Circulatory Metabolite Ratios as Indicators of Lifestyle Risk Factors Based on a Greek NAFLD Case-Control Study.

An ensemble of confounding factors, such as an unhealthy diet, obesity, physical inactivity, and smoking, have been linked to a lifestyle that increases one's susceptibility to chronic diseases and early mortality. The circulatory metabolome may provide a rational means of pinpointing the advent of metabolite variations that reflect an adherence to a lifestyle and are associated with the occurrence of chronic diseases. Data related to four major modifiable lifestyle factors, including adherence to the Mediterranean diet (estimated on MedDietScore), body mass index (BMI), smoking, and physical activity level (PAL), were used to create the lifestyle risk score (LS). The LS was further categorized into four groups, where a higher score group indicates a less healthy lifestyle. Drawing on this, we analyzed 223 NMR serum spectra, 89 MASLD patients and 134 controls; these were coupled to chemometrics to identify "key" features and understand the biological processes involved in specific lifestyles. The unsupervised analysis verified that lifestyle was the factor influencing the samples' differentiation, while the supervised analysis highlighted metabolic signatures. Τhe metabolic ratios of alanine/formic acid and leucine/formic acid, with AUROC > 0.8, may constitute discriminant indexes of lifestyle. On these grounds, this research contributed to understanding the impact of lifestyle on the circulatory metabolome and highlighted "prudent lifestyle" biomarkers.

Serum metabolomic profiling unveils distinct sex-related metabolic patterns in NAFLD.

Objective: Obesity poses an increased risk for the onset of Nonalcoholic fatty liver disease (NAFLD). The influence of other factors, such as sex in the incidence and severity of this liver disease has not yet been fully elucidated. Thus, we aimed to identify the NAFLD serum metabolic signatures associated with sex in normal, overweight and obese patients and to associate the metabolite fluctuations across the increasing liver steatosis stages. Methods and results: Using nuclear magnetic resonance (NMR) serum samples of 210 NAFLD cases and control individuals diagnosed with liver U/S, our untargeted metabolomics enquiry provided a sex distinct metabolic bouquet. Increased levels of alanine, histidine and tyrosine are associated with severity of NAFLD in both men and women. Moreover, higher serum concentrations of valine, aspartic acid and mannose were positively associated with the progression of NAFLD among the male subjects, while a negative association was observed with the levels of creatine, phosphorylcholine and acetic acid. On the other hand, glucose was positively associated with the progression of NAFLD among the female subjects, while levels of threonine were negatively related. Fluctuations in ketone bodies acetoacetate and acetone were also observed among the female subjects probing a significant reduction in the circulatory levels of the former in NAFLD cases. A complex glycine response to hepatic steatosis of the female subjects deserves further investigation. Conclusion: Results of this study aspire to address the paucity of data on sex differences regarding NAFLD pathogenesis. Targeted circulatory metabolome measurements could be used as diagnostic markers for the distinct stages of NAFLD in each sex and eventually aid in the development of novel sex-related therapeutic options.

Maternal Dietary Protein Patterns and Neonatal Anthropometrics: A Prospective Study with Insights from NMR Metabolomics in Amniotic Fluid.

This study aimed to characterize dietary protein patterns (DPPs) in a sample pool of 298 well-nourished pregnant women and explore potential associations between DPPs and neonatal anthropometrics. Maternal dietary data were collected using a validated food frequency questionnaire. Neonatal anthropometrics were abstracted from health booklets. A hierarchical cluster analysis identified three DPPs: "Dairy-focused", "Med-fusion", and "Traditional-inspired". The "Dairy-focused" DPP exhibited the highest protein intake (p < 0.001), predominantly animal protein (p < 0.001), while the "Traditional-inspired" DPP presented higher plant protein (p < 0.001) and fiber intakes (p < 0.001), and, therefore, a reduced carbohydrate-to-fiber quotient (p < 0.001). The "Med-fusion" DPP had the lowest protein-to-fat ratio (p < 0.001). Infants of women following the "Dairy-focused" DPP had the highest birth height centiles (p = 0.007) and the lowest ponderal index (p = 0.003). The NMR-metabolomics approach was implemented on a subset of women that provided amniotic fluid (AF) specimens (n = 62) to elucidate distinct metabolic signatures associated with DPPs. PCA and OPLS-DA models verified the adherence to three DPPs, revealing that the levels of several amino acids (AAs) were the highest in "Dairy-focused", reflecting its protein-rich nature. The "Traditional-inspired" DPP showed decreased AAs and glucose levels. This knowledge may contribute to optimizing maternal dietary recommendations. Further research is needed to validate these findings and better understand the relationships between maternal diet, AF metabolic signature, and neonatal anthropometrics.

Urine Metabolomic Profile of Breast- versus Formula-Fed Neonates Using a Synbiotic-Enriched Formula.

The aim of this study was to compare the urine metabolic fingerprint of healthy neonates exclusively breastfed with that of neonates fed with a synbiotic-enriched formula (Rontamil® Complete 1) at four time points (the 3rd and 15th days of life and the 2nd and 3rd months). The determination of urine metabolic fingerprint was performed using NMR metabolomics. Multivariate data analyses were performed with SIMCA-P 15.0 software and R language. Non-distinct profiles for both groups (breastfeeding and synbiotic formula) for the two first time points (3rd and 15th days of life) were detected, whereas after the 2nd month of life, a discrimination trend was observed between the two groups, which was further confirmed at the 3rd month of life. A clear discrimination of the synbiotic formula samples was evident when comparing the metabolites taken in the first days of life (3rd day) with those taken in the 2nd and 3rd months of life. In both cases, OPLS-DA models explained more than 75% of the metabolic variance. Non-distinct metabolomic profiles were obtained between breastfed and synbiotic-formula-fed neonates up to the 15th day of life. Discrimination trends were observed only after the 2nd month of the study, which could be attributed to breastfeeding variations and the consequent dynamic profile of urine metabolites compared to the stable ingredients of the synbiotic formula.

Uncontrolled Thyroid during Pregnancy Alters the Circulative and Exerted Metabolome.

Normal levels of thyroid hormones (THs) are essential for a normal pregnancy outcome, fetal growth and the normal function of the central nervous system. Hypothyroidism, a common endocrine disorder during pregnancy, is a significant metabolic factor leading to cognitive impairments. It is essential to investigate whether patients with thyroid dysfunction may present an altered circulative and excreted metabolic profile, even after receiving treatment with thyroxine supplements. NMR metabolomics was employed to analyze 90 serum and corresponding colostrum samples. Parallel analyses of the two biological specimens provided a snapshot of the maternal metabolism through the excretive and circulating characteristics of mothers. The metabolomics data were analyzed by performing multivariate statistical, biomarker and pathway analyses. Our results highlight the impact of hypothyroidism on metabolites' composition during pregnancy and lactation. Thyroid disorder causing metabolite fluctuations may lead to impaired lipid and glucose metabolic pathways as well as aberrant prenatal neurodevelopment, thus posing a background for the occurrence of metabolic syndrome or neurogenerative diseases later in life. This risk applies to not only untreated but also hypothyroid women under replacement therapy since our findings in both biofluids framed a different metabolic phenotype for the latter group, thus emphasizing the need to monitor women adequately after treatment initiation.

Linking the IL-17A immune response with NMR-based faecal metabolic profile in IBD patients treated with Mastiha.

Dysregulation of intestinal immune response plays a critical role in the pathogenesis of Inflammatory Bowel Disease (IBD). Mastiha's anti-inflammatory properties are well established. Our aim was to investigate Mastiha's regulatory effect on IL-17A serum levels in IBD patients. Alterations of the faecal metabolome as a functional readout of microbial activity were explored. A randomized, double-blind, placebo-controlled, parallel-group design was applied for a total of 3 months in active and 6 months in inactive IBD patients. Serum IL-17A increased significantly in Mastiha group (p = 0.006), and the mean change differed significantly between Mastiha and placebo (p = 0.003) even after adjusting for age, sex and BMI (p = 0.001) in inactive patients. In inactive UC patients IL-17A decreased significantly only in placebo (p = 0.033). No significant differences were detected in active disease. Faecal metabolomics indicated that intervention with Mastiha influenced considerably the metabolic profile of IBD patients in remission exhibiting, in between others, increased levels of glycine and tryptophan. Glycine has been proposed to have a therapeutic effect against IBD, while tryptophan derivatives are involved in immunoregalutory mechanisms, such as the Th17 cells differentiation. Thus, it is quite possible that the immunoregulatory role of Mastiha in quiescent IBD involves the regulation of Th17 cells function and differentiation.

Insights into intrauterine growth restriction based on maternal and umbilical cord blood metabolomics.

Intrauterine growth restriction (IUGR) is a fetal adverse condition, ascribed by limited oxygen and nutrient supply from the mother to the fetus. Management of IUGR is an ongoing challenge because of its connection with increased fetal mortality, preterm delivery and postnatal pathologies. Untargeted nuclear magnetic resonance (1H NMR) metabolomics was applied in 84 umbilical cord blood and maternal blood samples obtained from 48 IUGR and 36 appropriate for gestational age (AGA) deliveries. Orthogonal projections to latent structures discriminant analysis (OPLS-DA) followed by pathway and enrichment analysis generated classification models and revealed significant metabolites that were associated with altered pathways. A clear association between maternal and cord blood altered metabolomic profile was evidenced in IUGR pregnancies. Increased levels of the amino acids alanine, leucine, valine, isoleucine and phenylalanine were prominent in IUGR pregnancies indicating a connection with impaired amino acid metabolism and transplacental flux. Tryptophan was individually connected with cord blood discrimination while 3-hydroxybutyrate assisted only maternal blood discrimination. Lower glycerol levels in IUGR samples ascribed to imbalance between gluconeogenesis and glycolysis pathways, suggesting poor glycolysis. The elevated levels of branched chain amino acids (leucine, isoleucine and valine) in intrauterine growth restricted pregnancies were linked with increased insulin resistance.

Expanding the Role of Sub-Exploited DOE-High Energy Extraction and Metabolomic Profiling towards Agro-Byproduct Valorization: The Case of Carotenoid-Rich Apricot Pulp.

Traditional extraction remains the method-of-choice for phytochemical analyses. However, the absence of an integrated analytical platform, focusing on customized, validated extraction steps, generates tendentious and non-reproducible data regarding the phytochemical profile. Such a platform would also support the exploration and exploitation of plant byproducts, which are a valuable source of bioactive metabolites. This study deals with the incorporation of (a) the currently sub-exploited high energy extraction methods (ultrasound (UAE)- and microwave-assisted extraction (MAE)), (b) experimental design (DOE), and (c) metabolomics, in an integrated analytical platform for the extensive study of plant metabolomics and phytochemical profiling. The recovery of carotenoids from apricot by-products (pulp) is examined as a case study. MAE, using ethanol as solvent, achieved higher carotenoid yields compared to UAE, where 1:1 chloroform-methanol was employed, and classic extraction. Nuclear magnetic resonance (NMR)-based metabolomic profiling classified extracts according to the variations in co-extractives in relation to the extraction conditions. Extracts with a lower carotenoid content contained branched-chain amino acids as co-extractives. Medium carotenoid content extracts contained choline, unsaturated fatty acids, and sugar alcohols, while the highest carotenoid extracts were also rich in sugars. Overall, the proposed pipeline can provide different the phytochemical fractions of bioactive compounds according to the needs of different industrial sectors (cosmetics, nutraceuticals, etc.).

Early Human-Milk Metabolome in Cases of Intrauterine Growth-Restricted and Macrosomic Infants.

BACKGROUND: Abnormal fetal growth is associated with short-term and long-term metabolic dysregulation and susceptibility to obesity-related disorders. Maternal milk, the ideal source of infantile nutrition, protects from metabolic diseases in adulthood. By applying nuclear magnetic resonance (NMR) metabolomics, this study investigated the metabolic profile of early human milk/colostrum (EHM/C) at the extremes of fetal-growth conditions, which could affect its nutritional value. METHODS: From 98 mothers delivering 60 appropriate-for-gestational-age (AGA), 19 large-for-gestational-age (LGA), and 19 intrauterine growth-restricted (IUGR) full-term neonates, milk samples collected on the third to fourth day post partum were examined by NMR spectroscopy. Multivariate data analysis elicited information from NMR spectra and probed to metabolic signatures of EHM/C. RESULTS: LGA and IUGR EHM/C samples depicted increased content in lactose, citric acid, choline, phosphocholine, and N-acetylglutamine. AGA samples exhibited increased isoleucine and valine. Metabolic pathways involved were valine, leucine/isoleucine biosynthesis and degradation, glycerophospholipid metabolism, aminoacyl-transfer RNA biosynthesis, and citrate cycle. Orthogonal projections to latent structures discriminant analysis models were validated. CONCLUSION: This holistic metabolomics study framed an increased content of certain essential nutrients in EHM/C samples following the birth of LGA and IUGR infants prone to short- and long-term metabolic disorders, thus stressing additional benefits of early breastfeeding. Assessing the metabolic profile of EHΜ/C enables evaluation of its nutrition value, adjusted to fetal growth, and introduction of appropriate dietary interventions.

Evaluating the experimental cultivation of peppers in low-energy-demand greenhouses. An interdisciplinary study.

BACKGROUND: Photovoltaics (PV) provide an alternative solution to cover energy demands in greenhouses. This study evaluates the effect of PV panels installed on the roofs of greenhouses, and the partial shading that they cause, on the growth parameters and growth indicators of the experimental cultivation of peppers (Capsicum annuum cv. California Wonder). The growth of the plants, the antioxidant profile, radical scavenging activity, total phenolic content, and the phenolic and metabolic profiles (using LC-MS spectrometry and NMR spectroscopy) are evaluated. RESULTS: Data are presented from a full cultivation period. Results indicated that indoor temperatures were similar for both glass and glass-PV (glass with PV panels installed) greenhouses during the day and the night. The production yield was higher for the glass-PV greenhouses. The pepper fruits' weight, dimensions, and thickness were similar in both cases. Comparison of the pepper fruit extracts in terms of total phenolic content, antioxidant, and antiradical activities indicated differences that were not statistically significant. Photometric and spectroscopic studies both showed a smaller distribution of values in the case of the glass-PV greenhouse, probably indicating a more consistent phytochemical profile. CONCLUSION: Covering only a small proportion (ca. 20%) of the greenhouse roof with photovoltaic panels contributes considerably to its energy demands without affecting plant growth and quality. © 2018 Society of Chemical Industry.

On the Characterization and Correlation of Compositional, Antioxidant and Colour Profile of Common and Balsamic Vinegars.

Commercially available common and balsamic vinegars were examined, using a combination of spectrophotometric, chromatographic, colorimetric and spectroscopic methods. Total phenolic content, antioxidant activity, radical scavenging capacity, phenolic profile, colour parameters, Fourier Transform Infrared (FT-IR) absorbance spectra and Nuclear Magnetic Resonance (¹H NMR) spectra were comparatively studied. The main scope was the assessment of vinegar antioxidant and metabolic profiles and the identification of the most appropriate features influencing their type and subtypes. Red grape balsamic vinegars exhibited the strongest antioxidant profile. High total phenolic content and radical scavenging-antioxidant activity of vinegars was strongly correlated with high hue-angle and colour density values and low lightness and a* values. FT-IR spectra analysis confirmed the presence of organic acids and carbohydrates and, in combination with Gas Chromatography-Mass Spectrometry (GC-MS), the occurrence of phenolic compounds. NMR spectroscopy enabled the identification of 27 characteristic metabolites in each type of vinegar. The combination of all applied techniques provides critical information on compositional differences among the vinegars and could serve as an application tool for similar fermentation products.

1H NMR-based metabolomics reveals the effect of maternal habitual dietary patterns on human amniotic fluid profile.

Maternal diet may influence offspring's health, even within well-nourished populations. Amniotic fluid (AF) provides a rational compartment for studies on fetal metabolism. Evidence in animal models indicates that maternal diet affects AF metabolic profile; however, data from human studies are scarce. Therefore, we have explored whether AF content may be influenced by maternal diet, using a validated food-frequency questionnaire and implementing NMR-based metabolomics. Sixty-five AF specimens, from women undergoing second-trimester amniocentesis for prenatal diagnosis, were analysed. Complementary, maternal serum and urine samples were profiled. Hierarchical cluster analysis identified 2 dietary patterns, cluster 1 (C1, n = 33) and cluster 2 (C2, n = 32). C1 was characterized by significantly higher percentages of energy derived from refined cereals, yellow cheese, red meat, poultry, and "ready-to-eat" foods, while C2 by higher (P < 0.05) whole cereals, vegetables, fruits, legumes, and nuts. 1H NMR spectra allowed the identification of metabolites associated with these dietary patterns; glucose, alanine, tyrosine, valine, citrate, cis-acotinate, and formate were the key discriminatory metabolites elevated in C1 AF specimens. This is the first evidence to suggest that the composition of AF is influenced by maternal habitual dietary patterns. Our results highlight the need to broaden the knowledge on the importance of maternal nutrition during pregnancy.

Herbal distillates: A new era of grape marc distillates with enriched antioxidant profile.

Grape marc distillates are traditional alcoholic beverages, produced mostly in the Mediterranean countries. The present study proposes the enrichment of a Greek traditional grape marc distillate (tsikoudia) with selected herbs to enhance its natural antioxidants and functional properties. Total phenolic content, the antiradical and antioxidant activities, as well as the phenolic and sugar profiles using NMR and FT-IR spectroscopy were evaluated. The enrichment of distillates with Syzygium aromaticum L., Jasminum officinale L. and Eucalyptus globulus Labill. exhibited the highest total phenolic content as well as the highest antioxidant and antiradical activities, whereas the lowest values were observed with Hippophae rhamnoides L. and Lycium Barbarum Mill. The implementation of NMR and FT-IR spectroscopies attested to the presence of phenolic compounds and of specific carbohydrates in herbal distillates, postulating their migration from selected herbal species to tsikoudia and probably contributing to their organoleptic characteristics. The target of this approach leads to new added-value distillates with enhanced characteristics.

NMR Metabolomics Investigates the Influence of Flavonoid-Enriched Rations on Chicken Plasma.

The use of flavonoids as dietary supplements is well established, mainly due to their intense antioxidant and anti-inflammatory properties. In the present study, hesperidin, naringin, and vitamin E were used as additives at different concentrations in poultry rations in order to achieve meat of improved quality. NMR metabolomics was applied to chicken blood serum samples to discern whether and how the enriched rations affected the animals' metabolic profile. Variations in the metabolic patterns according to sustenance consumption were traced by orthogonal projections to latent structures discriminant analysis (OPLS-DA) models and were attributed to specific metabolites by using S-line plots. In particular, serum samples from chickens fed with vitamin E displayed higher concentrations of glycine and succinic acid compared to control samples, which were mainly characterized by betaine, formic acid, and lipoproteins. Samples from chickens fed with hesperidin were characterized by increased levels of lactic acid, citric acid, creatine, carnosine, creatinine, phosphocreatine, anserine, glucose, and alanine compared to control samples. Lastly, naringin samples exhibited increased levels of citric and acetic acids. Results verify the scalability of NMR metabolomics to highlight metabolite variations among chicken serum samples in relation to food rations.

Metabolic and antioxidant profiles of herbal infusions and decoctions.

This study implements NMR metabolomics and spectrophotometric studies (Folin-Ciocalteu, FRAP, ABTS) to infusions and decoctions of ten plant species in order to assess and compare the metabolic and antioxidant profiles for each botanical family. Multivariate and univariate data analyses highlighted the differences among the samples and pinpointed specific classes of compounds for each plant species as well as infusions and decoctions. The identified phenolic compounds by NMR, as well as the antioxidant profile, framed a trend of increased values in infusions compared to the decoctions. Moreover, the infusion procedure positively affected the extractability of the phenolic compounds compared to decoctions. The highest total phenolic content was found in Mentha spicata, while the lowest in Matricaria chamomilla preparations, irrespective of the preparation method. The preparation time for the decoctions was examined showing that the 15min preparations were generally found richer in phenolics and of higher antioxidant capacity.

NMR metabolic fingerprinting and chemometrics driven authentication of Greek grape marc spirits.

NMR metabolomics was used to investigate 57 Greek spirits of four indigenous and four international varieties from Macedonia (tsipouro) and Crete (tsikoudia) to establish their authenticity. The metabolic profile of Romeiko, Malvasia, Xinomavro, Sangiovese and Nebbiolo varieties was assessed for the first time. The WET1D sequence was used to improve sensitivity and unveil minor metabolites. PCA was applied to delineate the provenance of samples and associate metabolites with distinct varietal characteristics, such as the acidity of Sangiovese, the overripe grapes harvesting of Romeiko, the intense body of Cabernet Sauvignon, the light body of Xinomavro and the glutamic acid for Malvasia. The migration of Cabernet Sauvignon from north Greece to Crete was framed. Monitoring multi-varietal spirits introduced 2-vinylethanol as a marker for yeast selection. OPLS-DA was applied to samples from the same vineyard, thus highlighting genotypic markers. Consequently, the findings address the concepts of typicity and traceability in grape marc spirits.

Evaluation of total reducing power of edible oils.

The lipophilicity of untreated edible oils narrows the application of most published methods for the determination of antioxidant activity to hydrophilic extracts of oils. This research addresses the issue of the estimation of the total antioxidant properties of untreated edible oils by modifying two widely applied analytical methods, the Fe-Phenanthroline and the CUPRAC assays, to be used in untreated oils. The modifications pertain to the selection of mixture of solvents (ethanol-butanol in 3:1 v/v ratio), and the optimization of the reaction conditions (reagents concentration and reaction time). The developed methods were applied to a number of hydrophilic and lipophilic standard compounds and different types of commercial edible oils, as well as their corresponding aqueous or organic extracts. This implementation elucidated the differences in the antioxidant content of edible oils. All the results were compared to those of the DPPH and Folin-Ciocalteu methods and the analytical figures of merit for the methods have been estimated. Lastly, it was concluded that the modified CUPRAC assay has higher sensitivity compared to the Fe-Phenanthroline assay.

NMR metabolite profiling of Greek grape marc spirits.

This (1)H NMR based study profiles metabolites in Greek grape marc distillates, tsipouro and tsikoudia. Eightysix samples of indigenous and international varieties, stemming from major vine growing regions of Greece were investigated. The monitoring protocol addressed the global metabolic profile of untreated samples and accomplished the unambiguous assignment of 35 metabolites. NMR spectra were acquired by applying the robust, sensitive and rapid WET1D NMR pulse sequence, which succeeded to unveil the presence of minor compounds in a high ethanol matrix. PCA classified the samples according to their provenance, incorporating also information related to the variety, vintage year and production process within each formed regional assembly. Metabolites such as fusel alcohols, polyols, ethyl esters, mono- and di-saccharides were associated with the classification of samples. OPLS-DA ascribed to samples of common regional entity characteristic genotypic metabolites and probed to the potential influence of the vintage effect. Finally, metabolite profiling underlined the influence of the fermentation and distillation procedures.

Comparative study of the AT1 receptor prodrug antagonist candesartan cilexetil with other sartans on the interactions with membrane bilayers.

Drug-membrane interactions of the candesartan cilexetil (TCV-116) have been studied on molecular basis by applying various complementary biophysical techniques namely differential scanning calorimetry (DSC), Raman spectroscopy, small and wide angle X-ray scattering (SAXS and WAXS), solution ¹H and ¹³C nuclear magnetic resonance (NMR) and solid state ¹³C and ³¹P (NMR) spectroscopies. In addition, ³¹P cross polarization (CP) NMR broadline fitting methodology in combination with ab initio computations has been applied. Finally molecular dynamics (MD) was applied to find the low energy conformation and position of candesartan cilexetil in the bilayers. Thus, the experimental results complemented with in silico MD results provided information on the localization, orientation, and dynamic properties of TCV-116 in the lipidic environment. The effects of this prodrug have been compared with other AT1 receptor antagonists hitherto studied. The prodrug TCV-116 as other sartans has been found to be accommodated in the polar/apolar interface of the bilayer. In particular, it anchors in the mesophase region of the lipid bilayers with the tetrazole group oriented toward the polar headgroup spanning from water interface toward the mesophase and upper segment of the hydrophobic region. In spite of their localization identity, their thermal and dynamic effects are distinct pointing out that each sartan has its own fingerprint of action in the membrane bilayer, which is determined by the parameters derived from the above mentioned biophysical techniques.

Comparative biophysical studies of sartan class drug molecules losartan and candesartan (CV-11974) with membrane bilayers.

The interactions of the antihypertensive AT(1) antagonists candesartan and losartan with membrane bilayers were studied through the application of DSC, Raman, and solid state (31)P NMR spectroscopies. (1)H and (13)C NMR resonances of candesartan were assigned on the basis of 1D and 2D NMR spectroscopy. A (31)P CP NMR broadline fitting methodology in combination with ab initio computations was implemented and, in conjunction with DSC and Raman results, provided valuable information regarding the perturbation, localization, orientation, and dynamic properties of the drugs in membrane models. In particular, results indicate that losartan anchors in the mesophase region of the lipid bilayers with the tetrazole group oriented toward the polar headgroup, whereas candesartan has less definite localization spanning from water interface toward the mesophase and upper segment of the hydrophobic region. Both sartan molecules decrease the mobilization of the phospholipids alkyl chains. Losartan exerts stronger interactions compared with candesartan, as depicted by the more prominent thermal, structural, and dipolar (1)H-(31)P changes that are caused in the lipid bilayers. At higher concentrations, candesartan strengthens the polar interactions and induces increased order at the bilayer surface. At the highest concentration used (20 mol %), only losartan induces formation of microdomains attributed to the flexibility of its alkyl chain. These results in correlation to reported data with other AT(1) antagonists strengthen the hypothesis that this class of molecules may approach the active site of the receptor by insertion in the lipid core, followed by lateral diffusion toward the binding site. Further, the similarities and differences of these drugs in their interactions with lipid bilayers establish, at least in part, their pharmacological properties.