Differential Lipid Accumulation on HepG2 Cells Triggered by Palmitic and Linoleic Fatty Acids Exposure.

Lipid metabolism pathways such as ß-oxidation, lipolysis and, lipogenesis, are mainly associated with normal liver function. However, steatosis is a growing pathology caused by the accumulation of lipids in hepatic cells due to increased lipogenesis, dysregulated lipid metabolism, and/or reduced lipolysis. Accordingly, this investigation hypothesizes a selective in vitro accumulation of palmitic and linoleic fatty acids on hepatocytes. After assessing the metabolic inhibition, apoptotic effect, and reactive oxygen species (ROS) generation by linoleic (LA) and palmitic (PA) fatty acids, HepG2 cells were exposed to different ratios of LA and PA to study the lipid accumulation using the lipophilic dye Oil Red O. Lipidomic studies were also carried out after lipid isolation. Results revealed that LA was highly accumulated and induced ROS production when compared to PA. Lipid profile modifications were observed after LA:PA 1:1 (v/v) exposure, which led to a four-fold increase in triglycerides (TGs) (mainly in linoleic acid-containing species), as well as a increase in cholesterol and polyunsaturated fatty acids (PUFA) content when compared to the control cells. The present work highlights the importance of balancing both PA and LA fatty acids concentrations in HepG2 cells to maintain normal levels of free fatty acids (FFAs), cholesterol, and TGs and to minimize some of the observed in vitro effects (i.e., apoptosis, ROS generation and lipid accumulation) caused by these fatty acids.

Use of Various Sugarcane Byproducts to Produce Lipid Extracts with Bioactive Properties: Physicochemical and Biological Characterization.

Sugarcane, a globally cultivated crop constituting nearly 80% of total sugar production, yields residues from harvesting and sugar production known for their renewable bioactive compounds with health-promoting properties. Despite previous studies, the intricate interplay of extracts from diverse sugarcane byproducts and their biological attributes remains underexplored. This study focused on extracting the lipid fraction from a blend of selected sugarcane byproducts (straw, bagasse, and filter cake) using ethanol. The resulting extract underwent comprehensive characterization, including physicochemical analysis (FT-IR, DSC, particle size distribution, and color) and chemical composition assessment (GC-MS). The biological properties were evaluated through antihypertensive (ACE), anticholesterolemic (HMG-CoA reductase), and antidiabetic (alpha-glucosidase and Dipeptidyl Peptidase-IV) assays, alongside in vitro biocompatibility assessments in Caco-2 and Hep G2 cells. The phytochemicals identified, such as ß-sitosterol and 1-octacosanol, likely contribute to the extract's antidiabetic, anticholesterolemic, and antihypertensive potential, given their association with various beneficial bioactivities. The extract exhibited substantial antidiabetic effects, inhibiting α-glucosidase (5-60%) and DPP-IV activity (25-100%), anticholesterolemic potential with HMG-CoA reductase inhibition (11.4-63.2%), and antihypertensive properties through ACE inhibition (24.0-27.3%). These findings lay the groundwork for incorporating these ingredients into the development of food supplements or nutraceuticals, offering potential for preventing and managing metabolic syndrome-associated conditions.

Novel Lipids to Regulate Obesity and Brain Function: Comparing Available Evidence and Insights from QSAR In Silico Models.

Lipid molecules, such as policosanol, ergosterol, sphingomyelin, omega 3 rich phosphatidylcholine, α-tocopherol, and sodium butyrate, have emerged as novel additions to the portfolio of bioactive lipids. In this state-of-the-art review, we discuss these lipids, and their activity against obesity and mental or neurological disorders, with a focus on their proposed cellular targets and the ways in which they produce their beneficial effects. Furthermore, this available information is compared with that provided by in silico Absorption, Distribution, Metabolism, Excretion, and Toxicity (ADMET) models in order to understand the usefulness of these tools for the discovery of new bioactive compounds. Accordingly, it was possible to highlight how these lipids interact with various cellular targets related to the molecule transportation and absorption (e.g., α-tocopherol transfer protein for α-Tocopherol, ATP-binding cassette ABC transporters or Apolipoprotein E for sphingomyelins and phospholipids) or other processes, such as the regulation of gene expression (involving Sterol Regulatory Element-Binding Proteins for ergosterol or Peroxisome Proliferator-Activated Receptors in the case of policosanol) and inflammation (the regulation of interleukins by sodium butyrate). When comparing the literature with in silico Quantitative Structure-Activity Relationship (QSAR) models, it was observed that although they are useful for selecting bioactive molecules when compared in batch, the information they provide does not coincide when assessed individually. Our review highlights the importance of considering a broad range of lipids as potential bioactives and the need for accurate prediction of ADMET parameters in the discovery of new biomolecules. The information presented here provides a useful resource for researchers interested in developing new strategies for the treatment of obesity and mental or neurological disorders.

Suitability of Solvent-Assisted Extraction for Recovery of Lipophilic Phytochemicals in Sugarcane Straw and Bagasse.

Sugarcane is primarily harvested to meet up to 80% of global sugar demand. Recently, lipids recovered from their biomass (straw and bagasse) have attracted much attention due to their possible utilisation in biofuel production but also by the presence of health-promoting compounds as phytosterols (i.e., improvement of cardiovascular function) or 1-octacosanol (i.e., anti-obesity). Although this fraction is commonly obtained through solid-liquid isolation, there is scarce information about how different solvents affect the composition of the extracts. This research work aimed to study whether, in sugarcane straw and bagasse samples, Soxtec extraction with widely used dichloromethane (DCM) would be suitable to recover most of the lipid classes when compared to other available solvents such as food grade ethanol (EtOH) or solvents without regulation restrictions for food and drug applications (i.e., acetone and ethyl acetate). The obtained results allow concluding that sugarcane waxes from straw and bagasse are complex lipid mixtures of polar and non-polar compounds. According to the extraction yield, the best results were obtained with ethanol (5.12 ± 0.30% and 1.97 ± 0.31%) for both straw and bagasse, respectively. The extractant greatly influenced the lipid composition of the obtained product. Thus, DCM enriched the isolates in glycerolipids (mono-, di- and triglycerides), free fatty acids, fatty alcohols, fatty aldehydes, phytosterols and hydrocarbons. On the other hand, EtOH resulted in polar isolates rich in glycolipids. Therefore, depending on the application and objectives of future research studies, the solvent to recover such lipids needs to be carefully selected.

Bioactive Sugarcane Lipids in a Circular Economy Context.

Most of the global sugar and ethanol supply trade comes from the harvesting of Saccharum officinarum (i.e., sugarcane). Its industrial processing results in numerous by-products and waste streams, such as tops, straw, filter cake, molasses and bagasse. The recovery of lipids (i.e., octacosanol, phytosterols, long-chain aldehydes and triterpenoids) from these residues is an excellent starting point for the development of new products for various application fields, such as health and well-being, representing an important feature of the circular economy. By selecting green scalable extraction procedures, industry can reduce its environmental impact. Refluxed ethanol extraction methods have been demonstrated to meet these characteristics. On the other hand, effective non-solvent methodologies such as molecular distillation and supercritical CO2 extraction can fractionate lipids based on high temperature and pressure application with similar yields. Sugarcane lipophilic extracts are usually analyzed through gas chromatography (GC) and liquid chromatography (LC) techniques. In many cases, the identification of such compounds involves the development of high-temperature GC-MS/FID techniques. On the other hand, for the identification and quantification of thermolabile lipids, LC-MS techniques are suitable for the separation and identification of major lipid classes. Generically, its composition includes terpenes, phytosterols, tocopherol, free fatty acids, fatty alcohols, wax esters, triglycerides, diglycerides and monoglycerides. These compounds are already known for their interesting application in various fields such as pharma and cosmetics due to their anti-hypercholesterolemic, anti-hyperglycemic, antioxidant and anti-inflammatory properties.

Phytosterols and Novel Triterpenes Recovered from Industrial Fermentation Coproducts Exert In Vitro Anti-Inflammatory Activity in Macrophages.

The unstoppable growth of human population that occurs in parallel with all manufacturing activities leads to a relentless increase in the demand for resources, cultivation land, and energy. In response, currently, there is significant interest in developing strategies to optimize any available resources and their biowaste. While solutions initially focused on recovering biomolecules with applications in food, energy, or materials, the feasibility of synthetic biology in this field has been demonstrated in recent years. For instance, it is possible to genetically modify Saccharomyces cerevisiae to produce terpenes for commercial applications (i.e., against malaria or as biodiesel). But the production process, similar to any industrial activity, generates biowastes containing promising biomolecules (from fermentation) that if recovered may have applications in different areas. To test this hypothesis, in the present study, the lipid composition of by-products from the industrial production of ß-farnesene by genetically modified Saccharomyces cerevisiae are studied to identify potentially bioactive compounds, their recovery, and finally, their stability and in vitro bioactivity. The assayed biowaste showed the presence of triterpenes, phytosterols, and 1-octacosanol which were recovered through molecular distillation into a single fraction. During the assayed stability test, compositional modifications were observed, mainly for the phytosterols and 1-octacosanol, probably due to oxidative reactions. However, such changes did not affect the in vitro bioactivity in macrophages, where it was found that the obtained fraction decreased the production of TNF-α and IL-6 in lipopolysaccharide (LPS)-induced inflammation.

Automated analytical microsystem for the spectrophotometric monitoring of titratable acidity in white, rosé and red wines.

The design, construction and evaluation of a low-cost cyclic olefin copolymer (COC)-based continuous flow microanalyzer with optical detection to determine the titratable acidity content of wine is here presented. The analysis method is based on the monitoring of the blue coloration decrease of a buffered bromothymol blue (BTB) solution in the presence of the acidic compounds of wine. The microanalyzer monolithically integrates the required microfluidic motifs as well as an optical flow cell where the measurements are performed by using a miniaturized and versatile photometric detection system. Fluid management is totally automated by the use of computer-controlled microvalves, permitting the automatic calibration of the system as well as the automatic sampling, including in-line dilution and analysis. The reduced size of the whole system along with its high simplicity and automation make it suitable for its application to the on-line monitoring of titratable acidity during wine-making processes. With the optimal conditions, a linear range up to 0.50 g L-1 tartaric acid, a quantification limit (LOQ) of 0.01 g L-1 and a detection limit (LOD) of 0.004 g L-1 were obtained, covering the most common acidity content of musts and wines. A sampling rate up to 26 h-1 could be achieved, consuming less than 3 mL of inexpensive reagents per analysis and requiring no pretreatment of the sample. The microsystem has been successfully applied to the quantification of the titratable acidity content of several wine samples, being the results in excellent agreement with the ones obtained by the reference method.

A flow-based platform for measuring the acidity parameters in wine.

The present work describes a valuable tool for winemaking industry to measure the acidity parameters with rapid response, simple sample handling, with no or minimal pre-treatment. Thus, a sequential injection analysis (SIA) system with spectrophotometric detection was used as platform for the development of methodologies for the quantification of volatile and total acidity in wine samples. Both procedures make use of the same colour reagent, bromothymol blue (BTB) that changes its intrinsic colour in the presence of the acidity compounds. The volatile acidity value was attained with the separation of the volatile fraction of acids by means of a membrane separation technique, a gas-diffusion unit. For the total acidity value, the sample was merged with the colour reagent on the way towards detection. The fixed acidity is a result of the difference from the total and the volatile acidity. The presented tool displayed a low sample and reagent consumption, 346 and 102µL of sample and 37 and 32µg of BTB, for the volatile acidity (VA) and for the total acidity (TA), respectively. The observed limits of detection and quantification were 0.03 and 0.01gL-1 (VA) and 0.09 and 0.02gL-1 (TA) with high determination rates, 35 (VA) and 62 (TA) determinations per hour. The proposed system was successfully applied to the quantification of volatile, total and fixed acidity in white table wine samples. The obtained results were in good agreement with the ones obtained by the reference methods.

Sequential injection system for the enzymatic determination of ethanol in wine.

A sequential injection system was developed for the enzymatic determination of ethanol in wine. The spectrophotometric determination is based on the enzymatic reaction catalyzed by alcohol dehydrogenase in the presence of NAD+. The system was applied to the determination of ethanol in a range of 0.008-0.024% (v/v) with good repeatability; RSD(n=10) < 2.3%. The results obtained with the developed system showed good agreement with those obtained by using the reference method. The determination rate was 25 h(-1); 1 micromol of NAD+, 1.1 units of enzyme, and 50 microL of sample were consumed per determination; and the waste produced was 2.2 mL per assay.

A flow-injection system coupled to a micro-guard cartridge for monitoring a vinification process.

The present paper describes a flow-injection system coupled to a Micro-Guard cartridge and a miniaturized optical CCD detection system used to monitor the sugars (glucose/fructose) and ethanol content during alcoholic fermentation. The carrier stream (mobile phase) is composed of an aqueous sulfuric acid solution. The flow parameters were studied in order to obtain a good resolution and a wide dynamic concentration range, with a good repeatability. The relative standard deviation (RSD) obtained was inferior to 4%, for n = 3. It was possible to achieve a linear range of up to 12 g L(-1) of sugars and up to 2% (v/v) of ethanol with a detection limit of 2.3 g L(-1) and 0.4% (v/v), for sugars and ethanol concentrations, respectively. The proposed system was successfully applied to monitor a vinification process by the quantification of sugars and ethanol, and also in some finished port wines.

Determination of total protein content in white wines by solid phase spectrometry in a SI-LOV system.

Although present at low concentration in wine samples, proteins, have considerable technological importance, due to their capability of haze formation. The present work presents a methodology for the quantification of total protein in white wine in a sequential injection lab-on-valve system, exploiting the bead injection concept for solid phase extraction with spectrophotometric detection. The method is based on the retention of the proteins in the solid support, NTA (nitrilotriacetic acid) superflow beads, charged by Cu(2+). The change in the absorbance is monitored at 500nm at the surface of the beads after addition of the Folin-Ciocalteu's reagent (FCr). The developed method presented a sample consumption of 400µL per assay and a consumption of FCr and Cu(2+) solution of 25µL and 100µL per assay, respectively. It was possible to achieve a linear range up to 0.30g/L with a limit of detection and quantification of 0.03 and 0.10g/L, respectively. The proposed method was successfully applied to white wine samples.

Exploiting the bead injection LOV approach to carry out spectrophotometric assays in wine: application to the determination of iron.

A sequential injection lab-on-valve (SI-LOV) system was used to develop a new methodology for the determination of iron in wine samples exploiting the bead injection (BI) concept for solid phase extraction and spectrophotometric measurement. Nitrilotriacetic Acid (NTA) Superflow resin was used to build the bead column of the flow through sensor. The iron (III) ions were retained by the bead column and react with SCN(-) producing an intense red colour. The change in absorbance was monitored spectrophotometrically on the optosensor at 480 nm. It was possible to achieve a linear range of 0.09-5.0 mg L(-1) of iron, with low sample and reagent consumption; 500 µL of sample, 15 µmol of SCN(-), and 9 µmol of H(2)O(2), per assay. The proposed method was successfully applied to the determination of iron in wine, with no previous treatment other than dilution, and to other food samples.

Sequential injection lab-on-valve system for the determination of the activity of peroxidase in vegetables.

Horseradish peroxidase (HRP) has been broadly used and investigated for many analytical purposes; it is an enzyme that catalyzes the reduction of hydrogen peroxide in the presence of a reducing compound. The objective of this work was to develop a methodology for the spectrophotometric determination of the activity of peroxidase in vegetable extracts using a flow method with a sequential injection lab-on-valve format. The developed system is based on the reaction between hydrogen peroxide (H(2)O(2)) and 2,2-azinobis(3-ethylbenzothiazoline-6)sulfonic acid (ABTS) catalyzed by the enzyme (HRP). The method presented a sample consumption of 15 microL per assay and a consumption of ABTS and H(2)O(2) of 24 microg and 12 microg per assay, respectively. It was also possible to monitor online the thermal inactivation of peroxidase at different temperature ranges.