Small-angle x-ray scattering investigation of the integration of free fatty acids in polysorbate 20 micelles.

A critical quality attribute for liquid formulations is the absence of visible particles. Such particles may form upon polysorbate hydrolysis resulting in release of free fatty acids into solution followed by precipitation. Strategies to avoid this effect are of major interest for the pharmaceutical industry. In this context, we investigated the structural organization of polysorbate micelles alone and upon addition of the fatty acid myristic acid (MA) by small-angle x-ray scattering. Two complementary approaches using a model of polydisperse core-shell ellipsoidal micelles and an ensemble of quasiatomistic micelle structures gave consistent results well describing the experimental data. The small-angle x-ray scattering data reveal polydisperse mixtures of ellipsoidal micelles containing about 22-35 molecules per micelle. The addition of MA at concentrations up to 100 µg/mL reveals only marginal effects on the scattering data. At the same time, addition of high amounts of MA (>500 µg/mL) increases the average sizes of the micelles indicating that MA penetrates into the surfactant micelles. These results together with molecular modeling shed light on the polysorbate contribution to fatty acid solubilization preventing or delaying fatty acid particle formation.

Colour and pigment in raw ground meat incorporated crushed garlic during the refrigerated storage: Their relationship to lipolytic and volatilomic changes.

The effects of freshly crushed garlic incorporated in raw ground meat at different rates 0 % to 2 % on colour, pigment forms, TBARS, peroxide, free fatty acids and volatilomic were investigated during 96 h storage at 4 °C. With advancing storage time and increasing garlic rate from 0 % to 2 %, the redness (a*), colour stability, oxymyoglobin and deoxymyoglobin decreased, but metmyoglobin, TBARS, peroxide, free fatty acids (C6, C15-C17), and aldehydes and alcohols, especially hexanal, hexanol, benzaldehyde, increased. Principal component analysis based on pigment, colour, lipolytic and volatilomic changes classified successfully the meat samples. Metmyoglobin was positively correlated with lipid oxidation products (TBARS, hexanal), but the other pigment forms and colour parameters (a* and b* values) were negatively correlated. Increased metmyoglobin proportion and decreased redness and colour stability may be used as reliable indicators of lipid oxidation. Also, the incorporation of fresh garlic into ground meat was not promising to increase oxidative stability.

Biodiesel Production from Chlorella homosphaera by Two-Step Catalytic Conversion Using Waste Radish Leaves as a Source for Heterogeneous Catalyst.

The economic viability of algal biodiesel can be improved by enhancing the microalgal lipid accumulation and using agricultural waste as a cheap and sustainable source of catalysts. In the current study, the effect of various nitrogen concentrations on the growth and lipid of Chlorella homosphaera were investigated. Furthermore, two-step catalytic conversion was applied to convert the oil of C. homosphaera with high free fatty acids (FFA) to biodiesel using waste radish leaves as a source of a heterogeneous base catalyst. The result revealed that the maximum lipid productivity of 25.0 mg L-1 day-1 and lipid content of 30.83% were obtained under nitrogen-depleted and limited nitrogen conditions, respectively. The FFA was reduced from 18.79 to 0.76%, and the acid value was decreased from 37.4 to 1.52 mg KOH g-1 using a 15:1 methanol to oil molar ratio (MTOR), 1.5 wt.% H2SO4, at 60 °C for 150 min. Under the optimized conditions, i.e., MTOR of 10:1, 3 wt.% of catalyst ratio for 120 min at 60 °C, the highest oil conversion of 96.61% was obtained. The physicochemical properties of the produced biodiesel were in the range of the standard specification norms for biodiesel. Hence, the proposed two-step catalytic conversion using calcined radish leaves as a heterogeneous catalyst has thus exhibited good potential for biodiesel production using algal oil with high FFA.

Estradiol production of granulosa cells is unaffected by the physiological mix of nonesterified fatty acids in follicular fluid.

Ovarian cycle is controlled by circulating levels of the steroid hormone 17-ß-estradiol, which is predominantly synthesized by the granulosa cells (GCs) of ovarian follicles. Our earlier studies showed that unsaturated fatty acids (USFs) downregulate and saturated fatty acids (SFAs) upregulate estradiol production in GCs. However, it was unclear whether pituitary gonadotropins induce accumulation of free fatty acids (FFAs) in the follicular fluid since follicle-stimulating hormone induces and luteinizing hormone inhibits estradiol production in the mammalian ovary. Interestingly, we show here the gas chromatography analysis of follicular fluid revealed no differential accumulation of FFAs between pre- and post-luteinizing hormone surge follicles. We therefore wondered how estradiol production is regulated in the physiological context, as USFs and SFAs are mutually present in the follicular fluid. We thus performed in vitro primary GC cultures with palmitate, palmitoleate, stearate, oleate, linoleate, and alpha-linolenate, representing >80% of the FFA fraction in the follicular fluid, and analyzed 62 different cell culture conditions to understand the regulation of estradiol biosynthesis under diverse FFA combinations. Our analyses showed co-supplementation of SFAs with USFs rescued estradiol production by restoring gonadotropin receptors and aromatase, antagonizing the inhibitory effects of USFs. Furthermore, transcriptome data of oleic acid-treated GCs indicated USFs induce the ERK and Akt signaling pathways. We show SFAs inhibit USF-induced ERK1/2 and Akt activation, wherein ERK1/2 acts as a negative regulator of estradiol synthesis. We propose SFAs are vital components of the follicular fluid, without which gonadotropin signaling and the ovarian cycle would probably be shattered by USFs.

Two-stage continuous production process for fatty acid methyl ester from high FFA crude palm oil using rotor-stator hydrocavitation.

Two-stage continuous production process for fatty acid methyl ester (FAME) from crude palm oil was performed using the rotor-stator hydrocavitation reactor. The novel ABS filament printed rotor having spherical holes on the surface of the rotor which is an efficient, fast and cost-effective procedure, was installed in the stainless steel stator of hydrosonic reactor. The 3D printed hydrosonic reactor was used to treat the FFA-rich in MCPO by esterification and followed by transesterification to produce the methyl ester. The optimum conditions of both esterification and transesterification processes were determined using the response surface methodology (RSM). For the 1st step esterification, the conditions of methanol 17.7 vol%, sulfuric acid 2.9 vol%, 3000 rpm rotor speed, hole's diameter and depth 4 and 6 mm, and 25 L/h MCPO, were used for decreasing the FFA from 11.456 to 1.028 wt%. For the 2nd step, transesterification was employed with the optimal condition of 28.6 vol% methanol, 6.2 g/L of potassium hydroxide, 3000 rpm rotor speed, the dimension of the spherical holes on the rotor's surface having diameter of 6.4 mm and 6.2 mm in depth, and esterified oil flow rate 25 L/h, for producing the methyl ester to over 99.163 wt%. Moreover, the purified biodiesel yields and the average energy consumption for the entire two-stage continuous process between hydrosonic and ultrasonic clamp reactors were compared. The results of purified methyl ester clearly indicate that the methyl esters of 99.163 wt% and 97.814 wt% were achieved from hydrosonic and ultrasonic clamp reactors, respectively, under the same optimized conditions. The maximum yields of purified biodiesel were 97.51 vol% and 88.69 vol% using hydrosonic and ultrasonic clamp reactors, respectively. The average energy consumption for the entire continuous two-stage process for both hydrosonic and ultrasonic clamp reactors were 0.049 and 0.056 kW h/L, respectively. For practical industrial processes, stainless steel rotors inside the stator was manufactured by CNC machine, which was also verified under the optimum conditions. The results showed that 1.07 wt% FFA and 99.221 wt% methyl ester of were obtained from first step and second step, respectively.

Effects of enzymatic free fatty acid reduction process on the composition and phytochemicals of rice bran oil.

The effects of enzymatic free fatty acid reduction process (EFFARP) on the composition and phytochemicals of dewaxed and degummed rice bran oil (DDRBO) were investigated and compared with the effects observed using internal acyl acceptors. The acid value of DDRBO was effectively decreased from 16.99 mg KOH/g to approximately 0.36 mg KOH/g by EFFARP. EFFARP significantly decreased the moisture content and peroxide value of DDRBO and increased the induction period. The Sn-2 fatty acid comoposition of DDRBO after EFFARP was very reaching the total fatty acid composition. EFFARP significantly increased the triacylglycerol content compared to the control, while the oryzanol content was not obviously affected. The contents of free sterol, and total tocopherol and tocotrienol were increased slightly by EFFARP compared to the control. When conducted under vacuum with added nitrogen, EFFARP shows great application potential in the edible oil industry.

Effect of unsaturation of free fatty acids and phytosterols on the formation of esterified phytosterols during deodorization of corn oil.

BACKGROUND: Phytosterols are partly removed during oil refining, and the magnitude of phytosterols loss largely depends on the refining conditions applied and the molecular conformation. The aim of this research was to study the effect of deodorization conditions and molecular unsaturation on the esterification of phytosterols during deodorization of corn oil. RESULTS: In the chemical model, free fatty acids (FFAs) were the major provider of acyl groups during the formation of phytosteryl fatty acid esters (PEs) under deodorization conditions. Among the main parameters of the deodorization, temperature played a role in the formation of PEs with a time-dependent manner. In comparison, saturated palmitic acid had a higher capability of esterifying free phytosterols (FPs) to PEs than unsaturated oleic acid and linoleic acid. Moreover, the influence of FFA unsaturation on the degradation of FPs depended on temperature. Besides, the formation of stigmasteryl ester had a competitive advantage over that of sitosteryl ester by quantum chemistry simulation. CONCLUSION: For laboratory-scale deodorization of corn oil, saturated fatty acids and deodorization process with steam as stripping gas could obviously esterify FPs to PEs. FPs were abundantly enriched in distillate during the deodorization process with nitrogen as stripping gas, whereas FPs and PEs were distilled simultaneously during the deodorization process with steam. © 2020 Society of Chemical Industry.

Dietary oleuropein extract supplementation and its combination with α-tocopheryl acetate and selenium modifies the free fatty acid profile of pork and improves its stability.

BACKGROUND: Olive-derived antioxidants have been shown to affect the oxidative status of meat and have also been associated with greater consumption of glucose, which might affect glycogen stores and muscle characteristics. This study evaluated the effect of oleuropein extract supplementation (OLE) versus vitamin E + Se (VE), and their combination (VEOLE), in pig diets, on pH, drip loss, the proportion of free fatty acids, and meat stability, and their prediction by blood oxidative status markers. RESULTS: The drip loss of muscle was lower in antioxidant-supplemented groups when compared with controls. α-Tocopherol concentration and total fatty acids profile were not affected by dietary oleuropein supplementation. However, OLE and VEOLE had lower free n-3 polyunsaturated fatty acid (PUFA) levels when compared with VE and tended to have higher free monounsaturated fatty acid (MUFA) levels. Furthermore, the VEOLE group had lower free n-6 PUFA levels when compared with controls or VE, whereas the OLE group had intermediated values. Muscle samples from pigs subjected to the antioxidant-mixed supplementation (VEOLE) had lower malondialdehyde concentration when compared with the others. The VE and OLE groups showed intermediate malondialdehyde values. Chilled meat stability was highly correlated with antioxidant status in vivo. CONCLUSION: The administration of 96 mg oleuropein kg-1 feed produced similar meat quality characteristics as the use of 100 mg kg-1 α-tocopheryl acetate +0.26 mg kg-1 sodium selenite and it would be an interesting alternative in Mediterranean countries. The VEOLE group was the most effective for reducing lipid oxidation and for the production of polyunsaturated free fatty acids in meat, which would result in lower rancidity formation and better aroma development in products. © 2020 Society of Chemical Industry.

Pistachio Green Hull Extract Induces Apoptosis through Multiple Signaling Pathways by Causing Oxidative Stress on Colon Cancer Cells.

BACKGROUND: Pistachio is considered to be one of the fifty foods with the highest antioxidant effect. However, the anticancer effect mechanisms of this plant extracts are unknown. OBJECTIVE: The aim of this study was to investigate the anticancer effect of different extracts from the green hull of pistachio. METHODS: The cytotoxic effects of different solvent extracts on cancer and normal cells were examined by cell viability assay and flow cytometric analysis. The levels of the apoptotic gene and protein were investigated by Western Blot and ELISA, and qPCR. The intracellular free radical exchange was determined by oxidative and nitric oxide analyses. DNA damage level was measured by the 8-OHdG test. Phenolic and free fatty acid components were examined by LC-MS/MS and GC-MS, respectively. RESULTS: It was determined that the n-hexane fraction showed a higher cytotoxic effect on cancer cells. Oxidative and cell cycle analyses indicated that the n-hexane fraction arrested cell cycle of HT-29 at the sub-G1 phase by increasing DNA damage through oxidative stress. In addition, gene expression analysis of the HT-29 treated with the n-hexane fraction indicated that apoptotic and autophagic gene expressions were significantly upregulated. LC-MS/MS analysis of the n-hexane fraction revealed the presence of 15 phenolic compounds, containing mainly gallic acid and catechin hydrate, and GC-MS analysis determined the presence of the following fatty acids: 9-octadecenoic acid, 9,12-octadecadienoic acid and hexadecenoic acid. CONCLUSION: Based on these grounds, we suggest that the n-hexane fraction of pistachio green hull damages DNA, arrests the cell cycle at the G1 subphase, and induces apoptosis through oxidative pathways in colon cancer.

Preparation of nanofibrillated cellulose from grapefruit peel and its application as fat substitute in ice cream.

Grapefruit peel nanofibrillated cellulose (GNFC) was used as fat substitute in ice cream. GNFC was characterized by TEM, SEM, and XRD. The effects of GNFC on textural profiles, rheological properties, melting resistance, sensory properties, microstructure, and gross energy (GE) of ice cream were investigated. The results showed that GNFC was short rod-shaped crystal. Ice cream added with GNFC exhibited elastic-dominated behavior and better textural properties. The sensory evaluation score reached the highest level with desirable three-dimensional network structure at 0.4 % GNFC addition. GE of ice cream significantly decreased with reducing fat with maximal reduction rate of 17.90 %. Furthermore, the results of in vitro simulated digestion showed that GNFC addition and fat reduction significantly inhibited fat digestibility of ice cream due to coalescence of fat droplets on GNFC. This study provides new sustainable perspectives for the application of GNFC prepared from agricultural waste as fat substitute in food products.

Eucaloric diets enriched in palm olein, cocoa butter, and soybean oil did not differentially affect liver fat concentration in healthy participants: a 16-week randomized controlled trial.

BACKGROUND: Effects of dietary fat quality on liver fat remain to be elucidated. Inconsistent evidence may be influenced by fatty acid saturation, chain-length, and regio-specificity within triacylglycerol (TAG) molecules. OBJECTIVES: We aimed to compare eucaloric diets enriched in palm olein (POo), cocoa butter (COB), and soybean oil (SBO) on liver fat concentration in healthy participants. Secondary outcomes included visceral (VAT) and abdominal subcutaneous (aSCAT) adipose tissue, plus other obesity and cardiometabolic health outcomes. METHODS: Eighty-three healthy participants (20-45 y, BMI 18.5-27.5 kg/m2) commenced and 64 completed a 16-wk randomized parallel intervention, preceded by a 2-wk run-in. Participants consumed identical eucaloric background diets differing in test fats [contributing 20% total energy intake (%E)], providing 33%E total fat with the following ratios for PUFAs/SFAs/MUFAs: POo, 4.2/13.5/15%E; SBO, 14.4/8.8/9.4%E; COB, 2.3/19.5/11%E. Liver fat and abdominal adiposity were measured at weeks 0 and 16 using 1H-magnetic resonance spectroscopy/imaging; all other outcomes were measured at 0, 4, 8, 12, and 16 wk. RESULTS: Fat quality did not affect liver fat concentration, VAT, aSCAT, obesity indexes, blood pressure, liver enzymes, leptin, or fasting glucose. Body fat mass decreased with SBO and COB compared with POo. SBO decreased serum total cholesterol (TC), LDL cholesterol, and TC:HDL cholesterol relative to POo [estimated marginal mean (95% CI) differences: -0.57 (-0.94, -0.20) mmol/L; -0.37 (-0.68, -0.07) mmol/L; and -0.42 (-0.73, -0.11) mmol/L, respectively]. No diet differences were observed on HDL cholesterol, TAG, apoA1, apoB, apoB:apoA1, or fecal free fatty acids (FFAs), except for lower FFA pentadecanoic acid (15:0) with COB than with SBO and POo. CONCLUSIONS: In healthy adults, when consumed as part of eucaloric typical Australian diets, 3 different dietary fat sources did not differentially affect liver fat concentration and amounts of adipose tissue. Effects on serum lipids were inconsistent across lipid profiles. The findings must be confirmed in metabolically impaired individuals before recommendations can be made.

Borage Oil Treated with Immobilized Lipase Inhibits Melanogenesis.

In the present study, we demonstrated that borage (Borago officinalis L.) seed oil subjected to immobilized lipase pretreatment are enriched with linoleic acid (LNA, 18:2n-6), γ-linolenic acid (GLA, 18:3n-6), and oleic acid (OLA, 18:1n-9). We further showed that lipase-treated borage oil (LT-BOL) regulates the activity and degradation of tyrosinase, an important enzyme implicated in the synthesis of melanin in murine melanocytes, B16F10. LT-BOL and its free fatty acid components reduced the levels of melanin and tyrosinase in melanocytes with GLA exerting similar or stronger effects compared with LNA and OLA. The brightening efficacy of LT-BOL on melanin metabolism in humans was tested by an 8-week, double-blind, randomized clinical trial, which enrolled 21 Korean female adults (mean age 48.57 ± 3.28). Visual evaluation showed that cream containing 1% LT-BOL significantly decreased (p < 0.05) melasma on the treated skin area after 6 and 8 weeks. The analysis of the skin brightness using Chromameter CR-400 confirmed that the brightness of the treated area was significantly increased (p < 0.01) after 4, 6, and 8 weeks. Together, our results suggest that LT-BOL may be suitable as a natural skin whitening cosmeceutical product.

Effects of deodorization temperature and time on the formation of 3-MCPD, 2-MCPD, and glycidyl esters and physicochemical changes of palm oil.

This study verified the formation profile of esters of 3-monochloropropane-1,2-diol (3-MCPDE), 2-monochloropropane-1,2-diol (2-MCPDE), and glycidol (GE), and evaluated the physicochemical changes (free-fatty acid, acylglycerols, and colour) that occurred during the deodorization of palm oil in different conditions of time (30, 60, 90, and 120 min) and temperature (210, 230, 250, and 270 °C). Levels of 3-MCPD and 2-MCPD esters ranged from 1.91 to 2.70 mg/kg and 0.68 to 1.44 mg/kg, respectively, and were formed at the mildest tested condition (210 °C, 30 min). No correlation was observed between these contaminants and physicochemical changes. GE levels varied from 0.12 to 8.51 mg/kg and showed correlation with color and diacylglycerol content. While the temperature had little influence on the formation of esters of 3-MCPD and 2-MCPD, the content of GE considerably raised above 250 °C.

In Vitro Gastrointestinal Digestion of Palm Olein and Palm Stearin-in-Water Emulsions with Different Physical States and Fat Contents.

The impacts of lipid physical state and content on lipid digestion behavior were investigated using 4 and 20% palm olein-in-water emulsions (4% PO and 20% PO) and 4 and 20% palm stearin-in-water emulsions (4% PS and 20% PS). The changes of lipid physical state, particle size, and microstructure during gastrointestinal digestion; the free fatty acid (FFA) released in the intestinal phase; and the fatty acid composition of micellar phases were investigated. After gastric digestion, all emulsions underwent flocculation and coalescence, with 20% PS showing the most extensive aggregation. During intestinal digestion, the FFA release rate and level decreased as the lipid content increased from 4 to 20%, with 4% PO presenting the highest digestion rate and extent. Besides, the solid fat in 4% PS and 20% PS decreased and increased the maximum lipid digestibility, respectively. These results highlighted the combined roles of lipid physical state and content in modulating dietary lipid digestion.

Controlled polysorbate 20 hydrolysis - A new approach to assess the impact of polysorbate 20 degradation on biopharmaceutical product quality in shortened time.

Hydrolysis of polysorbate in biopharmaceutical liquid formulations upon long-term storage represents a risk factor, since reduction of the intact surfactant concentration may compromise protein stability. Moreover, accumulation of polysorbate degradation products is associated with the formation of particulates potentially affecting drug product stability and quality. These effects are conventionally assessed by real-time end-of-shelf life studies constituting an integral yet lengthy process of formulation development. To accelerate this procedure, we describe here a powerful tool to conduct shake stress studies based on the controlled hydrolysis of polysorbate 20 by beads-immobilized lipases. For this purpose, the production of stable, partially degraded material characterized by a representative presence of non-emulsifying degradants such as ethoxylated sorbitan and free fatty acids was monitored by state-of-the-art chromatographic methods ensuring realistic pharmaceutical conditions. Freeze-thaw, shaking and shipping stress studies of a mAb formulation did not only demonstrate that this approach is useful to determine the critical degradation level impairing drug product quality, but furthermore revealed significant differences in protective effects depending on the hydrolysis pattern. As these results emphasize, the outlined strategy may support formulation scientists to unveil the interrelationship between polysorbate hydrolysis products and stabilization of the active pharmaceutical ingredient in a holistic and time-saving manner.

Qualitative Phytochemical Fingerprint and Network Pharmacology Investigation of Achyranthes aspera Linn. Extracts.

Achyranthes aspera Linn. (Amaranthaceae), commonly known as the Prickly Chaff flower, is used as herbal medicine in the Ivorian's culture, Africa. Nonetheless, there is currently a paucity of scientific information on A. aspera from the Ivory Coast. Herein, the antioxidant activity of A. aspera extracts (methanol, dichloromethane, ethyl acetate and infusion) as well as the enzymatic inhibitory potentials towards key enzymes in human diseases, namely Alzheimer's disease, (cholinesterases: AchE and BChE), type 2 diabetes (α-glucosidase and α-amylase) and hyperpigmentation (tyrosinase) were assessed. The total phenolic (TPC) and flavonoid (TFC) content was determined using colorimetric methods and the individual compounds were characterized using ultra-high performance liquid chromatography coupled with hybrid quadrupole-Orbitrap high resolution mass spectrometry (UHPLC-HRMS). Furthermore, a network pharmacology analysis was conducted to predict putative targets of identified phenolic compounds. The highest TPC was observed in the infused extract (28.86 ± 0.12 mg GAE/g), while the dichloromethane extract (38.48 ± 1.48 mg RE/g) showed the highest level of TFC. UHPLC-HRMS analysis has revealed an abundance of fatty acids, flavonoids, phenols and acylquinic acids. Among tested extracts, the infused extract displayed the highest free radical quenching, reducing and metal-chelating ability. The extracts (except infusion) were effective as enzyme inhibitors against AChE, while only methanolic and infused extracts showed noteworthy anti-BChE effects. The methanolic extract showed a remarkable antityrosinase effect (56.24 ± 5.05 mg KAE/g), as well. Modest to moderate inhibitory activity was observed against α-amylase (all extracts) and α-glucosidase (only dichloromethane extract). Finally, the network pharmacology analysis suggested the carbonic anhydrase II enzyme as a putative target for explaining, at least in part, the traditional use of A. aspera preparations as diuretic and blood clotting agent. Data amassed herein tend to validate the use of A. aspera in traditional medicine, as well as act as a stepping stone for further studies in the quest for novel phytopharmaceuticals. In this context, it is desirable that this study will contribute to the validation of the traditional uses of this plant in the African herbal medicine, and to the valorization of the whole chain production of A. aspera, as a local and sustainable botanical resource.

Interaction of the Coffee Diterpenes Cafestol and 16-O-Methyl-Cafestol Palmitates with Serum Albumins.

The main coffee diterpenes cafestol, kahweol, and 16-O-methylcafestol, present in the bean lipid fraction, are mostly esterified with fatty acids. They are believed to induce dyslipidaemia and hypercholesterolemia when taken with certain types of coffee brews. The study of their binding to serum albumins could help explain their interactions with biologically active xenobiotics. We investigated the interactions occurring between cafestol and 16-O-methylcafestol palmitates with Bovine Serum Albumin (BSA), Human Serum Albumin (HSA), and Fatty Free Human Serum Albumin (ffHSA) by means of circular dichroism and fluorimetry. Circular Dichroism (CD) revealed a slight change (up to 3%) in the secondary structure of fatty-free human albumin in the presence of the diterpene esters, suggesting that the aliphatic chain of the palmitate partly occupies one of the fatty acid sites of the protein. A warfarin displacement experiment was performed to identify the binding site, which is probably close but not coincident with Sudlow site I, as the affinity for warfarin is enhanced. Fluorescence quenching titrations revealed a complex behaviour, with Stern-Volmer constants in the order of 103-104 Lmol-1. A model of the HSA-warfarin-cafestol palmitate complex was obtained by docking, and the most favourable solution was found with the terpene palmitate chain inside the FA4 fatty acid site and the cafestol moiety fronting warfarin at the interface with site I.

Effect of Almond Supplementation on Non-Esterified Fatty Acid Values and Exercise Performance.

Several studies have investigated the effects of fat intake before exercise on subsequent substrate oxidation and exercise performance. While some studies have reported that unsaturated fatty acid supplementation slightly increases fat oxidation, the changes have not been reflected in the maximum oxygen uptake or in other performance and physiological parameters. We selected almonds as a fatty acid (FA) source for acute supplementation and investigated their effect on non-esterified fatty acid (NEFA) values and exercise performance. Five physically active male subjects (age 32.9 ± 12.7 years, height 178.5 ± 3.3 cm, and weight 81.3 ± 9.7 kg) were randomly assigned to take an almond or placebo supplement 2 h before participating in two cycling resistance training sessions separated by an interval of 7-10 days. Their performance was evaluated with a maximal incremental test until exhaustion. Blood samples collected before, during, and after testing were biochemically analysed. The results indicated a NEFA value average increase of 0.09 mg·dL-1 (95% CI: 0.05-0.14; p < 0.001) after active supplement intake and enhanced performance (5389 ± 1795 W vs. placebo 4470 ± 2053 W, p = 0.043) after almond supplementation compared to the placebo. The almond supplementation did not cause gastrointestinal disturbances. Our study suggests that acute almond supplementation 2 h before exercise can improve performance in endurance exercise in trained subjects.

Application of flow cytometry as novel technology in studying lipid oxidation and mass transport phenomena in oil-in-water emulsions.

Flow cytometry was used to determine if lipid oxidation products could transfer between individual emulsion droplets. Medium chain triacyclglycerol oil-in-water emulsions containing an oxidizable fluorescent dye, BODIPY665/676, was blended with a soybean oil-in-water emulsion. Results showed that when the concentration of sodium dodecyl sulfonate (SDS) were lower than critical micelle concentration (CMC), lipid oxidation products of triacylglycerols were not able to escape out until emulsions were extremely oxidized. With surfactant micelles, oxidation of BODIPY665/676 was observed. In the presence of free fatty acids, the transfer of prooxidants between droplets was observed even when surfactant concentration was lower than CMC. The decomposition product, 2,4,-decadienal, was also found to be transferred between droplets. The effect of surfactant concentration on prooxidant transfer was investigated using the lipid-soluble radical generator, AMVN. Results showed that surfactants promoted AMVN-triggered degradation of fluorescence at low concentrations but inhibited degradation at high concentration. The CMC of SDS was decreased by NaCl thus affecting the transfer phenomenon. With flow cytometry, the phenomenon of mass transfer between individual droplets was revealed which provides a better understanding of lipid oxidation in emulsion system.

Free fatty acids promote transformation efficiency of yeast.

High transformation efficiency is essential in genetic engineering for functional metabolic analysis and cell factory construction, in particular in construction of long biosynthetic pathways with multiple genes. Here, we found that free fatty acid (FFA)-overproducing strain showed higher transformation efficiency in Saccharomyces cerevisiae. We then verified that external supplementation of FFAs, to the culture media for competent cell preparation, improved yeast transformation efficiency significantly. Among all tested FFAs, 0.5 g/L C16:0 FFA worked best on promoting transformation of S. cerevisiae and Komagataella phaffii (previously named as Pichia pastoris). Furthermore, C16:0 FFA improved the assembly efficiency of multiple DNA fragments into large plasmids and genome by 100%, which will facilitate the construction and optimization of multigene-containing long pathways.