Comparative Study on Assisted Solvent Extraction Techniques for the Extraction of Biologically Active Compounds from Sideritis raeseri and Sideritis scardica.

The plants in the Sideritis genus are postulated to exhibit several important medicinal properties due to their unique chemical composition. To isolate the targeted phytochemical compounds, the selection of a suitable extraction method is of primary importance. In this work, a comparative study on the phytochemical profiles of various Sideritis raeseri and Sideritis scardica extracts has been carried out. An untargeted metabolomics approach based on ultra-high performance liquid chromatography coupled with high-resolution mass spectrometry was applied to investigate the metabolic differences between extracts obtained by conventional extraction and extractions assisted by microwaves, ultrasounds and high pressure. Additionally, the influence of extraction solvents on HPLC antioxidant profiles obtained following the derivatization of analytes with ABTS reagent was evaluated. A total of 102 metabolites have been putatively identified. The major secondary metabolites groups were classified as flavonoids, terpenoids, phenylethanoid glycosides and phenolic acids. The main antioxidants in the extracts were isoscutellarein and hypolaetin derivatives as well as verbascoside and chlorogenic acid. The results showed that 70% ethanol was the most effective extractant for different classes of phytochemicals including antioxidants. In addition, extraction supported with microwaves, ultrasounds or high pressure improved the overall recovery of metabolites by about 3 times compared to the conventional extraction method.

Kombucha from alternative raw materials - The review.

Nowadays, people's awareness about the role of diet in maintaining well-being and good health has increased. Consumers expect that the products not only provide them with essential nutrients but will also be a source of biologically active substances, which are beneficial to their health. One of the "healthy trends," which has appeared among the consumers worldwide is kombucha, a tea drink with high antioxidant potential, obtained through the activity of a consortium of acetic acid bacteria and osmophilic yeast, which is also called "tea fungus." Kombucha obtained from tea is characterized by its health-promoting properties. Promising results in in vitro and in vivo studies have prompted research groups from around the world to search for alternative raw materials for tea fungus fermentation. Attempts are made to obtain functional beverages from leaves, herb infusions, vegetable pulp, fruit juices, or milk. This review focuses on describing the progress in obtaining a fermented beverage and bacterial cellulose using tea fungus on alternative raw materials.

The Effect of High Pressure and Subzero Temperature on Gelation of Washed Cod and Salmon Meat.

The objective of the present work is to examine the influence of pressure up to 193 MPa at subzero temperature (without freezing of water) on myofibrillar proteins of salmon and cod meat and on the properties of gels obtained from washed mince of these fish. The solubility of proteins from myofibrils of cod and salmon meat suspended in 100 mM KCl solution increased after treating the samples with pressure above 60 MPa. The results of SDS- -PAGE analysis showed that under these conditions two myosin light chains, tropomyosin and troponin T were released from myofibrils. The solubility of proteins in 0.9 M NaCl solution of washed fish meat after pressure treatment at 60 MPa and -5 °C decreased to about 80-90% and at 193 MPa and -20 °C to 60%. Pressurization of cod meat decreased only slightly the solubility of proteins in SDS and urea solution and the solubility of salmon meat was similar to that in the unpressurized sample. There were no differences in the electrophoretic pattern of proteins from untreated and pressurized cod and salmon meat in the range of 60 to 193 MPa and -5 to -20 °C. The pressure treatment of washed salmon and cod meat at a temperature below 0 °C induced gelation; on the other hand, hardness of gels was lower by 28 and 26%, respectively, than that of gels formed by heating. The salmon and cod gels pressurized at 193 MPa and -20 °C and then heated were much harder than only pressurized or heated gels.

The effect of high pressure at subzero temperature on proteins solubility, drip loss and texture of fish (cod and salmon) and mammal's (pork and beef) meat.

One of the possibilities of using high-pressure technique in inactivation of microorganism is conducting this process at subzero temperature. However, for its practical application in meat preservation the appropriate properties of meat should be maintained. Therefore, the aim of this work was to examine the effect of pressure at subzero temperature (without freezing of water) on proteins and texture of mammal's and cold-adapted fish meat. The data showed that cod and salmon meat proteins were more susceptible to pressure-induced denaturation/aggregation than beef and pork proteins. Glucose and saccharose exerted protective effect on fish meat proteins treated with pressure of 111 MPa(tc) and -10 degrees C but not at 193 MPa(tc) and -20 degrees C. The pressure treatment under the latter conditions increased cook loss of fish meat but not of mammal's meat. However, after cooking the hardness of all kinds of pressurized meat was at the same level as that for unpressurized cooked samples.

The Effect of Hyperbaric Storage on the Nutritional Value and Retention of Certain Bioactive Proteins in Human Milk.

Human milk (HM) contains the essential macronutrients and bioactive compounds necessary for the normal growth and development of newborns. The milk collected by human milk banks is stored frozen and pasteurized, reducing its nutritional and biological value. The purpose of this study was to determine the effect of hyperbaric storage at subzero temperatures (HS-ST) on the macronutrients and bioactive proteins in HM. As control samples, HM was stored at the same temperatures under 0.1 MPa. A Miris HM analyzer was used to determine the macronutrients and the energy value. The lactoferrin (LF), lysozyme (LYZ) and α-lactalbumin (α-LAC) content was checked using high-performance liquid chromatography, and an ELISA test was used to quantify secretory immunoglobulin A (sIgA). The results showed that the macronutrient content did not change significantly after 90 days of storage at 60 MPa/-5 °C, 78 MPa/-7 °C, 111 MPa/-10 °C or 130 MPa/-12 °C. Retention higher than 90% of LYZ, α-LAC, LF and sIgA was observed in the HM stored at conditions of up to 111 MPa/-10 °C. However, at 130 MPa/-12 °C, there was a reduction in LYZ and LF, by 39 and 89%, respectively. The storage of HM at subzero temperatures at 0.1 MPa did not affect the content of carbohydrates or crude and true protein. For fat and the energy value, significant decreases were observed at -5 °C after 90 days of storage.

Effect of convection and microwave heating on the retention of bioactive components in human milk.

Bioactive substances are very important components of human milk (HM), especially for premature newborns. The effects of convection (CH) and microwave heating (MWH) at 62.5 and 66 °C, on the level of selected bioactive components of HM: lysozyme (LZ), lactoferrin (LF), secretory immunoglobulin A (sIgA), basal lipase (BL), cytokine TGF-2, vitamin C and total antioxidant capacity (TAC) was compared. Regardless of the used heating methods the TAC of HM, determined by TEAC and ORAC-FL assay, proved to be insensitive to temperature pasteurization, in contrary to BL. MWH in the conditions of 62.5 for 5 min and 66 °C for 3 min are ensuring microbiological safety with a higher retention of most of the tested active HM proteins compared to CH. Only in the case of LZ the MWH had a more degradative effect on its concentration. Controlled conditions of MWH preserve the bioactive components of the HM better than CH.

Effect of microwave and convection heating on selected nutrients of human milk.

The aim of this study was to determine the influence of the microwave heating method (MWH) on selected determinants of the nutritional value of human milk (HM) and compare to the effect exerted by the standard convection heating (CH) method, including holder pasteurization (HoP). It was showed that using MWH under conditions assumed to ensure microbiological safety, changes in the level of the nutrients were not observed. In these conditions concentration of fatty acids (FAs), lipid peroxides (LP) and α-lactalbumin (α-La) were on a similar level as in raw milk, and furosine was not formed. MWH treatment of HM led to slight increase in the malonaldehyde (MDA) and protein carbonyls (PC) content. However, MDA content in HM as a result of MWH was lower than after application CH.

Effect of lysozyme or nisin on survival of some bacteria treated with high pressure at subzero temperature.

The aim of this work was to examine the inactivation of some Gram-positive and Gram-negative bacteria exposed to the pressure of 193 MPa at -20 °C in the presence of lysozyme or nisin at concentration of 400 µg/ml. The highest effect of pressure at subzero temperature and lysozyme was found with pressure sensitive Pseudomonas fluorescens; viable cells of this strain were not detected in 1 ml of sample after combined treatment. The action of pressure at subzero temperature and lysozyme or nisin against Escherichia coli led to synergistic reduction by 0.7 or 1.6 log cycles, respectively, while it was practically insignificant for two Staphylococcus aureus strains. Viability loss of E. coli and S. aureus occurred during storage for 20 h of the samples at 37 and 5 °C, which were previously pressurized with lysozyme or nisin. The synergistic effect of pressure and nisin at pH 5 against E. coli cells just after the pressure treatment was lower than that at pH 7, however, the extent of the lethal effect after storage was higher.

Assessment of the usefulness of bacterial cellulose produced by Gluconacetobacter xylinus E25 as a new biological implant.

Bionanocellulose (BNC) is a clear polymer produced by the bacterium Gluconacetobacter xylinus. In our current study, "Research on the use of bacterial nanocellulose (BNC) in regenerative medicine as a function of the biological implants in cardiac and vascular surgery", we carried out material analysis, biochemical analysis, in vitro tests and in vivo animal model testing. In stage 1 of the project, we carried out physical and biological tests of BNC. This allowed us to modify subsequent samples of bacterial bionanocellulose. Finally, we obtained a sample that was accepted for testing on an animal model. That sample we define BNC1. Patches of BNC1 were then implanted into pigs' vessel walls. During the surgical procedures, we evaluated the technical aspects of sewing in the bioimplant, paying special attention to bleeding control and tightness of the suture line and the BNC1 bioimplant itself. We carried out studies evaluating the reaction of an animal body to an implantation of BNC1 into the circulatory system, including the general and local inflammatory reaction to the bioimplant. These studies allowed us to document the potential usefulness of BNC as a biological implant of the circulatory system and allowed for additional modifications of the BNC to improve the properties of this new implantable biological material.

Changes in bacterial cells induced by high pressure at subzero temperature.

The aim of this study was to examine the effect of pressure treatment at 193MPa and -20°C on membrane damage, changes in activity of membrane-bound ATPases and degradation of nucleic acids. The experiments were carried out with three Escherichia coli strains, in the exponential and stationary phases of growth, and differing in sensitivity to pressure. All E. coli strains subjected to pressure in the exponential phase of growth were inactivated by 6 log cycles, independently of the strain, which was accompanied by a total loss of ability to plasmolyse, an increase in irreversible membrane permeability to PI, and a reduction of cellular ATP by more than 80%. After pressure treatment of stationary phase cells, the relationship between the inactivation level and the ability to plasmolyse was not as evident as in the case of exponential phase cells. Pressure treatment of two strains of E. coli K-12 and Ec160/59 in the stationary phase that decreased viability by no more than one log cycle led only to reversible permeabilization of bacterial membranes, while irreversible permeabilization was observed in the pressure sensitive E. coli IBA72 strain phase that was inactivated by 4.6 log cycles. The reduction of ATP and changes in ATPase activity after pressure treatment of tested E. coli strains in the stationary phase of growth depended on the stage of inactivation of the particular strain. Electrophoretic analysis showed degradation of RNA isolated after pressure treatment from cells of all E. coli strains tested in the exponential phase of growth. The changes of RNA induced by pressure were not visible in the case of cells in the stationary phase. The degradation of DNA isolated from pressure treated E. coli strains from the exponential as well as from the stationary phase of growth was not observed.

Effect of lysozyme or nisin on survival of some bacteria treated with high pressure at subzero temperature

The aim of this work was to examine the inactivation of some Gram-positive and Gram-negative bacteria exposed to the pressure of 193 MPa at -20 ºC in the presence of lysozyme or nisin at concentration of 400 mg/ml. The highest effect of pressure at subzero temperature and lysozyme was found with pressure sensitive Pseudomonas fluorescens; viable cells of this strain were not detected in 1 ml of sample after combined treatment. The action of pressure at subzero temperature and lysozyme or nisin against Escherichia coli led to synergistic reduction by 0.7 or 1.6 log cycles, respectively, while it was practically insignificant for two Staphylococcus aureus strains. Viability loss of E. coli and S. aureus occurred during storage for 20 h of the samples at 37 and 5 ºC, which were previously pressurized with lysozyme or nisin. The synergistic effect of pressure and nisin at pH 5 against E. coli cells just after the pressure treatment was lower than that at pH 7, however, the extent of the lethal effect after storage was higher.