Analysis of Hepatic Lipid Metabolism Model: Simulation and Non-Stationary Global Sensitivity Analysis.

Lipid metabolism is a complex process and it is extremely helpful to simulate its performance with different models that explain all the biological processes that comprise it, which then enables its better understanding as well as understanding the kinetics of the process itself. Typically, kinetic parameters are obtained from a number of sources under specific experimental conditions, and they are a source of uncertainty. Sensitivity analysis is a useful technique for controlling the uncertainty of model parameters. It evaluates a model's dependence on its input variables. In this work, hepatic lipid metabolism was mathematically simulated and analyzed. Simulations of the model were performed using different initial plasma glucose (GB) and plasma triacylglyceride (TAG) concentrations according to proposed menus for different meals (breakfast, lunch, snack and dinner). A non-stationary Fourier amplitude sensitivity test (FAST) was applied to analyze the effect of 78 kinetic parameters on 24 metabolite concentrations and 45 reaction rates of the biological part of the hepatic lipid metabolism model at five time points (tf = 10, 50, 100, 250 and 500 min). This study examined the total influence of input parameter uncertainty on the variance of metabolic model predictions. The majority of the propagated uncertainty is due to the interactions of numerous factors rather than being linear from one parameter to one result. Obtained results showed differences in the model control regarding the different initial concentrations and also the changes in the model control over time. The aforementioned knowledge enables dietitians and physicians, working with patients who need to regulate fat metabolism due to illness and/or excessive body mass, to better understand the problem.

Emulsification of Rosemary and Oregano Aqueous Extracts and Their In Vitro Bioavailability.

Due to their richness in phenolic compounds, Mediterranean plants such as rosemary and oregano are increasingly recommended for consumption for their numerous health benefits. The pH shift and the presence of digestive enzymes significantly reduce the bioavailability of these biochemicals as they pass through the gastrointestinal tract. To prevent this degradation of phenolic compounds, methods such as emulsification of plant aqueous extracts are used. The aim of this study was to investigate the effects of emulsification conditions on the chemical properties (total polyphenolic content and antioxidant activity) of emulsified rosemary and oregano extracts. Response surface methodology was applied to optimize sunflower oil concentration, rotational speed, and emulsifier concentration (commercial pea protein). The emulsions prepared under optimal conditions were then used in bioavailability studies (in vitro digestion). The antioxidant activity of the emulsified rosemary/oregano extracts, measured by the DPPH method, remained largely stable when simulating in vitro digestion. Analysis of antioxidant activity after in vitro simulation of the gastrointestinal system revealed a higher degree of maintenance (up to 76%) for emulsified plant extracts compared to aqueous plant extracts. This article contributes to our understanding of how plant extracts are prepared to preserve their biological activity and their application in the food industry.

Comprehensive Study of Traditional Plant Ground Ivy (Glechoma hederacea L.) Grown in Croatia in Terms of Nutritional and Bioactive Composition.

In the present study, ground ivy was harvested from different natural habitats in Croatia and subjected to screening analysis for nutritional and bioactive composition. To achieve maximum recovery of phenolic compounds, different extraction techniques were investigated-heat-assisted (HAE), microwave-assisted (MAE) and subcritical water (SWE) extraction. Prepared extracts were analysed by spectrophotometric methods, LC-MS/MS and HPLC-PAD methodologies. Results regarding nutritive analyses, conducted using standard AOAC methods, showed the abundance of samples in terms of insoluble dietary fibre, protein, calcium and potassium, while rutin, chlorogenic, cryptochlorogenic, caffeic and rosmarinic acid were the most dominant phenolic compounds. In addition, LC-MS/MS analysis revealed the presence of apigenin and luteolin in glycosylated form. Maximum recovery of target phenolic compounds was achieved with MAE, while SWE led to the formation of new antioxidants, which is commonly known as neoformation. Moreover, efficient prediction of phenolic composition of prepared extracts was achieved using NIR spectroscopy combined with ANN modelling.

Comparison of Drying Methods and Their Effect on the Stability of Grasevina Grape Pomace Biologically Active Compounds.

Valorisation of grape pomace, a by-product of the winery industry, has been pushed into the spotlight in recent years since it can enable lower environmental impact, but it can also bring an added value to the wine production process by recovering several grape pomace biologically active compounds. The first step that allows for grape pomace reuse is its drying, which should be carefully performed in order to preserve the biologically active compounds' stability. In this study, the effects of different drying methods on the stability of polyphenols, tannins and tartaric acid in grape pomace (Vitis vinifera) cv. Grasevina were investigated. In particular, vacuum drying (at different temperatures: 35, 50 and 70 °C), conventional drying at 70 °C and open sun drying were performed and the drying kinetics was described using Peleg's model. Considering the processing time and thermodynamics, vacuum drying at 70 °C was the most convenient processing method. Polyphenols were highly stable during drying, and slight degradation occurred during vacuum drying at 35 and 50 °C. Tannins and tartaric acid were more prone to degradation depending on the drying method applied and showed the greatest stability during vacuum drying at 70 °C.

Lipase catalysed biodiesel synthesis with integrated glycerol separation in continuously operated microchips connected in series.

Although the application of microreactors in different processes has been extensively explored in recent decades, microreactors continue to be underexplored in the context of the enzyme-catalysed process for biodiesel production. Due to their numerous advantages, microreactors could become the next step in the development of a biodiesel production process characterised by sustainability, cost-effectiveness and energy efficiency. In this investigation, biodiesel production was catalysed by lipase from Thermomyces lanuginosus (Lipolase L100). Edible sunflower oil was used as a model substrate in order to investigate the process. After optimal process conditions had been determined, waste-cooking oil was used for biodiesel production to make the production process more sustainable. Three different substrate-feeding strategies were investigated and finally an optimal strategy was proposed. In all the investigated systems, fatty acids methyl esters (FAME) content was higher than 95% and obtained in a significantly shorter time (less than 2 h) compared to the batch process in which biodiesel production was catalysed by lipase (C = 95%, t = 96 h). After the optimal biodiesel production system had been proposed, an integrated system with two microchips connected in series was developed. The first microchip was used for biodiesel production and the second for simultaneous purification i.e. glycerol separation. Finally, purified biodiesel was produced with glycerol content below the detection limit.

Catechol Removal from Aqueous Media Using Laccase Immobilized in Different Macro- and Microreactor Systems.

Laccase belongs to the group of enzymes that are capable to catalyze the oxidation of phenols. Since the water is only by-product in laccase-catalyzed phenol oxidations, it is ideally "green" enzyme with many possible applications in different industrial processes. To make the oxidation process more sustainable in terms of biocatalyst consumption, immobilization of the enzyme is implemented in to the processes. Additionally, when developing a process, choice of a reactor type plays a significant role in the total outcome.In this study, the use of immobilized laccase from Trametes versicolor for biocatalytic catechol oxidation was explored. Two different methods of immobilization were performed and compared using five different reactor types. In order to compare different systems used for catechol oxidation, biocatalyst turnover number and turnover frequency were calculated. With low consumption of the enzyme and good efficiency, obtained results go in favor of microreactors with enzyme covalently immobilized on the microchannel surface.

Adaptation of CHO cells in serum-free conditions for erythropoietin production: Application of EVOP technique for process optimization.

Mammalian cell cultures are the preferred expression systems for the production of biopharmaceuticals requiring posttranslational processing. Usually, cell cultures are cultivated in medium supplemented with serum, which supports cell proliferation, viability, and productivity. However, due to scientific and regulatory concerns, serum-free conditions are required in recombinant protein production. Cell lines that are intended for commercial recombinant protein production have to adapt to serum- or protein-free conditions early in their development. This is a labor- and time-consuming process because of the specific cell requirements related to their adaptation in new microenvironment. In the present study, a Chinese hamster ovary (CHO) cell line producing glycosylated recombinant human erythropoietin (rhEPO) was adapted for growth and rhEPO production in serum- and protein-free conditions. The physiology, growth parameters, and morphology of the CHO cells and rhEPO biosynthesis and structure were closely monitored during the adaptation process to avoid unwanted selection of cell subpopulations. The results showed that the CHO cells were successfully adapted to suspension growth and rhEPO production in the protein-free conditions and that the structure of rhEPO remained nearly unchanged. In addition, during rhEPO production in the protein-free suspension conditions, the agitation rate seem to be significant for optimal process performance in contrast to the initial cell concentration, evaluated through evolutionary operation method.