Effects of temperature and body size on radiocaesium retention in brown trout, Salmo trutta

1. The elimination rate of radiocaesium in brown trout Salmo trutta L. was determined in the laboratory at four water temperatures (range 4.4–15.6°C). In the experiments three or four homogenous size-groups of fish (mean weights 23–496 g) were studied at each temperature. 2. The brown trout received acute oral doses of ¹³⁴Cs and were killed at intervals for radioactivity counting. The retention versus time curves were composed of two distinct exponential components. The long-lived component was quantitatively the most important for retention of radiocaesium. Elimination rate increased with increasing water temperature and decreased with increasing body weight. 3. The biological half-life of ¹³⁴Cs (T_b, days) was related to fresh body weight (W, g) and water temperature (t, °C) by the equation: T_b= 290 ×W°.¹⁷⁶× e-°.^106×t. The elimination rate of Cs could be predicted from weight-specific metabolic rate as given by Elliott's equations for brown trout.     

Fate of sodium arsenate in dairy sheep and goats

This study followed the uptake, distribution, and elimination of sodium arsenate administered in a single dose and in multiple doses, respectively, to Iranian dairy sheep and goats. In the single dosing study, the blood concentration data fit an open two-compartment model of the form:C _b (t)=?(A+B)e ^?kat +Ae ^?αt +Be ^?βt Absorption distribution and elimination rate constants were statistically significantly different for the two animal species. In the multiple dosing study, arsenic accumulated in the blood of both animal species, as expressed by a one compartment model of the form:C _t =C _ss (1-e ^?kt ) Arsenic was eliminated rapidly at the termination of dosing, with the blood washout half-life being shorter in sheep than in goats. Urinary excretion was the major elimination route from the body of both species.     

Kinetics of homovanillic acid and determination of its production rate in the rhesus monkey.

Homovanillic acid (HVA) labelled with two deuterium atoms (d₂-HVA) was used to label the peripheral body pool of endogenous HVA (d₀-HVA). D₂-HVA was rapidly injected intravenously into Rhesus monkeys and concentrations of both d₂- and d₀-HVA in sequential samples of blood serum and urine determined by gas chromatography-mass spectrometry. Parameters describing the distribution and elimination of HVA, as well as its pool size, turnover, and synthesis rate were then calculated. Results indicate that the decline of serum d₂-HVA concentration with time is multiexponential, with a biological half-life ranging from 35.9 to 102 min in the four monkeys studied. The apparent volume of distribution of d₂-HVA in the body was 0.813–1.17 1/kg. Serum clearance was 7.28–18.2 ml/kg/min. For most animals, only about 50% of the administered dose of d₂-HVA was recovered unchanged in the urine. Renal clearance ranged from 3.79 to 17.0 ml/kg/min, and d₀-HVA excretion rates ranged from 19.5 to 64.1 μg/hr. The size of the peripheral body pool of HVA, calculated from serum kinetic parameters, was 63.3–80.7 μg; HVA turnover was 5.43–13.9 μg/1/hr; and HVA production rate was calculated to be 36.6–84.3 (5.23–12.6 μg/kg/hr).     

Daily Rhythms in Blood and Milk Lead Toxicokinetics Following Intravenous Administration of Lead Acetate to Dairy Cows in Winter

The objective was to investigate circadian variations of blood and milk lead toxicokinetics in dairy cows in winter. Twenty lactating Holstein animals were randomly assigned to 4 treatments, corresponding to different hours after onset of light (HALO): 2, 8, 14 and 20. Cows received a single intravenous administration of 2.5 mg/kg lead as lead acetate. Blood and milk samples were taken and analyzed by atomic absorption spectrophotometry. For each toxicokinetic parameter, a one-way analysis of variance was performed to outline the existence of daily variations. Significant differences as a function of HALO were detected in blood for the hybrid constant of elimination (β), half-life of elimination (t_1/2β), area under the curve (AUC) and clearance (Cl) (p &lt; 0.01) and volume of distribution at steady state (V_ss) (p < 0.05). Half-life of elimination was highest when lead acetatae was injected at 2 HALO, and lowest following the 14 HALO administration. Milk data showed significant differences for maximum concentration, AUC and Cl (p &lt; 0.001), volume of distribution and V_ss (p < 0.005). The ratio AUC_milk/ABC_blood was utilized to estimate penetration of lead in milk. It differed significantly along the day (p < 0.01). Blood daily variations could be fitted a circadian rhythms. No circadian rhythms were detected in milk parameters or in the ratio AUC_milk/ABC_blood. It was concluded that blood and milk lead toxicokinetics are distinctly affected by the time of the day.     

Cross-species analysis of Fc engineered anti-Lewis-Y human IgG1 variants in human neonatal receptor transgenic mice reveal importance of S254 and Y436 in binding human neonatal Fc receptor

IgG has a long half-life through engagement of its Fc region with the neonatal Fc receptor (FcRn). The FcRn binding site on IgG1 has been shown to contain I253 and H310 in the CH2 domain and H435 in the CH3 domain. Altering the half-life of IgG has been pursued with the aim to prolong or reduce the half-life of therapeutic IgGs. More recent studies have shown that IgGs bind differently to mouse and human FcRn. In this study we characterize a set of hu3S193 IgG1 variants with mutations in the FcRn binding site. A double mutation in the binding site is necessary to abrogate binding to murine FcRn, whereas a single mutation in the FcRn binding site is sufficient to no longer detect binding to human FcRn and create hu3S193 IgG1 variants with a half-life similar to previously studied hu3S193 F(ab')₂ (t_1/2β, I253A, 12.23 h; H310A, 12.94; H435A, 12.57; F(ab')₂, 12.6 h). Alanine substitutions in S254 in the CH2 domain and Y436 in the CH3 domain showed reduced binding in vitro to human FcRn and reduced elimination half-lives in huFcRn transgenic mice (t_1/2β, S254A, 37.43 h; Y436A, 39.53 h; wild-type, 83.15 h). These variants had minimal effect on half-life in BALB/c nu/nu mice (t_1/2β, S254A, 119.9 h; Y436A, 162.1 h; wild-type, 163.1 h). These results provide insight into the interaction of human Fc by human FcRn, and are important for antibody-based therapeutics with optimal pharmacokinetics for payload strategies used in the clinic.     

Influence of sodium selenite on203Hg absorption,distribution, and elimination in male mice exposed to methyl203Hg

To study the effects of long-term selenium supplementation on absorption, distribution, and elimination of methylmercury (MeHg) in mice, three groups of male mice (Balb/c CA) were exposed for 7 wk to 0, 0.6, and 3 ppm sodium selenite in tap water. They were then given a single oral dose of Me²⁰³Hg (2 μmol/kg) by gastric intubation, and elimination of²⁰³Hg was followed by whole-body counting for 49 d at the same Se exposure as previously. Twenty-four hours and 49 d after dosage, 6–7 animals/group were sampled for analysis of²⁰³Hg distribution in the body. Glutathione peroxidase (GSH-PX) activity in blood and selenium levels in the liver were used as measures of selenium status. Gastrointestinal absorption of Me²⁰³Hg was not influenced by the Se status of the animals. Selenium supplementation of MeHg-exposed mice caused an enhanced whole-body elimination of Hg, but selenium-supplemented animals did not have lower Hg levels in the brain and kidney than nonsupplemented animals. The effect of selenium on the accumulation, of Hg in the brain was dose-dependent, a high dose (3 ppm Se) causing a higher initial accumulation of Hg. The intracellular distribution of²⁰³Hg in the liver and kidney was not affected by Se. The results indicate that selenium treatment of MeHg-exposed mice may have a positive effection the health of the animals by decreasing the total body burden of MeHg.     

Daily rhythms in blood and milk lead toxicokinetics following intravenous administration of lead acetate to dairy cows in summer

The objective of this study was to investigate circadian variations of blood and milk lead toxicokinetics in dairy cows in summer. Twenty lactating Holstein animals were randomly assigned to four treatments corresponding to different hours after onset of light (HALO): 2, 8, 14, and 20. Cows received a single intravenous administration of 2.5 mg/kg lead as lead acetate. Blood and milk samples were taken and analyzed by atomic absorption spectrophotometry. For each toxicokinetic parameter, a one-way analysis of variance (ANOVA) was performed to outline the existence of daily variations. Significant blood differences as a function of HALO were found for the hybrid constant of distribution (α), hybrid constant of elimination (β), elimination half-life (), area under the curve (AUC), volume of distribution at steady state (V_ss) and clearance (Cl_B) (p<0.05). Half-life of elimination presented two peaks at 2 and 14 HALO. Milk data showed significant differences for maximum concentration and AUC (p<0.05). The ratio AUC_milk/AUC_blood was utilized to estimate penetration of lead in milk. It differed significantly throughout the day (p<0.05). Milk data for the significant parameters could be fitted to circadian rhythms. No circadian rhythms were detected in blood parameters or in the ratio AUC_milk/AUC_blood.     

Bioavailability of Astaxanthin in Haematococcus Algal Extract: The Effects of Timing of Diet and Smoking Habits

Astaxanthin is a caroteonid that possesses strong antioxidant activity. Recently, many studies on biological activity have been reported. In general, the absorption of carotenoids is affected greatly by diet and by smoking. In this report, we investigated astaxanthin pharmacokinetics after administration of Haematococcus algal extract, a source of astaxanthin, to smokers and nonsmokers before and after a meal; astaxanthin was given before the meal to nonsmokers (n=7), after the meal to nonsmokers (n=6), and after the meal to smokers (n=7), then serum samples were analyzed. The timing of administration greatly affected astaxanthin bioavailability including the area under the curve (AUC_0–168, 2,968±959 μg h/l in the before-meal group vs. 7,219±3,118 μg h/l in the after-meal group), indicating high availability in the after-meal group. Smoking also affected the pharmacokinetic parameters and reduced the half-life (t_1?2) of astaxanthin elimination significantly.     

Continued optimization of the M5 NAM ML375: Discovery of VU6008667, an M5 NAM with high CNS penetration and a desired short half-life in rat for addiction studies

This letter describes the continued optimization of M₅ NAM ML375 (VU0483253). While a valuable in vivo tool compound, ML375 has an excessively long elimination half-life in rat (t_1/2 = 80 h), which can be problematic in certain rodent addiction paradigms (e.g., reinstatement). Thus, we required an M₅ NAM of comparable potency to ML375, but with a rat t_1/2 of less than 4 h. Steep SAR plagued this chemotype, and here we detail aniline replacements that offered some improvements over ML375, but failed to advance. Ultimately, incorporation of a single methyl group to the 9b-phenyl ring acted as a metabolic shunt, providing (S)-11 (VU6008667), an equipotent M₅ NAM, with high CNS penetration, excellent selectivity versus M_1–4 and the desired short half-life (t_1/2 = 2.3 h) in rat.     

Methylmercury Alters the Activities of Hsp90 Client Proteins,Prostaglandin E Synthase/p23 (PGES/23) and nNOS

Methylmercury (MeHg) is a persistent pollutant with known neurotoxic effects. We have previously shown that astrocytes accumulate MeHg and play a prominent role in mediating MeHg toxicity in the central nervous system (CNS) by altering glutamate signaling, generating oxidative stress, depleting glutathione (GSH) and initiating lipid peroxidation. Interestingly, all of these pathways can be regulated by the constitutively expressed, 90-kDa heat shock protein, Hsp90. As Hsp90 function is regulated by oxidative stress, we hypothesized that MeHg disrupts Hsp90-client protein functions. Astrocytes were treated with MeHg and expression of Hsp90, as well as the abundance of complexes of Hsp90-neuronal nitric oxide synthase (nNOS) and Hsp90-prostaglandin E synthase/p23 (PGES/p23) were assessed. MeHg exposure decreased Hsp90 protein expression following 12 h of treatment while shorter exposures had no effect on Hsp90 protein expression. Interestingly, following 1 or 6 h of MeHg exposure, Hsp90 binding to PGES/p23 or nNOS was significantly increased, resulting in increased prostaglandin E₂ (PGE₂) synthesis from MeHg-treated astrocytes. These effects were attenuated by the Hsp90 antagonist, geldanmycin. NOS activity was increased following MeHg treatment while cGMP formation was decreased. This was accompanied by an increase in •O₂ ⁻ and H₂O₂ levels, suggesting that MeHg uncouples NO formation from NO-dependent signaling and increases oxidative stress. Altogether, our data demonstrates that Hsp90 interactions with client proteins are increased following MeHg exposure, but over time Hsp90 levels decline, contributing to oxidative stress and MeHg-dependent excitotoxicity.     

The Bioconjugation and Radiosynthesis of 89Zr-DFO-labeled Antibodies

The exceptional affinity, specificity, and selectivity of antibodies make them extraordinarily attractive vectors for tumor-targeted PET radiopharmaceuticals. Due to their multi-day biological half-life, antibodies must be labeled with positron-emitting radionuclides with relatively long physical decay half-lives. Traditionally, the positron-emitting isotopes ¹²⁴I (t_1/2 = 4.18 d), ⁸⁶Y (t_1/2 = 14.7 hr), and ⁶⁴Cu (t_1/2 = 12.7 hr) have been used to label antibodies for PET imaging. More recently, however, the field has witnessed a dramatic increase in the use of the positron-emitting radiometal ⁸⁹Zr in antibody-based PET imaging agents. ⁸⁹Zr is a nearly ideal radioisotope for PET imaging with immunoconjugates, as it possesses a physical half-life (t_1/2 = 78.4 hr) that is compatible with the in vivo pharmacokinetics of antibodies and emits a relatively low energy positron that produces high resolution images. Furthermore, antibodies can be straightforwardly labeled with ⁸⁹Zr using the siderophore-derived chelator desferrioxamine (DFO). In this protocol, the prostate-specific membrane antigen targeting antibody J591 will be used as a model system to illustrate (1) the bioconjugation of the bifunctional chelator DFO-isothiocyanate to an antibody, (2) the radiosynthesis and purification of a ⁸⁹Zr-DFO-mAb radioimmunoconjugate, and (3) in vivo PET imaging with an ⁸⁹Zr-DFO-mAb radioimmunoconjugate in a murine model of cancer.     

Tissue distribution,metabolism, and elimination of perfluorooctanoic acid in male and female rats

The elimination, tissue distribution, and metabolism of [1-¹⁴C]perfluorooctanoic acid (PFOA) was examined in male and female rats for 28 days after a single ip dose (9.4 μmol/kg, 4 mg/kg). A sex difference in urinary elimination of PFOA-derived ¹⁴C was observed. Female rats eliminated PFOA-derived radioactivity rapidly in the urine with 91% of the dose being excreted in the first 24 hr. In the same period, male rats eliminated only 6% of the administered ¹⁴C in the urine. The sex-related difference in urinary elimination resulted in the observed difference in the whole-body elimination half-life (t_1/2) of PFOA in males (t_1/2 = 15 days) and females (t_1/2 < 1 day). Analysis of PFOA-derived ¹⁴C in tissues showed that the liver and plasma of male rats and the liver, plasma, and kidney of female rats were the primary tissues of distribution. The relatively high concentration of PFOA in the male liver was further examined using an in situ nonrecirculating liver perfusion technique. It was shown that 11% of the PFOA infused was extracted by the liver in a single pass. The ability of the liver to eliminate PFOA into bile was examined in rats whose renal pedicles were ligated to alleviate sex differences in the urinary excretion of PFOA. In a 6-hr period following IP administration of PFOA, there was no apparent difference in biliary excretion, where both males and females eliminated less than 1% of the PFOA dose via this route. We hypothesized that the sex difference in the persistence of PFOA was due to a more rapid formation of a PFOA-containing lipid (i.e., a PFOA-containing mono-, di-, or triacylglycerol, cholesteryl ester, methyl ester, or phospholipid) in the male rat. Also, the increased urinary elimination of PFOA in females may have been due to increased metabolism to a PFOA-glucuronide or sulfate ester. However, no evidence that PFOA is conjugated to form a persistent hybrid lipid was obtained, nor were polar metabolites of PFOA in urine or bile detected. In addition, daily urinary excretion of fluoride in male and female rats before or after PFOA treatment were similar, suggesting that the parent compound is not defluorinated. Thus, the more rapid elimination of PFOA from female rats is not due to formation of a PFOA metabolite.     

Stability of a Fusarium solani pisi recombinant cutinase in phosphatidylcholine reversed micelles

Summary A Fusarium solani pisi recombinant cutinase solubilized in phosphatidylcholine/isooctane reversed micelles was used to catalyse the esterification reaction of butyric acid with 2-butanol at pH 10.7. The influence of temperature, Wo and substrates on lipase stability was evaluated. The enzyme displays a better stability, with a half-life over 125 days, at a temperature of 22°C and for a low water content (W_O= 6.5). Butyric acid increased the cutinase deactivation (t_1/2=0.56h), while 2-butanol led to a similar half-life (t_1/2=14h) as without substrate.     

Pharmacokinetics Study of Recombinant Hirudin in the Plasma of Rats Using Chromogenic Substrate,ELISA, and Radioisotope Assays

Aim

To compare the analytical methods used to study the pharmacokinetics of recombinant hirudin in the plasma of rats that had been injected with ¹²⁵I-recombinant hirudin.

Methods

2.0 mg/kg ¹²⁵I-recombinant hirudin were injected intravenously into rats. The recombinant hirudins in the plasma was analyzed by chromogenic substrate assay, enzyme-linked immunosorbent assay (ELISA), total radioisotope assay (RA) and trichloroacetic acid pre-treated total radioisotope assay (TCA-RA).

Results

The chromogenic substrate assay standard curve was linear over the concentration range from 3.12 to 40.00 ng/ml for the recombinant hirudin in plasma. The relative standard deviations (RSD) for the intra- and inter-day variation were 5.0 to 6.3% and 11.9 to 12.6%, respectively. The recoveries of recombinant hirudin was 89.8% to 100.7%. The limit of quantification (LOQ) was 3.12 ng/ml. The concentration-time curve of the recombinant hirudin in the plasma could be explained as a two-compartment model. Pharmacokinetic parameters, including the half-life of distribution phase (t_1/2 α), the half-life of elimination phase (t_1/2 β), volume of apparent distribution (Vd), and area under the concentration-time curve from zero to infinite time (AUC_0–t) were 7.59 min, 46.99 min, 0.17 L/kg, and 204.5 mg/L/min, respectively, as determined by chromogenic substrate assay; 6.41 min, 47.28 min, 1.24 L/kg, and 575.18 mg/L/min, respectively, as determined by ELISA; 3.69 min, 701.90 min, 0.04 L/kg, and 4189 mg/L/min, respectively as determined by RA; and 4.57 min, 724.9 min, 0.09 L/kg, and 2329 mg/L/min, respectively, as determined by TCA-RA.

Conclusions

The chromogenic substrate assay on the concentration dynamics of the recombinant hirudin in the plasma is a specific, sensitive, and accurate analytical method for pharmacokinetic studies. Moreover, the pharmacokinetic parameters determined by the chromogenic substrate assay and ELISA are congruent except for AUC.     

Factors Affecting Methylmercury Levels in Surficial Tailings from Historical Nova Scotia Gold Mines

Large quantities of Hg remain in tailings dumps from historical Nova Scotian gold mines. Depth profiles of total Hg (Hg_T) and methylmercury (MeHg) were compared with geochemical and microbiological variables, to identify factors influencing MeHg levels in tailings. Hg_T and MeHg were highly variable in tailings (0.2–73.5 μ mol kg^{? 1} and < dl-56.4 nmol kg^{? 1}, respectively), and were influenced by a complex set of in situ factors. Elevated MeHg was linked with > 5 μ mol kg^?1 Hg_T, organic matter, hydrology, abundance and activity of sulfate reducing bacteria, and demethylation processes. Methylmercury levels in tailings from a wet, bog-like site appeared to undergo seasonal fluctuations, with higher concentrations measured in September and October, and lower concentrations in May. Evaluations of amalgamation tailings should examine MeHg and Hg_T transport out of low-lying, saturated tailings dumps after snowmelt and major rainfall events, and should take into account the possibility of seasonal variation in MeHg levels in northern regions.     

Mortality as a mathematical function of organic growth and diameter structure

ABSTRACT

An indirect method to determine the exponential mortality coefficient (λ) as well as the annual mortality (m) is proposed. In this method, both parameters are deduced mathematically from a model of organic growth (von Bertalanffy) and from a model of uneven-aged population structure (De Liocourt and Meyer) used with forest trees, which relates the frequency of individuals by size classes. The resultant equations allow the determination of either the instantaneous or mean value for λ or m between any two ages t ₁ and t ₂. Both concepts of mortality were used to determine the half-life (t _0.5) as a function of λ and m . The method is applied to determine the curves of mortality and the half-life of Otoba gracilipes (Myristicaceae), a tropical tree that grows in the forested wetlands of the Colombian Pacific coast.     

EtHg is more toxic than MeHg to human peripheral blood mononuclear cells: Involvement of apoptotic,mitochondrial, oxidative and proliferative parameters

BackgroundMethylmercury (MeHg) and ethylmercury (EtHg) are potent toxicants affecting the environment and human healthy. In this way, the present study aimed to investigate and compare the effects of MeHg and EtHg exposure on human peripheral blood mononuclear cells (PBMCs), which are critical components of the mammalian immune system.MethodsPBMCs were exposed to 2.5 μM MeHg or 2.5 μM EtHg. The number of cells and incubation times varied according to each assay. After exposures, the PBMCs were subjected to different evaluations, including cell viability, morphological aspects, cell cycle phases, indices of apoptosis and necrosis, reactive species (RS) production, and mitochondrial functionality.ResultsPBMCs exposed to EtHg were characterized by decreased viability and size, increased granularity, RS production, and apoptotic indexes accompanied by an intensification of Sub-G1 and reduction in G0-G1 cell cycle phases. Preceding these effects, we found mitochondrial dysfunctions, namely a reduction in the electron transport system related to mitochondrial complex I. In contrast, PBMCs exposed to MeHg showed only reduced viability. By ICP-MS, we found that PBMCs treated with EtHg accumulated Hg + levels ∼1.8-fold greater than MeHg-exposed cells.Conclusions and significanceTaken together, our findings provide important insights about mercury immunotoxicity, showing that EtHg is more immunotoxic to human PBMCs than MeHg.     

Toxicokinetics and biotransformation of pentachlorophenol in the topsmelt (Atherinops affinis)

The toxicokinetics and biotransformation of pentachlorophenol (PCP) were determined in the topsmelt (Atherinops affinis) In a static system, topsmelt (n = 9) were exposed to 50 μg/L of [U-¹⁴C]PCP for 24 hours to determine the absorption rate constant (Ka), the whole-body bio-concentration (at steady-state conditions), the elimination rate constant UQ, and the elimination half-life (t_1/2). Kinetics were determined by direct quantitation of radioactivity in the exposure water. Following exposure, fish were placed in a flow-through metabolism chamber for 24 hours to allow depuration of retained residues, which were collected on XAD-4 resin. Excreted residues were identified and quantified by high-pressure liquid co-chromatography, fraction collection, and liquid scintillation counting. The Ka and Ke, calculated using a simplified model, were 0.012 M-1 0.005/h and 0.014±;0.003/h, respectively, while the 24 hour total concentration factor was 278.0×182.0 and the t_1/2 was 52.7±;11.2. During 24 hours of exposure to dean seawater, topsmelt depurated 32.9% of retained residues, and while PCP was primarily excreted unchanged (64.9%), significant amounts of both pentachlorophenylsulfate (18.9%) and pentachloro-β-D-glucuronide (16.2%) were also formed.     

The preparation and kinetic properties of multiple forms of chicken brain acetylcholinesterase

A method for preparing various forms of acetylcholinesterase (A ChE) from chicken brain has been developed and they have been characterized in terms of kinetic parameters such as K_m, rate constant (k), turnover number (k_p), specificity constant (k_sp), V_max and half-life (t_1/2). The solubility experiments show that, there are two major forms of A ChE i.e. water-soluble and membrane-bound A ChE (MBA ChE). The MBA ChE shows several subforms, and on the basis of percentage activity only three MBA ChE forms have been selected for complete characterization by various kinetic parameters. It was found that these three forms of MBA ChE demonstrate significant differences in their kinetic properties.     

An improved GC/MS-based procedure for the quantitation of the isoprostane 15-F2t-IsoP in rat plasma

This article describes a procedure for the quantitation of the isoprostane 15-F_2t-IsoP (9a,11a,15S-trihydroxy-(8b)-prosta-5Z,13E-dien-1-oic acid [CAS#27415-26-5] formerly known as 8-epi-PGF_2a or 8-iso-PGF_2a, and also as iPF_2a-III). We have combined features from several earlier methods for 15-F_2t-IsoP and prostaglandins, and identified and modified those steps that may lead to poor recoveries. The resulting protocol is precise and reliable, and was validated by a blind time-course study of plasma levels in rats treated with 120 and 1200 mg CCl₄/kg body weight. Plasma levels of 15-F_2t-IsoP, as measured according to the procedure described above, are good indicators of acute oxidative stress as induced by CCl₄. The precision of the measurements allows detection of elevated plasma 15-F_2t-IsoP levels as long as 16 h after an acute exposure of 120 mg CCl₄/kg body weight, and 2 h after an exposure of 1 mg CCl₄/kg body weight. The results of this low-dose, pilot study suggest that this method has sufficient analytical precision to allow the detection of the small changes in plasma isoprostane levels, which result from chronic and/or lower-level exposures to agents causing oxidative stress.