Use Of Isotopic Tracers Research Articles

Pregnancy-induced alterations of 1-carbon metabolism and significance for maternal nutrition requirements.

The pregnancy-induced alterations in 1-carbon (1C) metabolism, effects of advancing gestation on maternal plasma concentrations of methyl nutrients, and potential implications for maternal dietary intake and infant clinical outcomes are summarized in this narrative review. 1C metabolism encompasses a series of pathways where 1C units are transferred among nutrients such as B vitamins, choline, and amino acids (the methyl nutrients). Use of isotopic tracers and measuring methyl nutrients in maternal plasma and infant cord blood has advanced the understanding of 1C flux in pregnancy and kinetics of maternal-placental-fetal transfer. Methyl nutrients are supplied from maternal plasma to the placenta and fetus to support growth and 1C metabolism in these compartments. A literature review was completed in MEDLINE and Google Scholar using search terms related to 1C metabolism, methyl nutrients, and nutrition requirements in pregnancy. English-language articles were reviewed in which 1C metabolism in pregnancy, maternal-placental-fetal transfer of methyl nutrients, and determinants of maternal plasma concentrations of methyl nutrients among healthy pregnant women were assessed. Adaptations in 1C metabolism occur throughout a healthy pregnancy to support this unique period of accelerated growth. Studies report similar temporal changes in plasma concentrations of many methyl nutrients, including B vitamins, choline, betaine, methionine, and cysteine, among healthy pregnant women from diverse geographic regions. Other key findings discussed in this review include an apparent high degree of B vitamin transfer to the placenta and fetus, influence of choline supplementation on 1C flux and possible benefit of supplementation for infant cognitive development, and that glycine may be conditionally essential in pregnancy. Understanding the flux of 1C metabolism in pregnancy and methyl nutrient transfer from maternal plasma is needed to establish appropriate plasma references ranges and, ultimately, dietary recommendations that aim to prevent deficiency and associated adverse health outcomes for mother and baby.

Minimal and Maximal Models to Quantitate Glucose Metabolism: Tools to Measure, to Simulate and to Run in Silico Clinical Trials.

Several models have been proposed to describe the glucose system at whole-body, organ/tissue and cellular level, designed to measure non-accessible parameters (minimal models), to simulate system behavior and run in silico clinical trials (maximal models). Here, we will review the authors' work, by putting it into a concise historical background. We will discuss first the parametric portrait provided by the oral minimal models-building on the classical intravenous glucose tolerance test minimal models-to measure otherwise non-accessible key parameters like insulin sensitivity and beta-cell responsivity from a physiological oral test, the mixed meal or the oral glucose tolerance tests, and what can be gained by adding a tracer to the oral glucose dose. These models were used in various pathophysiological studies, which we will briefly review. A deeper understanding of insulin sensitivity can be gained by measuring insulin action in the skeletal muscle. This requires the use of isotopic tracers: both the classical multiple-tracer dilution and the positron emission tomography techniques are discussed, which quantitate the effect of insulin on the individual steps of glucose metabolism, that is, bidirectional transport plasma-interstitium, and phosphorylation. Finally, we will present a cellular model of insulin secretion that, using a multiscale modeling approach, highlights the relations between minimal model indices and subcellular secretory events. In terms of maximal models, we will move from a parametric to a flux portrait of the system by discussing the triple tracer meal protocol implemented with the tracer-to-tracee clamp technique. This allows to arrive at quasi-model independent measurement of glucose rate of appearance (Ra), endogenous glucose production (EGP), and glucose rate of disappearance (Rd). Both the fast absorbing simple carbs and the slow absorbing complex carbs are discussed. This rich data base has allowed us to build the UVA/Padova Type 1 diabetes and the Padova Type 2 diabetes large scale simulators. In particular, the UVA/Padova Type 1 simulator proved to be a very useful tool to safely and effectively test in silico closed-loop control algorithms for an artificial pancreas (AP). This was the first and unique simulator of the glucose system accepted by the U.S. Food and Drug Administration as a substitute to animal trials for in silico testing AP algorithms. Recent uses of the simulator have looked at glucose sensors for non-adjunctive use and new insulin molecules.

Correlation between ferromagnetic and isotopic tracers for sentinel lymph node detection in cutaneous melanoma: IMINEM study.

The usefulness of sentinel lymph node biopsy (SLNB) in staging cutaneous melanoma has been proven. Therefore, different tracers have been used to identify the sentinel lymph nodes (SLNs). The use of isotopic tracers together with radioactivity detectors allowed a much more precise and direct approach to the SLNs. However, not all centres have access to a Nuclear Medicine department hindering sentinel lymph node detection (SLND) and consequently, other markers such as ferromagnetic tracers have been evaluated looking for the same advantages and effectiveness as isotopic tracers. Ferromagnetic tracers have proven their usefulness in other cancer entities such as breast, prostate and thyroid cancer. The objective was to assess the detection and concordance rates between isotopic and ferromagnetic techniques for SLNB in cutaneous melanoma. Isotopic SLNB technique and ferromagnetic tracer were compared for cutaneous melanoma in a non-inferiority multicentre prospective study carried out in six Spanish hospitals. A total of 60 patients were recruited and 133 lymph nodes removed. The detection rate was slightly higher with ferromagnetic tracer in head-neck and trunk melanomas, and with isotopic tracer in limbs. The patients' and nodes' concordance rates between both techniques for ex vivo samples were 95% and 86% for head-neck and trunk tumours and 97% and 93% for limbs tumours, respectively. The concordance rates for involved nodes were 100% and 88.2% for patients and nodes, respectively. The intraoperative detection and biopsy of SLN in cutaneous melanoma using a ferromagnetic was a reliable alternative method to the isotopic technique in cutaneous melanomas.

Key Concepts Surrounding Studies of Stable Isotope-Resolved Metabolomics.

"Omics"-based analyses are widely used in numerous areas of research, advances in instrumentation (both hardware and software) allow investigators to collect a wealth of data and therein characterize metabolic systems. Although analyses generally examine differences in absolute or relative (fold-) changes in concentrations, the ability to extract mechanistic insight would benefit from the use of isotopic tracers. Herein, we discuss important concepts that should be considered when stable isotope tracers are used to capture biochemical flux. Special attention is placed on in vivo systems, however, many of the general ideas have immediate impact on studies in cellular models or isolated-perfused tissues. While it is somewhat trivial to administer labeled precursor molecules and measure the enrichment of downstream products, the ability to make correct interpretations can be challenging. We will outline several critical factors that may influence choices when developing and/or applying a stable isotope tracer method. For example, is there a "best" tracer for a given study? How do I administer a tracer? When do I collect my sample(s)? While these questions may seem straightforward, we will present scenarios that can have dramatic effects on conclusions surrounding apparent rates of metabolic activity.

Hydrology at Aberdeen – thinking about water locally and globally

ABSTRACTAlthough geographical hydrology research at Aberdeen contributed significant insights into the connections between river channel processes and landscape evolution in the early twentieth century it was not until the 1980s that hydrology became a discernible research theme in the Department of Geography. Whilst the research initially focused on understanding the linkages between fluvial geomorphology, hydrology and in-stream hydraulics, since the mid-1990s a burgeoning research community has made significant contributions to advancing physical hydrology, understanding water quality issues and elucidating inter-connections between hydrology and both terrestrial and aquatic ecosystems. A particular research strength has been the use of isotopic tracers (especially high-resolution, field-based data) to advance hydrological models. Work at Aberdeen has also been internationally leading in terms of understanding the role of hydrology in shaping the life cycle of Scottish Atlantic salmon and other aquatic organisms. Ideas and research approaches developed at local Scottish field sites are being increasingly applied and adapted to advance hydrological understanding in geographical regions as diverse as the rain forests of Costa Rica, the karst of south west China, aquifers in east Africa, the Chilean high Andes, Arctic Canada, peatlands in western Siberia and forests in northern Germany.

Nitrate addition stimulates microbial decomposition of organic matter in salt marsh sediments

Salt marshes sequester carbon at rates more than an order of magnitude greater than their terrestrial counterparts, helping to mitigate climate change. As nitrogen loading to coastal waters continues, primarily in the form of nitrate, it is unclear what effect it will have on carbon storage capacity of these highly productive systems. This uncertainty is largely driven by the dual role nitrate can play in biological processes, where it can serve as a nutrient-stimulating primary production or a thermodynamically favorable electron acceptor fueling heterotrophic metabolism. Here, we used a controlled flow-through reactor experiment to test the role of nitrate as an electron acceptor, and its effect on organic matter decomposition and the associated microbial community in salt marsh sediments. Organic matter decomposition significantly increased in response to nitrate, even at sediment depths typically considered resistant to decomposition. The use of isotope tracers suggests that this pattern was largely driven by stimulated denitrification. Nitrate addition also significantly altered the microbial community and decreased alpha diversity, selecting for taxa belonging to groups known to reduce nitrate and oxidize more complex forms of organic matter. Fourier Transform-Infrared Spectroscopy further supported these results, suggesting that nitrate facilitated decomposition of complex organic matter compounds into more bioavailable forms. Taken together, these results suggest the existence of organic matter pools that only become accessible with nitrate and would otherwise remain stabilized in the sediment. The existence of such pools could have important implications for carbon storage, since greater decomposition rates as N loading increases may result in less overall burial of organic-rich sediment. Given the extent of nitrogen loading along our coastlines, it is imperative that we better understand the resilience of salt marsh systems to nutrient enrichment, especially if we hope to rely on salt marshes, and other blue carbon systems, for long-term carbon storage.

Lactate as a key metabolic intermediate in cancer.

In 1985, George Brooks proposed the “Lactate Shuttle” (1), subsequently labeled the “Cell-Cell Lactate Shuttle” (2-4). In that initial treatise, relying largely on the use of isotopic tracers, he proposed a new concept for lactate (La − ) as a central player in metabolism during both rest and exercise.

Spatio‐temporal heterogeneity in soil water stable isotopic composition and its ecohydrologic implications in semiarid ecosystems

AbstractSpatio‐temporal heterogeneity in soil water content is recognized as a common phenomenon, but heterogeneity in the hydrogen and oxygen isotope composition of soil water, which can reveal processes of water cycling within soils, has not been well studied. New advances are being driven by measurement approaches allowing sampling with high density in both space and time. Using in situ soil water vapour probe techniques, combined with conventional soil and plant water vacuum distillation extraction, we monitored the hydrogen and oxygen stable isotopic composition of soil and plant waters at paired sites dominated by grasses and Gambel's oak (Quercus gambelii) within a semiarid montane ecosystem over the course of a growing season. We found that sites spaced only 20 m apart had profoundly different soil water isotopic and volumetric conditions. We document patterns of depth‐ and time‐explicit variation in soil water isotopic conditions at these sites and consider mechanisms for the observed heterogeneity. We found that soil water content and isotopic variability were damped under Q. gambelii, perhaps due in part to hydraulic redistribution of deep soil water or groundwater by Q. gambelii in these soils relative to the grass‐dominated site. We also found some support for H isotope discrimination effects during water uptake by Q. gambelii. In this ecosystem, the soil water content was higher than that at the neighbouring Grass site, and thus, 25% more water was available for transpiration by Q. gambelii compared with the Grass site. This work highlights the role of plants in governing soil water variation and demonstrates that they can also strongly influence the isotope ratios of soil water. The resulting fine‐scale heterogeneity has implications for the use of isotope tracers to study soil hydrology and evaporation and transpiration fluxes to improve understanding of water cycling through the soil–plant–atmosphere continuum.

Use of Isotope Tracers to Assess Lipid Absorption in Conscious Lymph Fistula Mice.

This protocol provides a comprehensive reference for the evolution of the lymph fistula model, the mechanism of lipid absorption, the detailed procedure for studying lipid absorption using the lymph fistula model, the interpretation of the results, and consideration of the experimental design. The lymph fistula model is an approach to assess the concentration and rate of a range of molecules transported by the lymph by cannulating lymph duct in animals. In this protocol, mice first undergo surgery with the implantation of cannulae in the duodenum and mesenteric lymph duct and are allowed to recover overnight in Bollman restraining cages housed in a temperature-regulated environment. To study in vivo lipid absorption, a lipid emulsion is prepared with labeled tracers, including [3 H]-triolein and [14 C]-cholesterol. On the day of the experiment, mice are continuously infused with lipid emulsion via the duodenum for 6 hr, and lymph is usually collected hourly. At the end of the study, gastrointestinal segments and their luminal contents are collected separately for determination of the digestion, uptake, and transport of exogenous lipids. © 2019 by John Wiley & Sons, Inc.

Tracing Mineral Reactions Using Confocal Raman Spectroscopy

Raman spectroscopy is a powerful tool used to identify mineral phases, study aqueous solutions and gas inclusions as well as providing crystallinity, crystallographic orientation and chemistry of mineral phases. When united with isotopic tracers, the information gained from Raman spectroscopy can be expanded and includes kinetic information on isotope substitution and replacement mechanisms. This review will examine the research to date that utilizes Raman spectroscopy and isotopic tracers. Beginning with the Raman effect and its use in mineralogy, the review will show how the kinetics of isotope exchange between an oxyanion and isotopically enriched water can be determined in situ. Moreover, we show how isotope tracers can help to unravel the mechanisms of mineral replacement that occur at the nanoscale and how they lead to the formation of pseudomorphs. Finally, the use of isotopic tracers as an in situ clock for mineral replacement processes will be discussed as well as where this area of research can potentially be applied in the future.

Isotope mixing models require individual isotopic tracer content for correct quantification of sediment source contributions

AbstractThe use of isotopic tracers for sediment source apportionment is gaining interest with recent introduction of compound‐specific stable isotope tracers. The method relies on linear mixing of source isotopic tracers, and deconvolution of a sediment mixture initially quantifies the contribution of sources to the mixture's tracer signature. Therefore, a correction to obtain real sediment source proportions is subsequently required. As far as we are aware, all published studies to date have used total isotopic tracer content or a proxy (e.g., soil carbon content) for this post‐unmixing correction. However, as the relationship between the isotopic tracer mixture and the source mixture is different for each isotopic tracer, post‐unmixing corrections cannot be carried out with one single factor.This contribution presents an isotopic tracer model structure—the concentration‐dependent isotope mixing model (CD‐IMM)—to overcome this limitation. Herein, we aim to clarify why the “conventional” approach to converting isotopic tracer proportions to source proportions using a single factor is wrong. In an initial mathematical assessment, error incurred by not using CD‐IMM (NCD‐IMM) in unmixing two sources with two isotopic tracers showed a complex relation as a function of relative tracer contents. Next, three artificial mixtures with different proportions of three soil sources were prepared and deconvoluted using 13C of fatty acids using CD‐IMM and NCD‐IMM. Using NCD‐IMM affected both accuracy (mean average error increased up to a threefold compared with the CD‐IMM output) and precision (interquartile range was up to 2.5 times larger). Finally, as an illustrative example, the proportional source contribution reported in a published study was recalculated using CD‐IMM. This resulted in changes in estimated source proportions and associated uncertainties.Content of isotopic tracers is seldom reported in published work concerning use of isotopic tracers for sediment source partitioning. The magnitude of errors made by miscalculation in former studies is therefore difficult to assess. With this contribution, we hope the community will acknowledge the limitations of prior approaches and use a CD‐IMM in future studies.

Drivers of nitrogen transfer in stream food webs across continents

Studies of trophic-level material and energy transfers are central to ecology.The use of isotopic tracers has now made it possible to measure trophic transfer efficiencies of important nutrients and to better understand how these materials move through food webs.We analyzed data from thirteen 15 N-ammonium tracer addition experiments to quantify N transfer from basal resources to animals in headwater streams with varying physical, chemical, and biological features. N transfer efficiencies from primary uptake compartments (PUCs; heterotrophic microorganisms and primary producers) to primary consumers was lower (mean 11.5%, range <1% to 43%) than N transfer efficiencies from primary consumers to predators (mean 80%, range 5% to >100%). Total N transferred (as a rate) was greater in streams with open compared to closed canopies and overall N transfer efficiency generally followed a similar pattern, although was not statistically significant. We used principal component analysis to condense a suite of site characteristics into two environmental components. Total N uptake rates among trophic levels were best predicted by the component that was correlated with latitude, DIN:SRP, GPP:ER, and percent canopy cover. N transfer efficiency did not respond consistently to environmental variables. Our results suggest that canopy cover influences N movement through stream food webs because light availability and primary production facilitate N transfer to higher trophic levels.

Isotopic Tracers for Combustion Research

ABSTRACTThis review article deals with the use of isotopic tracers in the field of combustion science. A number of researchers have reported the use of isotopic techniques, which have been employed to solve a wide range of combustion problems. Radioactive and stable isotopes have been utilized as tracers, including isotopes of carbon (13C and 14C), oxygen (18O), and deuterium (D). One of the main applications has been to quantitatively determine the propensity of a molecule in a mixture, or specific atom within a molecule, to form pollutant emissions. Tracer studies have also been used for the elucidation of combustion reaction pathways, and kinetic rate constant determination of elementary reactions. A number of analytical techniques have been used for isotope detection; and the merits of some of the different techniques are discussed in the context of combustion research. This article concludes by exploring emerging methods and potential future techniques and applications.

Stable isotopes reveal sources of precipitation in the Qinghai Lake Basin of the northeastern Tibetan Plateau

The use of isotopic tracers is an effective approach for characterizing the moisture sources of precipitation in cold and arid regions, especially in the Tibetan Plateau (TP), an area of sparse human habitation with few weather and hydrological stations. This study investigated stable isotope characteristics of precipitation in the Qinghai Lake Basin, analyzed moisture sources using data sets from NCEP–NCAR, and calculated vapor contributions from lake evaporation to the precipitation in the basin using a two-component mixing model. Results showed that the Local Meteoric Water Line (LMWL) was defined as δ2H=7.86 δ18O+15.01, with a slope of less than 8, indicating that some non-equilibrium evaporation processes occurred when the drops fell below the cloud base. Temperature effects controlled δ18O and δ2H in precipitation in the basin, with high values in summer season and low values in winter season. Moisture in the basin was derived predominantly from the Southeast Asian Monsoon (SEAM) from June to August and the Westerly Circulation (WC) from September through May. Meanwhile, the transition in atmospheric circulation took place in June and September. The SEAM strengthened gradually, while the WC weakened gradually in June, and inversely in September. However, the Southwest Asian Monsoon (SWAM) did not reach the Qinghai Lake Basin due to the barrier posed by Tanggula Mountain. High d-excess (>10‰) and significant altitude and lake effects of δ18O in precipitation suggested that the vapor evaporated from Qinghai Lake, strongly influenced annual precipitation, and affected the regional water cycle in the basin distinctly. The monthly contribution of lake evaporation to basin precipitation ranged from 3.03% to 37.93%, with an annual contribution of 23.42% or 90.54mm, the majority of which occurred in the summer season. The findings demonstrate that the contribution of evaporation from lakes to atmospheric vapor is fundamental to water cycling on the TP.

Use of isotopic tracers to characterize the interaction of water components and nitrate contamination in the arid Rasafeh area (Syria)

Chemical and isotopic data in atmospheric precipitation, surface water, and groundwater in arid Rasafeh area, northeast Syria, are used to clarify the status of groundwater quality, the interaction of water components, groundwater dating, and vulnerability to anthropogenic contamination. Interpretation of chemical data with thermodynamic calculation reveals that the dissolution of evaporate mineral is the main factor of high salinity. The δ18O and δ2H relationships indicate that the groundwater is fed by mixing water from Euphrates River and precipitation and the isotope balance equation were used to estimate the contribution of the Euphrates River to the aquifers recharge. High tritium content, together with high 14C activity in the majority of groundwater samples, indicate shorter residence times and consequently potentially greater recharge. The presence of high nitrate concentration associated with high tritium concentration in both shallow and deep aquifer units indicates the presence of high permeability, so that groundwater is highly susceptible to anthropogenic contamination. Nitrate seems to derive exclusively from the application of N fertilizers. The high nitrate values are characteristic of the areas with intensive agricultural activity, indicating the importance of irrigated return flow on the groundwater.

Linear compartmental systems. I. kinetic analysis and derivation of their optimized symbolic equations

The study of many biological systems requires the application of a compartmental analysis, together with the use of isotopic tracers, parameter identification and methods to evaluate the mean parameters. For all this, the kinetic equations of the compartmental system as a function of its parameters are needed. In this paper, we present some considerations on the diagrams of connectivity of linear compartmental systems and obtain new properties from the matrix corresponding to the ordinary first-order linear differential equation systems which describe their kinetic behaviour. Using these properties, symbolic equations are obtained in a simplified form. These equations provide the instantaneous amount of substance in any compartment of the system when zero input is injected into one or more of the system compartments, solely as a function of those parameters of compartmental systems which really have an influence on the sought expression. This is unlike what happens in the other symbolic equations obtained in a previous contribution that included all the fractional transfer coefficients involved in the compartmental system, regardless of whether or not they had an influence on the instantaneous amount of substance.

The use of stable-isotopically labeled oleic acid to interrogate lipid assembly in vivo: assessing pharmacological effects in preclinical species

The use of stable isotopically labeled substrates and analysis by mass spectrometry have provided substantial insight into rates of synthesis, disposition, and utilization of lipids in vivo. The information to be gained from such studies is of particular benefit to therapeutic research where the underlying causes of disease may be related to the production and utilization of lipids. When studying biology through the use of isotope tracers, care must be exercised in interpreting the data to ensure that any response observed can truly be interpreted as biological and not as an artifact of the experimental design or a dilutional effect on the isotope. We studied the effects of dosing route and tracer concentration on the mass isotopomer distribution profile as well as the action of selective inhibitors of microsomal tri-glyceride transfer protein (MTP) in mice and diacylglycerol acyltransferase 1 (DGAT1) in nonhuman primates, using a stable-isotopically labeled approach. Subjects were treated with inhibitor and subsequently given a dose of uniformly ¹³C-labeled oleic acid. Samples were analyzed using a rapid LC-MS technique, allowing the effects of the intervention on the assembly and disposition of triglycerides, cholesteryl esters, and phospholipids to be determined in a single 3 min run from just 10 μl of plasma.

Metabolomics in Drug Target Discovery

Most diseases result in metabolic changes. In many cases, these changes play a causative role in disease progression. By identifying pathological metabolic changes, metabolomics can point to potential new sites for therapeutic intervention. Particularly promising enzymatic targets are those that carry increased flux in the disease state. Definitive assessment of flux requires the use of isotope tracers. Here we present techniques for finding new drug targets using metabolomics and isotope tracers. The utility of these methods is exemplified in the study of three different viral pathogens. For influenza A and herpes simplex virus, metabolomic analysis of infected versus mock-infected cells revealed dramatic concentration changes around the current antiviral target enzymes. Similar analysis of human-cytomegalovirus-infected cells, however, found the greatest changes in a region of metabolism unrelated to the current antiviral target. Instead, it pointed to the tricarboxylic acid (TCA) cycle and its efflux to feed fatty acid biosynthesis as a potential preferred target. Isotope tracer studies revealed that cytomegalovirus greatly increases flux through the key fatty acid metabolic enzyme acetyl-coenzyme A carboxylase. Inhibition of this enzyme blocks human cytomegalovirus replication. Examples where metabolomics has contributed to identification of anticancer drug targets are also discussed. Eventual proof of the value of metabolomics as a drug target discovery strategy will be successful clinical development of therapeutics hitting these new targets.

Investigation of Hg species binding biomolecules in dolphin liver combining GC and LC-ICP-MS with isotopic tracers

Hg stable isotopically labeled species were used to assist the investigation of Hg species (i.e. IHg and MeHg) binding biomolecules in the aqueous soluble fraction of white-sided dolphin (Lagenorhynchus acutus) liver homogenate (QC04LH4). By using stable enriched isotopic tracers (199IHg and 201MeHg), the transformation yields of both mercury species (i.e. methylation and demethylation) were evaluated during the different sample preparation steps, such as the tissue lyophilization and the extraction by ultrasonication. Hg species partitioning after fractionation of the sample (cytosolic fraction and residual fraction) was also studied and showed that MeHg is mainly present in the cytosol (60%), while IHg is preferentially associated to the solid residue (88%). The analysis of the water-soluble fraction by size exclusion chromatography-ICP MS revealed the association of Hg species with biomolecules in a wide molecular weight range. The ratio of IHg and MeHg was successively determined in each SEC fraction by GC-ICP-MS. Furthermore the use of isotopic tracers allowed demonstration of specific Hg species affinity with different biomolecule molecular weight fractions. The results indicate a specific affinity of the IHg with a biomolecule at an elution time corresponding to a metallothionein whereas MeHg is associated to a biomolecule of larger molecular weight.

Chronic Ethanol‐Induced Insulin Resistance Is Associated With Macrophage Infiltration Into Adipose Tissue and Altered Expression of Adipocytokines

Chronic ethanol consumption disrupts glucose homeostasis and is associated with the development of insulin resistance. While adipose tissue and skeletal muscle are the two major organs utilizing glucose in response to insulin, the relative contribution of these two tissues to impaired glucose homeostasis during chronic ethanol feeding is not known. As other models of insulin resistance, such as obesity, are characterized by an infiltration of macrophages into adipose tissue, as well as changes in the expression of adipocytokines that play a central role in the regulation of insulin sensitivity, we hypothesized that chronic ethanol-induced insulin resistance would be associated with increased macrophage infiltration into adipose tissue and changes in the expression of adipocytokines by adipose tissue. Male Wistar rats were fed a liquid diet containing ethanol as 36% of calories or pair-fed a control diet for 4 weeks. The effects of chronic ethanol feeding on insulin-stimulated glucose utilization were studied using the hyperinsulinemic-euglycemic clamp technique, coupled with the use of isotopic tracers. Further, macrophage infiltration into adipose tissue and expression of adipocytokines were also assessed after chronic ethanol feeding. Hyperinsulinemic-euglycemic clamp studies revealed that chronic ethanol feeding to rats decreased whole-body glucose utilization and decreased insulin-mediated suppression of hepatic glucose production. Chronic ethanol feeding decreased glucose uptake in epididymal, subcutaneous, and omental adipose tissue during the hyperinsulinemic-euglycemic clamp, but had no effect on glucose disposal in skeletal muscle. Chronic ethanol feeding increased the infiltration of macrophages into epididymal adipose tissue and changed the expression of mRNA for adipocytokines: expression of mRNA for monocyte chemoattractant protein 1, tumor necrosis factor alpha, and interleukin-6 were increased, while expression of mRNA for retinol binding protein 4 and adiponectin were decreased in epididymal adipose tissue. These data demonstrate that chronic ethanol feeding results in the development of insulin resistance, associated with impaired insulin-mediated suppression of hepatic glucose production and decreased insulin-stimulated glucose uptake into adipose tissue. Chronic ethanol-induced insulin resistance was associated with increased macrophage infiltration into adipose tissue, as well as changes in the expression of adipocytokines by adipose tissue.