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Biomedical subjects

R E Forster

Publications and source records attributed to R E Forster.

At least 73 records · Page 4Linked to original sources
PubMed1977

Carbonic anhydrase activity in intact red blood cells measured with 18O exchange.

We have used a stirred, temperature-regulated, reaction vessel separated by a Teflon membrane from the ion source of a mass spectrometer to monitor continuously the time course of disappearance of C18O16O, mass 46, at chemical equilibrium as the 18O exchanges with 16O in water. This instrument is sensitive to less than 0.01 mm Hg of partial pressure of C18O16O with a response time of less than 3 s. The equation of Mills and Urey was used to calculate the hydration velocity constant for uncatalyzed or catalyzed homogenous solutions from the exponential disappearance of mass 46. Addition of red blood cells to the reaction mixture produces biphasic (double exponential) disappearance curve for mass 46. A theory of this process has been developed which describes the time course of [C18O16O] as a function of the catalytic factor for intracellular carbonic anhydrase (A) and the permeability of the cell membrane to HCO3- (P) in addition to the known values; water volume of the cells in the suspension, extracellular pH, the extracellular hydration reaction velocity constant, ku, and dehydration reaction velocity constant, ku. Using this theory, A and P were estimated from the disappearance curve for mass 46 at different values of hematocrit in the reaction mixture, both by a trial and error curve fitting procedure and by a more convenient graphical linearization method. The values of A and P obtained were very sensitive to small amounts of lysis (less than 1%), but the graphical method of analysis minimized this effect. For the blood cells of five normal subjects suspended in 24 mM bicarbonate in 145 mM NaCl at pH 7.4 and 37 degrees, using the graphical method we obtained an average value of 9,906 for A as compared to 19,900 for a comparable concentration of hemolysate. Correcting for a lower pH and chloride concentration inside the cell the latter figure would reduce to 17,500, still 80% higher than the intracellular value. The reason for this discrepancy is not clear. The average permeability of the red cell to bicarbonate ion was 3 X 10(-4) cm/s.

Adult↗
PubMed1977

Can alveolar pCO2 exceed pulmonary end-capillary CO2? No.

The exchanges and chemical reactions of CO2 within the blood are not complete during its transit time through the alveolar capillaries, so that theoretically alveolar PCO2 cannot exactly equal end-capillary blood PCO2. However, as a practical matter, the technical errors in determining alveolar and peripheral arterial PCO2 are so large in comparison with the difference between pulmonary arterial PCO2 and pulmonary venous PCO2, owing to the large effective solubility of CO2 in blood, that for this reason alone any difference between alveolar and end-capillary PCO2 can be neglected. The hypothetical mechanism that was originally proposed by Gurtner, Song, and Farhi for the production of an alveolar PCO2 greater than end-capillary PCO2 during rebreathing and depends on the existence of a radial gradient of H+ X HCO3- in the capillary blood near the wall appears unlikely because a) it would require a significant separation of charged ions over a large proportion of the capillary length and for a large proportion of the blood transit time and b) it would require a large radial gradient of PCO2 within the blood which on thermodynamic grounds would in turn require an unreasonable amount of energy to maintain.

Animals↗
PubMed1977

Postcapillary changes in blood pH in vivo during carbonic anhydrase inhibition.

A rapidly responding stopped-flow glass pH electrode apparatus was used to investigate pH changes in blood in vivo after it exits from an exchange capillary. Arterial blood was drawn from anesthetized animals through the apparatus. Temperature and pH of the blood in the electrode chamber were continuously recorded, both during withdrawal and after flow was stopped. Blood pH did not change after stopping flow in control experiments. When benzolamide (2 mg/kg) was given to inhibit carbonic anhydrase activity available to plasma (e.g., due to lysis) while having less effect on intracellular activity, pH increased 0.02-0.04 (t1/2 approximately 8 s) after stopping flow. Administration of acetazolamide (50 mg/kg) resulted in pH decreasing 0.07-0.10 (t1/2 approximately 15 s) after stopping flow. Ventilation for 1 min with N2 resulted in an increased rise in pH for the benzolamide-treated animals but a decreased fall in pH for the acetazolamide-treated animals. These shifts in arterial blood pH after gas exchange are largely due to disequilibrium of [H+] between red cells and plasma at the end of the pulmonary capillary.

Acetazolamide↗
PubMed1976

The carbamate reaction of glycylglycine, plasma, and tissue extracts evaluated by a pH stopped flow apparatus.

We have used a stopped flow rapid reaction pH apparatus to investigate the carbamate equilibrium in glycylglycine solutions and in three biological tissues, human plasma, sheep muscle, and sheep brain, as well as to investigate the kinetics of carbamate formation in glyclyglycine solution and in human plasma. The rapid reaction apparatus was equipped with a pH sensitive glass electrode in order to follow the time course of pH from 0.005 to 100 s after rapid mixing of a solution of amine or protein and CO2. Two phases of the pH curve were observed: a fast phase representing carbamate formation, and a slow phase due to the hydration of CO2 which was uncatalyzed since a carbonic anhydrase inhibitor was added to the biological solutions. From the time course of pH change during the fast phase K2, the R-NH2 ionization constant, and Kc, the carbamate equilibrium constant as well as the velocity constant for the formation of carbamate, ka could be calculated from data at different pH and pCO2. The carbamate formed in glycylglycine solutions over a wide range of pH and pCO2 was found consistent with the theory of carbamate formation and with published data. At ionic strength 0.16 and 37 degrees pK is 7.67. pKc 4.58. The heat of the carbamate reaction (deltaH) was calculated to be -3.2 kcal/mol between 20 degrees and 37 degrees. Kt of glycylglycine depends quantitatively on ionic strength as predicted by the Debye-Huckel theory. With ionic strength 0.16 ku was found to be 2,500 M1 S1 at 37 degrees. The activation energy of carbamate formation is 6.7 kcal/mol. Carbamate measurements in human plasma at pCO2 from 38 to 359 Torr. pH from 6.9 to 8.3, temperature 37 degrees, and ionic strength 0.15 provided evidence that two kinds of amino groups participate in carbamate formation. From the equilibrium constants computed for the two species they could be identified as alpha- and epsilon-amino groups. On the basis of a protein molecular weight of 69.000. 0.6 alpha-amino groups/molecule with pKz=7.0 and pKc=4.2, and 5.9 epsilon-amino groups/molecule with pKz=9.0 and pKc=4.3 contribute to carbamate formation. The velocity constant ka was estimated to be 4,950 M1 S1 for the alpha-amino groups and 13,800 M1 S1 for the epsilon-amino groups. Under physiological conditions (pCO2=40 Torr. pH=7.4). The concentration of carbamate in plasma is 0.6 mM and the half-time of carbamate formation is 0.05 s. In extracts prepared from sheep brain at 37 degrees pH=7 and pCO2=35 Torr. the carbamate formation was estimated to be 0.8 mM. With pCO2=70 Torr and the same pH and temperature the carbamate concentration in muscle approximates 0.3 mM and increases to 7 mM as pH rises to 8. It is concluded that, as in plasma, a considerable number of epsilon-amino groups appear to be available for carbamate formation in these tissues.

Animals↗
PubMed1976

The role of exchange transfusion in the management of low-birth-weight infants with and without severe respiratory distress syndrome. I. Initial observations.

Exchange transfusions within the first 8 hours of life, as an adjunct to conventional therapy, were evaluated in two groups of infants: (1) infants with birth weights of less than 1,250 gm without severe respiratory distress and (2) infants of any birth weight with evidence of severe respiratory distress syndrome. A total of 63 infants were studied in Group 1. Infants who received an exchange transfusion had a survival rate of 86% as contrasted with a survival rate of 57% in the control group (p less than 0.01). A total of 82 infants were studied in Group 2. Infants who received an exchange transfusion had a 59% survival rate as compared with a 39% survival rate for the control group (p less than 0.04). The mechanism by which early exchange transfusion improves survival rate is unknown.

Birth Weight↗
PubMed1976

Kinetics of bicarbonate-chloride exchange across the human red blood cell membrane.

The kinetics of bicarbonate-chloride exchange across the human red cell membrane was studied by following the time course of extracellular pH in a stopped-flow rapid-reaction apparatus during transfer of H+ into the cell by the CO2 hydration-dehydration cycle, under conditions where the rate of the process was determined by HCO3--Cl- exchange flux across the membrane. The flux of bicarbonate increased linearly with [HCO3-] gradient from 0.6 to 20 mM across the red cell membrane at both 37 degrees C and 2 degrees C, and decreased as transmembrane potential was increased by decreasing extracellular [Cl-]. An Arrhenius plot of the rate constants for the exchange indicates that the Q10 is strongly dependent on temperature, being about 1.7 between 24 degrees C and 42 degrees C and about 7 between 2 degrees C and 12 degrees C. These data agree well with the published values for Q10 of 1.2 between 24 degrees C and 40 degrees C and of 8 between 0 degrees C and 10 degrees C. The results suggest that different processes may determine the rate of HCO3--Cl- exchange at low vs. physiological temperatures, and that the functional (and/or structural) properties of the red cell membrane vary markedly with temperature.

Bicarbonates↗
PubMed1975

Effect of temperature on rate of CO2 uptake by human red cell suspensions.

The time course of carbon dioxide uptake by oxygenated supensions of human red cells was followed using a CO2 electrode in a Hartridge-Roughton continuous-flow rapid-reaction apparatus. Measurements were made at several temperatures from 1i to 42 degrees C, with the initial PCO2 in the reacting mixture from 40 to 60 mmHg. The initial part of the uptake curve is presumably rate limited by the intracellular hydration of CO2 with reaction-velocity constants in cell water from 280 to 960 S-1 at 42 degrees C and an activation energy of 2.4 kcal mol-1. The later stages of CO2 uptake were much slower, with half-times from greater than 1.5 s at 12 degrees C to 0.7 s at 42 degrees C, and were presumably rate limited by the chloride-bicarbonate shift and H+ interchanges. The results indicate that despite the acceleration of the hydration reaction in cell water by a factor of 5,000 at 37 degrees C and 3,800 at 42 degrees C, the later part of the exchange is too slow to permit blood to come intoC02, equilibrium with actively exercising muscles during its passage through the capillary bed.

Adult↗
PubMed1975

Time course of exchanges between red cells and extracellular fluid during CO2 uptake.

A stopped-flow rapid-reaction apparatus was used to follow the time course of extracellular pH in a human red cell suspension following a sudden increase in PCO2. The extracellular pH change was slow (t1/2 similar to 3.5 s) considering the presence of carbonic anhydrase in the cells. When carbonic anhydrase was added to the extracellular fluid, the half-time was reduced to less than 20 ms. The explanation for these phenomena is that the equilibration of H+ across the red cell membrane is rate-limited by the uncatalyzed reaction CO2 plus H2O formed from H2CO3 outside the cells. A theoretical model was developed which successfully reproduced the experimental results. When the model was used to simulate CO2 exchange in vivo, it was determined that blood PCO2 and pH require long times (greater than 50 s) to approach equilibrium between cells and plasma after leaving an exchange capillary. We conclude that cell-plasma equilibrium may never be reached in vivo, and that in vitro measurements of these quantities may not represent their true values at the site of sampling.

Carbon Dioxide↗