Extraction Of Sterols Research Articles

Development of Novel Cost Efficient Process for Extraction of Sterols and Tocopherols from Wild Plant Seed

The present work focuses on the compositional characterization of tamarind and sea buckthorn seed oils, which have a wide range of uses in the cosmetic, pharmaceutical and food sectors. The characterization includes the analysis of tocopherols, sterols and fatty acid profile in both oils. The tamarind and sea buckthorn oils were converted to fatty acid methyl ester and the fatty acid composition was analyzed by using gas chromatography technique. The quantitative and qualitative analysis of tocopherols performed using high performance liquid chromatography. This study reveals that the oil samples consist of a variety of saturated and unsaturated fatty acids, such as myristic acid (0 and 0.4%), palmitic acid (11 and 30.5%), palmitoleic (0 and 39.6%) stearic acid (0 and 0.5%), oleic acid (18.3 and 26.1%), linoleic acid (45.8 and 2.1%), linolenic acid (4.1% and 0.5%), arachidic acid (1.8% and 0%) behenic acid (9.6 and 0%) and lignoceric acid (9.4 and 0%). Additionally, the analysis detects various tocopherol components, including α-tocopherol (21.4 and 32.6%), β-tocopherol (14.49 and 7.24%), γ-tocopherol (53.72 and 45.66%), δ-tocopherol (9.64 and 9.11%) and α-tocotrienol (0 and 5.13%) in tamarind and sea buckthorn oil, respectively.

Biochemical and histological study of the effect of sterol extract from pistachio on the level of antioxidants in the liver tissue green

The effect of sterol extract of pistachio on the levels of anti- oxidant was carried out in liver tissue of adult male rabbit, twenty eight of local rabbite, weigh range(800-1500)g. The animals was divided randomly into 4 groups (7 rabbits in each group). three groups G1,G2 and G3 in was treated orally with (50,100,150) mg/kg of sterol extract of pistachio respectively for a period of 30 days, after the end of the experiment starved rabbits for 12 hours and then seduced by the use of ether after anesthesia animal transfer to a bowl anatomy as proven front and rear sides of the animal Baldbabis and then worked an opening in the the abdomen to the sternum was extracted liver tissue as it was saved in the solution Alveslchi was estimated antioxidants in liver tissue, including the glutathione(GSH) and malon dialdehyde (MDA). The results showed that the glutathion level significantly increased in liver tissue in all groups compared with control, while the MDA level significantly increased in liver tissue in G2,G3 compared with control, While the results of microscopic histological study showed that the treatment groups sterol extract raw green pistachios be free from the distortions in addition to the lack of a change in the nature of the liver tissue.

Analysis of Cholesterol Lipids Using Gas Chromatography Mass Spectrometry.

An optimized Bligh and Dyer protocol and subsequent derivatization is described in this chapter for the extraction of free cholesterol and cholesterol esters from tissue samples. Quantification analysis of lipid species is then described utilizing gas chromatography-mass spectrometry, the ideal method for analysis of volatile organic compounds and extraction of sterols.

Sterols Content of Fruiting Bodies of Medicinal Artist's Bracket Mushroom Ganoderma applanatum (Agaricomycetes) Collected in Armenia.

The qualitative analysis of hexane extracts obtained from different trama layers (WT, T1-T4) of dried fruiting bodies of medicinal bracket fungus Ganoderma applanatum collected in the Tavoush region of North-East Armenia was performed by GC-MS analysis. Three sterols [(7.22-ergostadienon, ergosterol and ergosta-14.22-diene-3-ol (3β, 5α, 22E)] have been identified. The results have shown that the content and ratio of sterols differ in analyzed trama samples. The highest amount of sterols was detected in middle parts of T2 and T3 layers, while content of sterols gradually decreased to the upper cortical (T4) and lower hymenial (T1) layers. The chromatographic profiles of identified compounds indicate that different sterols dominated in each layer: 7.22-ergostadienon in T4, ergosterol in T3, T2, and T1. The average weight loss of analyzed trama samples during six days of drying was about 40 wt.% (37.0-43.49 wt.%) of the total weight of basidiome, which decreased up to 5 wt.% in the next two days. The complete extraction of sterols lasted six days. Its further prolongation leads to stationary phase without an increase in the amount of extracted sterols.

Ginsenoside-Rg1 synergized with voluntary running exercise protects against glial activation and dysregulation of neuronal plasticity in depression.

Depression is a common psychological disease accompanied by mental disorders and somatic symptoms. However, the underlying mechanisms regarding the pathogenesis of depression are still not clear. Neuronal damage resulting from inflammation is considered to be one of the important risk factors for depression. Ginsenoside-Rg1, a sterol extract extracted from ginseng herbs, has been shown to have neuroprotective effects against neurodegenerative diseases. Moreover, running exercise, a simple behavioral therapy, has been recently shown to have antidepressant effects. However, whether these two synergized strategies are more efficient in depression treatment, especially the neural mechanisms underlying this practical and interesting treatment is unknown. In this study, we have shown that ginsenoside-Rg1 synergized with voluntary running exercise exerts more efficiency on suppressing neuroinflammation, up-regulating expression of neurotrophic factors, and synaptic-related proteins, ameliorating neuronal structural damages than that of ginsenoside-Rg1 or voluntary exercise alone, suggesting its better neuroprotective effects. More importantly, the antidepressant-like effect of this synergistic treatment was also significantly better than either of these two treatments. These results suggest that ginsenoside-Rg1, synergized with voluntary running, may have higher efficacy in the treatment of depression through anti-inflammation and the improvement of neuroplasticity. These findings may provide a new perspective for the development of a therapeutic strategy for depression.

Biogas digestate as a sustainable phytosterol source for biotechnological cascade valorization.

Every year, several million tonnes of anaerobic digestate are produced worldwide as a by-product of the biogas industry, most of which is applied as agricultural fertilizer. However, in the context of a circular bioeconomy, more sustainable uses of residual digestate biomass would be desirable. This study investigates the fate of the sterol lipids β-sitosterol and cholesterol from the feedstocks to the final digestates of three agricultural and one biowaste biogas plants to assess if sterols are degraded during anaerobic digestion or if they remain in the digestate, which could provide a novel opportunity for digestate cascade valorization. Gas chromatographic analyses showed that feedstock sterols were not degraded during anaerobic digestion, resulting in their accumulation in the digestates to up to 0.15% of the dry weight. The highest concentrations of around 1440 mg β-sitosterol and 185 mg cholesterol per kg dry weight were found in liquid digestate fractions, suggesting partial sterol solubilization. Methanogenic batch cultures spiked with β-sitosterol, cholesterol, testosterone and β-oestradiol confirmed that steroids persist during anaerobic digestion. Mycobacterium neoaurum was able to transform digestate sterols quantitatively into androstadienedione, a platform chemical for steroid hormones, without prior sterol extraction or purification. These results suggest that digestate from agricultural and municipal biowaste is an untapped resource for natural sterols for biotechnological applications, providing a new strategy for digestate cascade valorization beyond land application.

P063 Research on antifungal activity of a new azoles compound, YZJT-1903

Poster session 1, September 21, 2022, 12:30 PM - 1:30 PM ObjectivesFluconazole is widely used to treat invasive fungal infections, but the fluconazole resistance of pathogenic fungi undermines its antifungal efficacy. Although voriconazole has a good anti-Aspergillus fumigatus activity, it is not a first-line drug for treating infections caused by Candida and Cryptococcus species. To meet the demand for new azoles, in our previous studies, we identified a new azoles compound, YZJT-1903 (Fig. 1e), and found that YZJT-1903 has a longer in vivo half-life (˃10 days in rat) and a wider therapeutic window than voriconazole, indicating that YZJT-1903 is a promising azoles compound to treat invasive fungal infections. In this study, we further investigated antifungal activity of YZJT-1903 compared with fluconazole and voriconazole.MethodsWe investigated the antifungal spectrum of YZJT-1903 and minimum inhibitory concentration (MIC) and accumulative antifungal percentage of YZJT-1903 against various pathogenic fungi using microbroth dilution method. We quantitatively analyzed the inhibitory effect of YZJT-1903 on fungal growth by the Time-Growth Curve assay. Mutation Preventative Concentration (MPC) was carried out to verify the antifungal potential of YZJT-1903. Sterols of plasma membrane were extracted and the composition was analyzed through Gas Chromatograph-Mass Spectrometer (GC-MS) to verify the classical action mechanism of azoles.ResultsOur MIC assay results demonstrated that YZJT-1903 has a broad antifungal spectrum and can significantly inhibit the growth of A. fumigatus, Cryptococcus neoformans, Cryptococcus gatti, and Candida species (such as Candida albicans, C. tropicalis, C. parapsilosis, C. glabrata, C. guilliermondii, and C. auris) (Fig. 1a). We found that the antifungal activity of YZJT-1903 is equivalent to that of voriconazole, but superior to that of fluconazole (Fig. 1a). Accumulative antifungal percentage can directly reflect this relationship quantitatively among YZJT-1903, fluconazole, and voriconazole due to the curve of YZJT-1903 has no significance with voriconazole (Fig. 1b). YZJT-1903 (0.25 μg/ml) has similar inhibitory effect to voriconazole (0.25 μg/ml) on the growth of C. albicans and C. neoformans, but has the best inhibitory effect on C. auris [compared with voriconazole (0.25 μg/ml), and fluconazole (0.25 μg/ml)] (Fig. 1c). MPC of YZJT-1903 in C. neoformans is as low as 0.25 μg/ml which means YZJT-1903 at low concentration can already prevent the drug-resistant mutation of strains, therefore, avoid the occurrence of resistance in Cryptococcus spp. (Fig. 1d). Extraction of sterols showed that the fungal cell treated with YZJT-1903 at 0.063 μg/ml have completely no detectable ergosterol which is similar to the group of fluconazole at 4 μg/ml, and the group of voriconazole at 0.031 μg/ml (Fig. 2). This result gave a hint that maybe the property of YZJT-1903 is more similar to voriconazole which may matter with the structure of these two compounds. We found that feeding exogenous 100 μm ergosterol counteracted the antifungal activity of YZJT-1903, indicating that YZJT-1903 has an antifungal activity by inhibiting the synthesis of ergosterol.ConclusionOur study demonstrates that YZJT-1903 has a potent antifungal activity, which will contribute to addressing devastating global invasive fungal infections caused by Candida and Cryptococcus species.

Sterol Extraction from Isolated Plant Plasma Membrane Vesicles Affects H+-ATPase Activity and H+-Transport.

Plasma membrane H+-ATPase is known to be detected in detergent-resistant sterol-enriched fractions, also called “raft” domains. Studies on H+-ATPase reconstituted in artificial or native membrane vesicles have shown both sterol-mediated stimulations and inhibitions of its activity. Here, using sealed isolated plasma membrane vesicles, we investigated the effects of sterol depletion in the presence of methyl-β-cyclodextrin (MβCD) on H+-ATPase activity. The rate of ATP-dependent ∆µH+ generation and the kinetic parameters of ATP hydrolysis were evaluated. We show that the relative sterols content in membrane vesicles decreased gradually after treatment with MβCD and reached approximately 40% of their initial level in 30 mM probe solution. However, changes in the hydrolytic and H+-transport activities of the enzyme were nonlinear. The extraction of up to 20% of the initial sterols was accompanied by strong stimulation of ATP-dependent H+-transport in comparison with the hydrolytic activity of enzymes. Further sterol depletion led to a significant inhibition of active proton transport with an increase in passive H+-leakage. The solubilization of control and sterol-depleted vesicles in the presence of dodecyl maltoside negated the differences in the kinetics parameters of ATP hydrolysis, and all samples demonstrated maximal hydrolytic activities. The mechanisms behind the sensitivity of ATP-dependent H+-transport to sterols in the lipid environment of plasma membrane H+-ATPase are discussed.

Kinetics characteristics and thermodynamics analysis of soybean pods sterols extraction process

In order to make full use of biomass resources and extract sterols from the waste soybean pods, the mass transfer characteristic of the extraction process with ethanol aqueous solution was explored. The relationships between sterols mass concentration in extracting solution and extraction temperature and extraction time were studied under appropriate ethanol concentration and liquid-solid ratio. The kinetics model fitting, the mass transfer characteristic parameters research and thermodynamic analysis of extraction process were carried out. The result showed that the extraction process could be described by a second order kinetic model. Within the research scope, with the increase of extraction temperature, the mass transfer rate constant increased from 0.286 9 mL min−1 mg−1 to 0.848 5 mL min−1 mg−1, the half life was reduced from 20.9 min to 6.5 min, the activation energy of extraction process was 27.73 kJ mol−1. The Gibbs free energies were all less than zero, indicating that the sterols extraction process was spontaneous. The enthalpy change was 33.18 kJ mol−1 and entropy change was 122.2 J mol−1K−1 in the extraction process, the extraction process was an endothermic and entropy increases process. The research results could provide reference for the large-scale extraction of sterols from soybean pods.

Optimization of extraction technology of sterols from discarded soybean pod by response surface methodology

The ethanol solution extraction of sterols from discarded soybean pod was studied. The effects of factors on sterols yield was investigated by single factor experiment. On this basis, liquid to material ratio, ethanol concentration, extraction time and extraction temperature were selected, response surface method was used to optimize the extraction process of sterols. The model between extraction conditions and yield was established and analyzed for obtaining the optimal technological parameters and improve the sterols yield. The results showed that the sterols yield was significantly affected by the above four selected factors. The model obtained was significant. The optimum technological conditions of the ethanol extraction of soybean pod sterols were the liquid to material ratio of 26.8 mL g-1, ethanol concentration of 72.4%, extracting time of 1.86 h, extraction temperature of 68.4 °C. The theoretical maximum sterols yield was 7.82 mg g-1, the actual sterols yield was 7.74 mg g-1.

Sterols in red macroalgae from antarctica: extraction and quantification by Gas Chromatography–Mass spectrometry

Antarctica is considered a major reservoir of microorganisms and macroorganisms, including seaweed, which can biosynthesize secondary metabolites with unique structures and specific biological activities. Macroalgae are considered an interesting source of steroids, a class of metabolites involved in the control of membrane fluidity and permeability. This study reports the extraction and quantification of sterols in four red macroalgal species from Antarctica: Palmaria decipiens, Plocamium cartilagineum, Iridaea cordata, and Pyropia endiviifolia. The extraction of sterols involved alkaline hydrolysis from algal biomass. The quantification of cholesterol, brassicasterol, campesterol, stigmasterol, β-sitosterol, and fucosterol were performed by gas chromatography hyphenated with mass spectrometry (GC–MS). In general, cholesterol was the most abundant compound (216.32 to 3195.69 mg kg−1), followed by β-sitosterol (131.42 to 654.18 mg kg−1), stigmasterol (68.35 to 393.20 mg kg−1), brassicasterol (68.56 to 337.43 mg kg−1), campesterol (64.95 to 277.78 mg kg−1), and fucosterol (57.67 to 286.18 mg kg−1). Brassicasterol was not found in I. cordata, but it was identified in the other three samples. This work enriches the knowledge about the profile of steroids biosynthesized by Antarctic macroalgae and supports research to discover bioactive compounds with nutraceutical and pharmaceutical applications.

Response surface methodology (RSM) to evaluate both the extraction of triterpenes and sterols from jackfruit seed with supercritical CO2 and the biological activity of the extracts.

Jackfruit seeds are an underestimate residue having important biological activity such as anti-inflammatory, cytotoxicity and antimicrobial effects. However few researches have been done for this material using alternative extraction technologies, so this study aimed to evaluate the extraction of triterpenes and sterols from jackfruit seed by applying high- and low-pressure techniques. Response surface methodology (RSM) was used to determine the best conditions of pressure, temperature and CO2 flow rate for extraction with supercritical CO2. The yield and profile of these compounds were compared with the low pressure technique, which was considered as a reference. In vitro biological tests of anti-inflammatory activity and cytotoxicity in L929 and RAW 264.7 cells were also performed. The best extraction conditions in SFE for sterols were 40°C/20MPa/4mLmin-1 (0.832 ± 0.007 mgSR g-1 sample) and 40°C/20MPa/3 mLmin-1 (0.800 ± 0.009 mgSR g-1 sample), for triterpenes were 50°C/12MPa/4mLmin-1 (1.501 ± 0.004 mgTT g-1 sample) and 45°C/9.3MPa/3.5mLmin-1 (1.485 ± 0.004 mgTT g-1 sample). No cytotoxic activity was detected in L929 cells in the extracts obtained from ethanol up to concentration of 100μgmL-1 of extract. The Pearson's coefficient indicated that the reduction in cell viability was related to the concentration of triterpenes. Anti-inflammatory assays showed that some extracts could inhibit the inflammatory action induced in RAW 264.7 cells at concentration of 30μgmL-1 of extract. Our results justify the further exploration of these characteristics to obtain natural products for the pharmaceutical and food industries.

Ginsenoside-Rg1 Rescues Stress-Induced Depression-Like Behaviors via Suppression of Oxidative Stress and Neural Inflammation in Rats.

Depression is an inflammatory-related condition, with the progression in neuronal damage resulting in major depression disorder. Ginsenoside-Rg1, a sterol extract from the herb Panax ginseng, has been shown to exert neuroprotective effects upon neurodegeneration disorders. However, whether ginsenoside-Rg1 confers antidepressant-like effects on neuroinflammation as associated with depression, as well as the possible mechanism involved in these neuroprotective effects, is currently unclear. In the present report, we show that treatment with ginsenoside-Rg1 (40 mg/kg, i.p.) significantly ameliorated depressive-like behaviors as induced by chronic unpredictable mild stress (CUMS) in a rat model of depression. Moreover, these CUMS rats treated with ginsenoside-Rg1 showed reductions in the levels of the oxidative stress products and the activity in the antioxidant stress kinase. Furthermore, CUMS rats treated with ginsenoside-Rg1 showed ameliorated neuroinflammation and associated neuronal apoptosis along with a reduction in dendritic spine atrophy and display of depressive behaviors. Taken together, the results of this study suggest that ginsenoside-Rg1 produces antidepressant-like effects in CUMS-exposed rats; and one of the mechanisms for these antidepressant-like effects of ginsenoside-Rg1 appears to involve protection against oxidative stress and thus the neuronal deterioration resulting from inflammatory responses. These findings provide evidence for the therapeutic potential of ginsenoside-Rg1 in the treatment of stress-related depression.

Theoretical and experimental assessment of supercritical CO2 in the extraction of phytosterols from rapeseed oil deodorizer distillates

Phytosterols, a group of naturally occurring compounds found in plant cell membranes, are structurally similar to the body's cholesterol and compete with that for absorption in the digestive system leads to the reduction of its blood level. This work is focused on the applying supercritical fluid extraction to isolate phytosterols from rapeseed oil deodorizer distillate (RODD). A thermodynamic model, the Peng Robinson equation of state model, was applied to show phase equilibrium behavior of the multicomponent oil-type material CO2 mixture and optimize the supercritical extraction process parameters. The extraction of sterols in the saponified RODD sample at 35 MPa, 313 K, and solvent to feed mass ratio (S/F) of 5, showed a recovery of 76 wt% with a purity of 60 wt%. Furthermore, the use of ethanol (5 wt%) as co-solvent, increased the phytosterol efficiency and purity. By coupling experimental studies with thermodynamic modeling, supercritical extraction processes could be analyzed more efficiently and rapidly.

Sepiolite Clay Attenuates the Development of Hypercholesterolemia and Obesity in Mice Fed a High-Fat High-Cholesterol Diet.

Obesity reduces the quality of life and life expectancy, whereas nonoperative interventions have shown poor results so far. Statins effectively combat hypercholesterolemia but are not well tolerated at high doses, raising the need for coprescription with cholesterol sorbents and/or absorption inhibitors. Montmorillonite (MMT) clay was found to attenuate hypercholesterolemia and obesity by reducing cholesterol and fat absorption. However, acicular clay-like sepiolite may offer better results due to its more substantial adsorption of nonpolar molecules. We herein aimed at (1) assessing in vitro the capacity of sepiolite to adsorb edible oil and cholesterol compared with that of MMT and (2) assessing in vivo the effect of continuous feeding on a high-fat high-cholesterol diet (HFD) (53.6% w/w fat and 0.2% cholesterol) supplemented with 5% (w/w) edible sepiolite, on diet-induced obesity rate, hypercholesterolemia, and hyperlipidemia. Fourier transform infrared spectroscopy showed in vitro that sepiolite adsorption of olive oil and cholesterol was five to eight times greater than that of MMT clay. Sepiolite supplementation to HFD fed to mature mice for 12.5 weeks resulted in lower total blood cholesterol and triacylglycerol levels and attenuated body weight gain, by reducing fat gain. Sepiolite supplementation did not affect energy intake but increased fecal extraction of sterols and lipids, without notable side effects. These results demonstrate that supplementing a HFD with sepiolite attenuates gastrointestinal absorption of dietary lipids and sterols, thus mitigating obesity, hyperlipidemia, and hypercholesterolemia. Further exploration of the efficacy, mechanism of action, and safety of sepiolite as a food supplement for combating the metabolic syndrome is needed.

Recovery of sterols from vegetable oil distillate by enzymatic and non-enzymatic processes

Sterols are a group of molecules found in plants and animals, which have a number of valuable applications. The deodorization residue, referred to as “deodistillate”, was previously considered as a waste but its economical value nowadays increased due to the presence of high concentrations of sterols, tocopherols and other secondary metabolites attractive for the cosmetic, pharmaceutical or food industry. Sterols can be extracted from vegetable oil deodistillate through a variety of physical and chemical separation processes or their combination. Recently, the use of lipase enzymes has been demonstrated to separate sterols more selectively in higher yields and in milder conditions. This article reviews these lipase-assisted sterol extractions and their main advantages and drawbacks in economic and environmental terms. • Sterol extraction from vegetable oil deodistillate. • Reviewing on enzymatic process to involve extraction. • Different strategies are discussed in an industrial operation approach.

Valorization of spent shiitake substrate for recovery of antitumor fungal sterols by ultrasound-assisted extraction

The ongoing expansion of mushroom industries has generated enormous spent mushroom substrate with the potential for production of valuable chemicals. This study aimed to investigate ultrasound-assisted extraction (UAE) of fungal sterols from spent shiitake (Lentinula edodes) substrate (SSS) by response surface methodology. The results showed that three variables, including ultrasonic power, extraction time, and liquid–solid ratio, had considerable influence on the extraction efficiency of total fungal sterols with liquid–solid ratio being highly significant for the process. The optimal UAE conditions included liquid–solid ratio 22 mL/g, ultrasonic power 432 W, and extraction time 16 min for the maximum recovery of fungal sterols (151.62 mg ergosterol equivalents/100 g dry weight) from SSS. Further GC-MS analysis indicated that the extracts yielded under the optimal conditions contained ergosterol, ergosta-7,22-dienol, and β-sitosterol, in which ergosterol was the most abundant mycosterol. The extracts also showed a comparable antitumor effect against three cancer cell lines. Practical applications Spent mushroom substrate, a major by-product of mushroom industries, remains a big challenge for the profitability of mushroom industries. Our results indicated that spent shiitake substrate can be valorized by UAE for production of fungal sterols.

Fucosterol Causes Small Changes in Lipid Storage and Brassicasterol Affects some Markers of Lipid Metabolism in Atlantic Salmon Hepatocytes.

Several feeding trials with Atlantic salmon fed naturally high phytosterol concentrations due to dietary rapeseed oil inclusion have shown changes in lipid metabolism and increased hepatic lipid storage in the fish. An in vitro trial with Atlantic salmon hepatocytes was, therefore, performed to study the possible direct effects of phytosterols on lipid storage and metabolism. The isolated hepatocytes were exposed to seven different sterol treatments and gene expression, as well as lipid accumulation by Oil Red O dyeing, was assessed. Fucosterol, a sterol found in many algae species, had an effect on the size of individual lipid droplets, leading to smaller lipid droplets than in the control without added sterols. A sterol extract from soybean/rapeseed led to an increase in the percentage of hepatocytes with visible lipid droplets at 20× magnification, while hepatocytes of both the sterol extract-treated groups and fucosterol-treated groups had a larger proportion of their area covered with lipids compared to control cells. Brassicasterol, a sterol characteristic of rapeseed oil, was the only sterol treatment leading to a change in gene expression, affecting the expression of the nuclear receptors, peroxisome proliferator-activated receptor gamma (pparg) and retinoid X receptor (rxr). The current study thus shows that phytosterols can have direct, although subtle, effects on both hepatic lipid storage and gene expression of Atlantic salmon in vitro.

Quantitative Proteomics and Cytology of Rice Pollen Sterol-Rich Membrane Domains Reveals Pre-established Cell Polarity Cues in Mature Pollen.

Cell polarity is essential for generating diverse cell functions. The underlying mechanisms of how a cell establishes, maintains, and changes its polarity are poorly understood. Recently, sterol-rich membrane microdomains are found to be associated with these processes. However, both its exact characteristics and importance are still unclear. Here we show microdomains change dynamically in developing and germinating rice pollen with selective enrichment in the aperture and the tip of newly born pollen tubes by use of the sterol-specific probe filipin. Using the sterol extraction sensitivities of microdomain proteins and quantitative proteomics, we identified 237 microdomain-associated proteins from 934 identified pollen detergent resistant membrane proteins. This proteome includes almost all of the known key regulators comprising the polar growth network, and it shows more similarity to front-back polarized HeLa cells than nonpolarized Arabidopsis suspension cells. We immunolocalize flotilin-like protein, a representative of these sterol-dependent proteins and directly visualize microdomains in pollen. These results indicate the presence of microdomains in pollen and pre-established cell polarity around the aperture during pollen maturation. Our findings reveal an atlas of the microdomain-associated proteome in pollen. This work provides useful resources and knowledge needed to further dissect the mechanisms for the establishment and maintenance of cell polarity.

Oral Administration of CardioAid and Lunasin Alleviates Liver Damage in a High-Fat Diet Nonalcoholic Steatohepatitis Model

Background: Several of the drugs in development for treatment of nonalcoholic steatohepatitis (NASH) target liver fibrosis or have side effects that prohibit their long-term use in patients with mild to moderate disease. Lunasin is a soy-derived peptide with anti-inflammatory properties. ADM's CardioAid™ is a plant sterol extract that exerts cholesterol- and triacylglycerol-lowering effects. Aim: To determine the immunomodulatory effects of CardioAid and lunasin in a high-fat diet (HFD) animal model of NASH. Methods: C57BL/6 mice on an HFD were orally administered CardioAid or lunasin for 25 weeks. The effects on the immune system, liver function, insulin resistance and lipid profile were studied. Results: Treatment with CardioAid and lunasin was associated with a significant decrease in the CD4/CD8 ratio and an increase in CD4+CD25+ lymphocytes. A decrease in interleukin 1-alpha serum levels and an increase in transforming growth factor beta serum levels were noted. These were associated with alleviation of liver damage as indicated by a significant decrease in liver enzymes and improvement in the histological nonalcoholic fatty liver disease activity score (NAS). Decreases in both serum triglyceride and serum glucose levels were observed in treated mice. A decrease in total body fat measured by EchoMRI was also observed in treated mice. Conclusions: CardioAid and lunasin exerted hepatoprotective and glucose-protective effects in an HFD NASH model. These data and the high-safety profiles of CardioAid and Lunasin support their use in patients in the early stages of NASH to prevent deterioration due to the disease.