pH 7.4 reduced the degree of spike frequency adaptation and abolished depolarizing current-evoked burst-firing behaviour when this was present under standard conditions. 5. In tetrodotoxin (TTX)- and tetraethylammonium (TEA)-poisoned neurones, perfusion with HCO3(-)-CO2-free medium at pH 7.4 slightly raised the threshold for activation of Ca(2+)-dependent potentials and slightly reduced their duration, compared to responses obtained in HCO3(-)-CO2-buffered medium at the same pH. The AHP following the Ca2+ spike was, however, markedly attenuated. 6. Perfusion with a low-pH HCO3(-)-CO2-buffered medium (7 mM-HCO3-, 5% CO2, pH 6.9) produced changes qualitatively similar to those observed during perfusion with HEPES-buffered medium at pH 7.4. Raising the pH of the HEPES-buffered medium to 7.8 or 7.9 reversed inconsistently and then only in part the changes noted on the transition from a HCO3(-)-CO2- to a HEPES-buffered medium at the same pH (7.4). 7. The effects noted are unlikely to be due to a direct action of HEPES itself on neuronal membrane conductances. Rather, I suggest that they are likely to be caused by intracellular acidosis consequent upon the omission of HCO3- and CO2 from the extracellular medium.

Action Potentials↗

Occurrence and identification of microorganisms in compacted clay-based buffer material designed for use in a nuclear fuel waste disposal vault.

A full-scale nuclear fuel waste disposal container experiment was carried out 240 m below ground in an underground granitic rock research laboratory in Canada. An electric heater was surrounded by buffer material composed of sand and bentonite clay and provided heat equivalent to what is anticipated in a Canadian nuclear fuel waste repository. During the experiment, the heat caused a mass transport of water and moisture content gradients developed in the buffer ranging from 13% closest to the heater to 23% at the rock wall of the deposition hole. Upon decommissioning after 2.5 years, microorganisms could be cultured from all samples having a moisture content above 15% but not from samples with a moisture content below 15%. Heterotrophic aerobic and anaerobic bacteria were found in numbers ranging from 10(1) to 10(6) cells/g dry weight buffer. Approximately 10(2), or less, sulphate-reducing bacteria and methanogens per gram of dry weight buffer were also found. Identification of buffer population members was performed using Analytical Profile Index (API) strips for isolated bacteria and 16S rRNA gene sequencing for in situ samples. A total of 79 isolates from five buffer layers were identified with API strips as representing the beta, gamma and delta groups of Proteobacteria and Gram-positive bacteria. Sixty-seven 16S rRNA clones that were obtained from three buffer layers were classified into 21 clone groups representing alpha and gamma groups of Proteobacteria, Gram-positive bacteria, and a yeast. Approximately 20% of the population comprised Gram-positive bacteria. Members of the genera Amycolatopsis, Bacillus, and Nocardia predominated. Among Gram-negative bacteria, the genera Acinetobacter and Pseudomonas predominated. Analysis of lipid biomarker signatures and in situ leucine uptake demonstrated that the buffer population was viable. The results suggest that a nuclear fuel waste buffer will be populated by active microorganisms only if the moisture content is above a value where free water is available for active life.

Aluminum Silicates↗

Enhanced solubilization of rock phosphate by Penicillium bilaiae in pH-buffered solution culture.

Little is known about how pH-buffering capacity affects phosphorus (P) solubilization by Penicillium bilaiae. This study compared solubilization of rock phosphate (RP) by P. bilaiae in nonbuffered (pH 5.0) and buffered (pH 7.0) media. Fungal growth reached the stationary phase around day 12 and was slightly enhanced in the buffered medium. The fungus reduced solution pH from 5.0 to 4.1 in the nonbuffered medium and from 7.0 to 4.9 in the buffered medium by day 12. Phosphorus concentrations increased after day 9 more in the buffered than in the nonbuffered media (53 and 5 mg P x L(-1), respectively, on day 12). On day 12, higher concentrations of citric and oxalic acids were detected in the buffered (2.0 and 1.2 g x L(-1), respectively) than nonbuffered media (0.5 and 0.04 g x L(-1), respectively). Solubilization of RP was simulated without P. bilaiae in solutions equivalent to the nonbuffered and buffered cultures of P. bilaiae grown with RP. After a 24 h incubation, the P concentrations were of similar magnitudes to those observed in the P. bilaiae culture (18 and 47 mg P x L(-1), respectively, in the nonbuffered and buffered media). Under increased pH-buffering conditions, the enhanced production of citric and oxalic acids led to significant RP solubilization.

Calcium Phosphates↗

Parathyroid hormone and extrarenal acid buffering.

The role of parathyroid hormone (PTH) on the extrarenal buffering of an acid load was examined during HCl infusion (5 meq x kg-1 x h-1) to bilaterally nephrectomized rats. Thyroparathyroidectomized (TPTX) rats replaced with PTH had significantly higher blood pH and HCO3 values than TPTX rats not infused with PTH. Administration of EDTA, in a dose shown to release PTH, was associated with a significant increase in buffering capacity in intact but not in TPTX rats. Colchicine, given in a dose capable of stimulating PTH release, was also associated with enhanced buffering capacity in intact but not in TPTX rats. In TPTX rats infused with acetazolamide and PTH, the hormone failed to enhance extrarenal buffering of an acid load. Animals with chronic renal failure, induced by infarction of the kidney, also had an enhanced capacity to buffer an acid load. This enhanced buffering capacity in chronic renal failure was abolished by TPTX. Acute renal failure induced by bilateral ureteral ligation was also associated with increased buffering only in the presence of parathyroid glands. These data demonstrated that PTH, from either an exogenous or endogenous source, enhances extrarenal buffering capacity of an acid load. Chronic and acute renal failure are associated with increased buffering capacity, which is dependent on the presence of parathyroid glands. The data suggest that this effect is mediated through carbonic anhydrase.

Acid-Base Equilibrium↗

A comparison of buffered lidocaine versus ELA-Max before peripheral intravenous catheter insertions in children.

BACKGROUND: Peripheral intravenous catheter (PIV) insertion is a common, painful experience for many children in the pediatric emergency department. Although local anesthetics such as injected buffered lidocaine have been shown to be effective at reducing pain and anxiety associated with PIV insertion, they are not routinely used. ELA-Max, a topical local anesthetic, has the advantage of needle-free administration but has not been compared with buffered lidocaine for PIV insertion. OBJECTIVE: To compare the reduction of pain and anxiety during PIV insertion provided by subcutaneous buffered 1% lidocaine or topical ELA-Max in children. METHODS: A randomized trial in children 4 to 17 years old undergoing PIV insertion with 22-gauge catheters was conducted. Children received either buffered lidocaine or ELA-Max. Buffered lidocaine was administered by using 30-gauge needles to inject 0.1 to 0.2 mL subcutaneously just before PIV insertion. ELA-Max was applied to the skin and occluded with Tegaderm 30 minutes before PIV insertion. Self-reported Visual Analog Scale (VAS) questionnaires (rating on a scale of 1-10; 1 = no pain, anxiety) were completed by patients and their parents before PIV insertion to assess baseline perceptions about pain and anxiety associated with PIV insertion and immediately after PIV insertion to assess pain and anxiety associated with the experience. After PIV insertion, the nurse who inserted the PIV also completed a VAS questionnaire assessing technical difficulty and satisfaction with the local anesthesia. A blinded observer also completed a VAS questionnaire to assess pain and anxiety associated with the PIV insertion. Data were analyzed by using chi2 and t tests. RESULTS: Sixty-nine subjects were enrolled, and questionnaires were competed by all (mean age: 12.1 +/- 4.5 years; 61% female). There were no differences for buffered lidocaine and ELA-Max groups in age, gender, race, prior IV experience, or baseline pain and anxiety. There were no significant differences between buffered lidocaine and ELA-Max in mean pain and anxiety after PIV insertion by patient, parent, and blinded observer ratings. Nurse ratings of technical difficulty, number of PIV-insertion attempts, and satisfaction with local anesthesia also were not significantly different for buffered lidocaine and ELA-Max groups. CONCLUSIONS: ELA-Max provided similar pain and anxiety reduction during PIV insertion in children compared with injected buffered lidocaine. Technical difficulty and satisfaction by nurses inserting the PIV also were similar.

Administration, Topical↗

Potentiometric determination of bromate using an Fe(III)-Fe(II) potential buffer by circulatory flow-injection analysis.

A method for the potentiometric determination of bromate by circulatory flow injection analysis (CFIA) is described. The procedure involves the use of an Fe(III)-Fe(II) potential buffer solution, which is recycled via a reservoir. The analytical method is based on a linear relationship between the concentration of bromate and a very transient potential change in the electrode potential due to the generation of intermediate bromine during the reaction of bromate with the Fe(III)-Fe(II) potential buffer solution, which also contains NaBr, (NH4)6Mo7O24 and H2SO4. An aliquot (5 microl) of a bromate sample solution was injected into the stream of the potential buffer solution, 100 ml of which was circulated at a flow rate of 1 ml/min; the potential buffer solution stream was then returned to the reservoir after passing through a flow-through redox electrode detector. A potential change due to the reaction of the injected sample with the potential buffer in a reaction coil was measured with the detector in the form of a peak signal. The effects of the bromide, sulfuric acid and Fe(III)-Fe(II) concentrations in the potential buffer, and length of the reaction coil on the peak heights were examined in order to optimize the proposed CFIA method. The analytical sensitivities to bromate were 5.6 mV/microM for 1 x 10(-2) M and 30.9 mV/microM for 1 x 10(-3) M in the concentration of Fe(III)-Fe(II) in a potential buffer solution containing 0.35 M NaBr, 0.2% (NH4)6Mo7O24 and 1 M H2SO4. The detection limit of bromate obtained by a 1 x 10(-3) M Fe(III)-Fe(II) potential buffer solution was 0.02 microM (2.5 ppb). The numbers of repetitive determinations in which the relative sensitivities within 5% were regarded as being tolerated were ca. 4000 and 2000 for the use of only 100 ml of 1 x 10(-2) M and 1 x 10(-3) M Fe(III)-Fe(II) potential buffer solution, respectively.

Journal Article↗

Efficacy of natural grassland buffers for removal of Cryptosporidium parvum in rangeland runoff.

Our goal for this project was to estimate the retention efficiency of natural grassland buffers for Cryptosporidium parvum. Three sets of 16 plots (2.0 by 3.0 m) were established at 5, 20, and 35% slopes. Within each set of 16 plots, residual dry vegetation matter treatments of 225, 560, and 900 kg/ha were implemented, along with a noncut control averaging 4,500 kg/ ha. Buffer width treatments were implemented by placing cattle fecal material containing known loads of C. parvum 0.1, 1.1, or 2.1 m up-slope of the runoff collector. Grassland buffers of 1.1 and 2.1 m generated 3.2- to 8.8-log and 3.6- to 8.8-log retention of C. parvum, respectively, across the range of residual dry vegetation matter, land slope, rainfall, and runoff conditions examined during this project. Buffers with an increased percent land slope exhibited improved the retention efficiencies, whereas buffers experiencing larger maximum annual runoff events exhibited reduced retention efficiencies. Water-quality data from the 0.1-m-wide buffer plots (effectively no buffer) demonstrated that the majority of C. parvum oocysts (98 to 99.999%) were retained in the fecal matrix for the duration of the storm season, irrespective of the presence of a vegetated buffer. In conclusion, these results support the assertion that grassland buffers are an effective method for reducing animal agricultural inputs of waterborne C. parvum into drinking and irrigation water supplies.

Agriculture↗

A new buffering system and its use in electrophoresis and isoelectric focusing.

A novel buffer system with rather unique properties particularly well suited for preparative isoelectric focusing is described. The system was developed in response to the long standing need for pH gradients formulated using inexpensive components of known chemical composition. The system is binary consisting of buffer pairs covering the pH range of 3 to 10, in increments of less than 1 pH. The components are all readily available, nontoxic, and biologically acceptable, and are selected on the basis of their dissociation constants. The requirement for each buffer pair is that there be an overlapping pH zone where both components are largely non-ionized, having a ratio of neutral species to total buffer concentration between about 0.90 and 0.99. This ratio is defined as the 'electrophoretic reserve capacity' (ERC). Within the above ERC range a stable pH gradient is obtained, assuring sharply resolved protein focusing. Buffer parameters, mainly pH, buffering capacity, ERC, transport numbers, and conductivity, can be easily calculated and the buffer composition adjusted to meet requirements. All components are used at high concentration, typically of the order of up to 100 mM, yielding buffering capacities and conductivities analog to those customary in electrophoresis.

Acids↗