Double-tracer Study Research Articles

Release of nitrous oxide and dinitrogen from a transition bog under drained and rewetted conditions due to denitrification: results from a [15N]nitrate–bromide double-tracer study

Denitrification is well known being the most important nitrate-consuming process in water-logged peat soils, whereby the intermediate compound nitrous oxide (N2O) and the end product dinitrogen (N2) are ultimately released. The present study was aimed at evaluating the release of these gases (due to denitrification) from a nutrient-poor transition bog ecosystem under drained and three differently rewetted conditions at the field scale using a 15N-tracer approach ([15N]nitrate application, 30 kg N ha−1) and a common closed-chamber technique. The drained site is characterized by a constant water table (WT) of –30 cm (here referred to as D30), while rewetted sites represent a constant WT of –15 cm, a constant WT of 0 cm (i.e. waterlogged), and an initial WT of 0 cm (which decreased slightly during the experiment), respectively, (here referred to as R15, R0, and R0d, respectively). The highest N2O emissions were observed at D30 (291 µg N2O–N m−2 h−1) as well as at R0d (665 µg N2O–N m−2 h−1). At the rewetted peat sites with a constant WT (i.e. R15 and R0), considerably lower N2O emissions were observed (maximal 37 µg N2O–N m−2 h−1). Concerning N2 only at the initially water-logged peat site R0d considerable release rates (up to 3110 µg N2–N m−2 h−1) were observed, while under drained conditions (D30) no N2 emission and under rewetted conditions with a constant WT (R15 and R0) significantly lower N2 release rates (maximal 668 µg N2–N m–2 h−1) could be detected. In addition, it has been found that natural WT fluctuations at rewetted peat sites, in particular a rapid drop down of the WT, can induce high emission rates for both N2O and N2.

Nitrate turnover in a peat soil under drained and rewetted conditions: results from a [15N]nitrate–bromide double-tracer study

Under natural conditions, peatlands are generally nitrate-limited. However, recent concerns about an additional N input into peatlands by atmospheric N deposition have highlighted the risk of an increased denitrification activity and hence the likelihood of a rise of emissions of the greenhouse gas nitrous oxide. Therefore, the aim of the present study was to investigate the turnover of added nitrate in a drained and a rewetted peatland using a [15N]nitrate–bromide double-tracer method. The double-tracer method allows a separation between physical effects (dilution, dispersion and dislocation) and microbial and chemical nitrate transformation by comparing with the conservative Br− tracer. In the drained peat site, low NO3− consumption rates have been observed. In contrast, NO3− consumption at the rewetted peat site rises rapidly to about 100% within 4 days after tracer application. Concomitantly, the 15N abundances of nitrite and ammonium in soil water increased and lead to the conclusion that, besides commonly known NO3− reduction to nitrite (i.e. denitrification), a dissimilatory nitrate reduction to ammonium has simultaneously taken place. The present study reveals that increasing NO3− inputs into rewetted peatlands via atmospheric deposition results in a rapid NO3− consumption, which could lead to an increase in N2O emissions into the atmosphere.

Human monoclonal antibodies to domain C of tenascin-C selectively target solid tumors in vivo

We had previously reported that splice isoforms of tenascin-C containing the extra-domain C are virtually absent in normal adult tissues but are highly abundant in high-grade astrocytomas, with a prominent peri-vascular pattern of expression. We now report that the extra-domain C of tenascin-C is strongly expressed in the majority of lung cancers, with a vascular and stromal pattern of expression. Using antibody phage technology, we have generated a human monoclonal antibody (G11), with a dissociation constant K(D) = 4.2 nM for the human domain C. The G11 antibody, expressed in scFv and in mini-antibody (SIP) format, as well as a scFv-interleukin-2 fusion protein, was then characterized in quantitative biodistribution studies using mice grafted subcutaneously with U87 gliomas, revealing a selective tumor uptake, with tumor/blood ratios up to 11.8:1 at 24 h. A radioiodinated preparation of SIP(G11) was also investigated in a double tracer study using an orthotopic rat glioma model, confirming the antibody's ability to preferentially localize at the tumor site, with tumor/brain ratios superior to the ones observed with (18)F-fluorodeoxyglucose. These tumor-targeting properties, together with the strong immunohistochemical staining of human tumor sections, indicate that the G11 antibody may be used as a portable targeting moiety for the selective delivery of imaging and therapeutic agents to gliomas and lung tumors.

Plasma appearance of labeled β-carotene, lutein, and retinol in humans after consumption of isotopically labeled kale

The bioavailability of carotenoids from kale was investigated by labeling nutrients in kale with 13C, feeding the kale to seven adult volunteers, and analyzing serial plasma samples for labeled lutein, beta-carotene, and retinol. Ingested doses of labeled carotenoids were 34 micromol for beta-carotene and 33 micromol for lutein. Peak plasma concentrations, areas under the plasma concentration-time curves (AUCs), and percentages of dose recovered at peak plasma concentrations were calculated. Average peak plasma concentrations were 0.38, 0.068, and 0.079 microM for [13C]lutein, [13C]beta-carotene, and [13C]retinol, respectively. Average AUC values (over 28 days) were 42.8, 13.6, 13.2 microM h for [13C]lutein, [13C]beta-carotene, and [13C]retinol, respectively. Percentages of dose recovered at peak plasma concentrations were 3.6, 0.7, and 0.7% for [13C]lutein, [13C]beta-carotene, and [13C]retinol, respectively. A positive relationship was observed between baseline plasma retinol levels and [13C]retinol plasma response. It is possible that this relationship was mediated either through some aspect of beta-carotene absorption or via the common pathways of metabolism for postdose and endogenous retinoid.

Variability in conversion of β-carotene to vitamin A in men as measured by using a double-tracer study design

The vitamin A activity of beta-carotene is variable and surprisingly low in women. The reasons for this are not well understood. The vitamin A activity of beta-carotene in men is still uncertain. Contributions of dietary factors compared with individual traits are largely unknown. Our objective was to measure the intrinsic variability in the vitamin A activity of beta-carotene among healthy, well-fed men living in a controlled environment. We used a double-tracer test-retest design. We dosed 11 healthy men orally with 30 micromol hexadeuterated (D6) retinyl acetate (all-trans-19,19,19,20,20,20-[2H6]retinyl acetate) and then with 37 micromol D6 beta-carotene (19,19,19,19',19',19'-[2H6]beta-carotene) 1 wk later. Doses were taken with breakfasts containing 16 g fat. We measured D6 retinol, D6 beta-carotene, and trideuterated (D3) retinol (derived from D6 beta-carotene) concentrations in plasma. Areas under the plasma concentration x time since dosing curves (AUCs) were determined for D6 retinol, D6 beta-carotene, and D3 retinol. All men had detectable D6 retinol concentrations in plasma. The mean (+/-SE) absorption of D6 beta-carotene in all subjects was 2.235 +/- 0.925%, and the mean conversion ratio was 0.0296 +/- 0.0108 mol retinol to 1 mol beta-carotene. Only 6 of 11 men had sufficient plasma concentrations of D6 beta-carotene and D3 retinol that we could measure. The mean absorption of D6 beta-carotene in these 6 subjects was 4.097 +/- 1.208%, and the mean conversion ratio was 0.0540 +/- 0.0128 mol retinol to 1 mol beta-carotene. The vitamin A activity of beta-carotene, even when measured under controlled conditions, can be surprisingly low and variable.

Variability of the conversion of β-carotene to vitamin A in women measured by using a double-tracer study design

Blood beta-carotene and vitamin A responses to oral beta-carotene are variable in humans. Some individuals are characterized as responders and others as low- or nonresponders. A better understanding of the conditions that produce the variability is important to help design public health programs that ensure vitamin A sufficiency. Our objective was to assess variability in absorption and conversion of beta-carotene to vitamin A in vivo in humans by using a novel double-tracer ¿hexadeuterated (D(6)) beta-carotene and D(6) retinyl acetate approach. Eleven healthy women were housed at the US Department of Agriculture Western Human Nutrition Research Center metabolic unit for 44 d, where they consumed diets adequate in vitamins and minerals except for carotenoids. After an adaptation period, the women were given 30 micromol D(6) retinyl acetate orally, followed 1 wk later with 37 micromol D(6) beta-carotene (approximately equimolar doses). Time-dependent plasma concentration curves were determined for D(6) retinol, D(6) beta-carotene, and trideuterated (D(3)) retinol (derived from D(6) beta-carotene). Mean (+/-SE) absorption of D(6) beta-carotene was 3.3 +/- 1.3% for all subjects. The mean conversion ratio was 0.81 +/- 0.34 mol D(3) retinol to 1 mol D(6) beta-carotene for all subjects. However, only 6 of the 11 subjects had plasma D(6) beta-carotene and D(3) retinol concentrations that we could measure. The mean absorption of D(6) beta-carotene in these 6 subjects was 6.1 +/- 0.02% and their conversion ratio was 1.47 +/- 0.49 mol D(3) retinol to 1 mol D(6) beta-carotene. The remaining 5 subjects were low responders with </=0.01% absorption and a mean conversion ratio of 0.014 +/- 0.004 mol D(3) retinol to 1 mol D(6) beta-carotene. Variable absorption and conversion of beta-carotene to vitamin A both contribute to the variable response to consumption of beta-carotene. Our double-tracer approach is adaptable for identifying efficient converters of carotenoid to retinoid.

Comparative Turn-Over of Heparin Cofactor II and Antithrombin III in Baboons Influence of Heparin and Pentosan Polysulfate Administration

Heparin and pentosan polysulfate (PPS) interact in plasma with antithrombin III (AT III) and Heparin cofactor II (HC II) respectively. To assess the influence of heparin or PPS treatment on the metabolism of their respective cofactors, we performed a double tracer study in baboons receiving heparin or PPS. Purified AT III and HC II from human plasma were labelled with 131I and 125I respectively by the lactoperoxidase-glucose oxidase technique. The tracers had unchanged biological activities, were homogeneous in SDS-PAGE, migrated as native proteins by crossed immunoelectrophoresis in the presence of heparin or PPS and virtually coeluted with endogenous baboon proteins from heparin-agarose. Nine animals were randomly allocated to receive, during the metabolic study, heparin (500 IU/kg/d, n = 3), PPS (5 mg/kg,d, n = 3) or a placebo (n = 3) given in 2 daily subcutaneous injections. Heparin levels and anticoagulant effects were similar in extent and duration to those usually achieved in man. The plasma concentrations of AT III and HC II did not vary under treatment. The half-life of the elimination phase in the placebo group ranged from 1.95 to 2.33 d for AT III and from 1.96 to 2.21 d for HC II. There was no significant difference in the half-lives of the 2 inhibitors between the placebo group and the animals receiving heparin or PPS. This suggest that clinical conditions associated with heparin treatment may be important for the effect of heparin on AT III metabolism previously reported in patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Double-tracer study of the fine regional blood-brain glucose transfer in the rat by computer-assisted autoradiography.

Glucose is transported from blood to brain tissue by facilitated diffusion of limited capacity. The regional variation of the glucose transport capacity is not known in detail because methods used previously have been too gross to detect fine regional differences. Therefore, it is not known to what extent the glucose transport capacity varies in proportion to regional blood flow and metabolic rate. To resolve this question, we used double-tracer, dual-label autoradiography to measure blood-brain glucose clearance and blood flow simultaneously in hypo-, normal, and hyperglycemic rats. From the values of glucose clearance and blood flow at various plasma glucose levels, we calculated the affinity constant and maximal transport capacity of the glucose transport system. The transport capacity (Tmax) varied in approximate proportion to the blood flow and, by inference, the metabolic rate. In contrast, the affinity constant (Kt) did not vary systematically between the regions (mean value 7.1 mM). The variation of Tmax from 270 to 890 mumol hg-1 min-1 reflected a parallel variation of total regional capillary length and surface. We conclude from the study that the cerebral capillary is a fixed unit, i.e., that the number of glucose transporters per unit of capillary surface area is the same in all regions. Regional differences are the result of different capillary densities in the regions of the brain.

A comparison between the balance method and radiotracer methods for measuring calcium absorption in treated and untreated patients with osteoporosis

Measurements of calcium absorption by the balance method and by the double tracer, multiple blood sampling radiocalcium tracer method were synchronized in 48 studies in 35 patients with osteoporosis. In the patients who were studied twice, results were obtained before and during treatment. Significant correlations were obtained between fractional dietary calcium absorption and three measures of the rate of 47Ca absorption : the maximum rate of absorption (M.A.R.) and mean rate of absorption ( AR ), both calculated by deconvolution analysis from the results of the double tracer study ; and the mean rate of absorption calculated from the oral tracer appearance curve using Marshall and Nordin's (1969) method (L.A.R.). None of these correlations were sufficiently good for the radiocalcium absorption test to be able to replace the expensive and time-consuming balance technique in the detailed study of absorption of calcium from the diet or calcium balance in individual patients with osteoporosis. Only a small part of this lack of correspondence could be accounted for by methodological uncertainties. The most sensitive radiocalcium absorption tests appear to be those which measure rates of absorption and they appear best suited to studying patients in homogeneous groups rather than individually.

A double tracer study on iron metabolism and red cell life span in various diseases using radioactive iron andchromium

A double tracer study on iron metabolism and red cell life span in various diseases using radioactive iron andchromium

The Influence of Dietary Potassium on the Retention of Chronically Ingested Cesium-137 in the Rat

The influence of dietary K on the retention of chronically ingested Cs/ sup 137/ by muscle, liver, testes, femur, whole blood, and brain was determined by long-term isotope studies and a short-term double-tracer study with K/sup 42/ and Cs/sup 137/. At 35 days, it was observed that a ninefold elevation in dietary K caused a twofold reduction in Cs/sup 137/ retention. This is probably the maximum effect. Tissue K levels were not altered by dietary K levels. The pattern of Cs/sup 137/ accumulation by the selected tissues at 1 day was different from that at35 days. At 1 day, the levels of Cs/sup 137/, in decreasing order, were liver, muscle, testes, femur, whole blood, brain. At 35 days, the levels, in decreasing order, were muscle, liver, testes, brain, femur, whole blood. It was calculated that the Cs/K ratio of the tissues divided by the Cs/K ratio of diet varied almost proportionally with dietary K levels; a ninefold increase in dietary K caused about a fivefold increase in Cs/K ratio (diet -- tissue). Therefore, it appears that the concept of discrimination factor has severe limitations in theoretical and practical application for this ion pair. In the shortterm double-tracer study, dietary Kmore » decreased Ku retention but did not influence Cs/sup 137/ retention. It was concluded that potassium does not effectively compete with cesiunn for membrane transport sites or intraceliular binding sites. (auth)« less

Strontium Studies in Beagles

Previous skeletal irradiation from bone-deposited radionuclides does not seem to influence the metabolism of a Sr/sup 85/ tracer in beagles under the conditions, dose rates, and times studied. Confinement immediately after the injection of a Sr/sup 85/ tracer reduces Its retention. As the age of the beagles increases from 6 months to 10 years, their retentlon of a Sr/sup 85/ tracer decreases. The fact that strontium retention in beagles is less than radium retention was verified in a Sr/sup 85/-Ra/sup 226/ double-tracer study. (auth)