Lipophilicity Profiles Research Articles

Chromatographic HILIC indexes to characterize the lipophilicity of zwitterions

Reverse phase high pressure liquid chromatography (RP-HPLC) is widely employed in drug discovery for lipophilicity measurements. Hydrophilic interaction liquid chromatography (HILIC) may represent a good alternative to RP-HPLC in the determination of the lipophilicity of hydrophilic compounds like zwitterions. In this paper three different HILIC stationary phases (ZIC®-HILIC, ZIC®-pHILIC and ZIC®-cHILIC) and two different mobile phases (80%ACN/20%buffer and 90%ACN/10%buffer) were combined to set-up six chromatographic systems. A computational tool named Block Relevance (BR) analysis was firstly used to deconvolute the balance of intermolecular forces governing retention in the six systems. Then the lipophilicity profiles (log k vs pH) of ten model ampholytes were determined. Results support that the lipophilicity of zwitterions at any pH can be successfully determined with a ZIC®-cHILIC stationary phase and an 80%ACN/20%buffer mobile phase. To extend the dataset and confirm results, a second series of zwitterionic drugs was also analyzed.

Botanical Origin Authentication of Polish Phacelia Honey Using the Combination of Volatile Fraction Profiling by HS-SPME and Lipophilic Fraction Profiling by HPTLC

Eleven samples of Polish Phacelia tanacetifolia Benth., three Brassica napus and one Salix spp. honeys were characterized by melissopalynology and analysis of the compositions of their volatile fractions. Headspace solid-phase microextraction coupled with gas chromatography mass spectrometry (HS-SPME/GC–MS) using PDMS/CAR/DVB fiber was used for the isolation of low-molecular weight compounds which create a volatile fraction. To differentiate and indicate the most representative unifloral samples, chemometric techniques such as principal component analysis (PCA) and hierarchical-tree clustering (HTC) were applied to the dataset of the chromatographic fingerprints. Based on the obtained results, a unique chemical fingerprint of phacelia honey was generated. This study allows us to discriminate the botanical origin of the phacelia honeys based on the GC–MS and HPTLC analysis. In case of the GC–MS analysis trans-linalool oxide, hotrienol, cis-linalool oxide and cis-epoxylinalool were identified as a predominant compound. Additionally lipophilic fractions obtained by ultrasound-assisted extraction (UAE) and solid-phase extraction (SPE) were subjected to the HPTLC analysis. It allowed the construction of a barcode-type identifier that could be used to differentiate the honey samples even without identifying the individual components of the obtained fraction.

Fatty acid and lipophilic vitamin composition of seaweeds from Antalya and Çanakkale (Turkey)

The lipophilic vitamin and fatty acid profiles were determined in five edible seaweeds: Corallina elongata J. Ellis & Solander, 1786; Cystoseira barbata (Stackhouse) C. Agardh, 1820; Jania rubens (Linnaeus) J.V. Lamouroux, 1816; Laurencia obtusa (Hudson) J.V. Lamouroux, 1813 and Sargassum vulgare C. Agardh, 1820. Saturated fatty acids (SFAs) were the major fatty acid group, and16:0 formed the highest SFA content (34–40%). 16:0 was higher in the brown seaweeds (C. barbata, 40.55%; S. vulgare, 37.11%) than in the red seaweeds (C. elongata, 36.5%; L. obtusa, 34.57%; J. rubens, 34.22%). The other major fatty acids in the seaweeds were 18:1n-9 and 16:1n-7 from MUFA in the analyzed species. In addition, EPA was found in significant levels in the red seaweeds, whereas DHA was not detected in the analyzed species. The findings showed that difference among species were not statistically significant (p = 0.09–0.11), yet differences between two families (Sargassaceae and Corallinaceae) were significant (p = 0.006) in the fatty acid profile (p < 0.01). Also, differences between the Corallinaceae and Rhomomelaceae families were partially significant (p = 0.011–0.013) (p < 0.01). K1 and K2 vitamins as well as δ-tocopherol, α-tocopherol and retinol acetate were determined to be present in the seaweed extracts. Finally, ergosterol, stigmasterol and ß-sitosterol were found in all samples in differing ratios per species.

Heat shock and UV-B episodes modulate olive leaves lipophilic and phenolic metabolite profiles

Olea europaea L. is the basis of the olive oil chain value. Olive by-products remain economically underexplored despite their richness in polyphenols and fatty acids. Heat waves and raised UV-B radiation episodes (that increasingly affect Mediterranean orchards) may influence these compounds. Herein we characterize the most relevant leaf lipophilic and phenolic compounds of an important Portuguese cultivar Cobrançosa, immediately and 30 days after exposure to high UV-B radiation or heat shock (HS). Plants were exposed to either UV-B (12 kJ m−2 d-1) or HS (40 °C for 2 h) for two consecutive days. Hydroxycinnamic acids, triterpenes, sterols and fatty acids are less influenced by UV-B and HS, while the amounts of flavonoids, secoiridoids and monoterpenes increased. In particular, after both treatments, quercetin-3-O-glucoside levels increased in leaves, and UV-B stimulated oleuropein levels. HS reduced the amounts of verbascoside, oleuropein and luteolin-7-O-glucoside, while UV-B only decrease apigenin-7-O-rutinoside levels. Thirty days after UV-B and HS relief, these compounds showed different profiles. Whereas UV-B recovering plants showed increased amounts of thymol-β-d-glucopyranoside, in HS recovering plants, the increase was more relevant for oleuropein, chrysoeriol-7-O-glucoside and luteolin-7-O-glucoside. Data evidence that Cobrançosa leaves are particularly rich in luteolin-7-O-glucoside, oleuropein, and oleic and palmitic acids, and that their levels may be stimulated by climate-change related conditions, contributing to the economical valorisation of leaves.

Comparison of amnesic, paralytic and lipophilic toxins profiles in cockle (Acanthocardia tuberculata) and smooth clam (Callista chione) from the central Adriatic Sea (Croatia)

Searching for Amnesic (ASP), Paralytic (PSP) and Lipophilic (LT) toxins in seafood is of great importance for consumer protection. Studies are usually focused on the most aquacultured species, the mussel. But, there are a number of potentially commercially important shellfish species as rough cockle Acanthocardia tuberculata (Linnaeus, 1758) and smooth clam Callista chione (Linnaeus, 1758) which are common in the Croatian Adriatic Sea. Investigation of marine biotoxins accumulation in these two species of shellfish from the Adriatic Sea has not been conducted up to now. In order to detect the potential marine biotoxin profile of A. tuberculata and C. chione wild populations, samples were taken monthly during one-year survey from the estuarine area in the central Adriatic Sea. HPLC-FLD with pre-column oxidation and HPLC-UV-DAD methods were employed for PSP and ASP toxins determination, respectively, while LTs were determined by LC-MS/MS. This research had revealed the differences in the accumulation of ASP, PSP and LT toxins between the two studied species, as Acanthocardia tuberculata showed more diverse profile with higher concentrations of analysed toxins. Both investigated shellfish species had shown levels of these biotoxins under the legal limits set by the European Commission.

Toward steroidal anticancer drugs: Non-parametric and 3D-QSAR modeling of 17-picolyl and 17-picolinylidene androstanes with antiproliferative activity on breast adenocarcinoma cells

The present study is aimed to analyze lipophilicity and ADMET profiles, and to develop field based 3D-QSAR and ligand-based pharmacophore hypothesis for a series of 17α-picolyl and 17(E)-picolinylidene androstane derivatives in order to give detailed structural insights and to highlight important binding features of novel androstane derivatives, as compounds with antiproliferative activity toward breast adenocarcinoma cells. This study can provide guidelines for the rational design of novel potent compounds. Sum of ranking differences (SRD), as a non-parametric method, was applied for compounds ranking. 3D-QSAR methods, including comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA), were applied to predict the antiproliferative effect on breast adenocarcinoma cells and provide the regions in space where interactive fields may influence the activity. The compounds are ranked so the compounds with the most favorable ADME and lipophilicity features together with significant anticancer activity can be distinguished. The established 3D-QSAR model could be used for design of new compounds with antiproliferative activity on the human ER– breast adenocarcinoma cells. The pharmacophore model is able to accurately predict antiproliferative activity. Generally, the present study provides significant guidelines for further selection, synthesis and rational design of new highly potential androstane derivatives as anticancer drugs.

Study of pH and temperature effect on lipophilicity of catechol-containing antioxidants by reversed phase liquid chromatography

Lipophilicity of selected antioxidant phytochemicals, including flavonoids, phenolic acids and xanthonoids, was determined by reversed phase high performance liquid chromatography with UV detection (RP-HPLC-UV). The analyses run at different temperature and pH conditions in isocratic mode suggested that lipophilicity as distribution coefficient (logD) between aqueous and organic phase decreases with increasing temperature. For all studied compounds, logD changes slightly at lower pH and much decreases at a given pH range corresponding to the strongest dissociation by which dissociation constant (pKa) was estimated experimentally as well as their bioavailability in gastrointestinal pH conditions were predicted. Thermodynamic parameters (ΔtrH°, ΔtrS°, ΔtrG°) demonstrated the increase in energy of transfer from the aqueous to the organic phase at higher pH thus decrease in lipophilicity. Experimental models of pH and temperature effect obtained by multiple regressions underlined their significant impact (p < 0.05) on lipophilicity. The provided lipophilicity profiles and pKa data are useful descriptors to assess the affinity of the studied compounds for biological membranes, enzymes, carriers and target sites. Furthermore, the obtained result may be useful in the design of functional and medical foods, especially in order to increase the bioavailability of bioactive constituents, such as phenolic compounds, based on their lipophilic and acid-base character.

Immunoassay and amperometric biosensor approaches for the detection of deltamethrin in seawater.

The study of an enzyme-linked immunosorbent assay (ELISA) and an amperometric biosensor for the detection of the pyrethroid deltamethrin in seawater is reported. The preparation of specific polyclonal antibodies is addressed using two immunizing haptens based on deltamethrin and cypermethrin compounds, with a spacer arm placed at the cyano residue in the pyrethroid structure. Different conjugates based on bovine serum albumin and aminodextran are prepared depending on the lipophilic profile of the competitor haptens studied. A reproducible and sensitive indirect competitive ELISA is developed, reaching a limit of detection of 1.2 ± 0.04μgL-1 and an IC50 value of 21.4 ± 0.3μgL-1 (both n = 3). For validation of the assays described, artificial seawater samples fortified with deltamethrin are analyzed. For the ELISA assay, these accuracy studies reported a slope of 0.904. An amperometric immunosensor is developed using the same immunoreagents and achieving a comparable detectability in terms of LOD of 4.7μgL-1, measuring seawater without any pretreatment. These results suggest that both techniques can be used as rapid and simple analytical methods for deltamethrin quantification in seawater samples, which are great candidates for initial environmental screening programs. Graphical abstract ᅟ.

Investigation of Hydro-Lipophilic Properties of N-Alkoxyphenylhydroxynaphthalenecarboxamides †.

The evaluation of the lipophilic characteristics of biologically active agents is indispensable for the rational design of ADMET-tailored structure–activity models. N-Alkoxy-3-hydroxynaphthalene-2-carboxanilides, N-alkoxy-1-hydroxynaphthalene-2-carboxanilides, and N-alkoxy-2-hydroxynaphthalene-1-carboxanilides were recently reported as a series of compounds with antimycobacterial, antibacterial, and herbicidal activity. As it was found that the lipophilicity of these biologically active agents determines their activity, the hydro-lipophilic properties of all three series were investigated in this study. All 57 anilides were analyzed using the reversed-phase high-performance liquid chromatography method for the measurement of lipophilicity. The procedure was performed under isocratic conditions with methanol as an organic modifier in the mobile phase using an end-capped non-polar C18 stationary reversed-phase column. In the present study, a range of software lipophilicity predictors for the estimation of clogP values of a set of N-alkoxyphenylhydroxynaphthalenecarboxamides was employed and subsequently cross-compared with experimental parameters. Thus, the empirical values of lipophilicity (logk) and the distributive parameters (π) were compared with the corresponding in silico characteristics that were calculated using alternative methods for deducing the lipophilic features. To scrutinize (dis)similarities between the derivatives, a PCA procedure was applied to visualize the major differences in the performance of molecules with respect to their lipophilic profile, molecular weight, and violations of Lipinski’s Rule of Five.

Discovery of novel 2-[(4-hydroxy-6-oxo-2,3-dihydro-1H-pyridine-5-carbonyl)amino]acetic acid derivatives as HIF prolyl hydroxylase inhibitors for treatment of renal anemia

Prolyl hydroxylase domain-containing protein (PHD) inhibitors are useful as orally administered agents for the treatment of renal anemia. Based on the common structures of known PHD inhibitors, we found novel PHD inhibitor 1 with a 2-[(4-hydroxy-6-oxo-2,3-dihydro-1H-pyridine-5-carbonyl)amino]acetic acid motif. The PHD2-inhibitory activity, lipophilicity (CLogP), and PK profiles (hepatocyte metabolism, protein binding, and/or elimination half-life) of this inhibitor were used as the evaluation index to optimize the structure and eventually discovered clinical candidate 42 as the suitable compound. Compound 42 was demonstrated to promote the production of erythropoietin (EPO) following oral administration in mice and rats. The predicted half-life of this compound in humans was 1.3–5.6 h, therefore, this drug may be expected to administer once daily with few adverse effects caused by excessive EPO production.

UV-B radiation modulates physiology and lipophilic metabolite profile in Olea europaea

Ultraviolet-B (UV-B) radiation plays an important role in plant photomorphogenesis. Whilst the morpho-functional disorders induced by excessive UV irradiation are well-known, it remains unclear how this irradiation modulates the metabolome, and which metabolic shifts improve plants’ tolerance to UV-B. In this study, we use an important Mediterranean crop, Olea europaea, to decipher the impacts of enhanced UV-B radiation on the physiological performance and lipophilic metabolite profile. Young olive plants (cv. ‘Galega Vulgar’) were exposed for five days to UV-B biologically effective doses of 6.5 kJ m−2 d−1 and 12.4 kJ m−2 d−1. Cell cycle/ploidy, photosynthesis and oxidative stress, as well as GC–MS metabolites were assessed. Both UV-B treatments impaired net CO2 assimilation rate, transpiration rate, photosynthetic pigments, and RuBisCO activity, but 12.4 kJ m−2 d−1 also decreased the photochemical quenching (qP) and the effective efficiency of PSII (ΦPSII). UV-B treatments promoted mono/triperpene pathways, while only 12.4 kJ m−2 d−1 increased fatty acids and alkanes, and decreased geranylgeranyl-diphosphate. The interplay between physiology and metabolomics suggests some innate ability of these plants to tolerate moderate UV-B doses (6.5 kJ m−2 d−1). Also their tolerance to higher doses (12.4 kJ m−2 d−1) relies on plants’ metabolic adjustments, where the accumulation of specific compounds such as long-chain alkanes, palmitic acid, oleic acid and particularly oleamide (which is described for the first time in olive leaves) play an important protective role. This is the first study demonstrating photosynthetic changes and lipophilic metabolite adjustments in olive leaves under moderate and high UV-B doses.

DTPA-(PABn)-Leu5]-des-acyl ghrelin(1-5) as a new carrier of radionuclides and potential precursor of radiopharmaceuticals.

Ghrelin is a peptide consisting of 28 aminoacids and an octadecyl side chain (acyl group) binding to the growth hormone secretagogue receptor type 1a (GHS-R1a). Its des-acylated form, des-acyl ghrelin (DAG) binds to the corticotropin releasing factor receptor type 2a (CRF2a) located on endocrine cancer cells such as the prostate carcinoma cell line DU 145. The aim of this study is to develop a new DAG-based carrier of radionuclides with potential application in therapy. Trunctated C-terminal five aminoacids chain of the DAG peptide (H2N-Gly-Ser-Ser-Phe-Leu-COOH) was linked to DTPA to obtain [DTPA-(PABn)-Leu5]-DAG(1-5). For therapeutic application the lutetium-177 (177Lu) radionuclide was coordinated to the peptide. To determine biological and chemical properties of newly synthesized radiopharmaceutical, two iodine-131 (131I)-labelled compounds were used: [131I]-Tyr4-DAG(1-5) and full length [131I]-DAG(1-28) together with their nonradioactive forms: DAG(1-28) and DAG(1-5). Identical HPLC elution profiles of [177Lu-DTPA-(PABn)-Leu5]-DAG(1-5) before and after incubation with human serum proved its stability. The lipophilicity profile of [177Lu-DTPA-(PABn)-Leu5]-DAG(1-5) was log DO/W=-2.68±0.05, pH 7.4. Receptor affinity of the nonradioactive conjugate [Lu-DTPA-(PABn)-Leu5]-DAG(1-5) was IC50 (21.06 nmol/l), as shown against the [131I]-DAG(1-28) used as a competitor. The 3-(4,5-dimethyldiazol-2-yl)-2,5-diphenyltetrazolium bromide assay indicated the significant cytotoxicity of the newly synthesized compounds, similar to that of [131I]-Tyr4-DAG(1-5). The results obtained suggest the potency of the [DTPA-(PABn)-Leu5]-DAG(1-5) as a new carrier of radionuclides in radiopharmacy.

Passive Membrane Penetration of the Serotonin Precursor 5-Hydroxytryptophan is Controlled by Its Zwitterion.

Species-specific partition coefficients in the octanol/water system were determined for the neurotransmitter serotonin (5-HT) and its precursor 5-hydroxytryptophan (5-HTP). The pH-independent partition coefficients (p) of the individual microspecies were determined by combination of experimentally measured distribution constants and a custom-tailored evaluation method, using highly similar auxiliary compounds. Experimental microscopic partition coefficients for triprotic molecules have only been reported before for thyroxine and its derivatives. The parabolic pH-distribution profile of 5-HT shows the dominance of the lipophilic non-charged microspecies, with a log p of 0.66. However, the most lipophilic non-charged form of 5-HTP, with a log p of 0.31, has no significant contribution to the distribution coefficient at any pH value. Instead, the less lipophilic zwitterionic protonation isomer dominates the distribution in the pH range 2.10-11.11. Although the non-charged microspecies of 5-HTP is 151 times more lipophilic than its zwitterionic protonation isomer, the overwhelming dominance of the zwitterionic form ensures that its contribution to the overall lipophilicity exceeds 1320 times that of the non-charged one. This fact is another counter-example of the widespread belief that passive diffusion into lipophilic media is predominated by the non-charged species. The lipophilicity profile of 5-HT and 5-HTP is depicted in terms of species-specific lipophilicities.

In silico estimation of basic activity-relevant parameters for a set of drug absorption promoters

Finding a balance between a desired drug’s potency and its physicochemical properties that are important for its molecule pharmacokinetic or pharmacodynamics profile is still a challenging issue in rational drug discovery. Quantitative assessment of the lipophilic characteristics of potential drug molecules is indispensable for efficient development of Absorption, Distribution, Metabolism, Excretion, Toxicity-tailored structure–activity models; therefore reliable procedures for deriving log P from molecular structure are desirable. In the current work a range of various software log P predictors for estimation of the numerical lipophilic values for a set of cholic acid derivatives were employed and subsequently cross-compared with the experimental parameters. Thus, the empirical lipophilicity (RM) was compared with the corresponding log P characteristics calculated using alternative methods for deducing the lipophilic features. The mean values of the selected molecular descriptors that were averaged over the chosen calculation methods (consensus clog P) were subsequently correlated with the RM parameter. As an additional experiment, the iterative variable elimination partial least squares (IVE-PLS) methodology for an ensemble of descriptors retrieved from Dragon 6.0 software was applied for a set of drug transporters. To investigate the variations within the ensemble of cholic acid derivatives principal component analysis (PCA) and self-organizing neural network (SOM) procedures were used to visualize the major differences in the performance of drug promoters with respect to their lipophilic profile.

A comparative study of chromatographic behavior and lipophilicity of selected natural styryl lactones, their derivatives and analogues

This study is based on the analyses of the retention behavior of selected natural styryl lactones and their synthetic analogues in reversed-phase high-performance liquid chromatography. Chromatographic separations were achieved applying ZORBAX SB-C18 column and two different mobile phases: methanol-water and acetonitrile-water. Chromatographic lipophilicity of the analyzed compounds was defined by logk0 constant and correlated with in silico molecular descriptors. According to the statistical validation parameters, obtained results indicate that the presented linear and multiple quantitative structure-retention relationship models can successfully predict the chromatographic lipophilicity of structurally similar compounds. Hierarchical cluster analyses (HCA) was applied in order to group similar compounds according to their chromatographic and in silico lipophilicity. It can be concluded that chromatographic systems with methanol-water were better for modelling of logk0. Modelling was performed in order to characterize compounds regarding their lipophilicity profiles as future drug candidates.

Lipophilic Toxin Profile in Mytilus galloprovincialis from the North Atlantic Coast of Morocco: LC-MS/MS and Mouse Bioassay Analyses

Forthe Moroccan Phycotoxins Monitoring that is part of theSafety of theCoastal Monitoring Network(RSSL),shellfish sampleswereharvestedfromdifferentlocations at NorthAtlantic of Morocco whereharmful algaeblooms were known to have occurred. Forallshellfishsamples found positive by themouse bioassay fordiarrheicshellfishpoisoning(DSP)toxins,liquid chromatography coupled to tandem massspectrometry(LC-MS-MS) in order to search thefollowinglipophilictoxins: okadaic acid (OA), dinophysistoxins(DTXs), pectenotoxins(PTXs), azaspiracids(AZAs), yessotoxins(YTXs), spirolides(SPXs) and gymnodimines (GYMs).Theresultsrevealeddifferentlipophilictoxinprofiles as a function of theshellfish sampling location. It has beennoticed that allthesamples contained OA and its derivativesnamed dinophysistoxins(DTXs). In addition, other lipophilictoxinswerefound in shellfishsamples:(YTXs), and pectenotoxins(PTX-2, PTX-2-seco-acid and 7- epi-PTX-2-seco-acid) on theNorthAtlantic coast .This paper reports on thefirstdetection of YTXs,PTXsGYMs and their derivatives in Moroccan shellfish.

Lipophilicity estimation and characterization of selected steroid derivatives of biomedical importance applying RP HPLC

The present paper deals with chromatographic lipophilicity determination of twenty-nine selected steroid derivatives using reversed-phase high-performance liquid chromatography (RP HPLC) combined with two mobile phase, acetonitrile-water and methanol-water. Chromatographic behavior of four groups (triazole and tetrazole, toluenesulfonylhydrazide, nitrile and dinitrile and dione) of selected steroid derivatives was studied. Investigated compounds were grouped using principal component analysis (PCA) according to their logk values for both mobile phases. Grouping was in the very good accordance with the polarity and lipophilicity of the investigated compounds. QSRR (quantitative structure-retention relationship) approach was used to model chromatographic lipophilicity behavior using molecular descriptors. Modeling was performed using linear regression (LR) and multiple linear regression (MLR) methods. The most influential molecular descriptors were lipophilicity descriptors that are important for molecules ability to pass through biological membranes and geometrical descriptors. All established LR-QSRR and MLR-QSRR models were statistically validated by standards, cross- and external validation parameters as well as with two graphical methods. According to all these assessments, MLR models were better for chromatographic lipophilicity prediction. It was shown that chromatographic systems with methanol-water were better for modeling of logk than systems with acetonitrile-water, as well as the systems that contained lower volume fractions of organic component in mobile phase. Modeling was performed in order to obtain lipophilicity profiles of investigated compounds as future drug candidates of biomedical importance.

Distribution of Dinophysis species and their association with lipophilic phycotoxins in plankton from the Argentine Sea

Dinophysis is a cosmopolitan genus of marine dinoflagellates, considered as the major proximal source of diarrheic shellfish toxins and the only producer of pectenotoxins (PTX). From three oceanographic expeditions carried out during autumn, spring and late summer along the Argentine Sea (∼38–56°S), lipophilic phycotoxins were determined by liquid chromatography coupled to tandem mass spectrometry (LC–MS/MS) in size-fractionated plankton samples. Lipophilic toxin profiles were associated with species composition by microscopic analyses of toxigenic phytoplankton. Pectenotoxin-2 and PTX-11 were frequently found together with the presence of Dinophysis acuminata and Dinophysis tripos. By contrast, okadaic acid was rarely detected and only in trace concentrations, and dinophysistoxins were not found. The clear predominance of PTX over other lipophilic toxins in Dinophysis species from the Argentine Sea is in accordance with previous results obtained from north Patagonian Gulfs of the Argentine Sea, and from coastal waters of New Zealand, Chile, Denmark and United States. Dinophysis caudata was rarely found and it was confined to the north of the sampling area. Because of low cell densities, neither D. caudata nor Dinophysis norvegica could be biogeographically related to lipophilic toxins in this study. Nevertheless, the current identification of D. norvegica in the southern Argentine Sea is the first record for the southwestern Atlantic Ocean. Given the typical toxigenicity of this species on a global scale, this represents an important finding for future surveillance of plankton-toxin associations.

An efficient and rapid method for enrichment of lipophilic proteins from Mycobacterium tuberculosis H37Rv for two-dimensional gel electrophoresis.

Lipophilic proteome profiling is crucial because they have an anticipated role in biological processes and pathogenesis of Mycobacterium tuberculosis. These lipophilic proteins might be used as potential targets for the development of newer diagnostic markers and drug targets due to their association with membranes and drugs. We developed an efficient and rapid method to enrich the lipophilic proteins extraction from M. tuberculosis H37Rv for 2DE. In the extraction of lipophilic proteins, nonionic detergent (Triton X-100) was added in sonication buffer that augmented the solubilization of the proteins at the time of sonication. Enriched whole cell lysate was subjected to direct phase separation using Triton X-114, without the need for preisolation of membranes. In this study, we report that our optimized extraction buffer increased the lipophilic proteins extraction and their improved resolution on 2D gel up to two- to threefolds (quantitatively and qualitatively) as compared to standard extraction buffer. Some proteins were identified by MALDI-TOF/MS.

Metabolome Analyses Uncovered a Novel Inhibitory Effect of Acyclic Retinoid on Aberrant Lipogenesis in a Mouse Diethylnitrosamine-Induced Hepatic Tumorigenesis Model.

Acyclic retinoid (ACR) is a promising drug under clinical trials for preventing recurrence of hepatocellular carcinoma. The objective of this study was to gain insights into molecular basis of the antitumorigenic action of ACR from a metabolic point of view. To achieve this, comprehensive cationic and lipophilic liver metabolic profiling was performed in mouse diethylnitrosamine (DEN)-induced hepatic tumorigenesis model using both capillary electrophoresis time-of-flight mass spectrometry and liquid chromatography time-of-flight mass spectrometry. ACR significantly counteracted against acceleration of lipogenesis but not glucose metabolism in DEN-treated mice liver, suggesting an important role of lipid metabolic reprogramming in the initiation step of hepatic tumorigenesis. Knowledge-based pathway analysis suggested that inhibition of linoleic acid metabolites such as arachidonic acid, a proinflammatory precursor, played a crucial role in the prevention by ACR of DEN-induced chronic inflammation-mediated tumorigenesis of the liver. As a molecular mechanism of the ACR's effect to prevent the aberrant lipogenesis, microarray analysis identified that a key transcription regulator of both embryogenesis and tumorigenesis, COUP transcription factor 2, also known as NR2F2, was associated with the metabolic effect of ACR in human hepatocellular carcinoma cells. Our study provided potential therapeutic targets for the chemoprevention of hepatocellular carcinoma as well as new insights into the mechanisms underlying prevention of hepatic tumorigenesis.