Multiple Plasma Samples Research Articles

Limited Sampling Strategy to Predict AUC of the CYP3A Phenotyping Probe Midazolam in Adults: Application to Various Assay Techniques

Midazolam clearance is used to phenotype hepatic CYP3A activity but requires multiple plasma samples following a single intravenous dose. The authors evaluated the use of a limited sampling scheme, using different assay techniques to determine the reproducibility of such a strategy in estimating midazolam AUC. Seventy-three healthy adults received midazolam as a single intravenous bolus dose. At least eight plasma samples were collected from each subject and were assayed using either LC/MS/MS or electron capture gas chromatography. Eleven subjects were randomly selected for the training set using stepwise linear regression to determine relationships between midazolam plasma concentrations and AUC. Validation of the predictive equations was done using the remaining 62 subjects. Mean percent error (MPE), mean absolute error (MAE), and root mean square error (RMSE) were calculated to determine bias and precision. Based on the training set, five models were generated with coefficients of determination ranging from 0.87 to 0.95. Validation showed that MPE, MAE, and RMSE values were acceptable for three of the models. Intrasubject reproducibility was good. In addition, training set datafrom one institution were able to predict data from the other two institutions using other assay techniques. Minimized plasma sampling mayprovide a simpler method for estimating midazolam AUC for CYP3A phenotyping. A limited sampling strategy is more convenient and cost-effective than standard sampling strategies and is applicable to more than one assay technique.

Exchangeable Zinc Pool Masses and Turnover Are Maintained in Healthy Men with Low Zinc Intakes

Previous studies suggest that rapidly exchanging zinc pools (EZP), thought to supply the zinc required by tissues, are smaller and turn over more rapidly in individuals with lower zinc intakes. We studied the effects of low dietary zinc (4.6 mg/d) on EZP mass and turnover in seven healthy men confined during a 20-wk clinical study. Supplements of 9.1 mg zinc were given during the 5-wk baseline and repletion periods, and placebos were given during a 10-wk zinc-restriction period. Stable 70Zn tracers were administered intravenously at the end of baseline, 3 and 10 wk after the start of zinc restriction and at the end of repletion. Multiple plasma samples were collected over an 8-d period after tracer administration. 70Zn:66Zn ratios were measured using inductively coupled plasma mass spectrometry, and tracer-tracee data were analyzed by compartmental modeling. Activities of the zinc-dependent enzymes, alkaline phosphatase and 5'nucleotidase, were unchanged during the study. There were no significant changes in EZP masses or kinetic parameters. A three-compartment model indicated that the masses of plasma zinc and total EZP averaged 3.25 +/- 0.58 and 147.8 +/- 33.2 mg, respectively, at the four time points studied. Plasma zinc mass turned over at an average of 5.3 times per hour. There was an 11% reduction (P = 0.06) in plasma zinc flux 3 wk after the start of the low zinc diet period, but it returned to baseline values after 10 wk of zinc restriction. The results suggest that total EZP mass is maintained when dietary zinc is reduced to 4.6 mg/d over a 10-wk period.

Proteomic comparison of human and great ape blood plasma reveals conserved glycosylation and differences in thyroid hormone metabolism

Most blood plasma proteins are glycosylated. These glycoproteins typically carry sialic acid-bearing sugar chains, which can modify the observed molecular weights and isoelectric points of those proteins during electrophoretic analyses. To explore changes in protein expression and glycosylation that occurred during great ape and human evolution, we subjected multiple blood plasma samples from all these species to high-resolution proteomic analysis. We found very few species-specific differences, indicating a remarkable degree of conservation of plasma protein expression and glycosylation during approximately 12 million years of evolution. A few lineage-specific differences in protein migration were noted among the great apes. The only obvious differences between humans and all great apes were an apparent decrease in transthyretin (prealbumin) and a change in haptoglobin isoforms (the latter was predictable from prior genetic studies). Quantitative studies of transthyretin in samples of blood plasma (synthesized primarily by the liver) and of cerebrospinal fluid (synthesized locally by the choroid plexus of the brain) confirmed approximately 2-fold higher levels in chimpanzees compared to humans. Since transthyretin binds thyroid hormones, we next compared plasma thyroid hormone parameters between humans and chimpanzees. The results indicate significant differences in the status of thyroid hormone metabolism, which represent the first known endocrine difference between these species. Notably, thyroid hormones are known to play major roles in the development, differentiation, and metabolism of many organs and tissues, including the brain and the cranium. Also, transthyretin is known to be the major carrier of thyroid hormone in the cerebrospinal fluid, likely regulating delivery of this hormone to the brain. A potential secondary difference in retinoid (vitamin A) metabolism is also noted. The implications of these findings for explaining unique features of human evolution are discussed.

Determinants of ceftriaxone clearance by continuous venovenous hemofiltration and hemodialysis.

To guide individual ceftriaxone dosages in patients receiving continuous renal replacement therapy. Prospective, outpatient study. University-affiliated general clinical research center. Eight patients receiving hemodialysis. We performed controlled clearance studies with three hemofilters: an acrylonitrile copolymer 0.6 m2 (AN69), polymethylmethacrylate 2.1 m2 (PMMA), and polysulfone 0.65 m2 (PS). Subjects received ceftriaxone 1000 mg intravenously before the start of a clearance study. The concentration of ceftriaxone in multiple plasma and dialysate-ultrafiltrate samples was determined by high-performance liquid chromatography. The diffusional clearances (Cl(diffusion)) and sieving coefficients (SC) of ceftriaxone, urea, and creatinine were compared by a mixed-model repeated-measures analysis of variance with filter and blood, dialysate inflow, or ultrafiltration rate as the main effect and patient as a random effect. The fraction of ceftriaxone bound to plasma proteins was 43 +/- 15% (range 13-92%). Concentration dependence was evident in all three groups. The fraction unbound to plasma proteins (f(up)) at the time that SCs were determined was significantly lower in the PS group (0.16 +/- 0.07) than the AN69 group (0.30 +/- 0.17, p<0.01), but similar to that in the PMMA group (0.27 +/- 0.12). Despite the higher f(up), the SC of unbound ceftriaxone with the AN69 filter (0.48 +/- 0.13) was significantly lower than values for the PMMA (0.86 +/- 0.33) and PS (0.82 +/- 0.22) groups (p<0.05). Continuous venovenous hemofiltration clearance of urea and unbound ceftriaxone increased significantly only for the PMMA (p=0.006) and PS (p=0.015) filters when the ultrafiltration rate was increased. Significant linear relationships (p<0.0001) were observed between Cl(diffusion) of unbound ceftriaxone and clearance of urea for all three filters: AN69 slope = 0.57, PMMA slope = 0.90, and PS slope = 1.02. The slope of this relationship for the AN69 filter was significantly lower than for the other two filters. Ceftriaxone clearance was significantly increased and membrane dependent during continuous venovenous hemofiltration and continuous venovenous hemodialysis. Thus individual ceftriaxone dosages for patients receiving continuous renal replacement therapies should incorporate extracorporeal clearance.

Pharmacokinetic modeling of M6G formation after oral administration of morphine in healthy volunteers.

Morphine is metabolized to two major metabolites, morphine-3-glucuronide and morphine-6-glucuronide (M6G). Under the conditions of long-term oral morphine administration, the accumulation of M6G may contribute to the analgesic effects, but it may also cause respiratory depression. Five healthy male volunteers (ages 25-34 yr) received 90 mg MST (morphine sulfate 5H2O sustained-released tablet, equivalent to 67.8 mg oral morphine). Multiple plasma and urine samples were taken for as long as 14 and 36 h, respectively. Individual pharmacokinetics after intravenous administration of morphine and M6G were available from a previous investigation. A new model that considers the M6G-plasma profile as a sum of the input from the first-pass metabolism of morphine and the input from systemically available morphine was applied to the plasma concentration versus time curves of M6G. The concentrations of M6G at the effect site after long-term morphine administration were simulated. The fraction of morphine absorbed from the gut was 82+/-14%. Of this, 42+/-8% passed through the liver, resulting in an oral bioavailability of morphine of 34+/-9%. Of the total amount of M6G, 71+/-7% was formed during the first-pass metabolism, and 29+/-7% was formed by metabolism of systemic morphine. After 36 h, the amounts of M6G and morphine excreted in the urine were 92+/-17% and 9+/-3%, respectively. Simulation of effect-site concentrations of M6G indicated that after multiple oral dosing of morphine in patients with normal liver and renal function, M6G might reach concentrations two times greater than that of morphine. M6G may contribute to the analgesic and side effects seen with long-term morphine treatment. The current model of morphine and M6G pharmacokinetics after oral administration of morphine may serve as a pharmacokinetic basis for experiments evaluating the analgesic contribution of M6G with long-term oral dosing of morphine.

The influence of oral lipid loads on acylation stimulating protein (ASP) in healthy volunteers.

To examine the hypothesis that a sustained rise in plasma acylation stimulating protein (ASP, C3a desarg) accompanies the elevation in triacylglycerol that follows the ingestion of an oral fat load. Following an overnight fast, blood samples were obtained from healthy volunteers while fasting and 15 min, 1, 2, 4, 6 and 8 h following ingestion of: (i) a liquid meal, rich in dairy fat (eight subjects) and (ii) a semi-liquid meal, with higher total fat content and rich in polyunsaturated fat (six subjects). Four male and four female volunteers (age range: 22-51 y; body mass index (BMI): 17.9-26.9 kg/m2) received the first meal. Six subjects (age range: 32-60 y; BMI: 18.0-28.4 kg/m2), including three from the first study, received the second meal using the same protocol. ASP and C5a were measured by radioimmunoassay (RIA) and the complement proteins C3, factor B and C5 by radial immunodiffusion or nephelometry. Tumour necrosis factor (TNF)-alpha was measured by enhanced ELISA, and plasma cholesterol and triacylglycerol by an automated enzymatic method. The presence of chylomicrons was assessed in post-prandial plasma samples taken after the second meal. There was no significant change in mean ASP concentration in either group at any time point, following ingestion of either meal. However, there was a significant positive linear trend in ASP following the second fat challenge (ANOVA; P < 0.05). There was also no change in complement proteins, plasma cholesterol or TNF-alpha. Plasma triacylglycerol rose significantly after the first and second meals (P < 0.05 and P < 0.001 at 2 h post-prandially); the mean maximum rise above the fasting level was 58 +/- 41% and 89 +/- 38% respectively (mean +/- s.d.). Chylomicrons were detected in samples taken from each subject after the second meal. Analysis of individual ASP data showed a sustained rise in one subject after the first meal and two subjects after the second meal. Substantial variation in ASP concentration was observed in samples taken in the first 2 h post-prandially. There was no significant change in ASP nor other complement proteins for either group of subjects following ingestion of the lipid loads. Individual data showed substantial variation in post-prandial ASP, but multiple plasma sampling did not define the basis for this variation.

Glomerular filtration rate determination in diabetic patients using iohexol clearance — comparison of single and multiple plasma sampling methods

Use of iohexol clearance has been described as the gold standard for the measurement of glomerular filtration rate (GFR). It is suggested that multiple plasma sampling following iohexol injection is required to accurately determine GFR by area under plasma clearance curve. The aim of this study was to determine whether single plasma sampling 4 h after injection of iohexol could accurately determine GFR in diabetic patients with mild to moderate renal failure, compared to multiple plasma sampling. A total of 120 GFR determinations in 36 patients with non-insulin dependent diabetic renal disease were done over 1 year. No acute deterioration was seen in renal function following injection of contrast in any patient. Strong correlation in GFR measurement was observed between the multiple plasma sampling method and the single plasma sampling method ( r 2=0.975). Single plasma sampling 4 h after bolus injection of iohexol is a safe and accurate method of determining GFR and change in GFR in diabetic subjects with mild to moderate renal impairment.

A compartmental model of zinc metabolism in healthy women using oral and intravenous stable isotope tracers

A mathematical model of zinc metabolism in six healthy women (average age: 30 +/- 11 y) was developed by using stable isotopes of zinc. After equilibration on a constant diet containing 7.0 mg Zn/d, an oral tracer highly enriched in 67Zn and an intravenous tracer highly enriched in 70Zn were administered simultaneously. Multiple plasma and 24-h urine samples were collected for the next 7 d with complete fecal collections for 11 d. Tracer-trace ratios in plasma, urine, and feces were calculated from isotope ratios of 67Zn to 66Zn and 70Zn to 66Zn measured by using inductively coupled plasma-mass spectrometry. An a priori identifiable model composed of seven compartments was developed to describe the kinetics of both tracers as well as that of naturally occurring zinc. The parameters of the model were fitted to the data by using the SAAM-CONSAM modeling software and were estimated with good precision. Several important, not directly measurable zinc variables were estimated (mean +/- SEM) from the model including the fractional absorption from the gastrointestinal tract (0.279 +/- 0.043), the rates of endogenous secretion (2.79 +/- 0.49 mg/d) and excretion (2.01 +/- 0.35 mg/d), the fractional turnover rate of the plasma pool (131 +/- 20/d), and the sizes (7.2 +/- 1.2 and 77.1 +/- 6.4 mg) and fractional turnover rates (22.3 +/- 7.1 and 1.49 +/- 0.18/d) of the fast and slow tissue pools equilibrating with the plasma, respectively.

PLASMA FVIII LEVELS MEASURED AFTER INFUSION OF RECOMBINANT F VIII (rFVIII) VARY SIGNIFICANTLY WITH DIFFERENT ASSAY METHODS - (WILL THE REAL FVIII LEVEL PLEASE STAND UP!). † 934

As part of a bioequivalence study comparing two lots of rFVIII(Kogenate®), 9 persons with hemophilia A were infused with rFVIII from each lot, and multiple plasma samples were then obtained from each subject for determination of FVIII assays over 48 hrs. Samples were assayed by 4 different methods: chromogenic, and one stage APTT method using 3 different types of activators (A)-micronized silica, ellagic acid, and kaolin. All samples were immediately centrifuged, snap frozen and stored at -70°C until assayed in duplicate. The same reference plasma standard was used throughout. Results, shown below, indicate a consistent difference in F VIII assay values, with those run by chromogenic substrate method being roughly twice as high as those run by one-stage APTT method with kaolin as A. One stage results with micronized silica or ellagic acid as A were comparable, and were consistently higher than those with kaolin. Most U.S. clinical labs use a one-stage assay, with micronized silica or ellagic acid A. Because significantly different results can be obtained depending on F VIII assay method used, multicenter studies must carefully standardize assay techniques and reagents in order to combine data for analysis. Additionally, such differences have significant cost implications (partially funded by Bayer). Figure

Iohexol clearance for GFR-determination in renal failure--single or multiple plasma sampling?

Iohexol clearance for GFR-determination in renal failure--single or multiple plasma sampling?

Iohexol clearance for GFR-determination in renal failure—single or multiple plasma sampling?

The present study examined the agreement between single and multiple sample plasma clearance of iohexol, a non-ionic contrast agent, in renal failure. Sixty-five patients with varying degree of renal insufficiency received 10 ml iohexol (300 mg I/ml) i.v. and plasma samples were collected four times during the following 3-24 h. Plasma-iodine concentrations were determined by X-ray fluorescence. Predicted creatinine clearance was used to choose one of the samples for determination of single sample clearance. A single plasma specimen collected at 4 h for GFR above 50 ml/min, at 7 h for GFR between 20 and 50 ml/min, and at 24 h for GFR below 20 ml/min gave values in good agreement with those based on a four sample slope clearance. No sign of nephrotoxicity was noted after administration of the contrast agent. It is concluded that single sample plasma clearance after single injection of iohexol gives a good estimate of GFR in renal failure and is advantageous in clinical practice.

Elimination of n-butylated hydroxytoluene methylation during fatty acid analysis by gas chromatography

An improved method for fatty acids analysis with optimum recovery of highly polyunsaturated fatty acids methyl esters in biological systems is presented. The method is based on transesterification of phospholipid and triacylglycerols to fatty acid methyl esters using a commercially available reagent, Methyl-Prep II. Without proper precautions, as much as 50% of n-butylated hydroxytoluene (BHT) added to prevent oxidation of polyunsaturated fatty acids, could be methylated during the transesterification step. Methylated BHT elutes close to 14:0 (myristic acid) and no longer functions as an antioxidant, but the modified conditions virtually eliminate the methylation of BHT. Sample extraction and methylation was completed in 30 min at room temperature. A chelator (diethylenetriamine-pentaacetic acid; DTPA) is also added to prevent peroxidation of metal catalyzed free radical chain reactions. The standard deviations of the major fatty acids from multiple human plasma samples prepared on different days were less than 5%. The recovery of arachidonic acid, 20:4, from plasma was improved using the new method, and the recovery for docosahexaenoic acid, 22:6, spiked to human plasma was found to be 99%.

Diagnostic serologic testing of cage and aviary birds for chlamydiosis and suggested confirmatory testing.

A 2 x 2 contingency table was constructed to demonstrate the relationships between detectable chlamydial antibody activity and clinical health status of tested birds. The table revealed that 65.5% of clinically ill birds were antibody positive by elementary body agglutination (EBA) (> or = 10 titers) and 59.0% were antibody positive by latex agglutination (LA). Thus, EBA was slightly more sensitive than LA in detecting antibody activity. Of the clinically normal birds, 96.7% were antibody negative (< 10 titers) by EBA and 98.3% were antibody negative by LA. Individual serum or plasma samples from a group of mixed types of psittacine birds and cockatiels were tested as a separate group, and relationships between EBA-detectable antibody activity and health status were obtained from a 2 x 2 contingency table. Sixty-six percent of birds clinically ill with signs of chlamydiosis in the mixed-type group were antibody positive, whereas only 32.3% of clinically ill cockatiels were antibody positive. Statistical analysis of the contingency table using a chi-square test demonstrated that the EBA test differentiates between individual birds on the basis of health status (P < 0.001). When testing paired serum or plasma samples by EBA, LA, and direct complement fixation (DCF), the highest percentage of significant (> or =4-fold change) titer decreases was detected by LA, and the highest percentage of significant titer increases was detected by DCF. Examples of EBA, LA, and DCF titers in paired and multiple serum or plasma samples are presented to show the variety of responses that can occur. Results reflected variations seen in individual testing of birds with titer variability seen in the first sample tested. Additional types of testing believed necessary for confirming or ruling out an infectious process in birds are outlined. The current interpretations of serologic results are given.

Determination of robust ocular pharmacokinetic parameters in serum and vitreous humor of albino rabbits following systemic administration of ciprofloxacin from sparse data sets by using IT2S, a population pharmacokinetic modeling program.

Robust determination of the concentration-time profile of anti-infective agents in certain specialized compartments is often limited by the inability to obtain more than a single sample from such a site in any one subject. Vitreous humor and cerebrospinal fluid are obvious examples for which the determination of concentrations of anti-infective agents is limited. Advances in pharmacodynamics have pointed out the importance of understanding the profiles of drugs in the plasma and in specialized compartments in order to dose the drugs to obtain the best patient outcomes. Advances in population pharmacokinetic modeling hold the promise of allowing proper estimation of drug penetration into the vitreous (or other specialized compartment) with only a single vitreous sample, in conjunction with plasma sampling. We have developed a rabbit model which allows multiple samples of vitreous to be obtained without breaking down the blood-vitreous barrier. We have employed this model to test the hypothesis that robust estimates of vitreous penetration by the fluoroquinolone ciprofloxacin can be obtained from a traditional intensive plasma sampling set plus a single vitreous sample. We studied 33 rabbits which were receiving 40 mg of ciprofloxacin per kg of body weight intravenously as short infusions and from which multiple plasma and vitreous samples were obtained and assayed for ciprofloxacin content by high-performance liquid chromatography. Data were analyzed by the iterative two-stage population modeling technique (IT2S), employing the iterative two-stage program of Forrest et al. (Antimicrob. Agents Chemother. 37:1065-1072, 1993). Two data sets were analyzed: all plasma and vitreous samples versus all plasma samples and the initially obtained single vitreous sample. The pharmacokinetic parameter values identified were used to calculate the percent vitreous penetration as the ratio of the area under the concentration-time curve for the vitreous to that for the plasma. The values identified, 4% penetration for the full data set versus 3% penetration for the single vitreous sample data set, and their corresponding estimates were not statistically significantly different. We conclude that population modeling holds promise for the analysis of penetration of antimicrobiol agents into specialized spaces from which only single samples can be obtained, particularly for patients with whom robust plasma sampling can be performed.

Chronic glucocorticoid excess and impaired osmoregulation of vasopressin release in dogs with hepatic encephalopathy

Chronic liver disease may be accompanied by disturbed sodium and water homeostasis. There is usually sodium retention and ascites. However, spontaneous natriuresis has also been reported in humans and experimental animals with liver cirrhosis. Chronic hypercortisolism, which may occur in dogs with advanced liver disease, is known to induce the inhibition of the osmostimulation of vasopressin (AVP) release. We have therefore investigated the osmoregulation of AVP release in 11 dogs with chronic hypercortisolism associated with advanced liver dysfunction and hepatic encephalopathy and in 10 control dogs. Basal pituitary-adrenocortical activity was investigated by measuring the concentration in multiple plasma samples of adrenocorticotropin (ACTH), α-melanocyte-stimulating hormone (MSH), and cortisol and the cortisol:creatinine ratio in 24-hr urine. Urine specific gravity was also measured. The feedback regulation of the system was investigated by measuring these hormones in plasma after an intravenous (iv) injection of 0.01 mg/kg of dexamethasone. The osmoregulation of the release of AVP was investigated by the intravenous infusion of a 20% NaCl solution at a flow rate of 0.03 ml/kg for 2 hr and the measurement of AVP in plasma sampled at 20-min intervals. The AVP release was analyzed in terms of the threshold osmolality at which it commenced and the sensitivity, which reflects the magnitude of the response. All dogs had highly increased urinary cortisol:creatinine ratios, ranging from 21 to 210 × 10 −6 (normally <10 × 10 −6). The mean basal plasma concentrations of the three pituitary-adrenocortical hormones were significantly increased. ACTH values were 35 to 146 ng/l (normally, 14 to 68), MSH values were 26 to 118 ng/l (normally, 10 to 36), and cortisol values were 88 to 194 nmol/l (normally, 23 to 112). The feedback inhibition of the secretion of ACTH and cortisol in response to dexamethasone was unaffected. Urine specific gravity was significantly decreased. The regulation of AVP release was found to be abnormal in all dogs with hepatic encephalopathy. The osmotic threshold at which the release of AVP was induced was abnormally high in seven of the dogs with liver disease and in the normal range in one. It could not be determined in three dogs. The sensitivity of AVP release in response to increasing plasma hypertonicity was normal in two dogs and decreased in nine. In three dogs, there was no increase in AVP release. None of the dogs had normal values for both the sensitivity and the threshold. We conclude that dogs with liver disease associated with hepatic encephalopathy have hyper-adrenocorticism due to a resetting of the pituitary-adrenocortical axis, associated with profoundly impaired osmoregulation of the release of AVP by the posterior pituitary. These neuroendocrine derangements may contribute to impaired electrolyte homeostasis in chronic liver disease.

Pharmacokinetics and transplacental passage of imipenem during pregnancy.

Imipenem pharmacokinetics were studied in early pregnancy (n = 7; length of gestation, 8.6 +/- 1.5 weeks, mean +/- standard deviation), in late pregnancy (n = 7; length of gestation, 38.7 +/- 1.4 weeks), and in the nonpregnant state (n = 6). A single dose of 500 mg of imipenem-cilastatin (1:1) was administered as a 20-min infusion. Multiple plasma and urine samples, as well as specimens of umbilical plasma and amniotic fluid from the pregnant subjects, were collected at frequent intervals for 8 h. Imipenem concentrations were assayed by specific microbiologic assay. The mean peak concentrations in plasma were 14.7 +/- 4.9, 14.9 +/- 5.2, and 43 +/- 28.3 micrograms/ml in early pregnancy, late pregnancy, and the nonpregnant state, respectively. The volumes of distribution were significantly larger during early pregnancy (0.98 +/- 0.45 liter/kg of body weight, P < 0.005) and late pregnancy (0.59 +/- 0.19 liter/kg, P < 0.05) than in the nonpregnant state (0.33 +/- 0.10 liter/kg), and total clearances from plasma were faster in early pregnancy (12.7 +/- 7.8 ml min-1 kg-1, P < 0.05) and late pregnancy (10.7 +/- 4.6 ml min-1 kg-1, P < 0.05) than in the nonpregnant state (5.77 +/- 1.19 ml min-1 kg-1). The mean concentrations in amniotic fluid were 0.07 +/- 0.01 and 0.72 +/- 0.85 micrograms/ml in early and late pregnancy. The mean umbilical venous and arterial drug concentrations were 1.72 +/- 1.22 and 1.64 +/- 1.22 micrograms/ml. The feto-maternal ratio at the time of cesarean section was 0.33 +/- 0.12. These results indicate that an adjustment of doses of imipenem may be required when treating pregnant women because of considerable changes in imipenem pharmacokinetics during pregnancy.

Simultaneous determination of glomerular filtration rate and effective renal plasma flow

A method is described for the simultaneous determination of GFR and ERPF within the normal range using a single injection of 51Cr-EDTA and 125I-OIH. Reference values of GFR (range 59-176 ml min-1) and ERPF (268-810 ml min-1) were calculated from bi-exponential analysis of curves based on multiple plasma samples. A subset of the samples (44, 120, 180 and 240 min) was used to estimate GFR by fitting a bi-exponential curve. Regression analysis demonstrated that the reference GFR value could be predicted from this four-sample estimate with a standard error of estimate (SEE) of 2.9 ml min-1, which compared favourably with methods based on mono-exponential analysis (minimum SEE = 6.3 ml min-1) or single samples (minimum SEE = 7.7 ml min-1). The effective volume of distribution of 125I-OIH calculated from the 44 min sample was used to estimate ERPF using a published equation with a SEE of 52 ml min-1. The improved precision of this simplified method for the simultaneous measurement of GFR and ERPF in the normal range will facilitate longitudinal studies in large patient groups in the evaluation of the causative factors of renal disease.

A Comparison of Chloride, Bromide, and Sucrose Dilution Volumes in Neonatal Pigs

Use of 36Cl, 82Br, and [3H]sucrose to estimate extracellular water volume was evaluated in 14 piglets (7-14 days old). 36Cl and 82Br were distributed in approximately the same volume, but a period of 5-6 hr after injection was required to reach equilibrium in the neonatal pig. Dilution volumes calculated before equilibration (2-5 hr) for 36Cl (326 +/- 11 ml/kg) and 82Br (328 +/- 13 ml/kg) were different from equilibration (6-8 hr) phase volumes (356 +/- 13 ml/kg and 355 +/- 13 ml/kg, respectively; P less than 0.001). A 3-hr sample estimated the same volume distribution calculated by extrapolation of the 6- to 8-hr period because of the relationship between the two slopes of the plasma clearance curves. After the 82Br and 36Cl had achieved equilibration, each was distributed in a volume equivalent to total body chloride space (362 +/- 29 ml/kg) measured by neutron activation; no statistical differences were found (P = 0.6). The early equilibration phase measured a 10% smaller, faster exchangeable fraction of total body Cl. Sucrose dilution volume (332 +/- 19 ml/kg) required multiple plasma samples for extrapolation and measured a dilution volume 7% smaller (P less than 0.05) than total body chloride space.

Prostaglandin F1α levels during and after neonatal extracorporeal membrane oxygenation

Infants receiving extracorporeal membrane oxygenation therapy undergo long-term cardiopulmonary bypass, are systemically heparinized, and frequently receive platelet transfusions. Prostacyclin is a powerful inhibitor of platelet aggregation as well as a potent vasodilator. The levels of its stable metabolite prostaglandin F1 alpha increase significantly in children undergoing cardiopulmonary bypass during heart operations but decrease to preoperative levels after bypass. To determine the effect of long-term bypass on prostacyclin levels, multiple plasma samples were analyzed in 10 human neonates both during extracorporeal membrane oxygenation therapy and within 24 hours after extracorporeal membrane oxygenation. Prostaglandin F1 alpha, the stable metabolite of prostacyclin, was quantitated by radioimmunoassay in picograms per milliliter. Prostaglandin F1 alpha levels were elevated while the patients received extracorporeal membrane oxygenation therapy but decreased with duration of extracorporeal membrane oxygenation. In most infants, prostaglandin F1 alpha levels rose again during weaning from extracorporeal membrane oxygenation and remained elevated for 24 hours after extracorporeal membrane oxygenation. Extracorporeal membrane oxygenation course influenced circulating prostaglandin F1 alpha levels. Fluctuating prostaglandin F1 alpha levels are of clinical significance in the management of vasomotor tone and platelet function, common problems in the care and the prevention of hemorrhage in these critically ill infants.

Growth factor activity in the blood of women in whom preeclampsia develops is elevated from early pregnancy

Preeclampsia is a pregnancy-specific disorder of uncertain cause and pathophysiology that appears to be associated with endothelial cell injury. Our current studies demonstrate that a pregnancy growth factor activity is elevated compared with postpartum values in the blood of women with preeclampsia months before the onset of clinical manifestations of toxemia. A cohort of primigravid women was followed throughout pregnancy and multiple serial plasma samples from six women with preeclampsia and six matched normal women were assayed for mitogenic activity. The data indicated that the ratio of predelivery/postdelivery plasma mitogenic activity was greater in women predestined to meet strict criteria for the diagnosis of preeclampsia compared with matched normal primigravid women. Growth factor activity could distinguish women in whom preeclampsia would develop from their normal peers throughout pregnancy, and as early as the first trimester of gestation (p<0.05). Similar studies performed with plasma obtained >6 weeks post partum, when the two groups of patients were clinically indistinguishable, revealed no differences in this index of mitogenic activity. Our results indicate that elevated mitogenic activity ratios of prepartum versus postpartum plasma antedate the clinical recognition of preeclampsia, and return to normal with the resolution of the syndrome.