Changes In Plasma Protein Concentration Research Articles

Facing the Facts of Altered Plasma Protein Binding: Do Current Models Correctly Predict Changes in Fraction Unbound in Special Populations?

Accounting for variability in plasma protein binding of drugs is an essential input to physiologically-based pharmacokinetic (PBPK) models of special populations. Prediction of fraction unbound in plasma (fu) in such populations typically considers changes in plasma protein concentration while assuming that the binding affinity remains unchanged. A good correlation between predicted vs observed fu data reported for various drugs in a given special population is often used as a justification for such predictive methods. However, none of these analyses evaluated the prediction of the fold-change in fu in special populations relative to the reference population. This would be a more appropriate assessment of the predictivity, analogous to drug-drug interactions. In this study, predictive performance of the single protein binding model was assessed by predicting fu for alpha-1-acid glycoprotein and albumin bound drugs in hepatic impairment, renal impairment, paediatric, elderly, patients with inflammatory disease, and in different ethnic groups for a dataset of >200 drugs. For albumin models, the concordance correlation coefficients for predicted fu were >0.90 for 16 out of 17 populations with sub-groups, indicating strong agreement between predicted and observed values. In contrast, concordance correlation coefficients for predicted fold-change in fu for the same dataset were <0.38 for all populations and sub-groups. Trends were similar for alpha-1-acid glycoprotein models. Accordingly, the predictions of fu solely based on changes in protein concentrations in plasma cannot explain the observed values in some special populations. We recommend further consideration of the impact of changes in special populations to endogenous substances that competitively bind to plasma proteins, and changes in albumin structure due to posttranslational modifications. PBPK models of special populations for highly bound drugs should preferably use measured fu data to ensure reliable prediction of drug exposure or compare predicted unbound drug exposure between populations knowing that these will not be sensitive to changes in fu.

PSVIII-21 Modulation of Goat Plasma Proteins Using Different Preparations of Garlic

Abstract The main objective of this study was to evaluate the effect of different garlic preparations on plasma protein concentration and Galectin secretion in goat blood. Inflammatory diseases caused by pathogens including gastrointestinal parasites impact goat gut health and productivity. Changes in plasma protein concentration are indicators of health status with application in inflammatory disease diagnosis and prognosis. Galectins (Gals) are small soluble proteins that contain one or two carbohydrate recognition domains (CRDS). ‘Prototype’ galectin-1, -2, -7, contain one CRD, Galectin 3 is chimeric, and galectin-4, -8, -9 and -12, are tandem repeat galectins with two CRDs. Galectins regulate biologic functions, immunity and disease pathogenesis and can serve as biomarkers. Garlic (Allium sativum) is a medicinal herb that has anti-parasitic and immuno-modulatory effects. Six (N=6) adult female clinically healthy Boer x Spanish goats were used. Blood was collected from the jugular vein and treated with different concentrations of fresh garlic, allicin, garlic barrier, or aged garlic (Sigma aldich). Total plasma protein secretion was evaluated using the BCA assay. Secretion of Gals 1(Prototype), Gal 3 (chimera type) and Gal 9 (tandem-repeat type) determined using commercial ELISA kits as recommended by the manufacturer. All variables were assessed using SAS analysis software (version 9.0, Sas Institute Cary, p≤0.05). Garlic treatment had a dose dependent effect on total plasma protein and Gals secretion. The protein secretion was affected by different types of garlic. Allicin (72.86μg/ml) reduced the concentration of plasma proteins. No change was observed with fresh garlic (156.334μg/ml) and garlic barrier (160.73μg/ml). Garlic barrier treatment increased protein secretion (6.3%) in goat blood. All Gals were detected (Gals 1 &amp;gt;9 &amp;gt;3), with a variable response to Garlic preparation (Garlic barrier &amp;gt; fresh garlic Gals secretion). Garlic modulates protein secretion. Studies are needed to determine the effect of different preparations on leukocytes and molecular targets in goat blood.

Characterization of Plasma Protein Alterations in Pregnant and Postpartum Individuals Living With HIV to Support Physiologically-Based Pharmacokinetic Model Development.

Background: Alterations in plasma protein concentrations in pregnant and postpartum individuals can influence antiretroviral (ARV) pharmacokinetics. Physiologically-based pharmacokinetic (PBPK) models can serve to inform drug dosing decisions in understudied populations. However, development of such models requires quantitative physiological information (e.g., changes in plasma protein concentration) from the population of interest.Objective: To quantitatively describe the time-course of albumin and α1-acid glycoprotein (AAG) concentrations in pregnant and postpartum women living with HIV.Methods: Serum and plasma protein concentrations procured from the International Maternal Pediatric Adolescent AIDS Clinical Trial Protocol 1026s (P1026s) were analyzed using a generalized additive modeling approach. Separate non-parametric smoothing splines were fit to albumin and AAG concentrations as functions of gestational age or postpartum duration.Results: The analysis included 871 and 757 serum albumin concentrations collected from 380 pregnant (~20 to 42 wks gestation) and 354 postpartum (0 to 46 wks postpartum) women, respectively. Thirty-six and 32 plasma AAG concentrations from 31 pregnant (~24 to 38 wks gestation) and 30 postpartum women (~2–13 wks postpartum), respectively, were available for analysis. Estimated mean albumin concentrations remained stable from 20 wks gestation to term (33.4 to 34.3 g/L); whereas, concentrations rapidly increased postpartum until stabilizing at ~42.3 g/L 15 wk after delivery. Estimated AAG concentrations slightly decreased from 24 wks gestation to term (53.6 and 44.9 mg/dL) while postpartum levels were elevated at two wks after delivery (126.1 mg/dL) and subsequently declined thereafter. Computational functions were developed to quantitatively communicate study results in a form that can be readily utilized for PBPK model development.Conclusion: By characterizing the trajectory of plasma protein concentrations in pregnant and postpartum women living with HIV, our analysis can increase confidence in PBPK model predictions for HIV antiretrovirals and better inform drug dosing decisions in this understudied population.

The Predictive Value of Glomerular Filtration Rate-Based Scaling of Pediatric Clearance and Doses for Drugs Eliminated by Glomerular Filtration with Varying Protein-Binding Properties.

IntroductionFor drugs eliminated by glomerular filtration (GF), clearance (CL) is determined by GF rate (GFR) and the unbound fraction of the drug. When predicting CL of GF-eliminated drugs in children, instead of physiologically based pharmacokinetic (PBPK) methods that consider changes in both GFR and protein binding, empiric bodyweight-based methods are often used. In this article, we explore the predictive value of scaling using a GFR function, and compare the results with linear and allometric scaling methods for drugs with different protein-binding properties.MethodsFirst, different GFR maturation functions were compared to identify the GFR function that would yield the most accurate GFR predictions across the pediatric age range compared with published pediatric inulin/mannitol CL values. Subsequently, the accuracy of pediatric CL scaling using this GFR maturation function was assessed and compared with PBPK CL predictions for hypothetical drugs binding, to varying extents, to serum albumin or α-acid glycoprotein across the pediatric age range. Additionally, empiric bodyweight-based methods were assessed.ResultsThe published GFR maturation functions yielded comparable maturation profiles, with the function reported by Salem et al. leading to the most accurate predictions. On the basis of this function, GFR-based scaling yields reasonably accurate (percentage prediction error ≤ 50%) pediatric CL values for all drugs, except for some drugs highly bound to AGP in neonates. Overall, this method was more accurate than linear or 0.75 allometric bodyweight-based scaling.ConclusionWhen scaling CL and dose by GFR function, maturational changes in plasma protein concentrations impact GF minimally, making this method a superior alternative to empiric bodyweight-based scaling.Electronic supplementary materialThe online version of this article (10.1007/s40262-020-00890-2) contains supplementary material, which is available to authorized users.

Changes in the Plasma Protein Composition in Cosmonauts after Space Flight and its Significance for Endothelial Functions

Spaceflight (SF) conditions have a significant impact on the functioning of the human cardiovascular system. The endothelium plays an important role in the process of adaptation to SF factors. Therefore, the detection of biomarkers of endothelial dysfunction is necessary for understanding the molecular mechanisms involved in changes caused by SFs. For this purpose, the blood plasma proteins of 18 Russian cosmonauts were used for quantitative analysis by liquid chromatography with a UPLC 1290 Infinity chromatograph coupled to an Agilent 6490 triple quadrupole mass spectrometer. It was found that a decrease in the circulating plasma volume during the flight followed by the activation of fluid retention at the final stage of flight contributed to the changes in plasma protein concentrations on the first day after landing. We observed a significant increase in the concentration of the S100A9 protein that plays an important role in endothelium functioning and angiogenesis and can serve as a marker of inflammatory reactions. On the first day after landing, the complement system and acute-phase protein concentrations tended to increase, which can adversely affect the functioning of the endothelium.

Extravascular Lung Water, B-Type Natriuretic Peptide, and Blood Volume Contraction Enable Diagnosis of Weaning-Induced Pulmonary Edema*

We tested whether the changes in extravascular lung water indexed for ideal body weight could detect weaning-induced pulmonary edema. We also studied the diagnostic value of blood volume contraction indices and B-type natriuretic peptide variations. Prospective study. Twenty-one patients who failed a first spontaneous breathing trial. None. We performed a second 60-minute T-tube spontaneous breathing trial. Before and at the end of spontaneous breathing trial, we recorded pulmonary artery occlusion pressure, the extravascular lung water indexed for ideal body weight, plasma B-type natriuretic peptide level, hemoglobin, and plasma protein concentrations. Weaning-induced pulmonary edema was defined by the association of signs of clinical intolerance and a pulmonary artery occlusion pressure greater than or equal to 18 mm Hg at the end of spontaneous breathing trial. Because some patients performed several spontaneous breathing trial, a primary analysis included all spontaneous breathing trial and a secondary analysis included only the first spontaneous breathing trial of each patient. In primary analysis, 36 spontaneous breathing trials were analyzed, 21 spontaneous breathing trial with weaning-induced pulmonary edema and 15 without. During spontaneous breathing trial, extravascular lung water indexed for ideal body weight increased only in cases with weaning-induced pulmonary edema (25% ± 23%). Plasma protein concentration, hemoglobin concentration, and B-type natriuretic peptide also significantly increased only in cases with weaning-induced pulmonary edema (9% ± 3%, 9% ± 4%, 21% ± 23%, respectively). The areas under the receiver operating characteristics curves to detect weaning-induced pulmonary edema were 0.89 (95% CI, 0.78-0.99) for extravascular lung water indexed for ideal body weight, 0.97 (0.93-1.01) for spontaneous breathing trial-induced changes in plasma protein concentration, 0.96 (0.90-1.01) for changes in hemoglobin concentration, and 0.76 (0.60-0.93) for changes in B-type natriuretic peptide. An increase in extravascular lung water indexed for ideal body weight greater than or equal to 14% diagnosed weaning-induced pulmonary edema with a sensitivity of 67% (95% CI, 43-85%) and a specificity of 100% (95% CI, 78-100%). The secondary analysis confirmed these results. Spontaneous breathing trial-induced increases in extravascular lung water indexed for ideal body weight, plasma protein concentrations, hemoglobin concentration, and B-type natriuretic peptide are reliable alternatives to the pulmonary artery catheter for diagnosing weaning-induced pulmonary edema.

Body Sodium Overload Modulates the Firing Rate and Fos Immunoreactivity of Serotonergic Cells of Dorsal Raphe Nucleus

In order to determine whether serotonergic (5HT) dorsal raphe nucleus (DRN) cells are involved in body sodium status regulation, the effect of a s.c. infusion of either 2 M or 0.15 M NaCl on 5HT DRN neuron firing was studied using single unit extracellular recordings. In separate groups of 2 M and 0.15 M NaCl-infused rats, water intake, oxytocin (OT) plasma concentration, urine and plasma sodium and protein concentrations were also measured. Also, to determine the involvement of particular brain nuclei and neurochemical systems in body sodium overload (SO), animals from both groups were perfused for brain immunohistochemical detection of Fos, Fos-OT and Fos-5HT expression. SO produced a significant increase in serotonergic DRN neuron firing rate compared to baseline and 0.15 M NaCl-infused rats. As expected, 2 M NaCl s.c. infusion also induced a significant increase of water intake, diuresis and natriuresis, plasma sodium concentration and osmolality, even though plasma volume did not increase as indicated by changes in plasma protein concentration. The distribution of neurons along the forebrain and brainstem expressing Fos after SO showed the participation of the lamina terminalis, extended amygdala, supraoptic and paraventricular hypothalamic nuclei in the neural network that controls osmoregulatory responses. Both Fos-OT immunoreactive and plasma OT concentration increased after s.c. hypertonic sodium infusion. Finally, matching the “in vivo” electrophysiological study, SO doubled the number of Fos-5HT immunolabeled cells within the DRN. In summary, the results characterize the behavioral, renal and endocrine responses after body sodium overload without volume expansion and specify the cerebral nuclei that participate at different CNS levels in the control of these responses. The electrophysiological approach also allows us to determine in an “in vivo" model that DRN 5HT neurons increase their firing frequency during an increase in systemic sodium concentration and osmolality, possibly to modulate sodium and water intake/excretion and avoid extracellular volume expansion.

Comparison of two analyzers for measurement of plasma total carbon dioxide concentration in horses

To determine the degree of agreement between 2 analyzers for measurement of total CO₂ concentration (ctCO₂) in equine plasma. 6 healthy untrained horses, 6 trained Standardbreds undergoing a simulated race protocol, and 135 trained Standardbreds at a racetrack. Jugular venous blood samples were obtained from all horses. Two analyzers (commonly used analyzer A and less expensive analyzer B) were used to measure plasma ctCO₂ in each sample. Validation of both analyzers was conducted in accordance with guidelines established by the Clinical and Laboratory Standards Institute and involved characterization of linearity, total analytic error, and bias estimation. Total analytic error (instrument SD) was 0.58 mmol/L (coefficient of variation, 1.6%) and 0.49 mmol/L (coefficient of variation, 1.4%) for analyzers A and B, respectively, when measuring an aqueous standard containing 36.0 mmol of CO₂/L. A 1 g/L decrease in plasma protein concentration corresponded to an increase in ctCO₂ measured with analyzer B of 0.065 mmol/L. A difference plot indicated that analyzer B produced values 2.7% higher than analyzer A for 103 samples from the 6 trained and exercised Standardbreds (mean plasma protein concentration, 67 g/L). Analyzer B provided adequate precision and linearity for measurement of ctCO₂ from 5 to 40 mmol/L and was therefore suitable for measuring ctCO₂ in equine plasma, provided allowances are made for changes in plasma protein concentration.

Alkalinizing effect of NaHCO3 with and without glucose when administered orally to euhydrated neonatal dairy calves

The use of oral rehydration solutions (ORS) is well established as an effective treatment to correct water-, electrolyte-, and acid-base balance in diarrheic calves. The main ingredients of a commercial ORS are Na, glucose, and alkalinizing agents, such as NaHCO3. Particular importance is attributed to the combination of glucose and Na at a specific ratio to optimize intestinal sodium, and thereby water uptake, through the sodium-glucose co-transport. Enhancing intestinal Na absorption by combining glucose and Na in an ORS has the potential to improve the alkalinizing effect of an ORS according the strong ion theory. The objective of this study was to investigate the effect of glucose on the alkalinizing effect of NaHCO3 when administered orally. Nine healthy neonatal Holstein-Friesian calves underwent 3 oral treatments with 2-L solutions of NaHCO3 (150mmol/L), glucose (300mmol/L), and glucose + NaHCO3 (300mmol/L + 150mmol/L, respectively) in randomized order. Arterial and venous blood was obtained before treatment and in 30-min intervals thereafter for blood gas analysis and determination of plasma protein and electrolyte concentrations. Urine was collected volumetrically to determine urine volume, osmolality, pH, net acid excretion, and renal Na excretion after treatment. Plasma volume changes were extrapolated from plasma protein concentration changes. Treatment and time effects were tested with repeated measures ANOVA. Only subtle differences between oral administration of NaHCO3, with and without glucose, were observed for the change of the standard HCO3 concentration relative to baseline. No differences in plasma Na, plasma volume expansion, renal Na, net base excretion, urine volume, or pH could be identified between animals treated orally with NaHCO3 with and without glucose. Similarly, no differences in blood glucose concentration, plasma volume expansion, urine volume, or renal glucose excretion were observed in the 8h after treatment when comparing oral glucose treatment with and without NaHCO3. Our results indicate that combination of NaHCO3 with glucose in a hypertonic ORS only had a minor effect on the alkalinizing effect of NaHCO3, which is unlikely to be of clinical relevance. The combination of NaHCO3 and glucose neither improved Na, glucose, nor water absorption in euhydrated neonatal dairy calves, questioning the relevance of a specific ratio between Na and glucose in ORS for calves.

Plasma potassium-lowering effect of oral glucose, sodium bicarbonate, and the combination thereof in healthy neonatal dairy calves

Hyperkalemia is a common complication in neonatal diarrheic dairy calves and is corrected by administration of glucose or sodium bicarbonate. Although the hypokalemic effect of glucose is well established in other species, controversial results are reported for sodium bicarbonate. Our objective was to study the effect and mechanism of action of glucose and sodium bicarbonate on the potassium homeostasis of healthy neonatal dairy calves. Nine healthy neonatal Holstein-Friesian calves underwent 3 oral treatments with 2L of NaHCO3 (150mmol/L), glucose (300mmol/L), and glucose+NaHCO3 solution (300mmol/L+150mmol/L) in randomized order. Blood was obtained before treatment (T0) and at 30-min intervals thereafter. Changes between each time point and T0 were determined for all parameters. Urine was collected volumetrically to determine total renal potassium excretion over an 8-h posttreatment period. Plasma volume changes were extrapolated from changes in plasma protein concentration. Treatment and time effects were tested with repeated-measures ANOVA. Multivariate stepwise regression analysis using dummy variable coding was conducted to identify associations between changes in plasma potassium concentration ([K]) and changes in plasma glucose concentration ([glucose]), blood base excess, and plasma volume. Oral glucose and sodium bicarbonate treatments decreased [K] by 25 and 19%, respectively, whereas the combination of both compounds caused an intermediate [K] decline (22%). For the glucose treatment, the decline in [K] was only associated with changes in plasma [glucose] (partial R2=0.19). In NaHCO3-treated calves, [K] decline was associated with change of extracellular volume (partial R2=0.31) and blood base excess (partial R2=0.19). When glucose and NaHCO3 were combined, [K] decline was associated with changes in plasma volume (partial R2=0.30), BE (R2=0.22), and [glucose] (partial R2=0.03). Our results indicate that glucose lowers plasma [K] mainly through an insulin-dependent intracellular translocation of K, whereas NaHCO3 causes hypokalemia through hemodilution followed by intracellular translocation of K caused by the strong ion effect. The combination of glucose and NaHCO3 at the dosage used in this study does not have an additive hypokalemic effect. When combined, hemodilution and strong ion effect have the strongest effect on plasma [K], whereas the insulin-dependent effect of glucose appears to be blunted.

Changes in Clinical Enzyme Activity and Bile Acid Levels in Psittacine Birds With Altered Liver Function and Disease

The diagnosis of hepatic disease in psittacine birds by using biochemical enzyme and bile acid levels is problematic. In this study, we looked at changes in clinical enzyme concentrations, bile acid levels, and plasma protein electrophoresis results to determine whether these analytes correlate with hepatic disease. A total of 442 samples representing 8 species of psittacine birds were analyzed for levels of bile acids, aspartate aminotransferase (AST), and creatine kinase (CK) and for changes in plasma protein concentrations determined by electrophoresis. Follow-up questionnaires were received from the submitting veterinarian detailing the diagnosis of the patient and whether liver disease had been confirmed by histologic examination. When the data were examined without regard to diagnosis, high clinical enzyme concentrations correlated poorly with high levels of bile acids. These results suggest that the often used corollary of high AST and normal CK concentrations as a sole screening test to evaluate hepatocellular injury is not valid. High bile acid levels had the highest association with confirmed hepatic disease, followed by high concentrations of alpha2-globulins, AST, lactate dehydrogenase (LDH), and alpha1-globulins.

Increase in plasma protein concentration for diagnosing weaning-induced pulmonary oedema

Weaning-induced cardiogenic pulmonary oedema is a cause of weaning failure that is classically diagnosed by an increase in pulmonary artery occlusion pressure during a spontaneous breathing trial. During cardiogenic pulmonary oedema, a hypo-oncotic fluid is filtered toward the interstitial space. Thus, we tested whether the changes in plasma protein concentration during a weaning trial could diagnose weaning-induced pulmonary oedema. Prospective study. The 24-bed medical intensive care unit of a university hospital. Forty-six patients who had failed two weaning trials. Weaning-induced pulmonary oedema was diagnosed if a respiratory failure associated with an increase in pulmonary artery occlusion pressure above 18 mmHg occurred during a third weaning trial on a T-tube. The plasma protein concentration was measured before and at the end of the spontaneous breathing trial. During the weaning trial, pulmonary oedema was observed in 24 patients. In these patients, the plasma protein concentration increased by 11% (3-25%). The plasma protein concentration did not change significantly in patients who did not experience weaning-induced pulmonary oedema. An increase in the plasma protein concentration greater than 6% from baseline to the end of the weaning trial allowed detecting a weaning-induced pulmonary oedema with a sensitivity of 87% and a specificity of 95%. The acute changes in plasma protein concentration during a weaning trial represent an alternative method to right heart catheterisation for assessing weaning-induced pulmonary oedema.

Time course and magnitude of fluid and electrolyte shifts during recovery from high-intensity exercise in Standardbred racehorses

AbstractThe present study characterized the fluid and electrolyte shifts that occur in Standardbred racehorses during recovery from high-intensity exercise. Jugular venous blood was sampled from 13 Standardbreds in racing condition, at rest and for 2 h following a high-intensity training workout. Total body water (TBW), extracellular fluid volume (ECFV) and plasma volume (PV) were measured at rest using indicator dilution techniques (D2O, thiocyanate and Evans Blue, respectively). Changes in TBW were assessed from measures of body mass, and changes in PV and ECFV were calculated from changes in plasma protein concentration. Exercise resulted in a 26.9% decrease in PV. At 10 min of recovery TBW and ECFV were decreased by 2.2% and 16.5% respectively, while intracellular fluid volume was increased by 7.1%. There was a continued loss of fluid due to sweating throughout the recovery period such that TBW was decreased by 3.9% at 90 min of recovery. This decrease in TBW was nearly equally partitioned between the extracellular and intracellular fluid compartments. Plasma Na+and Cl−contents were decreased at 1 min of recovery, but not different from rest by 40 min of recovery. Plasma K+content at 1 min post exercise was not different from the pre-exercise value; however, by 5 min of recovery K+content was significantly decreased and it remained decreased throughout the recovery period. It is concluded that there are very rapid and large fluid and electrolyte shifts between body compartments during and after high-intensity exercise, and that full recovery of these shifts requires 90–120 min.

Acute phase mediator oncostatin M regulates affinity of α 1-protease inhibitor for concanavalin A in hepatoma-derived but not lung-derived epithelial cells

Quantitative changes in plasma protein concentrations during tissue injury or inflammation (acute phase response) are often accompanied by specific alterations in the carbohydrate moieties of these proteins. The glycosylation changes comprise alterations in the type of branching of the carbohydrate structures as revealed by modulated reactivity of acute phase glycoproteins with the lectin concanavalin A. Interestingly, inflammation-induced changes in the glycosylation of acute phase proteins have been shown to affect the functional properties of these proteins. In this study we demonstrate that synthesis of acute phase protein α 1-PI, the controlling inhibitor of neutrophil elastase, is significantly up-regulated in hepatic and lung-derived epithelial cells by the inflammatory mediator oncostatin M. Although oncostatin M markedly altered the concanavalin A reactivity of hepatic α 1-PI, lung-derived epithelial cells did not change the pattern of α 1-PI glycan branching upon stimulation with oncostatin M. These results indicate that inflammation-induced changes in glycosylation of α 1-PI may have different impacts on functional properties of liver and lung-synthesized α 1-PI.

Time course and magnitude of changes in total body water, extracellular fluid volume, intracellular fluid volume and plasma volume during submaximal exercise and recovery in horses

AbstractThe purpose of the present study was to determine the time course and magnitude of changes in extracellular and intracellular fluid volumes in relation to changes in total body water during prolonged submaximal exercise and recovery in horses. Seven horses were physically conditioned over a 2-month period and trained to trot on a treadmill. Total body water (TBW), extracellular fluid volume (ECFV) and plasma volume (PV) were measured at rest using indicator dilution techniques (D2O, thiocyanate and Evans Blue, respectively). Changes in TBW were assessed from measures of body mass, and changes in PV and ECFV were calculated from changes in plasma protein concentration. Horses exercised by trotting on a treadmill for 75–120 min incurred a 4.2% decrease in TBW. During exercise, the entire decrease in TBW (mean±standard error: 12.8±2.0 l at end of exercise) could be attributed to the decrease in ECFV (12.0±2.4 l at end of exercise), such that there was no change in intracellular fluid volume (ICFV; 0.9±2.4 l at end of exercise). PV decreased from 22.0±0.5 l at rest to 19.8±0.3 l at end of exercise and remained depressed (18–19 l) during the first 2 h of recovery. Recovery of fluid volumes after exercise was slow, and characterized by a further transient loss of ECFV (first 30 min of recovery) and a sustained increase in ICFV (between 0.5 and 3.5 h of recovery). Recovery of fluid volumes was complete by 13 h post exercise. It is concluded that prolonged submaximal exercise in horses favours net loss of fluid from the extracellular fluid compartment.

Atrial distension, haemodilution, and acute control of renin release during water immersion in humans.

We tested the hypothesis that atrial distension (stimulation of cardiopulmonary baroreceptors) is not the single pivotal stimulus for the acute suppression of renin release during water immersion in humans and that immersion-induced haemodilution constitutes an important additional stimulus. In nine healthy male subjects, identical increases in atrial distension were induced by two immersion procedures (of 30 min each); one without (WI) and one with attenuation (WI + cuff) of the concomitant haemodilution (estimated from changes in plasma protein concentration) by inflating thigh cuffs during immersion. During WI, central venous pressure (CVP) and left atrial diameter (LAD) increased (P < 0.05) by 5.5 +/- 0.4 mmHg and 4.6 +/- 0.5 mm, respectively, and plasma protein concentration and plasma renin activity (PRA) progressively decreased (P < 0.05) by 4.8 +/- 0.5 g L(-1) and 1.6 +/- 0.2 ng mL(-1) h(-1) (to 49 +/- 4% of baseline values), respectively. The WI + cuff caused similar atrial distension as WI (CVP and LAD increased by 6.9 +/- 0.5 mmHg and 5.5 +/- 0.5 mm, respectively), attenuated haemodilution (plasma protein concentration decreased by 1.9 +/- 0.4 g L(-1), P < 0.05 vs. WI), and markedly inhibited suppression of PRA, which decreased by 0.4 +/- 0.1 ng mL(-1) h(-1) (to 87 +/- 4% of baseline values, P < 0.05 vs. WI). Differences in renin release could not be accounted for by differences in mean arterial pressure. In conclusion, baroreceptor stimulation induced by atrial distension is not the single pivotal stimulus for the acute suppression of renin release in response to intravascular volume expansion by water immersion in humans. Haemodilution constitutes a significant and conceivably the principal stimulus for the acute immersion-induced suppression of renin-angiotensin system activity.

Recent advances in the pharmacokinetics of local anaesthetics. Long-acting amide enantiomers and continuous infusions.

The most widely used long-acting amide local anaesthetic is bupivacaine, a racemic mixture of 2 stereoisomers. However, there is evidence that the use of single enantiomer compounds offers advantages over racemic agents. Ropivacaine, the recently introduced propyl homologue of bupivacaine, is a pure S-(-)-enantiomer. It is associated with a reduced incidence of both cardiovascular and central nervous system toxicity, a concern with racemic bupivacaine, in preclinical studies. The relevant pharmacokinetic differences include a lower lipid solubility, a slightly higher plasma clearance and shorter elimination half-life (t 1/2 beta) compared with racemic bupivacaine, with a similar degree of plasma protein binding. More recently levobupivacaine, the pure S-(-)-enantiomer of bupivacaine, has been produced. Stereoselective differences have been observed between the 2 enantiomers and the racemic mixture, with levobupivacaine exhibiting a slightly higher degree of plasma protein binding, a lower volume of distribution, a higher plasma clearance, and a shorter t 1/2 beta than the R-(+)-enantiomer. In common with ropivacaine, levobupivacaine has been shown to have a reduced incidence of toxicity in comparison the R-(+)-enantiomer in preclinical studies, explained in part by a reduced affinity to both brain and myocardial tissue. Racemic bupivacaine is increasingly administered by continuous infusion to provide prolonged postoperative analgesia. The pharmacokinetic profile of the drug administered in this manner has only recently been elucidated and indicates a slow rise in total plasma concentration with increasing duration of infusion, mitigated by changes in plasma protein concentrations during the postoperative period. This appears to be the predominant reason why complications related to systemic toxicity are rarely observed with this technique. However, continuous administration of individual enantiomers may potentially serve as a safer option in the future.

Changes in plasma protein concentrations in ponies with experimentally induced alimentary laminitis

Abstract Objective To determine whether plasma protein concentrations were altered in ponies with alimentary laminitis. Animals 12 adult ponies. Procedure Acute laminitis was induced in 6 ponies by oral administration of carbohydrate (85% corn starch, 15% wood flour); the other 6 ponies were used as controls. A physical examination was performed and blood samples were collected immediately before and 4, 8, 12, 24, and 28 hours after administration of carbohydrate. Plasma protein concentrations were determined by means of sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Results 19 plasma proteins ranging from a molecular weight of 24,000 to a molecular weight of 350,000 were identified in all 12 ponies. Plasma concentrations of proteins with molecular weights of 350,000 (fibrinogen), 130,000 (ceruloplasmin), 118,000 (c-reactive protein), 67,000 (α1-antitrypsin I), 65,000 (α1-antitrypsin II), 50,000 (haptoglobulin), and 45,000 (acid glycoprotein) were significantly increased in ponies with laminitis, compared with concentrations in control ponies. Conclusion Changes in plasma protein concentrations are detectable within 4 hours after the onset of alimentary laminitis in ponies. Clinical Relevance Measurement of plasma protein concentrations may be useful in monitoring the progression of laminitis in ponies. (Am J Vet Res 1998;59:1234–1237)

A finite-element model of oxygen diffusion in the pulmonary capillaries.

We determined the overall pulmonary diffusing capacity (DL) and the diffusing capacities of the alveolar membrane (Dm) and the red blood cell (RBC) segments (De) of the diffusional pathway for O2 by using a two-dimensional finite-element model developed to represent the sheet-flow characteristics of pulmonary capillaries. An axisymmetric model was also considered to assess the effect of geometric configuration. Results showed the membrane segment contributing the major resistance, with the RBC segment resistance increasing as O2 saturation (SO2) rises during the RBC transit: RBC contributed 7% of the total resistance at the capillary inlet (SO2 = 75%) and 30% toward the capillary end (SO2 = 95%) for a 45% hematocrit (Hct). Both Dm and DL increased as the Hct increased but began approaching a plateau near an Hct of 35%, due to competition between RBCs for O2 influx. Both Dm and DL were found to be relatively insensitive (2-4%) to changes in plasma protein concentration (28-45%). Axisymmetric results showed similar trends for all Hct and protein concentrations but consistently overestimated the diffusing capacities (approximately 2.2 times), primarily because of an exaggerated air-tissue barrier surface area. The two-dimensional model correlated reasonably well with experimental data and can better represent the O2 uptake of the pulmonary capillary bed.

Influence of atrial natriuretic factor on fluid efflux from the splenic circulation of the rat.

1. Atrial natriuretic factor (ANF) causes a reduction in plasma volume that is abolished by splenectomy. Experiments were conceived to investigate whether ANF acts within the spleen to increase efflux of fluid from the intravascular to the extravascular space. 2. ANF, infused into the splenic artery of anaesthetized rats at rates of 1, 5 and 20 ng min -1, caused a dose-dependent increase in the arteriovenous difference in haematocrit as blood flowed through the spleen (basal difference, 0.18 +/- 0.10%; difference after 10 min at 20 ng min -1 ANF, 1.5 +/- 0.18%; n = 6). There was no such change in plasma protein concentration. 3. ANF (20 ng min -1) did not alter splenic arterial blood flow. However, splenic venous blood flow fell so that the arteriovenous difference increased significantly (basal difference, 0.34 +/- 0.19 ml min -1; difference at 60 min, 1.1 +/- 0.20 ml min-1, n = 7). There was no change in mean arterial pressure. 4. These data confirm our hypothesis that ANF acts within the spleen to increase fluid efflux from the intravascular to the extravascular space. Since there is no change in total splenic blood flow, we propose that the effects of ANF are mediated by dilatation of the splenic afferent arterioles and constriction of the efferent venules, thus increasing filtration pressure.