Development of Cyclodextrin-Based Mono and Dual Encapsulated Powders by Spray Drying for Successful Preservation of Everlasting Flower Extract.

The study aimed to develop encapsulation systems to maintain the preservation of everlasting (Helichrysum plicatum) flower extract polyphenols. Spray-dried encapsulates were formulated using ß-cyclodextrin (BCD) and 2-hydroxypropyl-ß-cyclodextrin (HPBCD) as supramolecular hosts, and their macromolecule mixtures with the conventional carriers, maltodextrin (MD) and whey protein (WP). The obtained microparticles were comparatively assessed regarding technological, physicochemical, and phytochemical properties. The highest yields were achieved by combining cyclodextrins with whey protein (73.96% for WP+BCD and 75.50% for WP+HPBCD compared to 62.48% of pure extract). The extract-carrier interactions and thermal stability were evaluated by FTIR and DSC analysis, suggesting successful entrapment within the carriers. Carriers reduced the particle diameter (3.99 to 4.86 µm compared to 6.49 µm of pure extract), classifying all encapsulates as microsystems. Carrier blends made the particle size distribution uniform, while SEM analysis revealed the production of more spherical and less aggregated particles. The HPBCD provided the highest encapsulation efficiency, with the highest content of detected aglycones and slightly lower values of their glycosylated forms. An analysis of the dual macromolecule encapsulation systems revealed the highest bioactive preservation potential for SHE+MD+BCD and SHE+WP+HPBCD. Overall, macromolecule combinations of cyclodextrins and conventional biopolymers in the spray-drying process can enhance the functional properties of H. plicatum extract.

Spasmolytic, Antimicrobial, and Antioxidant Activities of Spray-Dried Extracts of Gentiana asclepiadea L. with In Silico Pharmacokinetic Analysis.

This study aimed to evaluate the spasmolytic activity of an underground parts extract of Gentiana asclepiadea L. (Gentianaceae), assess its antioxidant and antimicrobial activities, and explore the impact of extract encapsulation on the aforementioned bioactivities. An extract encapsulated by spray drying with whey protein, pure extract, and pure whey protein were comparatively tested. The main compounds identified via HPLC-DAD analysis underwent in silico ADME assessment. The spasmolytic effect was tested on a model of spontaneous rat ileum contractions, and the mechanism of action was further evaluated on acetylcholine-, KCl-, CaCl2-, BaCl2-, histamine-, N(ω)-nitro-L-arginine methyl ester-, and glibenclamide-modified contractions. The most abundant compounds were secoiridoids (dominantly gentiopicroside), followed by C-glycosylated flavonoids and xanthones. Both pure and encapsulated extracts achieved significant spasmolytic effects, despite the spasmogenic activity of pure whey protein. The extract may exert its spasmolytic effect through multiple pathways, predominantly by antagonizing the Ca2+ channel and opening the K+ channel, while the nitric oxide pathway appears not to be involved. The antimicrobial and antioxidant activities of the pure extract were moderate. The extract stabilized by encapsulation retained all of the tested bioactivities of the unencapsulated extract. The obtained results suggest that G. asclepiadea has potential for use in the treatment of some gastrointestinal complaints and that the encapsulated extract could be a valuable functional ingredient in pharmaceutical and food products.

Microencapsulated Bilberry and Chokeberry Leaf Extracts with Potential Health Benefits.

The aim of the research was to develop microencapsulated powders of bilberry and chokeberry extracts via the spray drying technique. Two biopolymers, pectin alone and in combination with HP-ß-CD, were used to preserve the antioxidant, hypoglycemic, photoprotective, and antimicrobial bioactivity of the berry leaf extracts. Moreover, the formed powders were characterized in terms of technological, chemical, and several biological properties. The obtained micro-sized powders (mean average particle diameter from 3.83 to 5.94 µm) demonstrated a process yield of up to 73%. The added biopolymers improved the flowability and cohesive properties of the powders and increased their thermal stability to 170 °C. The total content of polyphenolics in the powders ranged from 323.35 to 367.76 mg GAE/g DW for bilberry and from 186.85 to 227.59 mg GAE/g DW for chokeberry powders; meanwhile, chlorogenic acid was the predominant compound in powders. All samples showed stronger α-glucosidase inhibitory activity (IC50 values ranged from 5.00 to 19.59 µg/mL) compared with the reference standard. The study confirmed that spray drying is a suitable method for the preservation of the polyphenolic-rich extracts, while the addition of carriers has a positive effect on the improvement of microencapsulated powders' properties.

Leaves micromorphology, chemical profile, and bioactivity of in vitro-propagated Nepeta cyrenaica (Lamiaceae).

INTRODUCTION: The endemic species Nepeta cyrenaica Quézel & Zaffran, native to northeastern Libya, is valued as an important honey-bearing plant. OBJECTIVES: This study was aimed to examine the micromorphology, phytochemistry, and bioactivity of in vitro-propagated N. cyrenaica for the first time. MATERIALS AND METHODS: The leaf indumentum was examined using light and scanning electron microscopy and further characterised for histochemistry. The chemical composition of essential oil (EO) was performed using GC-MS analysis, while dichloromethane (DCM), methanol (ME), ethanol (ET), and aqueous (AQ) extracts were analysed using qualitative and quantitative LC/MS analyses. The antioxidant activities of EO and extracts were assessed using three parallel assays, while enzyme-inhibiting effects were evaluated against four enzymes. RESULTS: The leaves bear various types of glandular trichomes, with lipophilic secretion predominating. The main EO component of EO was 1,8-cineole. A considerable number of phenolics and iridoids were tentatively identified in the ME extract. Quantitative LC/MS analysis confirmed that ferulic acid, rosmarinic acid, and epigallocatechin gallate were present in the highest amount in the extracts, in which three iridoids were also quantified. Although the ME extract contained the highest amount of polyphenolics and iridoids, the DCM extract showed the best overall biological potential. Additionally, EO exerted the strongest acetylcholinesterase and tyrosinase inhibition. CONCLUSION: This study demonstrated that the endemic N. cyrenaica can be efficiently grown under in vitro conditions, where it develops various glandular trichomes that are thought to secrete and/or accumulate bioactive compounds with valuable medicinal potential.

Pomegranate Peel Extract Attenuates Isoprenaline-Induced Takotsubo-like Myocardial Injury in Rats.

Takotsubo syndrome (TTS) is an acute heart failure syndrome characterised by catecholamine-induced oxidative tissue damage. Punica granatum, a fruit-bearing tree, is known to have high polyphenolic content and has been proven to be a potent antioxidant. This study aimed to investigate the effects of pomegranate peel extract (PoPEx) pre-treatment on isoprenaline-induced takotsubo-like myocardial injury in rats. Male Wistar rats were randomised into four groups. Animals in the PoPEx(P) and PoPEx + isoprenaline group (P + I) were pre-treated for 7 days with 100 mg/kg/day of PoPEx. On the sixth and the seventh day, TTS-like syndrome was induced in rats from the isoprenaline(I) and P + I groups by administering 85 mg/kg/day of isoprenaline. PoPEx pre-treatment led to the elevation of superoxide dismutase and catalase (p < 0.05), reduced glutathione (p < 0.001) levels, decreased the thiobarbituric acid reactive substances (p < 0.001), H2O2, O2- (p < 0.05), and NO2- (p < 0.001), in the P + I group, when compared to the I group. In addition, a significant reduction in the levels of cardiac damage markers, as well as a reduction in the extent of cardiac damage, was found. In conclusion, PoPEx pre-treatment significantly attenuated the isoprenaline-induced myocardial damage, primarily via the preservation of endogenous antioxidant capacity in the rat model of takotsubo-like cardiomyopathy.

Microencapsulates of Blue Maize Polyphenolics as a Promising Ingredient in the Food and Pharmaceutical Industry: Characterization, Antioxidant Properties, and In Vitro-Simulated Digestion.

An anthocyanin-rich blue maize waste product was used for anthocyanin extraction. To preserve bioactive phenolic compounds, a spray-drying technique was employed using conventional wall material maltodextrin (MD), with novel one, hydroxypropyl-ß-cyclodextrin (HPBCD). The obtained spray-dried maize extract (SME) and microencapsulates were analyzed based on physicochemical powder properties, chemical analysis, antioxidant activity, and digestibility. The examined microencapsulates demonstrated good powder properties, exhibited a high powder yield (up to 83%), and had a low moisture content (less than 5%). HPBCD and MD + HPBCD combinations demonstrated superior powder properties in the terms of decreasing the time necessary for rehydration (133.25 and 153.8 s, respectively). The mean average particle diameter ranged from 4.72 to 21.33 µm. DSC analyses signified high powder thermal stability, around 200 °C, related to the increasing preservation with biopolymer addition. The total phenolic and anthocyanin compounds ranged from 30,622 to 32,211 mg CE/kg (CE-catechin equivalents) and from 9642 to 12,182 mg CGE/kg (CGE-cyanidin 3-glucoside equivalents), respectively, associated with good bioactive compound protection. Microencapsulates with both carriers (15% MD and 15% HPBCD) had the highest digestibility (73.63%). Our results indicated that the microencapsulates created with the active ingredient and the wall materials (MD and HPBCD) could protect phenolic compounds/anthocyanins against ABTS radicals (63.53 and 62.47 mmol Trolox Eq/kg, respectively).

The Application of Ultrasonic Waves and Microwaves to Improve Antihyperglycaemic and Antimicrobial Activities of Marrubium vulgare Extracts.

Having scarce information about ultrasound assisted extraction (UAE) and microwave assisted extraction (MAE) of white horehound (Marrubium vulgare L.), the idea has emerged to determine the optimal process parameters for the maximization of polyphenols and to compare the efficiency of these green extraction technologies. The optimal UAE parameters are temperature of 73.6 °C, extraction time of 40 min and ultrasound power of 30.3 W/L, while the optimal MAE parameters are 63.8% ethanol, extraction time of 15 min and microwave power of 422 W. Extract obtained at optimal UAE parameters shows the highest antihyperglycemic activity (α-amylase inhibition: 50.63% and α-glucosidase inhibition: 48.67%), which can potentially be explained by the presence of chlorogenic acid and quercetin, which were not identified in the macerates. The most sensitive bacterial strain to optimal ultrasonic extract is Bacillus cereus, whereas the most sensitive fungal strain is Saccharomyces cerevisiae.

Ultrasound-Assisted Water Extraction of Gentiopicroside, Isogentisin, and Polyphenols from Willow Gentian "Dust" Supported by Hydroxypropyl-ß-Cyclodextrin as Cage Molecules.

The residue after sieving ("dust") from the willow gentian underground parts is an unexploited herbal tea by-product, although it contains valuable bioactive compounds. Cyclodextrins as efficient green co-solvents, cage molecules, and multifunctional excipients could improve the extraction and contribute to the added value of the resulting extracts. The objective of this study was to determine the optimal conditions for the extraction of gentiopicroside, isogentisin, and total phenolics (TPC) from willow gentian "dust" using ultrasound-assisted water extraction coupled with hydroxypropyl-ß-cyclodextrin (HPßCD). The influence of extraction temperature (X1: 20-80 °C), time (X2: 20-50 min), and HPßCD concentration (X3: 2-4% w/v) was analyzed employing the response surface methodology (RSM). The optimal extraction conditions for simultaneously maximizing the extraction yield of all monitored responses were X1: 74.89 °C, X2: 32.57 min, and X3: 3.01% w/v. The experimentally obtained response values under these conditions (46.96 mg/g DW for gentiopicroside, 0.51 mg/g DW for isogentisin, and 12.99 mg GAE/g DW for TPC) were in close agreement with those predicted, thus confirming the suitability and good predictive accuracy of the developed RSM models. Overall, the developed extraction system could be an applicable alternative strategy to improve the extraction of bioactive compounds from the underutilized "dust" of willow gentian underground parts.

Black Elderberry Press Cake as a Source of Bioactive Ingredients Using Green-Based Extraction Approaches.

To study the efficiency of two green-based extraction techniques for the isolation of bioactive constituents from black elderberry press cake, changes in phenolic compounds and main anthocyanin contents were analyzed. Polyphenolic content was correlated with antioxidant and antidiabetic potential by radical-scavenging activity and monitoring of α-amylase inhibition. Black elderberry press-cake extracts were obtained by ultrasound-assisted (UAE) and microwave-assisted (MAE) extractions under different extraction conditions. High-performance liquid chromatography (HPLC) analysis revealed that cyanidin-3-sambubioside and cyanidin-3-glucoside were the principal anthocyanins in all the obtained extracts, with their content being highest in MAE obtained at 80 °C over 5 min. The same extract induced two-fold higher antioxidant activity (IC50 6.89 µg/mL) and α-amylase inhibitory potential (IC50 2.18 mg/mL) in comparison to UAE extracts. The main compositional differences between extracts obtained by the same extraction technique were assigned to the extraction temperature. A principal component analysis confirmed that the antidiabetic feature is to be attributed to the rich content of anthocyanins in black elderberry press cake. Our results indicate the great potential of underutilized black elderberry press cake for the development of novel food and herbal formulations due to notable anthocyanin contents highly correlated with antidiabetic activity.

A prospective, randomized, double-blind, placebo-controlled trial of polyphenols on the outcomes of inflammatory factors and oxidative stress in patients with type 2 diabetes mellitus.

BACKGROUND: Type 2 diabetes mellitus (T2DM) is commonly associated with hyperglycemia, dyslipidemia, oxidative stress and inflammation which are well known cardiovascular risk factors. Pomegranate peel polyphenols have a proven hypolipemic, antioxidant and anti-inflammatory activity. However, there is a lack of clinical studies that would confirm its antioxidant and anti-inflammatory effects in diabetic patients. The potential of pomegranate peel extract (PoPEx) to counteract inflammation and oxidative stress in T2DM patients was investigated. For this purpose, a randomized, double-blind placebo-controlled study involving adult T2DM patients treated with PoPEx or placebo for eight-weeks was conducted. METHODS: Patients were randomly divided into two groups: the first group (n = 30) received capsules containing PoPEx 250 mg twice daily, while the placebo group (n = 30) received placebo capsules twice daily. Plasma concentration of inflammatory factors (interleukin 6 (IL-6), tumor necrosis factor α (TNF-α) and high sensitivity C reactive protein (hsCRP)), oxidative stress biomarkers (thiobarbituric acid reactive substances (TBARS), nitrites (NO2-), superoxide anion radical (O2-), hydrogen peroxide (H2O2), total antioxidant capacity (TAC)), homocysteine and lipid profile were analyzed. RESULTS: The PoPEx treatment showed a significant reduction of inflammatory factors (IL-6, TNF-α, hsCRP), oxidative stress biomarkers (TBARS, NO2-, O2-) and homocysteine, while the TAC was increased. Moreover, a significant improvement in lipid profile was observed in the PoPEx group. Additional analysis showed a significant inverse correlation between the decrements of all measured inflammatory markers and TAC in the PoPEx group. CONCLUSIONS: The study demonstrated that eight-week-long PoPEx administration had favorable effects on inflammatory status and oxidative stress biomarkers in diabetic patients.

Liposomal Bilayer as a Carrier of Rosa canina L. Seed Oil: Physicochemical Characterization, Stability, and Biological Potential.

Rosa canina L. seeds are rich in bioactive components that can add value to the various formulations. The focus of the study was the development of liposomes for R. canina oil to protect its sensitive compounds and prolong their shelf-life. Oil-loaded liposomes were characterized via the determination of the particle size, polydispersity index (PDI), zeta potential, conductivity, mobility, density, surface tension, viscosity, and stability. Raman and FT-IR spectroscopy were employed to investigate the chemical composition of the non-treated and UV-treated samples, and the presence of different interactions. Antioxidant and antimicrobial activities were examined as well. The liposome size was 970.4 ± 37.4 nm, the PDI 0.438 ± 0.038, the zeta potential -32.9 ± 0.8 mV, the conductivity 0.068 ± 0.002 mS/cm, the mobility -2.58 ± 0.06 µmcm/Vs, the density 0.974 ± 0.004 g/cm3, the surface tension 17.2 ± 1.4 mN/m, and the viscosity 13.5 ± 0.2 mPa•s. The Raman and FT-IR spectra showed the presence of lipids, fatty acids, polyphenols, and carotenoids. It was approved that the oil compounds were distributed inside the phospholipid bilayer and were combined with the membrane interface of the bilayer. The UV irradiation did not cause any chemical changes. However, neither the pure oil nor the oil-loaded liposomes showed any antimicrobial potential, while the antioxidant capacity of the oil-loaded liposomes was significantly low. The sizes of the liposomes did not change significantly during 60 days of storage. Due to the proven stability of the oil-loaded liposomes, as well as the liposome's ability to protect the sensitive oil compounds, their potential application in the pharmaceutical and cosmetic formulations could be investigated with a focus on the skin regeneration effects.

A Study of Phytochemistry, Genoprotective Activity, and Antitumor Effects of Extracts of the Selected Lamiaceae Species.

This study was designed to evaluate the genoprotective, antigenotoxic, as well as antitumor potential of methanolic, ethanolic, and aqueous extracts of Melissa officinalis, Mentha × piperita, Ocimum basilicum, Rosmarinus officinalis, Salvia officinalis, and Satureja montana (Lamiaceae), in different model systems. The polyphenols in these extracts were quantified both spectrophotometrically and using HPLC-DAD technique, while DPPH assay was used to assess the antioxidant activity. The genoprotective potential was tested on pUC19 Escherichia coli XL1-blue, and the antigenotoxicity on Salmonella typhimurium TA1535/pSK1002 and human lung fibroblasts, while the antitumor activity was assessed on colorectal cancer cells. Rosmarinic acid, quercetin, rutin, and luteolin-7-O-glucoside were among the identified compounds. Methanolic extracts had the best DPPH-scavenging and SOS-inducing activities, while ethanolic extracts exhibited the highest antigenotoxicity. Additionally, all extracts exhibited genoprotective potential on plasmid DNA. The antitumor effect was mediated by modulation of reactive oxygen species (ROS), nitric oxide (NO) production, and exhibition of genotoxic effects on tumor cells, especially with O. basilicum ethanolic extract. Generally, the investigated extracts were able to provide antioxidant protection for the acellular, prokaryotic, and normal human DNA, while also modulating the production of ROS and NO in tumor cells, leading to genotoxicity toward these cells and their decrease in proliferation.

Effect of Type and Concentration of Carrier Material on the Encapsulation of Pomegranate Peel Using Spray Drying Method.

This study aimed to establish a procedure for pomegranate peel (PP) valorization and attainment of stable extracts with preserved bioactive compounds. The technology applied was spray drying with carbohydrate-based (maltodextrin, MD) and protein-based (whey protein, WP) carrier materials in different concentrations (80, 100, and 120%). What was analyzed was the impact of the type and concentration of carrier material on the stability and quality of the final encapsulated powder. The best results were achieved when the PP extract was microencapsulated with the carbohydrate-based carrier (100%), where it had the highest encapsulation efficiency (EE) (88.63%), hygroscopicity (15.17%), and water solubility index (87.04%). The moisture content was in the range of 3.69-4.60% and 4.21-5.84% for MD and WP, respectively, indicating that both are suitable for long-term storage. It was observed that changes in carrier concentration significantly influenced most of the powders' physicochemical properties. Microencapsulation using MD yielded a higher content of punicalin, punicalagin, gallic, and ellagic acid than those with WP. Overall results demonstrated that carbohydrate-based microencapsulation can be utilized efficiently for the protection of powder stability and phytochemical characteristics.

Pomegranate peel extract polyphenols attenuate the SARS-CoV-2 S-glycoprotein binding ability to ACE2 Receptor: In silico and in vitro studies.

The novel coronavirus disease (Covid-19) has become a major health threat globally. The interaction of SARS-CoV-2 spike (S) glycoprotein receptor-binding domain (RBD) with ACE2 receptor on host cells was recognized as the first step of virus infection and therefore as one of the primary targets for novel therapeutics. Pomegranate extracts are rich sources of bioactive polyphenols that were already recognized for their beneficial health effects. In this study, both in silico and in vitro methods were employed for evaluation of pomegranate peel extract (PoPEx), their major polyphenols, as well as their major metabolite urolithin A, to attenuate the contact of S-glycoprotein RBD and ACE2. Our results showed that PoPEx, punicalin, punicalagin and urolithin A exerted significant potential to block the S-glycoprotein-ACE2 contact. These in vitro results strongly confirm the in silico predictions and provide a valuable insight in the potential of pomegranate polyphenols for application in SARS-CoV-2 infection.

Optimization of Ultrasound-Assisted Extraction of Phenolics from Sideritis raeseri Using Response Surface Methodology.

In this study we define the optimal conditions for ultrasound-assisted extraction of bioactive polyphenols from S. raeseri aerial parts using response surface methodology. The influence of ethanol concentration (10-90%), extraction temperature (20-80 °C), extraction time (10-60 min), and solid-to-solvent ratio (1:10-1:50) on total phenolic content as well as on content of individual flavonoids, and hypolaetin and isoscutellarein derivatives was studied. For the experimental design, a central composite design was chosen. In the obtained extracts, the following ranges of targeted compounds were detected: total phenol from 19.32 to 47.23 mg GAE/g dw, HYP from 1.05 to 11.46 mg/g dw, ISC 1 from 0.68 to 10.68 mg/g dw, and ISC 2 from 0.74 to 15.56 mg/g dw. The optimal extraction conditions were set as: ethanol concentration of 65%, extraction time of 50 min, extraction temperature of 63 °C, and solid-to-solvent ratio of 1:40. Contents of TP, HYP, ISC 1, and ISC 2 in optimal extracts were 47.11 mg GAE/g dw, 11.73 mg/g dw, 9.54 mg/g dw, and 15.40 mg/g dw, respectively. Experimentally set values were in good agreement with those predicted by the response surface methodology model, indicating suitability of the used model, as well as the success of response surface methodology in optimizing the conditions of the extraction.

A comprehensive assessment of the chemical composition, antioxidant, genoprotective and antigenotoxic activities of Lamiaceae species using different experimental models in vitro.

This research was aimed to assess the potential of Glechoma hederacea, Hyssopus officinalis, Lavandula angustifolia, Leonurus cardiaca, Marrubium vulgare and Sideritis scardica (Lamiaceae) methanolic, ethanolic and aqueous extracts against the damaging effects of oxidative stress using different experimental models. The chemical characterization was done spectrophotometrically by quantifying total phenolics, phenolic acids, flavonoids and flavonols in the extracts, as well as by employing HPLC-DAD technique. Moreover, DPPH assay was used to assess the extracts' radical scavenging potential. Genoprotective properties of the extracts were evaluated using plasmid pUC19 Escherichia coli XL1-Blue, whereas their antigenotoxic potential was determined using Salmonella typhimurium TA1535/pSK1002 and normal human lung fibroblasts. All of the extracts showed antioxidant activity in DPPH assay. Furthermore, the results have shown that aqueous extracts provided the best protection for plasmid DNA, while alcoholic extracts most effectively contributed to the preservation of prokaryotic DNA. Additionally, each of the tested samples significantly protected the eukaryotic cells against genomic damages. Finally, despite not showing exceptional results in DPPH assay, S. scardica extracts are regarded as the most favorable in maintaining the integrity of DNA, which might be due to high quantities of phenolics such as quercetin (up to 17.95 mg g-1), naringin (up to 5.07 mg g-1) and luteolin-7-O-glucoside (up to 3.54 mg g-1). Overall, this comprehensive concept highlights the ability of these Lamiaceae species to safeguard the DNA from reactive oxygen species, to curtail the inflicted damage and also improve the efficiency of the DNA repair mechanisms, while emphasizing the importance of polyphenols as their active principles.

Computational study of pomegranate peel extract polyphenols as potential inhibitors of SARS-CoV-2 virus internalization.

The search for effective coronavirus disease (COVID-19) therapy has attracted a great deal of scientific interest due to its unprecedented health care system overload worldwide. We have carried out a study to investigate the in silico effects of the most abundant pomegranate peel extract constituents on the multi-step process of serious acute respiratory syndrome coronavirus 2 (SARS-CoV-2) internalization in the host cells. Binding affinities and interactions of ellagic acid, gallic acid, punicalagin and punicalin were studied on four selected protein targets with a significant and confirmed role in the process of the entry of virus into a host cell. The protein targets used in this study were: SARS-CoV-2 spike glycoprotein, angiotensin-converting enzyme 2, furin and transmembrane serine protease 2. The results showed that the constituents of pomegranate peel extracts, namely punicalagin and punicalin had very promising potential for significant interactions with the selected protein targets and were therefore deemed good candidates for further in vitro and in vivo evaluation.

Mahonia aquifolium Extracts Promote Doxorubicin Effects against Lung Adenocarcinoma Cells In Vitro.

Mahonia aquifolium and its secondary metabolites have been shown to have anticancer potential. We performed MTT, scratch, and colony formation assays; analyzed cell cycle phase distribution and doxorubicin uptake and retention with flow cytometry; and detected alterations in the expression of genes involved in the formation of cell-cell interactions and migration using quantitative real-time PCR following treatment of lung adenocarcinoma cells with doxorubicin, M. aquifolium extracts, or their combination. MTT assay results suggested strong synergistic effects of the combined treatments, and their application led to an increase in cell numbers in the subG1 phase of the cell cycle. Both extracts were shown to prolong doxorubicin retention time in cancer cells, while the application of doxorubicin/extract combination led to a decrease in MMP9 expression. Furthermore, cells treated with doxorubicin/extract combinations were shown to have lower migratory and colony formation potentials than untreated cells or cells treated with doxorubicin alone. The obtained results suggest that nontoxic M. aquifolium extracts can enhance the activity of doxorubicin, thus potentially allowing the application of lower doxorubicin doses in vivo, which may decrease its toxic effects in normal tissues.

Immunomodulatory activity and protective effects of chokeberry fruit extract on Listeria monocytogenes infection in mice.

Chokeberry (Aronia melanocarpa) fruit extracts (CE) are rich in polyphenols and usually exhibit immunomodulatory, anti-viral and anti-bacterial effects. We have previously shown that the CE used in this study activated macrophages and stimulated effector T cell differentiation in vitro. When applied orally to healthy mice, CE increased the proportion of CD11c+ dendritic cells in the gut-associated lymphoid tissue. CE-pretreated BALB/c mice readily eradicated orally ingested Listeria monocytogenes as evidenced by a slighter decrease in body weight and number of bacteria recovered from the spleen and reduced spleen size compared to the control infected mice. CE pretreatment in infected mice resulted in higher proportions of CD11b+ macrophages and CD8+ cytotoxic T cells both in the gut and the spleen. Phagocytosis, reactive oxygen species production and the proportions of activated CD86+ macrophages (CD11b+) and dendritic cells (CD11c+) were also enhanced in CE-pretreated infected mice. Furthermore, the expression of inducible nitric oxide synthase and IL-6 was increased in CE-pretreated infected mice and similar results were obtained in peritoneal macrophages in vitro. This effect of CE was associated with increased phosphorylation of IκB and Notch1 production. Finally, CE pretreatment elevated the proportion of perforin-producing cells in the spleen compared to control infected mice. This study demonstrates that prophylactic treatment with CE leads to more rapid eradication of bacterial infection with L. monocytogenes predominantly through increased activity of myeloid cells in the gut and in the spleen.

Effect of pomegranate peel extract on the oxidative stability of pomegranate seed oil.

Pomegranate seed oil is valuable source of bioactive compounds with health-beneficial effects, but it is sensitive to oxidation due to high content of PUFA. In order to improve stability of pomegranate seed oil, the influence of natural antioxidant (0.1% pomegranate peel extract) and a combination of synthetic antioxidant (0.01% butylhydroxytoluene) and pomegranate peel extract (0.05%) was investigated. The oxidation process was followed by monitoring fatty acid composition and measuring the quality and stability parameters of oil (acid value, peroxide value, anisidine value and thiobarbituric acid reactive substances value) during storage for 12 days at 65 °C. Furthermore, total phenolic content and DPPH radical scavenging activity was measured. Pomegranate peel extract as well as a combination of pomegranate peel extract and BHT can have a significant positive impact (P < 0.05) on improvement of the quality and stability parameters of pomegranate seed oil. Furthermore, such activity was better than BHT used alone.