Sterol Compounds Research Articles

Inhibition of Virulence Associated Traits by β-Sitosterol Isolated from Hibiscus rosa-sinensis Flowers Against Candida albicans: Mechanistic Insight and Molecular Docking Studies.

The emerging drug resistance and lack of safer and more potent antifungal agents make Candida infections another hot topic in the healthcare system. At the same time, the potential of plant products in developing novel antifungal drugs is also in the limelight. Considering these facts, we have investigated the different extracts of the flowers of Hibiscus rosa-sinensis of the Malvaceae family for their antifungal efficacy against five different pathogenic Candida strains. Among the various extracts, the chloroform extract showed the maximum zone of inhibition (26.6 ± 0.5mm) against the Candida albicans strain. Furthermore, the chloroform fraction was isolated, and a sterol compound was identified as β-sitosterol. Mechanistic studies were conducted to understand the mechanism of action, and the results showed that β-sitosterol has significant antifungal activity and is capable of interrupting biofilm formation and acts by inhibiting ergosterol biosynthesis in Candida albicans cells. Microscopic and molecular docking studies confirmed these findings. Overall, the study validates the antifungal efficacy of Candida albicans due to the presence of β-sitosterol which can act as an effective constituent for antifungal drug development individually or in combination.

Hyperproduction of 7-dehydrocholesterol by rewiring the post-squalene module in lipid droplets of Saccharomyces cerevisiae

Lipid droplets (LDs) are specialized organelles that store neutral lipids to reduce the negative effects of lipotoxicity on cells. However, many neutral lipids are precursors for the synthesis of sterols and complex terpenoids, and this sequestration often greatly limits the efficient biosynthesis of sterols and complex terpenoids. In this study, taking 7-dehydrocholesterol (7-DHC) synthesis in Saccharomyces cerevisiae as an example, we revealed the blocking mechanism of LD sequestration on the efficient synthesis of metabolic products and found that LDs can sequester a significant amount of squalene, the precursor of 7-DHC, effectively preventing it from being directed toward the post-squalene pathway. Based on this, a post-squalene pathway was reconstructed on LDs, which resulted in a 28.7% increase in the 7-DHC titer, reaching 684.1 mg/L, whereas the squalene titer was reduced by approximately 97%. Subsequently, the triacylglycerol degradation pathway was weakened to release the storage space in LDs, and the esterification pathway was concurrently strengthened to guide 7-DHC storage within LDs, which further increased 7-DHC production, reaching 792.9 mg/L. Finally, by reducing the NADH/NAD+ ratio to alleviate the redox imbalance, the 7-DHC titer reached 867.6 mg/L in shake flask and 5.1 g/L in a 3-L bioreactor, which is the highest reported titer to date. In summary, this study provides new insights into the important role of LDs in sterol synthesis and offers a novel strategy for constructing cell factories for the efficient synthesis of sterol compounds.

UPLC-ESI-MS\MS Phytochemicals Profiling of n-Butanol Fraction of Xanthium strumarium Fruit Extract

The work aimed to explore the phytochemicals of various fractions of Xanthium strumarium fruit extract since this plant is found to be toxic for both humans and animals in Iraq. The extracts underwent phytochemical screening, which indicated the occurrence of flavonoids, alkaloids, phenols, coumarin, saponin, tannins, and terpenoids. After extraction, fractionation was done using hexane, chloroform, and N-butanol respectively, for identifying the phytochemicals found in each fraction, UPLC-electrospray ionization–tandem mass spectroscopy was used which revealed the presence of 6 sterol and terpenes compound in hexane fraction, Nine compounds were detected in the chloroform fraction, among which 2-acetyl-atractyligenin and Artemisinin were the primary components. Additionally, this particular plant has been found to contain these compounds for the first time. Furthermore, Pungiolide C, dihydroartemisinin, and atractylenolide II were also detected for the first time in this plant. For the n-Butanol fraction, eight diterpene glycosides were recognized, with Atractyloside and Carboxyatractyloside and their desulphated derivatives as the major compounds responsible for the plant's toxicity. The butanol fraction also showed the presence of eight phenolic compounds, among which caffeoylquinic acid derivatives and flavonoid Syringetin were the major compounds in this fraction.

Bio-efficacy of mustard seed extracts against Bihar hairy caterpillar Spilosoma obliqua (Erebidae: Lepidoptera) & assessment of mustard allelo-chemicals in response to mustard aphid Lipaphis erysimi (Aphididae: Hemiptera) infestation

Mustard seed contains glucosinolate-myrosinase system of substrate-enzyme metabolism, accompanied with compartmentalized cells. Upon rupture, enzyme myrosinase leads to breakdown of glucosinolates and its metabolized products like allyl, benzyl, phenyl type of isothiocyantes and many other types like nitriles are released. The myrosinase activity is tested in three globally utilized different seed coat color mustards viz., brown mustard (Brassica juncea), white mustard (Sinapis alba) and black mustard (Brassica nigra) to demonstrate the release of glucosinolates which are known to deter insect pests. Aqueous seed extracts of these mustard types were subjected to anti-feedant activity analysis and oil extracted from these mustard seeds was subjected to insecticidal activity analysis against Spilosoma obliqua 4th instars on castor leaves. Meanwhile, the mustard leaves in open field were allowed for natural aphid infestation during early months from January to March. Thereafter, the leaf twigs of white, black and brown mustard, infested and uninfested by aphids, were collected and myrosinase activity of plants was analyzed with varying levels of important pest inflicted biochemicals in plant, viz., Total phenols, ortho-dihydric phenols and flavonols. Gas Chromatography–Mass Spectrometry analysis of mustard oil used in insecticidal activity was performed to elucidate the role of biochemicals involved in action. These compounds along with fatty acids and sterols compounds are known to have anti-feedant & insecticidal activities. Aqueous extract components were also subjected to total glucosinolate analysis, spectrophotometrically for knowing their concentration, with highest in brown (85.81 µmol/g) followed by black (59.32 µmol/g) and lowest in white mustard (47.63 µmol/g). The amount of glucosinolates, free fatty acids & sterol compounds present in different coat color mustard seed types are being correlated here with their potential bioactivities against biological plant pests. Bio-control of potentially harmful insect pests is possible by utilizing anti-feeding & insecticidal potential of mustard seed extracts.

Soybean steroids improve crop abiotic stress tolerance and increase yield.

Sterols have long been associated with diverse fields, such as cancer treatment, drug development, and plant growth; however, their underlying mechanisms and functions remain enigmatic. Here, we unveil a critical role played by a GmNF-YC9-mediated CCAAT-box transcription complex in modulating the steroid metabolism pathway within soybeans. Specifically, this complex directly activates squalene monooxygenase (GmSQE1), which is a rate-limiting enzyme in steroid synthesis. Our findings demonstrate that overexpression of either GmNF-YC9 or GmSQE1 significantly enhances soybean stress tolerance, while the inhibition of SQE weakens this tolerance. Field experiments conducted over two seasons further reveal increased yields per plant in both GmNF-YC9 and GmSQE1 overexpressing plants under drought stress conditions. This enhanced stress tolerance is attributed to the reduction of abiotic stress-induced cell oxidative damage. Transcriptome and metabolome analyses shed light on the upregulation of multiple sterol compounds, including fucosterol and soyasaponin II, in GmNF-YC9 and GmSQE1 overexpressing soybean plants under stress conditions. Intriguingly, the application of soybean steroids, including fucosterol and soyasaponin II, significantly improves drought tolerance in soybean, wheat, foxtail millet, and maize. These findings underscore the pivotal role of soybean steroids in countering oxidative stress in plants and offer a new research strategy for enhancing crop stress tolerance and quality from gene regulation to chemical intervention.

Cytotoxic Activity of Cambodian Leaves Extract (Plumeria acuminate) on Breast Cancer Cells and COX-2 Targeted Prediction of Its Chemical Contents

Cambodian leaves are suspected to contain stigma sterol which may target Cyclo-Oxygenase-2 (COX-2) or Estrogen Receptor (ER) to contribute to its cytotoxic activity on breast cancer cells. This study aimed to determine the potential of Cambodian leaf compounds and extracts as chemopreventive agents for luminal breast cancer with a molecular target of COX-2. Ethanol was used to extract the active compound of Cambodian leaves. The study on chemical activity against COX-2 employed molecular docking with Molecular Operating Environment (MOE) and the cytotoxic property of Cambodian leaf extract (CLE) on T47D was determined using the trypan blue exclusion method. The extraction yielded as 4.87% w/w CLE. Thin layer chromatography showed that Cambodian leaves contain sterol. Molecular docking confirmed that several sterol compounds have greater affinity to COX-2 than native ligands indicating that they are potent as COX-2 inhibitors. They are Stigmast-7-en-3-ol, Lupeol Acetate, and Lupeol carboxylic acid with docking scores of -14.3874, -13.8098, and -14.1045 kcal/mol respectively. The CLE exhibited cytotoxic activity on T47D cells with an IC50 value of 18 μg/mL. Therefore, CLE has a potential effect as a chemopreventive agent for breast cancer and potentially as a COX-2 inhibitor.Keywords: Cambodian leaf extract, breast cancer, COX-2 inhibitor, chemopreventive.

A Brain-Protective Sterol from Soft Coral Inhibits Lipopolysaccharide-Induced Matrix Metalloproteinase-9-Mediated Astrocytic Migration.

Matrix metalloproteinases (MMPs), which are proteolytic enzymes, promote blood-brain barrier (BBB) disruption, leading to neuronal damage and neuroinflammation. Among them, MMP-9 upregulation serves as an inflammatory biomarker in the central nervous system (CNS). Currently, the development of marine organism-derived bioactive compounds or metabolites as anti-inflammatory drugs has received considerable attention. The 9,11-secosteroid, 3β,11-dihydroxy-9,11-secogorgost-5-en-9-one (4p3f), is a novel sterol compound extracted from the soft coral Sinularia leptoclado with potential anti-inflammatory activity. However, the effect of and potential for brain protection of 4p3f on brain astrocytes remain unclear. Herein, we used rat brain astrocytes (RBAs) to investigate the effects and signaling mechanisms of 4p3f on lipopolysaccharide (LPS)-induced MMP-9 expression via zymographic, quantitative reverse transcription-polymerase chain reaction (qRT-PCR), Western blot, immunofluorescence staining, promoter-reporter, and cell migration analyses. We first found that 4p3f blocked LPS-induced MMP-9 expression in RBAs. Next, we demonstrated that LPS induced MMP-9 expression via the activation of ERK1/2, p38 MAPK, and JNK1/2, which is linked to the STAT3-mediated NF-κB signaling pathway. Finally, 4p3f effectively inhibited LPS-induced upregulation of MMP-9-triggered RBA cell migration. These data suggest that a novel sterol from soft coral, 4p3f, may have anti-inflammatory and brain-protective effects by attenuating these signaling pathways of MMP-9-mediated events in brain astrocytes. Accordingly, the soft coral-derived sterol 4p3f may emerge as a potential candidate for drug development or as a natural compound with neuroprotective properties.

Physicochemical Properties and Bioactive Compounds of Three Varieties of Melon (Cucumis melo) Seed Oils

The physicochemical composition of the oil extracted from three different melon seed varieties (named Uzbek-Russian, Soski, and Soski Green), which were collected from three various geographical parts of Iran (north, northeast, and center), was determined in 2021 and 2022. The macronutrient components, fatty acid profile, sterol compounds, phenolic compounds, tocopherols, and oxidative stability of the melon seed oils were determined. The oil content of melon seeds ranged from 43.3% to 47.3%. Linoleic (54.48% to 60.35%), oleic (24.35% to 28.03%), and palmitic acids (10.15% to 12.65%) were the predominant fatty acids. No significant differences were observed between the three cultivars and between the 2 years of the study in terms of macronutrient components and fatty acid profiles (p > 0.05). Twelve different phenolic compounds were identified, of which caffeic acid was the major compound, ranging from 39.74% ± 1.70% (Soski Green) to 63.45% ± 1.25% (Uzbek-Russian). The amounts of caffeic acid and oxidative stability of Uzbek‐Russian were significantly higher than those in the two other cultivars. β‐Sitosterol (approximately 60%) and γ‐tocopherol (71.6 to 84.07 mg/100 g) were the major sterol and tocopherol compounds, respectively. The results indicated that the physical and chemical specifications of the three investigated melon seed oils were stable for two consecutive years. Therefore, melon seed oil is a rich source of bioactive compounds and could be introduced as a healthy oil for human consumption.

Comprehensive characterization of physicochemical, thermal, compositional, and sensory properties of cold-pressed rosehip seed oil

In this study, cold-pressed rosehip seed oil was fully characterized. Acidity and oxidation levels were near the limit values or slightly exceeded them and improvement in the storage conditions was suggested. The oil started to crystallize at -45.25 °C, and melt at -25.56 °C. Linoleic acid (51.1%), β-sitosterol (84.6%), γ-tocopherol (773.76 µg/g) and rosmarinic acid (31.38 µg/g) were determined as major fatty acid, sterol, tocopherol and phenolic compound, respectively. For the first time, aromatic volatile compounds and sensory descriptive terms were determined for cold-pressed rosehip seed oil. Sixty-seven volatile compounds were detected and L-limonene was found to be a major volatile compound. According to the sensory analysis, timber/kindling and raw vegetable tastes/aromas were found to be relatively dominant. Consequently, it is thought that rosehip seeds can be used as a raw material for edible and nutritionally-rich cold-pressed oil production and/or as source oil for functional food preparations.

Oxy- and Phytosterols as Biomarkers: Current Status and Future Perspectives.

Oxysterols and phytosterols are sterol compounds present at markedly low levels in tissues and serum of healthy individuals. A wealth of evidence suggests that they could be employed as biomarkers for human diseases or for cholesterol absorption.An increasing number of reports suggest circulating or tissue oxysterols as putative biomarkers for cardiovascular and neurodegenerative diseases or cancers. Thus far most of the studies have been carried out on small study populations. To achieve routine biomarker use, large prospective cohort studies are absolutely required. This, again, would necessitate thorough standardization of the oxysterol analytical methodology across the different laboratories, which now employ different technologies resulting in inconsistencies in the measured oxysterol levels. Routine use of oxysterol biomarkers would also necessitate the development of a new targeted analytical methodology suitable for high-throughput platforms.The most important use of phytosterols as biomarkers involves their use as markers for cholesterol absorption. For this to be achieved, (1) their quantitative analyses should be available in routine lipid laboratories, (2) it should be generally acknowledgment that the profile of cholesterol metabolism can reveal the risk of the development of atherosclerotic cardiovascular diseases (ASCVD), and (3) screening of the profile of cholesterol metabolism should be included in the ASCVD risk surveys. This should be done e.g. in families with a history of early onset or frequent ASCVD and in young adults aged 18-20years, to exclude the presence of high cholesterol absorption. Individuals in high cholesterol absorption families need preventive measures from young adulthood to inhibit the possibledevelopment and progression of atherosclerosis.

The Fungus Beauveria bassiana Alters Amounts of Sterols, Fatty Acids, and Hydroxycinnamic Acids in Potato Solanum tuberosum

The entomopathogenic endophytic fungus Beauveria bassiana can colonize plants resulting in growth promotion and protection against phytopathogenic microorganisms. However, physiological changes in potato plants (Solanum tuberosum) during this interaction are poorly understood. In the present work, gas chromatography–mass spectrometry and high-performance liquid chromatography were used to analyze sterol, fatty acid, and phenolic acid concentrations in potato plants inoculated with B. bassiana conidia in soil. We showed an increase in amounts of stigmasterol, minor sterol compounds, and some hydroxy fatty acids in leaves after the fungal treatment. Moreover, levels of hydroxycinnamic acids, especially chlorogenic acid, were elevated in roots following the B. bassiana inoculation. We propose that these changes could have been caused by oxidative reactions, and the alterations may have resulted in growth-stimulatory and protective effects of B. bassiana on the plants.

Molecular dynamics study of stigmasterol and beta-sitosterol of Morinda citrifolia L. towards α-amylase and α-glucosidase

Previous in vivo studies of Morinda citrifolia (Rubiaceae) reported that the extract inhibited α-amylase and reduced blood glucose levels in streptozotocin-induced diabetes mice. Moreover, molecular docking studies confirmed that ursolic acid and sterol compounds contained in the fruit interacted with important residues in the binding site of α-amylase and α-glucosidase. Our work aimed to study the complex stability of stigmasterol (which has been isolated from the M. citrifolia fruit for the first time) and beta-sitosterol towards α-amylase and α-glucosidase by employing molecular dynamics simulation on GROMACS 2016.3 embedded with the AMBER99SB-ILDN force field. The simulation was carried out for 100 ns at 310 oK. Based on the RMSD and RMSF graphs, the complexes of stigmasterol/α-amylase and stigmasterol/α-glucosidase are more stable compared to acarbose, the known inhibitor of both enzymes. Moreover, beta-sitosterol indicates a better stability complex with α-glucosidase compared to that of acarbose. Interestingly, the affinity of stigmasterol and beta-sitosterol to both enzymes, in terms of the total binding energy, is stronger than that of acarbose. Taken together, stigmasterol and beta-sitosterol in M. citrifolia fruit may have the potency to be developed as α-amylase and α-glucosidase inhibitors. Communicated by Ramaswamy H. Sarma

Calamus caesius (Rattan) wood: Chemical constituents, biological activities’ relative medicinal properties from Thai medicinal scriptures, and in silico antioxidant activity

Calamus caesius is known as Rattan. It was found as a component of many formulas from evidence-based Thai medicinal scriptures but no research about their medicinal properties. We investigated the literature review analysis from Thai medicinal textbooks for proposed biological activity relatives including antioxidant, cytotoxic, anti-inflammatory, and antimicrobial activities, and chemical profiles using gas chromatography-mass spectrometry (GC–MS). In silico studies were inspected on tyrosinase and NAD(P)H oxidase actions. Thirty formulas from Thai medicinal textbooks found C. caesius as a component with a percent ratio in the range of 1.43 to 14.99, the formula's properties are antipyretic, followed by antidiarrhea, and cure abscesses related to inflammation and infection. Both water extracted and ethanol extracted showed high antioxidant activities in all assays and showed no toxicity in macrophage-like cells and cancer cell lines. The ethanol extracted showed slightly bactericidal better than the water extracted, none of them inhibited against C. albicans. From GC–MS analysis, the highest components of water and ethanol extract are 3-tert-Butylamino-acrylonitrile and β-Sitosterol, respectively. Five chemical compounds revealed in both water and ethanol extracted of C. caesius are 1,3-di-tert-butylbenzene; 2,6-dimethoxyphenol; 2-propylphenol; 2,4-di-tert-butylphenol; methyl palmitate. Sterol compounds such as stigmasterol, beta-sitosterol, and campesterol from ethanol extracted showed outstanding interaction with both tyrosinase and NADPH oxidase in silico molecular docking study. All outcomes proven that C. caesius has potentially antioxidant effects to support health problems. Additionally, this is the first report on the scientific data of Calamus caesius wood related to its medicinal properties in the formula from Thai medicinal scriptures.

Inhibition of the mevalonate pathway causes accumulation of CD14 in the endoplasmic reticulum of macrophages

Abstract Mevalonate kinase deficiency (MKD) is an autoinflammatory disease caused by mutations in the mevalonate kinase enzyme, which acts early in the mevalonate pathway to produce sterol and non-sterol isoprenoid compounds. Patients with MKD have episodes of inflammatory symptoms accompanied by increased plasma levels of pro-inflammatory cytokines. CD14 is a receptor that binds to lipopolysaccharide (LPS) and induces the release of cytokines. Isoprenoid depletion was previously shown to increase cellular and decrease extracellular levels of CD14 in macrophages after LPS stimulation. The objective of this study is to characterize the cellular localization of CD14 following isoprenoid depletion. RAW 264.7 macrophages were treated with lovastatin to deplete isoprenoids and CD14 localization was examined using immunofluorescence and western blotting after LPS stimulation. CD14 co-localized with the endoplasmic reticulum (ER) protein calnexin following lovastatin treatment. To quantitate ER CD14, cell lysates were digested with endoglycosidase (Endo) H, which removes high mannose glycans found on ER proteins, but not complex glycans added in the Golgi apparatus. Western blot results showed that the level of Endo H sensitive CD14 was higher in lovastatin as compared to carrier-treated cells. Co-incubation of lovastatin-treated cells with mevalonate or geranylgeranyl pyrophosphate (GGPP), but not cholesterol, prevented the ER accumulation of CD14 indicating that GGPP depletion may cause this effect. Understanding how mevalonate pathway products contribute to protein trafficking in macrophages will expand our understanding of inflammatory diseases.

Skrining Fitokimia Hasil Ekstraksi Bertingkat Daun Jengkol (Archidendron jiringa) dan Aplikasinya Sebagai Zat Antijamur

Phytochemical screening and antifungal activity test of jengkol leaf (Archidendron jiringa) extract have been carried out. The purpose of this research is determine secondary metabolite content and antifungal activities of jengkol leaf extract. Extraction of jengkol leaf was conducted with sequential extraction technique with various solvent such as hexane, ethyl acetate and methanol respectively. FTIR characterization of jengkol leaf extract show the presence of O-H, C-H C-O and C=O groups. Based on phytochemical sceening test, jengkol leaf extract contains phenolic, flavonoid, tannin, sterol, and alkaloid compound. Antifungal test was investigated on Candida albicans by disc diffusion methods. Jengkol leaf extract that was obtained from methanol extraction show inhibiton zone to Candida albicans of (1,42 ± 0,16) mm, while extraction with ethyl acetate and hexane aren’t show inhibition zone. Antifungal test result showed that jengkol leaf extract has potential as antifungal agent

Comparative analysis of the composition of sterols in embryogenic and nonembryogenic cell lines of <i>Larix sibirica</i> Ledeb.

The cell lines of Siberian larch (Larix sibirica Ledeb.), possessing different embryogenic potential, were studied. Qualitative and quantitative content of sterol components was comparatively analyzed in their tissues with the help of gas chromatography-mass spectrometry. Significant differences in these parameters were found between both groups of the lines in fractions of free sterols and their esters. In addition to sterol compounds, squalen, which is a triterpenic intermediate in sterol biosynthesis, was revealed. Of free sterols, -sitosterol, campesterol, isofucosterol, and stigmasterol dominated in the embryogenic lines, while -sitosterol, campesterol, and stigmasterol in nonembryogenic ones. The campesterol concentration was 1.31.9 times higher in the embryogenic than nonembryogenic lines. Since campesterol is a precursor of brassinosteroids, it is likely that its level is high due to an embryogenic state of these cell lines. Only the embryogenic lines contained appreciable amounts of isofucosterol. The fraction of sterol esters exhibited wider component diversity in the nonembryogenic than the embryogenic lines. In all the cell lines, among the identified sterol esters, the substances without double bonds, bearing, as a structural fragment, a sterane coreperhydrocyclopenta[a]phenanthrenewere the most abundant. Their content ranged from 52 to 71% of the total amount of sterol esters. The revealed differences in the compositions of sterols and their esters, between the L. sibirica cell lines with different embryogenic potential, witness to the considerable rearrangements in sterol metabolism in the course of embryogenesis. The line-specific differences may indicate the involvement of these metabolites in formation of embryos.

Chemical composition of endemic and endangered Gypsophila pilulifera produced by in vitro micropropagation method

This article contains the result of the first study on the chemical composition of six methanol extracts of different plant parts of the Gypsophila pilulifera, which were grown in vitro and transferred to field conditions according to their distribution location. The methanol extracts of the leaves, branches, and roots of the G. pilulifera were obtained using the maceration method. All methanol extracts were derivatized with trimethylsilyl (-TMS) for gas chromatography-mass spectrometry (GC-MS) analysis. Fatty acid derivatives were the dominant group in all the extracts. Palmitic acid, (Z,Z)− 9,12-octadecadienoic acid, and stearic acid were common fatty acids in all extracts. Sterols were detected in all methanol extracts. The predominant sterol was (3β,22E)-Ergosta-7,22-dien-3-ol in the analyzed extracts. The determination of sterol compounds contributes to greater knowledge about the G. pilulifera and brings more attention to this plant species. G. pilulifera is an endemic and critically endangered (CR) species. The plant has potential use in different areas, especially in the pharmaceutical industry. In the current study, the chemical content of the underground and aboveground parts of G. pilulifera produced under in vitro conditions from the seeds collected from two different locations in nature was investigated comparatively according to their locations for the first time.

The Effect of Extraction Methods on Phytochemicals and Biological Activities of Green Coffee Beans Extracts.

The objectives of the present study consisted of identifying the impact of extraction methods and parameters held over the phytochemistry and biological activities of green coffee beans. Extraction processes belonging to two categories were performed: classical methods-maceration, Soxhlet extraction, and such innovative methods as turboextraction, ultrasound-assisted extraction, and a combination of the latter two. Total polyphenolic and flavonoid content, as well as in vitro antioxidant activity of the resulted extracts were spectrophotometrically determined. Extracts displaying the highest yields of bioactive compounds were subjected to High Performance Liquid Chromatography-Mass Spectrometry analysis. The extracts with the best phytochemical profiles were selected for biological activity assessment. In vivo, a model of plantar inflammation in Wistar rats was used to determine antioxidant activity, by evaluating the oxidative stress reduction potential, and anti-inflammatory activity. In vitro antimicrobial activity was also determined. The Soxhlet extraction and ultrasound-assisted extraction gave the highest bioactive compound yields. The highest total polyphenolic content was 2.691 mg/mL gallic acid equivalents and total flavonoid content was 0.487 mM quercetin equivalents for the Soxhlet extract subjected to 60 min extraction time. Regarding the antioxidant activity, ultrasound-assisted extraction reached the highest levels, i.e., 9.160 mg/mL Trolox equivalents in the DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) assay and 26.676 mM Trolox equivalents in the FRAP (Ferric Reducing Antioxidant Power) assay, at a 30 min extraction time and 50 °C extraction temperature. The 60 min Soxhlet extract reached the highest level for the ABTS+ (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) assay, 16.136 mM Trolox equivalents, respectively. Chlorogenic acid was present in the highest concentration in the same Soxhlet extract, 1657.179 µg/mL extract, respectively. Sterolic compounds were found in high concentrations throughout all the analyzed extracts. A proportional increase between yields and extraction parameter values was observed. Increased inhibition of Gram-negative bacteria was observed. The finally selected Soxhlet extract, that of 60 min extraction time, presented a significant in vivo antioxidant activity, with a slight anti-inflammatory activity. Antioxidant levels were elevated after 2 h of extract administration. Pro-inflammatory cytokine secretion was not influenced by the administration of the extract.

Effect of extraction process on quality of oil from Asphodelus tenuifolius seeds

In this study, Asphodelus tenuifolius (ATO) seeds oils were extracted by three different extraction methods, namely Soxhlet extraction (SE), microwave-assisted extraction (MAE) and ultrasonic-assisted extraction (UAE). The physicochemical properties, fatty acids, sterols, tocols, total phenol content and antioxidant activity of ATO seed were examined and compared. The best extraction yield was from SE (21.97 ± 0.01%), followed by MAE (19.28 ± 0.13%) while that of UAE was the lowest (16.5 ± 0.1%). Analysis of the chemical composition of these oils showed that linoleic acid and β-sitosterol were the main fatty acid and sterol compounds, respectively. A large amount of fatty acids and sterols were not significantly affected by the three extraction methods. Nevertheless, the tocols, the total phenols contents and the antioxidant power showed a variation between these three extracted oils. Based on these results, it was found that this specie has great potential for industrial use.

Human cytosolic steroid sulfotransferases: Versatile and rapid activity assays.

Conjugation of steroids and sterol compounds with a sulfonate group is a major pathway in the regulation of their activity, synthesis and excretion. Three human cytosolic sulfotransferases are highly involved in the sulfonation of sterol compounds. SULT1E1 has a low nM affinity for estrogen sulfonation and also conjugates non-aromatic steroids with a significantly lower affinity. SULT2A1 is responsible for the high levels of fetal and adult dehydroepiandrosterone (DHEA) sulfate synthesis in the adrenal gland as well as many 3α and 3ß-hydroxysteroids and bile acids. SULT2B1b is responsible for the majority of cholesterol sulfation in tissues as well as conjugating 3ß-hydroxysteroids. Although there are multiple methods for assaying cytosolic SULT activity, two relatively simple, rapid and versatile assays for steroid sulfonation are described. The first method utilizes radiolabeled substrates and organic solvent extraction to isolate the radiolabeled product from the aqueous phase. The second assay utilizes 35S-3'-phosphoadenosine 5'-phosphosulfate (PAPS) to generate 35S-conjugated products that are resolved by thin layer chromatography. Both assays useful in situations requiring measurement of SULT activity in a timely manner.