Marker Sterols Research Articles

Antifungal Activities of cis-trans Citral Isomers against Trichophyton rubrum with ERG6 as a Potential Target.

Trichophyton rubrum causes ringworm worldwide. Citral (CIT), extracted from Pectis plants, is a monoterpene and naturally composed of geometric isomers neral (cis-citral) and geranial (trans-citral). CIT has promising antifungal activities and ergosterol biosynthesis inhibition effects against several pathogenic fungi. However, no study has focused on neral and geranial against T. rubrum, which hinders the clinical application of CIT. This study aimed to compare antifungal activities of neral and geranial and preliminarily elucidate their ergosterol biosynthesis inhibition mechanism against T. rubrum. Herein, the disc diffusion assays, cellular leakage measurement, flow cytometry, SEM/TEM observation, sterol quantification, and sterol pattern change analyses were employed. The results showed geranial exhibited larger inhibition zones (p < 0.01 or 0.05), higher cellular leakage rates (p < 0.01), increased conidia with damaged membranes (p < 0.01) within 24 h, more distinct shriveled mycelium in SEM, prominent cellular material leakage, membrane damage, and morphological changes in TEM. Furthermore, geranial possessed more promising ergosterol biosynthesis inhibition effects than neral, and both induced the synthesis of 7-Dehydrodesmosterol and Cholesta-5,7,22,24-tetraen-3β-ol, which represented marker sterols when ERG6 was affected. These results suggest geranial is more potent than neral against T. rubrum, and both inhibit ergosterol biosynthesis by affecting ERG6.

A gas chromatography-mass spectrometry-based whole-cell screening assay for target identification in distal cholesterol biosynthesis.

Distal cholesterol biosynthesis (CB) has recently taken center stage as a promising drug target in several diseases previously not linked to this biochemical pathway, including cardiovascular disease, cancer, multiple sclerosis and Alzheimer's disease. Most enzymes involved in this pathway are hard to isolate, warranting dedicated analytical tools for biochemical screening. We describe the use of gas chromatography-electron ionization mass spectrometry (GC-MS) in a whole-cell screening assay aimed at monitoring interactions with all enzymes of distal CB in a single experiment. Following cell culture and lipid extraction, the trimethylsilyl ethers of sterols are analyzed by GC-MS. Analytical data for 23 relevant sterols (intermediates) are provided, allowing their unambiguous identification. Sterol pattern analysis reveals the target enzyme on the basis of characteristic marker sterols, whereas quantification of 2-13C-acetate incorporation correlates with the inhibitory activity of drug candidates. The protocol can be used by both experienced scientists and newcomers to the field, allowing detection and quantification of small molecule-enzyme interactions in distal CB. The entire protocol can be carried out within two working days.

Insights about sources, distribution, and degradation of sewage and biogenic molecular markers in surficial sediments and suspended particulate matter from a human-impacted subtropical estuary

The molecular markers sterols and linear alkylbenzenes (LABs) were analyzed in the surficial sediments and suspended particulate matter (SPM) of a subtropical estuary in South Atlantic (Paranaguá Estuarine System). The purpose of this study was identify the spatial distribution of sewage and the input of biogenic organic matter (OM) and to provide comparative insights about their behavior, compositions, and sources. The concentration of coprostanol ranged from < DL (detection limit) to 2.67 μg g−1 in SPM and from < DL to 0.94 μg g−1 in sediments. Total LABs ranged from 43.8 to 480.0 ng g−1 in SPM and from < DL to 21.0 ng g−1 in sediments. LABs homologs composition varied between the two matrices. The local hydrodynamic pattern may promote water column homogenization, dispersion, and dilution of sewage particles, and preferential sedimentation in fluvial and mixture zones. Results suggest that SPM is a good matrix for larger spatial and short time scale evaluation while sediments may help to define hot spot areas of input and final deposition of sewage particles. Marine sterols predominated in SPM while no dominance patterns of marine/terrestrial sterols occurred in surficial sediments. The higher degradation rates of sterols and LABs in the water column must be the main factor for the sharp drop in concentration towards the sediment and the variation of the preferential composition of these markers between compartments.

Sedimentary Steryl Chlorin Esters (SCEs) and Other Photosynthetic Pigments as Indicators of Paleolimnological Change Over the Last 28,000 Years from the Buguldeika Saddle of Lake Baikal

Detailed depth profiles of photosynthetic pigments in a sediment core (G-12) collected at the BDP93 site, the Buguldeika saddle, of south Lake Baikal, along with depth profiles of total organic carbon (TOC) and biogenic silica, were studied to elucidate the temporal changes of phytoplankton assemblages in the lake during the past 28 kyr. In addition to the quantification of carotenoids by high-performance liquid chromatography with photodiode-array detection (HPLC-PDA), steryl chlorin esters (SCEs) were analyzed by HPLC-PDA, HPLC-mass spectrometry (LC-MS) and sterols in SCEs by gas chromatography–mass spectrometry (GC–MS) to enrich the taxonomical information on the phytoplankton composition. Allochthonous input of organic matter from the Selenga River resulted in the higher TOC contents in core G-12 than in a previously reported core (G-6) collected at another site from the southern basin. The poorer correlation in core G-12 than in G-6 between TOC and chlorophyll-a-originating pigments, which are indicative of autochthonous production, also indicated a significant allochthonous input at the site. The abundance of lutein among the carotenoids detected, and the good correlation of total chlorophyll a and b shows that green algae represented a significant portion of the phytoplankton, accompanying the diatoms at the G-12 site, after the last glacial period. The presence of cryptomonads and cyanobacteria were confirmed from marker carotenoids in the sediment core. GC–MS analysis of sterols in SCEs detected marker sterols of diatoms, green algae, chrysophytes and dinoflagellates. The depth profiles of the measured indicators gave consistent features for temporal changes in phytoplankton assemblage at the G-12 site of Lake Baikal after the last glacial maximum.

Effect of Statins on Noncholesterol Sterol Levels: Implications for Use of Plant Stanols and Sterols

Normal serum contains small amounts of noncholesterol sterols, including those reflecting cholesterol absorption and those that are markers of cholesterol synthesis. Absorption marker sterols include serum plant sterols, whereas cholesterol precursor sterols correlate with whole-body synthesis of cholesterol. Thus, serum noncholesterol sterols, and especially their ratios to cholesterol, can be used to evaluate the major features of cholesterol metabolism (ie, synthesis and absorption). Statin treatment reduces serum cholesterol precursors but increases serum plant sterols severalfold, especially in subjects with high-absorption marker sterol levels indicative of efficient cholesterol and sterol absorption in general. Statin therapy is most effective in subjects with high serum cholesterol precursor levels. In subjects with high-absorption sterol markers, dietary cholesterol absorption inhibition (eg, with plant stanol and sterol ester margarine) needs to be combined with a statin to achieve effective serum cholesterol reduction. However, whereas dietary plant stanol esters reduce statin-induced elevations of serum plant sterol levels, serum plant sterol levels remain elevated during dietary plant sterol ester consumption. The clinical implication of high serum plant sterol levels is under active investigation.

Alkenone temperature records and biomarker flux at the subtropical front on the chatham rise, SW Pacific Ocean

Alkenones and a suite of sterol biomarkers were examined in two sediment trap arrays deployed at 300 m depth in subtropical and subantarctic waters to the east of New Zealand from late winter to autumn in 1996–1997. The two traps were located within 200 km of one another and the main difference between the two sites are the differential physical, chemical, and biological characteristics of the different water masses in which they were situated. The alkenone-based reconstructions of water temperatures ( U 37 K ' ) were compared to the COADS monthly averaged satellite and real-time weekly temperatures for the deployment period. The records correlate well with seasonal sea surface temperatures (SST) for the 9 months of the deployment, with temperature reconstructions within 2 °C of regional monthly averages for most of the year. There are a few short periods of poorer agreement where alkenone-based reconstructions deviate by up to 4 °C in both traps. Weekly averages of satellite SST obtained during the time of the deployment indicate that these deviations were not associated with short-term changes in surface temperatures overlying the traps. These instances of poor correlation are not due to lateral advection of particles, but rather seem to reflect differences in environmental controls on alkenone-derived SSTs in the two water masses. Subantarctic traps showed deviations only to warmer than average temperatures. These occurred in early winter and late summer, during times of low lipid fluxes, suggesting that slow growth associated with light limitation may have affected unsaturation levels in the alkenones. The subtropical traps showed deviations only to cooler temperatures, which occurred in the late summer to early autumn. These biases occurred during times of highest lipid fluxes and lowest nutrients in the surface mixed-layer. Alkenone temperatures during maximum flux periods were too cool to be caused by subsurface production alone, suggesting that nutrient limitation effects may have significantly depressed alkenone unsaturation levels. Sterol biomarker associations indicate very different timing and trophic interactions between phytoplankton in the respective water masses. In subtropical waters, fluxes of dinosterol (a biomarker for dinoflagellates) peak first in the spring bloom while alkenones and diatom marker sterols respond synchronously during the main bloom event. In subantarctic waters sterol fluxes indicate a succession of phytoplankton export production in the spring bloom, whereas lipid fluxes normalized to organic flux indicate that alkenone producers were a relatively small proportion of the main spring bloom and proportionally more important when organic fluxes were lower in summer. We infer that the difference in nutrient concentrations between the water masses may drive these trophic differences and the inverse relationship between flux and the temperature response of U 37 K ' between the two sites.

Cholesterol metabolism in type 1 diabetes.

Little information is available on cholesterol absorption and synthesis in human type 1 diabetes. We studied these variables using serum cholesterol precursor sterol ratios to cholesterol as surrogate markers of cholesterol synthesis and those of cholestanol and plant sterols to reflect cholesterol absorption in seven type 1 diabetic subjects and in five age- and body weight-matched control subjects. Total and lipoprotein cholesterol levels were similar, but triglycerides in intermediate-density lipoprotein (IDL) and LDL were higher in type 1 diabetic than in control subjects. Most of the marker sterols were transported by LDL and HDL in both groups. The percentage of esterified cholesterol was lower in triglyceride-rich lipoproteins in diabetic patients than in control subjects. The ratios of the absorption marker sterols in serum were higher, and those of the synthesis markers were lower in type 1 diabetic than in control subjects. The increased cholestanol ratios were seen in all lipoproteins, and those of free and total plant sterols were mainly in LDL, whereas the decreased free and total synthesis markers were mainly in all lipoproteins. In conclusion, high absorption and low synthesis marker sterols seem to characterize human type 1 diabetes. These findings could be related to low expression of ABC G/5 G/8 genes, resulting in high absorption of cholesterol and sterols in general and low synthesis of cholesterol compared with type 2 diabetes.

Synthesis and absorption markers of cholesterol in serum and lipoproteins during a large dose of statin treatment.

Serum contains noncholesterol sterols, which are reliable markers of cholesterol metabolism, but their presence and importance in different lipoproteins have been insufficiently studied. Serum and lipoprotein cholesterol precursors squalene, cholestanol, desmosterol and lathosterol (markers of cholesterol synthesis) and cholestanol and plant sterols (markers of cholesterol absorption), and absorption efficacy and absolute synthesis of cholesterol were studied at baseline and during 6-month atorvastatin (80 mg day(-1)) treatment by the sterol balance technique in men with type 2 diabetes. At baseline, approximately 14% of serum squalene was transported by VLDL, 12% by IDL, 40% by LDL and 30% by HDL. The respective values for the noncholesterol sterols were approximately 8, 4, 61 and 26%. The squalene to cholesterol ratios were highest in VLDL and IDL, those of cholestanol, desmosterol and absorption marker sterols were gradually higher, and that of lathosterol lower from VLDL to HDL. Atorvastatin reduced LDL cholesterol by approximately 50%, decreased the absolute cholesterol synthesis and turnover by approximately 40%, but increased significantly the fractional and mass absorption of cholesterol. In accordance with the fecal data, the ratios of the precursor sterols to cholesterol were reduced (-50%), but those of squalene (+48%) and the absorption sterols increased (e.g. 2.6-fold for sitosterol) similarly in each lipoprotein, but progressively from VLDL to HDL. Effective lowering of LDL cholesterol by large dose of statin is associated with decreased synthesis and turnover of cholesterol and increased fractional and mass absorption of cholesterol. These changes are detectable by noncholesterol sterols in serum and in different lipoprotein fractions.

Ineffective decrease of serum cholesterol by simvastatin in a subgroup of hypercholesterolemic coronary patients

We measured serum cholesterol concentrations and synthesis markers (e.g. serum lathosterol to cholesterol ratio), and absorption markers (e.g. serum campesterol to cholesterol ratio) of cholesterol in 319 good responders (GR; dose 20 mg up to 1 year) and in 115 poor responders (PR; dose increased at 6 weeks to 40 mg) among Finnish participants in the Scandinavian Simvastatin Survival Study at baseline, 6 weeks and 1 year of the simvastatin treatment. The baseline cholesterol level and the ratios of the absorption markers were higher and those of the synthesis markers lower in PR than GR. The ratios of the precursor sterols were negatively related to the baseline cholesterol in GR only ( P=0.003). The cholesterol levels, and the ratios of the precursor sterols were decreased and those of the absorption marker sterols increased less consistently in PR than GR by 20 mg, the group differences being only slightly lessened by the dose addition to 40 mg. One-year differences were still frequently significant. The baseline cholesterol concentrations were negatively related to the reduction of the precursor sterol ratios in GR, the change of cholesterol being positively related to those of the synthesis markers and negatively to those of the absorption markers only in PR. Thus, patients needing large statin dose for cholesterol normalization have high absorption and low synthesis of cholesterol, yet baseline synthesis is inversely related to cholesterol level only in GR. The synthesis rate is less markedly reduced by the large than by the small statin dose in the PR, and the reduction is related, in contrast to that in the GR, to lowering of cholesterol.

Detection of olive oils authenticity by determination of their sterol content using LC/GC

The content of selected sterols as declared in the EU Commission Regulation was used to prove the authenticity of olive oils. A modified method using the preparative LC with silica gel packed column and gradient elution with three mixtures of hexane and diethyl ether was used to separate undesirable interfering compounds in the unsaponifiable fraction before the determination of sterols using GC. Model experiments based on the determination of D-7-stigmastenol and campesterol (addition of sunflower and soybean oils), or brassicasterol (addition of rapeseed oil) were used to verify that this method is capable of identifying adulteration of olive oils by additions of sunflower, soybean or rapeseed oils. An elevated content of these marker sterols, in comparison with their permitted contents, enables the identification of an addition of 5&amp;ndash;10% of the above oils to the olive oil. This method was also used to evaluate the authenticity of five samples of olive oils from the SIAL exhibition (Paris) and ten samples of virgin olive oils obtained on thePrague markets. It was revealed that none of the samples showed the signs of adulteration.