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PubMed1995

Effects of chronic hyperfiltration on proximal tubule bicarbonate transport and cell electrolytes.

The compensatory response to unilateral nephrectomy (UNX) was investigated by a combination of renal clearance, microperfusion, electron microprobe, and morphological techniques. Filtration rate was significantly elevated 21 days following UNX and associated with a marked stimulation of bicarbonate and fluid absorption in the proximal tubule. Analysis of kinetic data of bicarbonate transport demonstrated strong flow-dependent activation of bicarbonate absorption in both control and experimental condition. The bicarbonate level at which half-saturation (apparent Kd) of transport occurred decreased uniformly at higher flow rates, but maximal transport rates (apparent Vmax) in the proximal tubule doubled in the remnant kidney. The flow dependence of bicarbonate transport in control and experimental conditions can be explained by an apparent unstirred layer effect modifying radial bicarbonate gradients in the tubule. Both Na/H-exchange and electrogenic H secretion contribute to bicarbonate absorption, but only Na/H-exchange increased significantly in proximal tubules of UNX rats. Cell ion concentrations after UNX were unchanged in cortical tubules, consistent with proportionately enhanced apical and basolateral ion transport. Proximal tubule cell rubidium concentration measured after a 30-second rubidium infusion as an index of basolateral Na,K-ATPase activity was unchanged in UNX rats. Inasmuch as cell volume increased significantly (25%), these data are consistent with a proportionate and similar stimulation of rubidium uptake and Na,K-ATPase activity.

Animals↗
PubMed1974

Effects of bicarbonate on intercompartmental reducing-equivalent translocation in isolated parenchymal cells from rat liver.

1. Incubation of isolated liver cells in a medium containing bicarbonate raises malate concentrations almost sixfold compared with values obtained in a bicarbonate-free phosphate medium. The malate concentration of about 0.3mm in bicarbonate medium is of the same order as the K(m) for malate dehydrogenase. 2. The utilization of ethanol, glyercol and sorbitol was increased by 20-35% in bicarbonate medium. 3. Fluoromalate, a specific inhibitor of malate dehydrogenase and the malate carrier, inhibited or ethanol oxidation by 23%, glycerol uptake by 20% and sorbitol uptake by 42% in bicarbonate medium, but had a much smaller inhibitory action in phosphate medium. In consequence fluoromalate almost abolished the stimulatory effects of bicarbonate on substrate utilization. 4. Difluoro-oxaloacetate, a specific inhibitor of aspartate aminotransferase, had about one-half the inhibitory activity of fluoromalate. The two inhibitors in combination were less effective than fluoromalate by itself. 5. It is concluded that bicarbonate stimulates the utilization of reduced substrates, which are oxidized in the cytoplasmic compartment of the liver cell, by increasing the activity of rate-limiting malate dehydrogenase-dependent intercompartmental hydrogen shuttles. Both malate-oxaloacetate and malate-aspartate systems are involved in these hydrogen-translocation processes.

Animals↗
PubMed1981

Effect of bicarbonate on the arginine-induced insulin and glucagon secretion in vitro.

The isolated perfused rat pancreas was used to investigate the effect of extracellular bicarbonate concentration (25 and 40 mmol/l) on the arginine-induced insulin and glucagon release with or without 5.5 mmol/l glucose in the perfusate. In the absence of glucose, the insulin response to arginine was increased at 40 mmol/l bicarbonate, while no potentiation by glucose of the arginine-induced insulin release, at this bicarbonate concentration, was observed. At 40 mmol/l bicarbonate, glucose inhibition of basal glucagon release was abolished. In contrast, the glucagon response to arginine was not affected by the bicarbonate concentration in the perfusion medium. These results confirm that the bicarbonate concentration in the medium markedly influences the insulin release from the isolated perfused rat pancreas, while the glucagon response to arginine seems to be less sensitive to changes in extracellular bicarbonate concentration.

Animals↗
PubMed1987

Impaired proximal duodenal mucosal bicarbonate secretion in patients with duodenal ulcer.

The defensive factors that prevent the human duodenal mucosa from acidic and peptic damage have not been fully evaluated. To determine whether duodenal mucosal bicarbonate production was altered in patients with inactive duodenal ulcer, we measured basal and acid-stimulated bicarbonate output from the duodenal bulb and the distal duodenum in healthy subjects and patients with inactive duodenal ulcer. As compared with 16 normal subjects, the 12 patients had significantly less mean (+/- SE) basal proximal duodenal mucosal bicarbonate secretion (185 +/- 13 vs. 107 +/- 18 mumol per centimeter per hour; P less than 0.001). Moreover, in response to a physiologic amount of hydrochloric acid (2 mmol per five minutes) instilled directly into the duodenal bulb, peak proximal duodenal bicarbonate output in the patients was 41 percent of the normal response (263 +/- 65 vs. 642 +/- 77 mumol per centimeter per hour; P less than 0.01). There was little overlap between groups. In contrast, bicarbonate outputs in the distal duodenum were similar in the two groups. We conclude that most patients with duodenal ulcer disease have decreased proximal duodenal mucosal bicarbonate production at rest, in response to hydrochloric acid, and in relation to peak gastric acid secretion. Impaired proximal duodenal mucosal bicarbonate secretion may be an important factor in the development and natural history of duodenal ulcer.

Adult↗
PubMed2003

Kinetic properties of "soluble" adenylyl cyclase. Synergism between calcium and bicarbonate.

"Soluble" adenylyl cyclase (sAC) is a widely expressed source of cAMP in mammalian cells that is evolutionarily, structurally, and biochemically distinct from the G protein-responsive transmembrane adenylyl cyclases. In contrast to transmembrane adenylyl cyclases, sAC is insensitive to heterotrimeric G protein regulation and forskolin stimulation and is uniquely modulated by bicarbonate ions. Here we present the first report detailing kinetic analysis and biochemical properties of purified recombinant sAC. We confirm that bicarbonate regulation is conserved among mammalian sAC orthologs and demonstrate that bicarbonate stimulation is consistent with an increase in the V(max) of the enzyme with little effect on the apparent K(m) for substrate, ATP-Mg(2+). Bicarbonate can further increase sAC activity by relieving substrate inhibition. We also identify calcium as a direct modulator of sAC activity. In contrast to bicarbonate, calcium stimulates sAC activity by decreasing its apparent K(m) for ATP-Mg(2+). Because of their different mechanisms, calcium and bicarbonate synergistically activate sAC; therefore, small changes of either calcium or bicarbonate will lead to significant changes in cellular cAMP levels.

Adenosine Triphosphate↗
PubMed2006

The structure of a cyanobacterial bicarbonate transport protein, CmpA.

Cyanobacteria, blue-green algae, are the most abundant autotrophs in aquatic environments and form the base of the food chain by fixing carbon and nitrogen into cellular biomass. To compensate for the low selectivity of Rubisco for CO2 over O2, cyanobacteria have developed highly efficient CO2-concentrating machinery of which the ABC transport system CmpABCD from Synechocystis PCC 6803 is one component. Here, we have described the structure of the bicarbonate-binding protein CmpA in the absence and presence of bicarbonate and carbonic acid. CmpA is highly homologous to the nitrate transport protein NrtA. CmpA binds carbonic acid at the entrance to the ligand-binding pocket, whereas bicarbonate binds in nearly an identical location compared with nitrate binding to NrtA. Unexpectedly, bicarbonate binding is accompanied by a metal ion, identified as Ca2+ via inductively coupled plasma optical emission spectrometry. The binding of bicarbonate and metal appears to be highly cooperative and suggests that CmpA may co-transport bicarbonate and calcium or that calcium acts a cofactor in bicarbonate transport.

ATP-Binding Cassette Transporters↗
PubMed2004

Bicarbonate promotes dye coupling in the epithelium and endothelium of the rabbit cornea.

PURPOSE: Examine the mechanism of bicarbonate maintenance of cell-to-cell coupling in rabbit corneal epithelium and endothelium. METHODS: Carboxyfluorescein was microinjected into rabbit corneal epithelial and endothelial cells. Adjacent cells were observed for fluorescence. Bathing solutions were buffered with bicarbonate, HEPES, phosphate, or acetate-citrate. The influence of intracellular pH and transmembrane voltage (V m ) were examined. RESULTS: Bicarbonate was the only buffer to increase dye coupling. Substitution of bicarbonate structural analogs bisulfite and carbamate in a HEPES-buffered solution increased dye coupling in both cell types. Intracellular pH and V m alterations in corneal epithelial cells bathed in HEPES vs. bicarbonate buffered media had no significant effects on dye coupling. CONCLUSIONS: Bicarbonate increases intercellular communication in the corneal epithelium and endothelium. This effect appears to result from an interaction of the bicarbonate molecule (or one of its structural analogs) with either gap junction proteins or an intermediary. We also demonstrate the presence of Cx43 in the rabbit corneal endothelium.

Animals↗
PubMedDate not supplied

Variation of urinary pH and bicarbonate concentrations of students in metropolitan and rural areas of Japan.

Changes in urinary pH and bicarbonate levels were studied in urine collected before and after final examinations taken by 91 medical students (65 males, 26 females) in Tokyo and from 87 students (73 males, 14 females) in Kanazawa, a rural municipality. Bicarbonate levels and pH in the urine of students in both cities increased significantly after attending 2 h of final examinations. The highest pH value and highest bicarbonate level were 7.66 and 80.3 mM, respectively, found in the urine of a student in Kanazawa. Urinary pH and bicarbonate levels averaged 6.48 and 11.1 mM, respectively, for students in Tokyo and 6.26 and 8.91 mM, respectively, for students in Kanazawa. Given that the concentrations of carbon dioxide in the air of the classrooms in both cities increased drastically during the final examinations (i.e., from 700 ppm to 1 200 ppm in Tokyo and from 700 ppm to 1 500 ppm in Kanazawa), it appears that the increases in urinary pH and bicarbonate levels were correlated with the increase of carbon dioxide (known to be convertible to bicarbonate in blood) in the rooms. This was also supported by the increase in urinary pH and bicarbonate levels found in urine collected from 20 students who stayed in a cinema for 3 h, at which location carbon dioxide levels in air reached 1 200 ppm.

Adult↗
PubMed2004

Electrolyte mass balance during CVVH: lactate vs. bicarbonate-buffered replacement fluids.

OBJECTIVE: To compare the effect of lactate vs. bicarbonate-buffered replacement fluids on electrolyte mass balance during isovolemic continuous veno-venous hemofiltration (CVVH). DESIGN: Randomized controlled study with double cross over. SETTING: Intensive care unit of a tertiary university hospital. PATIENTS AND PARTICIPANTS: Eight patients with acute renal failure (ARF). INTERVENTIONS: Isovolemic CVVH (2L/hr of replacement fluid) was performed in random order with either bicarbonate or lactate-buffered replacement fluid delivered pre-filter. MEASUREMENTS AND RESULTS: Sodium, potassium, chloride, magnesium, and phosphate, were measured in each sample. There was a mass gain of sodium, which was similar under both conditions (bicarbonate: 23.3+/-4.9 mmol/hr, lactate: 22.7+/-3.5 mmol/hr). Mass chloride gains occurred with bicarbonate-buffered replacement fluid only (12.8+/-5.3 mmol/hr), while there was an overall net loss of chloride with lactate fluids (-2.5+/-5.2 mmol/hr), resulting in a significant difference in chloride mass balance (p<0.0001). Magnesium mass balance was negative with bicarbonate buffer only (-0.6+/-0.2 mmol/hr) and also differed significantly from that obtained with lactate fluids (-0.1+/-0.2 mmol/hr, p<0.0001). Phosphate losses (bicarbonate: -1.7+/-0.7 mmol/hr, lactate: -1.7+/-0.5 mmol/hr) were equivalent with both buffers. Potassium mass balance was neutral. CONCLUSIONS: Mass balance during isovolemic CVVH is significantly affected by the type of replacement fluid administered prefilter. Isovolemic CVVH is not isonatremic and the use of bicarbonate-buffered fluid results in a significant accumulation of chloride and a loss of magnesium.

Acute Kidney Injury↗
PubMed1992

Serum bicarbonate declines with sample size in vacutainer tubes.

Serum bicarbonate testing has been reported to be inaccurate in several clinical studies. This prompted a search for systemic errors in bicarbonate testing devices, which has been inconclusive. The effect on serum bicarbonate level measurement of partially filling plain red-top (clot) vacutainer tubes was studied in healthy volunteers. First, 10-mL vacutainers were filled with 9.5 mL, 6 mL, 3 mL, and 1 mL of identical blood. Second, pediatric (3 mL) vacutainers were filled with 3 mL, 1 mL, and 0.5 mL of identical blood. The serum bicarbonate level was measured on an automated stat analyzer. Linear regression with dummy variables for subjects showed a significant reduction in bicarbonate reading as volume in the vacutainer decreased. Consistent decrease was noted regardless of whether samples were analyzed immediately after clotting (10 minutes) or at 20, 30, and 40 minutes after withdrawal of blood. This decrease was evident regardless of whether blood was aspirated directly into a vacutainer or first aspirated into a syringe and injected into a vacutainer. Underfilling of vacutainers may account for low bicarbonate readings and thus vacutainers should be filled completely to obtain accurate measurements of serum bicarbonate levels.

Bicarbonates↗
PubMed1997

In vivo and in vitro effects of amino-acid-based and bicarbonate-buffered peritoneal dialysis solutions with regard to peritoneal transport and cytokines/prostanoids dialysate concentrations.

BACKGROUND: Increasing evidence suggests that conventional PD solutions are detrimental to host defence mechanisms of peritoneal cells. We tested a new amino-acid-based and bicarbonate-buffered PD solution under in vivo and in vitro conditions. METHODS: During a prospective, cross-over randomized, intraindividual study 10 CAPD patients were investigated with three different solutions: Amino/Bic, 1% amino acid, 34 mmol/l bicarbonate; Glu/Bic, 1.5% glucose, 34 mmol/l bicarbonate; and Glu/Lac, 1.5% glucose, 35 mmol/l lactate. A PET was performed and transport properties (clearance, D/P ratio, MTAC) were calculated. Prostanoid and cytokine concentrations were measured in serum and the 6 h effluent. Using an in vitro model, mononuclear leukocytes of healthy donors were also incubated with the test fluids. In vivo results. Peritoneal clearance and MTAC of small solutes (creatinine, urea) were not significantly altered by amino acids or bicarbonate. Peritoneal permeability and transperitoneal excretion of higher-weight protein molecules (beta 2-microglobulin, albumin, IgG) were increased with Amino/Bic compared to Glu/Lac (P < 0.05) (D/P ratio albumin: Amino/Bic, 0.027 +/- 0.003; Glu/Bic, 0.023 +/- 0.003; Glu/Lac, 0.022 +/- 0.002). Application of Amino/Bic was accompanied by an increased effluent concentration of Il-6, Il-8, TNF alpha, PGE2, and 6-keto-PGF1a (P < 0.05). Dialysate nitrite/nitrate and cGMP concentrations (as indicators of NO generation) did not differ between the solutions. In vitro results. Both bicarbonate fluids demonstrated a better preservation of the mitochondrial dehydrogenases activity (MTT assay) compared to Glu/ Lac (P < 0.01) (Amino/Bic: 80.6 +/- 3.2%; Glu/Bic: 86.0 +/- 1.8%; Glu/Lac, 64.9 +/- 2.3%, referred to RPMI as control). Constitutive and LPS stimulated release of Il-1 beta and Il-6 was less suppressed with both bicarbonate fluids (P < 0.05) (LPS-stim. Il-6 release: Amino/Bic, 33.0 +/- 6.6%; Glu/Bic, 65.5 +/- 10.3%; Glu/Lac, 1.5 +/- 0.7% referred to RPMI). CONCLUSION: Application of an amino-acid/bicarbonate solution resulted in a small but significant increase in peritoneal permeability. Also increased concentrations of various cytokines/prostanoids were measured in the effluent. According to in vitro testing with mononuclear phagocytes both bicarbonate-buffered fluids were to the same extent less inhibitory to certain cell functions than lactate-buffered solution.

Adult↗
PubMed1993

New reinfusate composition in high UF haemodiafiltration: electrolyte solution combined with bicarbonate.

When high-permeability membranes are employed, high UF should be used in order to obtain optimal uraemic toxin removal and to avoid backfiltration. A high UF requires the infusion of an electrolyte solution including Ca2+ and Mg2+ which cannot be associated with bicarbonate in prepackaged solutions because of the risk of precipitation; therefore acetate or lactate are used as buffers. This study evaluated whether bicarbonate can be infused together with an electrolyte solution in high UF HDF, and if so, the clinical advantages that could be obtained by substituting acetate with bicarbonate in the reinfusate. In 12 patients on postdilutional high UF (121 +/- 10 ml/min) HDF (Qb 400, Qd 500 ml/min, dialysate containing Na, 141 +/- 2; K, 2.5; Ca, 3.5; Mg, 0.7; Cl, 111 +/- 2; acetate, 3; bicarbonate, 34 mEq/l; TMP 400 mmHg), an acid bag (Na, 128; K, 4; Ca, 7; Mg, 2; Cl, 141; acetate, 8 mEq/l), and a basic bag (Na, 150; HCO3, 80; Cl, 70 mEq/l), each containing 2 litres, were simultaneously infused through a Y connection. The final composition of reinfusate at the drip-chamber, combined with the above dialysate, allowed a negative intradialytic mass balance for Na, K, Mg and a positive one for Ca, acetate, to maintain prepostdialytic plasma values of these ions as well as bicarbonate close to normal limits. Furthermore, in five high-risk patients, clinical data were evaluated on high UF HDF, infusing a solution containing either acetate or bicarbonate, and an improvement of vascular stability was observed with the bicarbonate reinfusate.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetates↗
PubMed1990

Intravenous bicarbonate and sodium chloride both prolong endurance during intense cycle ergometer exercise.

To determine the effects of neutralizing exercise systemic acidosis via the intravenous route upon endurance and metabolic responses, eight lean, normal, postabsorptive men exercised to exhaustion at about 80% of their VO2 max (69 +/- 3%, mean +/- SEM, of maximum power output) on a cycle ergometer. Exercise studies were performed either with no infusion (control) or with a total infusion volume of about 1.5 L, mainly as 1.3% sodium bicarbonate or as 0.9% sodium chloride (NaCl), infused (double-blind) throughout exercise. The sodium bicarbonate was to prevent acid-base change, the sodium chloride was as a control for the volume infused. Arterialized venous blood and breath-by-breath analysis of expired gases were obtained. [H+] (nmol.L-1) and [HCO3-] (mmol.L-1) at exhaustion were similar in control and NaCl (46.5 +/- 1.8, 19.9 +/- 0.9), but remained unchanged from rest values with bicarbonate (38.4 +/- 0.9, 24.8 +/- 1.5, p less than 0.005 vs control and NaCl). At exhaustion, VO2, VCO2, RER, heart rate, and systolic BP as well as FFA, glycerol, alanine, insulin, norepinephrine, and epinephrine did not differ among protocols. Endurance was markedly prolonged (p less than 0.01) with bicarbonate (31.9 +/- 5.8 min) and NaCl (31.8 +/- 4.1 min) compared with the control (19.0 +/- 2.9 min) condition. Plasma glucose at exhaustion was higher (p less than 0.025) in the control compared to bicarbonate and NaCl experiments, while lactate was higher (p less than 0.025) in the bicarbonate than in the control and NaCl experiments. Thus, the prolonged endurance with sodium bicarbonate infusion could not be explained either by its effect of maintaining blood acid-base equilibrium or concomitant metabolic changes.

Acid-Base Equilibrium↗
PubMed1999

Does bicarbonate therapy improve the management of severe diabetic ketoacidosis?

OBJECTIVE: The use of bicarbonates in the treatment of severe diabetic ketoacidosis remains controversial, especially regarding the benefit/risk ratio. The aim of this study was to assess the efficacy of bicarbonate therapy during severe diabetic ketoacidosis (pH <7.10). DESIGN: Retrospective study. SETTING: The emergency unit of a teaching hospital. PATIENTS: The records of 39 patients consecutively admitted for severe diabetic ketoacidosis were analyzed (pH <7.10). The patients were divided into two groups: group 1 (n = 24; patients with bicarbonate treatment) and group 2 (n = 15; patients without bicarbonate treatment). INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: We compared two groups of patients presenting with severe diabetic ketoacidosis (pH values between 6.83 and 7.08) treated with or without bicarbonate. A group of 24 patients received 120+/-40 mmol sodium bicarbonate. The two groups were similar at admission with regard to clinical and biological parameters. No difference could be demonstrated between the two groups concerning the clinical parameters or the normalization time of biochemical parameters. If the number of patients with hypokalemia was comparable between the two groups, the potassium supply was significantly more important in group 1 compared with group 2 (366+/-74 mmol/L vs. 188+/-109 mmol/L, respectively; p < .001). CONCLUSIONS: Data from the literature and this study are not in favor of the use of bicarbonate in the treatment of diabetic ketoacidosis with pH values between 6.90 and 7.10.

Adult↗
PubMed1991

Evidence for a direct effect of bicarbonate on the rabbit corneal stroma.

Currently prevailing opinion on the mechanism of corneal deturgescence (stromal thinning) is that it is a process mediated by an endothelium-sited fluid pump. This pump is considered to be bicarbonate-dependent and balanced by the passive tendency of the stroma to absorb fluid that leaks from the aqueous across the endothelium. A case is presented that the utility of older key experiments as supportive evidence for this bicarbonate pump is limited by methodological errors in the formulation and use of bicarbonate-containing solutions. Recent studies reveal the pump to be generally unaffected by exogenous bicarbonate levels. Several recent studies confirm that the deturgescence of stroma-endothelium preparations is remarkably sensitive to exogenous bicarbonate and further indicate that the actual hydration of the stroma is inversely related to exogenous bicarbonate levels. These studies also show that it is possible to observe deturgescence in stroma-endothelial preparations which have no apparent net fluid pump activity. Conversely, net fluid pump activity has been measured in preparations which nonetheless fail to show deturgescence. Therefore, there does not appear to be an obligate coupling between deturgescence and endothelial fluid pump activity. The possible role of the endothelium in regulating the bicarbonate movement into the collagen-keratocyte matrix of the corneal stroma (rather than pumping it out) is discussed as a possible interpretation of more recent data.

Animals↗
PubMed1999

Role of CCK-A receptor in the regulation of pancreatic bicarbonate secretion in conscious rats: a study in naturally occurring CCK-A receptor gene knockout rats.

Whether cholecystokinin (CCK) has a direct action on duct cells and the role of CCK-A receptor in bicarbonate secretion were examined by comparing the results obtained from OLETF (CCK-A receptor-deficient rats) and control (LETO) rats. Rats were prepared with cannulae for draining bile and pancreatic juice separately, with two duodenal cannulae and an external jugular vein cannula. The experiments were conducted without anesthesia. The responses of bicarbonate secretion to intravenous infusion of CCK, acetyl-beta-methylcholine (Ach), and 2-deoxy-D-glucose (2DG), and to intraduodenal infusion of HCl and a liquid meal were examined. To examine the synergistic effect between CCK and secretin, the effect of CCK during a background secretin infusion was examined in LETO rats. CCK did not stimulate bicarbonate secretion in either strain, nor in LETO rats with secretin infusion. When gastric acid secretion was prevented by administration of omeprazole, Ach did not increase bicarbonate secretion, but 2DG did in both strains. Intraduodenal infusion of HCI and a liquid meal significantly increased bicarbonate secretion in both strains; however, the responses were much less in OLETF than LETO rats. In conclusion, intravenous injection of CCK did not stimulate bicarbonate secretion, and the lack of CCK-A receptor decreased bicarbonate secretion in response to luminal stimulants.

Animals↗
PubMed1989

Effect of bicarbonate on the growth of Actinobacillus actinomycetemcomitans in anaerobic fructose-limited chemostat culture.

The effect of bicarbonate on the growth and product formation by a periodontopathic bacterium, Actinobacillus actinomycetemcomitans, was examined in an anaerobic chemostat culture with fructose as the limiting nutrient. The chemostat cultures were run at dilution rates between 0.04 and 0.25 h-1 and the maximum growth yield (Ymax fructose) was estimated to be 40.3 and 61.7 g dry wt (mol fructose)-1 in the absence and presence of bicarbonate, respectively. The major fermentation products in the absence of bicarbonate were formate, acetate, ethanol and succinate, with small amounts of lactate. The addition of bicarbonate to the medium resulted in a marked decrease in ethanol production and in a significant increase in succinate production. Washed cells possessed activity for the cleavage of formate to CO2 and H2, which seemed to play a role in supplying CO2 for the synthesis of succinate in the absence of bicarbonate. The study of enzyme activities in cell-free extracts suggested that fructose was fermented by the Embden-Meyerhof-Parnas pathway. The values of Ymax ATP and the efficiency of ATP generation (ATP-Eff) during fructose catabolism were estimated and higher values were obtained for the culture in the presence of bicarbonate: 20.2 g dry wt (mol ATP)-1 and 3.0 mol ATP (mol fructose)-1, respectively, versus Ymax ATP = 15.1 and ATP-Eff = 2.7 in the absence of bicarbonate.

Actinobacillus↗
PubMed1996

The relationships between leukotoxin production, growth rate and the bicarbonate concentration in a toxin-production-variable strain of Actinobacillus actinomycetemcomitans.

Actinobacillus actinomycetemcomitans, a Gram-negative periodontopathic bacterium, produces a leukotoxin belonging to the RTX family. The production of leukotoxin varies greatly among different strains of this species. In this paper the effects of growth rate and bicarbonate on the leukotoxin production by a toxin-production-variable strain (301-b) during growth in a chemostat were examined. When the bacterium was grown in anaerobic fructose-limited chemostat cultures (pH 7.0 and 37 degrees C) at dilution rates (D) ranging from 0.04 to 0.20 h-1 in the absence and presence of 10 mM bicarbonate, it produced leukotoxin as a cluster of two polypeptides (M(r) 113,000 and 120,000) and complexed with nucleic acids on the bacterial cell surface. The relationship between leukotoxin production and specific growth rate was analysed by plotting the specific rate of leukotoxin production [qLT, in microgram (mg dry wt)-1 h-1] against D. The plots were approximated to the linear relationships qLT = 2.7D-0.058 and qLT = 9.3D-0.407 without and with bicarbonate, respectively. These relationships suggest that the apparent leukotoxin production is a result of both growth-rate-dependent production and growth-rate-independent decomposition. The cellular leukotoxin level was also followed after the change from chemostat to batch culture in the same fermenter. In batch culture leukotoxin production stopped immediately and the cellular toxin level rapidly decreased, suggesting toxin decomposition. From the slopes of the approximated linear relationships between qLT and D, a theoretical maximum leukotoxin yield (YLT) was estimated as 2.7 and 9.3 micrograms (mg dry wt)-1 in the absence and presence of 10 mM bicarbonate, respectively. The increased YLT value in the cultures containing bicarbonate indicated that the addition stimulated the efficiency of leukotoxin synthesis up to about threefold. Further increases of bicarbonate concentration to between 20 and 40 mM had no effect on the total leukotoxin production, but the amount of extracellular leukotoxin increased with higher bicarbonate concentrations.

Actinobacillus Infections↗