The Effect of Peppermint and Thyme Oils on Stabilizing the Fatty Acid Profile of Sunflower Oil.

Presently, there is an increasing shift towards the utilization of natural antioxidants and compounds with protective attributes for fatty acids in order to replace synthetic counterparts that may pose health risks. This transition aligns with the growing emphasis on promoting healthy and organic food choices. Essential oils stand out in this context due to scientific validations of their antioxidant properties. There are few published research results concerning changes in the fatty acid composition in model systems with the addition of essential oils. This study aims to investigate the impact of incorporating peppermint and thyme oils on inhibiting changes in the fatty acid profile of sunflower oil stored at both room temperature with exposure to daylight and in a thermostat set at 40 °C. The experimental procedure involved the addition of peppermint and thyme oils, along with butylated hydroxyanisole (BHA), to batches of sunflower oil. The samples were then stored for 11 months. The study observed a detrimental influence of storage conditions on the quantitative changes in the fatty acid profile of the sunflower oil. The addition of BHA stabilized the content of linoleic acid in the sunflower oil (approximately 53 g/100 g of linoleic acid compared to approximately 58 g/100 g in the control sample). Meanwhile, the model system of sunflower oil with the addition of peppermint and thyme oils (40 °C) exhibited a statistically significant decrease in the concentration of linoleic acid to approximately 8 g/100 g after eleven months of thermostating. Similar trends to those observed for linoleic acid were noted for the total fatty acid content in the sunflower oil. Notably, the efficacy of the selected substances in inhibiting adverse transformations in fats was contingent upon their concentration and the storage temperature.

Antinociceptive Effect of a p-Cymene/ß-Cyclodextrin Inclusion Complex in a Murine Cancer Pain Model: Characterization Aided through a Docking Study.

Pain is one of the most prevalent and difficult to manage symptoms in cancer patients, and conventional drugs present a range of adverse reactions. The development of ß-cyclodextrins (ß-CD) complexes has been used to avoid physicochemical and pharmacological limitations due to the lipophilicity of compounds such as p-Cymene (PC), a monoterpene with antinociceptive effects. Our aim was to obtain, characterize, and measure the effect of the complex of p-cymene and ß-cyclodextrin (PC/ß-CD) in a cancer pain model. Initially, molecular docking was performed to predict the viability of complex formation. Afterward, PC/ß-CD was obtained by slurry complexation, characterized by HPLC and NMR. Finally, PC/ß-CD was tested in a Sarcoma 180 (S180)-induced pain model. Molecular docking indicated that the occurrence of interaction between PC and ß-CD is favorable. PC/ß-CD showed complexation efficiency of 82.61%, and NMR demonstrated PC complexation in the ß-CD cavity. In the S180 cancer pain model, PC/ß-CD significantly reduced the mechanical hyperalgesia, spontaneous nociception, and nociception induced by non-noxious palpation at the doses tested (p < 0.05) when compared to vehicle differently from free PC (p > 0.05). Therefore, the complexation of PC in ß-CD was shown to improve the pharmacological effect of the drug as well as reducing the required dose.

Antibacterial Activity of the Essential Oil of Piper tuberculatum Jacq. Fruits against Multidrug-Resistant Strains: Inhibition of Efflux Pumps and ß-Lactamase.

Antimicrobial resistance has become a growing public health concern in recent decades, demanding a search for new effective treatments. Therefore, this study aimed to elucidate the phytochemical composition and evaluate the antibacterial activity of the essential oil obtained from the fruits of Piper tuberculatum Jacq. (EOPT) against strains carrying different mechanisms of antibiotic resistance. Phytochemical analysis was performed using gas chromatography-mass spectrometry (GC/MS). The antibacterial activity of EOPT and its ability to inhibit antibiotic resistance was evaluated through the broth microdilution method. The GC-MS analysis identified 99.59% of the constituents, with ß-pinene (31.51%), α-pinene (28.38%), and ß-cis-ocimene (20.22%) being identified as major constituents. The minimum inhibitory concentration (MIC) of EOPT was determined to assess its antibacterial activity against multidrug-resistant strains of Staphylococcus aureus (IS-58, 1199B, K2068, and K4100). The compound showed a MIC of ≥ 1024 µg/mL, suggesting a lack of intrinsic antibacterial activity. However, when the EOPT was associated with antibiotics and EtBr, a significant decrease in antibiotic resistance was observed, indicating the modulation of efflux pump activity. This evidence was corroborated with the observation of increased fluorescent light emission by the bacterial strains, indicating the involvement of the NorA and MepA efflux pumps. Additionally, the significant potentiation of ampicillin activity against the S. aureus strain K4414 suggests the ß-lactamase inhibitory activity of EOPT. These results suggest that the essential oil from P. tuberculatum fruits has antibiotic-enhancing properties, with a mechanism involving the inhibition of efflux pumps and ß-lactamase in MDR S. aureus strains. These findings provide new perspectives on the potential use of EOPT against antibiotic resistance and highlight the importance of Piper species as sources of bioactive compounds with promising therapeutic activities against MDR bacteria. Nevertheless, further preclinical (in vivo) studies remain necessary to confirm these in vitro-observed results.

Antibacterial Activity and Anxiolytic Effect in Adult Zebrafish of Genus Lippia L. Species.

Species belonging to the genus Lippia are used worldwide as foods, beverages, and seasonings. Studies have demonstrated that these species have antioxidant, sedative, analgesic, anti-inflammatory, and antipyretic activities. This work aimed to evaluate the antibacterial activity and anxiolytic effect by different pathways of essential oils and ethanolic extracts of three species of Lippia (Lippia alba, Lippia sidoides, and Lippia gracilis). The ethanolic extracts were characterized by HPLC-DAD-ESI-MSn and their phenolics were quantified. The antibacterial activity was evaluated by determining the minimal inhibitory concentration and modulation of antibiotic activity, and toxic and anxiolytic effects were evaluated in the zebrafish model. The extracts showed compositions with a low ratio and shared compounds. L. alba and L. gracilis showed higher amounts of phenols and flavonoids, respectively. All extracts and essential oils presented antibacterial activity, especially those obtained from L. sidoides. On the other hand, L. alba extract presented the most significant antibiotic-enhancing effect. The samples were not toxic after 96 h of exposure, but showed an anxiolytic effect through modulation of the GABAA receptor, while L. alba extract acted via modulation of the 5-HT receptor. This new pharmacological evidence opens horizons for therapeutic approaches targeting anxiolytic and antibacterial therapies and food conservation using these species and their constituents.

Synergistic Antioxidant Activity of Four-Component Mixture of Essential Oils: Basil, Cedarwood, Citronella and Thyme for the Use as Medicinal and Food Ingredient.

Mixture design is a statistical tool used to obtain the maximum desired effect using the minimum number of experiments. The aim of the presented work was the optimization of the composition of a mixture of essential oils from basil, citronella, cedarwood and thyme using simplex-lattice mixture design method. The optimized parameter was an antioxidant activity measured in DPPH assay and expressed as effective concentration (EC50). The test results showed an interesting synergy between the components of essential oils. The prepared binary and quaternary mixtures were characterized by higher activity than simple average activity. The designed mixture with approximated highest antioxidant activity was composed of: 54.4% citronella essential oil, 33.0% thyme essential oil, 9.2% cedarwood essential oil and 3.4% basil essential oil and its approximated activity was in agreement with experimental values. This work confirmed that it is possible to approximate the best antioxidant composition of four essential oils used as a potential medicinal and food ingredient.

LC-MS Analysis and Antifungal Activity of Turnera subulata Sm.

Fungi of the Candida genus are responsible for invasive candidiasis, which affects people all over the world and has high mortality rates. This is due to their virulence factors, which give them great resistance and pathogenicity. In addition, the emergence of multidrug-resistant strains makes it difficult to treat these infections. In this way, natural products have emerged as an alternative to standard drugs, where plants known for their medicinal properties such as Turnera subulata become attractive to research. The present work aimed to analyze the ethanol extract of Turnera subulata leaves against standard strains of Candida albicans, Candida krusei and Candida tropicalis using broth microdilution techniques. The identification of the compounds in T. subulata leaves by LC-MS revealed the presence of a wide variety of substances such as carboxylic acids and terpenes, with flavonoids and fatty acids being more evident. The antifungal assays showed that the extract was not able to inhibit the growth of the tested strains at concentrations with a clinical relevance. However, at higher concentrations, it was able to inhibit the fungal dimorphism of C. albicans and C. tropicalis. It is possible that the T. subulata extract has potential as an inhibitor of fungal virulence factors without affecting the cell viability. Further research should be carried out in order to assess its inhibitory potential for other fungal virulence factors.

Antifungal Effect and Inhibition of the Virulence Mechanism of D-Limonene against Candida parapsilosis.

Yeasts from the Candida parapsilosis complex are clinically relevant due to their high virulence and pathogenicity potential, such as adherence to epithelial cells and emission of filamentous structures, as well as their low susceptibility to antifungals. D-limonene, a natural compound, emerges as a promising alternative with previously described antibacterial, antiparasitic, and antifungal activity; however, its mechanisms of action and antivirulence activity against C. parapsilosis complex species have not been elucidated. Therefore, in the present study, we aimed to evaluate the antifungal and antivirulence action, as well as the mechanism of action of D-limonene against isolates from this complex. D-limonene exhibited relevant antifungal activity against C. parapsilosis complex yeasts, as well as excellent antivirulence activity by inhibiting yeast morphogenesis and adherence to the human epithelium. Furthermore, the apoptotic mechanism induced by this compound, which is not induced by oxidative stress, represents an important target for the development of new antifungal drugs.

Anxiolytic-like Effect of Quercetin Possibly through GABA Receptor Interaction Pathway: In Vivo and In Silico Studies.

Scientific evidence suggests that quercetin (QUR) has anxiolytic-like effects in experimental animals. However, the mechanism of action responsible for its anxiolytic-like effects is yet to be discovered. The goal of this research is to assess QUR's anxiolytic effects in mouse models to explicate the possible mechanism of action. After acute intraperitoneal (i.p.) treatment with QUR at a dose of 50 mg/kg (i.p.), behavioral models of open-field, hole board, swing box, and light-dark tests were performed. QUR was combined with a GABAergic agonist (diazepam) and/or antagonist (flumazenil) group. Furthermore, in silico analysis was also conducted to observe the interaction of QUR and GABA (α5), GABA (ß1), and GABA (ß2) receptors. In the experimental animal model, QUR had an anxiolytic-like effect. QUR, when combined with diazepam (2 mg/kg, i.p.), drastically potentiated an anxiolytic effect of diazepam. QUR is a more highly competitive ligand for the benzodiazepine recognition site that can displace flumazenil (2.5 mg/kg, i.p.). In all the test models, QUR acted similar to diazepam, with enhanced effects of the standard anxiolytic drug, which were reversed by pre-treatment with flumazenil. QUR showed the best interaction with the GABA (α5) receptor compared to the GABA (ß1) and GABA (ß2) receptors. In conclusion, QUR may exert an anxiolytic-like effect on mice, probably through the GABA-receptor-interacting pathway.

Chemical Profile and Biological Activities of Essential Oil from Piper arboreum for Development and Improvement of Mouthwash.

Studies show that more consumers are using natural health products in the modern world. We have noticed a growing demand in markets and the professional community for mouthwashes that contain natural compounds. The objective of this study was to assess the chemical characterization and microbiological potential of the essential oil Piper arboreum (EOPa) to provide data to enable the development of a low-cost mouthwash. The evaluation of the antibacterial and bacterial resistance modulating activity was performed by the microdilution method to determine the minimum inhibitory concentration. The chemical components were characterized by gas chromatography coupled with mass spectrometry, which identified 20 chemical constituents, with caryophyllene oxide being one of the major compounds. The EOPa showed an MIC ≥ 1024 µg/mL for all bacterial strains used in the tests. When evaluating the modulating activity of EOPa combined with chlorhexidine, mouthwash and antibiotics against the bacterial resistance, the oil limited synergistic activity between the MIC of the products tested in combination (37% to 87.5%). Therefore, we recommend expanding the tests with greater variation in the EOPa concentration and the products used, as well as toxicity assessments and in vivo testing, with the purpose of the development of a possible low-cost mouthwash base that is accessible to the most vulnerable populations.

GC-MS Chemical Characterization and Antibacterial Effect of the Essential oil of Piper mosenii.

Commercialized mouthwashes are generally expensive for the most financially vulnerable populations. Thus, several studies evaluate the antimicrobial potential of herbal products, such as essential oils, to reduce the activity of microorganisms in the mouth. The objective of this research was to carry out the chemical characterization and antibacterial activity of the essential oil of Piper mosenii (EOPm), providing data that enable the development of a low-cost mouthwash formulation aimed at vulnerable communities. The analysis of the antibacterial potential and modulator of bacterial resistance was verified by the microdilution method to determine the minimum inhibitory concentration-MIC. The chemical components were characterized by gas chromatography coupled to mass spectrometry, where 23 chemical constituents were detected, with α-pinene, being the major compound. The EOPm showed a MIC ≥ 1024 µg/mL for all bacterial strains used in the tests. When the EOPm modulating activity was evaluated together with chlorhexidine, mouthwash and antibiotics against bacterial resistance, the oil showed a significant synergistic effect, reducing the MIC of the products tested in combination, in percentages between 20.6% to 96.3%. Therefore, it is recommended to expand the tests with greater variation of EOPm concentration and the products used in this research, in addition to the evaluation of toxicity and in vivo tests, seeking the development of a possible formulation of mouthwash accessible to the vulnerable population.

UPLC-MS-ESI-QTOF Analysis and Antifungal Activity of Aqueous Extracts of Spondias tuberosa.

This study aimed to identify the chemical composition of the Spondias tuberosa aqueous leaf and root extracts (EALST and EARST) and to evaluate their effect, comparatively, against opportunistic pathogenic fungi. Ultra-Performance Liquid Chromatography Coupled to a Quadrupole/Time of Flight System (UPLC-MS-ESI-QTOF) was employed for chemical analysis. Candida albicans and C. tropicalis standard strains and clinical isolates were used (CA INCQS 40006, CT INCQS 40042, CA URM 5974, and CT URM 4262). The 50% Inhibitory Concentration for the fungal population (IC50) was determined for both the intrinsic action of the extracts and the extract/fluconazole (FCZ) associations. The determination of the Minimum Fungicidal Concentration (MFC) and the verification of effects over fungal morphological transitions were performed by subculture in Petri dishes and humid chambers, respectively, both based on micro-dilution. UPLC-MS-ESI-QTOF analysis revealed the presence of phenolic and flavonoid compounds. The association of the extracts with fluconazole, resulted in IC50 values from 2.62 µg/mL to 308.96 µg/mL. The MFC of the extracts was ≥16,384 µg/mL for all tested strains, while fluconazole obtained an MFC of 8192 µg/mL against C. albicans strains. A reduction in MFC against CA URM 5974 (EALST: 2048 µg/mL and EARST: 1024 µg/mL) occurred in the extract/fluconazole association.

Trace Elements and Heavy Metal Contents in West Algerian Natural Honey.

Analysis of trace elements and heavy metals in honey is essential for honey quality and safety and also monitoring environmental pollution. This study aimed to evaluate the composition of thirty-seven honey samples of different botanical origins (14 multifloral and 23 unifloral) obtained from beekeepers located in the west region of Algeria. Inductively coupled plasma-mass spectrometry (ICP-MS) and atomic absorption spectroscopy (AAS) methods were used to determine the levels of 19 elements in honey (K, Na, Ca, Mg, Mn, Cu, Fe, Zn, V, Cr, Co, As, Ru, Rh, Cd, W, Pt, Au, and Pb). Ru, Rh, Pt and, Au were not detected in any of the tested honey samples. The most abundant minerals were K, Ca, Na, and Mg ranging within 153.00-989.00 mg/kg, 33.10-502.00 mg/kg, 13.30-281.00 mg/kg, and 20.80-162.00 mg/kg, respectively. Fe, Mn, Zn, and Cu were the most abundant heavy metals while Pb, V, Cr, W, Co, and Cd were the lowest ones (<1 mg/kg) in the honey samples surveyed. Several honey types, lavender, rosemary, mild white mustard, thyme, milk thistle, carob tree, orange tree, Euphorbia, Eucalyptus, camphor, jujube tree, sage, and harmal, were studied, and the statistical analysis was carried out using principal component analysis (PCA) and hierarchical cluster analysis (HCA) techniques to evaluate the data. The results showed that the analyses of mineral content were sufficient to determine the floral origin and their variability may be related to geochemical and geographical differences. On other hand, all elements detected were at levels below safe thresholds.

Physical Activity and Its Determinants among Senior Residents of Podlasie, a Green Region of Poland, Based on the National PolSenior Study.

Physical activity is essential at all stages of life, and particularly so in the later years. The main objectives of the present work was to evaluate the physical activity of seniors, aged 65 years and older, living in the area of the Podlaskie voivodeship (Podlasie), Poland, based on the results of the nationwide PolSenior project, and to formulate recommendations for health policies implemented by both national and local government units. The study was performed as part of the national PolSenior project, whose aim was to evaluate various aspects of aging in Poland. The physical activity of a group of 186 randomly-selected people aged 65 years and above, 94 men and 92 women, was evaluated by questionnaire. The analysis included participants who took part in physical activity at least several times a week. Although all participants reported a decline in physical activity with age, the men remained physically active for longer. Among the respondents, 68.3% of women and 62.7% of men took short walks around the house as the main form of exercise, with working on the allotment or garden being another frequent activity; however, this was more common among men (53.9%) than women (34.7%). In addition, men were nearly twice as likely to take part in cycling (31.5%) than women (13.1%). The greatest motivation for physical activity given by the respondents was health, as noted by 73.8% of the men and 77.7% of the women. The physical activity of seniors in Podlasie is unsatisfactory and does not fulfil the WHO recommendations regarding the prophylaxis and prevention of chronic illness. This level does not, however, significantly differ from that reported in the nationwide PolSenior study or in other European countries in the Eurobarometer study. The decline in physical activity with age highlights the need for its greater promotion among seniors by local authorities. Such initiatives will help maintain the physical fitness and independence of this age group and contribute to a greater quality of life.

Cyclosporine A, in Contrast to Rapamycin, Affects the Ability of Dendritic Cells to Induce Immune Tolerance Mechanisms.

Following organ transplantation, it is essential that immune tolerance is induced in the graft recipient to reduce the risk of rejection and avoid complications associated with the long-term use of immunosuppressive drugs. Immature dendritic cells (DCs) are considered to promote transplant tolerance and may minimize the risk of graft rejection. The aim of the study was to evaluate the effects of immunosuppressive agents: rapamycin (Rapa) and cyclosporine A (CsA) on generation of human tolerogenic DCs (tolDCs) and also to evaluate the ability of these cells to induce mechanisms of immune tolerance. tolDCs were generated in the environment of Rapa or CsA. Next, we evaluated the effects of these agents on surface phenotypes (CD11c, MHC II, CD40, CD80, CD83, CD86, CCR7, TLR2, TLR4), cytokine production (IL-4, IL-6, IL-10, IL-12p70, TGF-ß), phagocytic capacity and resistant to lipopolysaccharide activation of these DCs. Moreover, we assessed ability of such tolDCs to induce T cell activation and apoptosis, Treg differentiation and production of Th1- and Th2-characteristic cytokine profile. Data obtained in this study demonstrate that rapamycin is effective at generating maturation-resistant tolDCs, however, does not change the ability of these cells to induce mechanisms of immune tolerance. In contrast, CsA affects the ability of these cells to induce mechanisms of immune tolerance, but is not efficient at generating maturation-resistant tolDCs.

Endometriotic Peritoneal Fluid Stimulates Recruitment of CD4+CD25highFOXP3+ Treg Cells.

Endometriosis is a common gynecological disorder characterized by the presence of endometrial-like tissue outside the uterus. The disease is associated with disturbed local and systemic immunity. It has been reported that the proportion of CD4+CD25highFOXP3+ Treg cells may be significantly increased in the peritoneal fluid of patients with endometriosis. Therefore, the aim of our study was to investigate whether the proportions of Treg cells in the peritoneal cavity of patients with endometriosis are related to the chemotactic and stimulatory activity of the local peritoneal milieu. The peritoneal fluid was collected from 13 women with ovarian endometriosis and 12 control women without the disease. T cell populations were analyzed by flow cytometry, cytokines and chemokines were evaluated using the cytometric bead kit, and cell chemotaxis was studied by cell migration assay. We confirmed that the proportions of Treg cells are increased in the peritoneal fluid of women with endometriosis as compared to the control women. Endometriosis was also associated with elevated concentrations of IL-6, IL-10, and TGF-ß1/2 as well as CCL20, CXCL8, CXCL9, and CXCL10. We did not reveal any changes in the proportion of peritoneal Th17 cells and concentrations of IL-17A. Peritoneal Treg cells positively correlated with concentrations of TGF-ß, IL-10, and CCL20. Endometriotic peritoneal fluid stimulated chemotaxis of both CD4+ and Treg cells. This chemotactic activity positively correlated with concentrations of CCL20. CCL20 stimulated the migration of Treg cells, and the chemotactic activity of the endometriotic peritoneal fluid was inhibited by neutralizing anti-CCL20 antibodies. These results imply that increased proportions of the peritoneal Treg cells in women with endometriosis may result from attraction and activation by local chemokines and cytokines, especially CCL20 and TGF-ß. Since Treg cells contribute to the immunopathogenesis of endometriosis, their chemotaxis and activation may be considered as a target for therapeutic intervention.

Peritoneal Fluid from Patients with Ovarian Endometriosis Displays Immunosuppressive Potential and Stimulates Th2 Response.

Endometriosis is a common gynaecological disorder characterized by the ectopic growth of endometrial tissue outside the uterine cavity. It is associated with chronic pelvic inflammation and autoimmune reactivity manifesting by autoantibody production and abrogated cellular immune responses. Endometriotic peritoneal fluid contains various infiltrating leucocyte populations and a bulk of proinflammatory and immunoregulatory cytokines. However, the nature and significance of the peritoneal milieu in women with endometriosis still remains obscure. Therefore, the aim of the present study was to investigate the immunoregulatory activity of the peritoneal fluid (PF) from women with endometriosis. The peritoneal fluid samples were collected during laparoscopic surgery from 30 women with and without endometriosis. Immunoregulatory cytokines (IL-2, IL-4, IL-6, IL-10, IL-17A, IFN-γ and TNF) and chemokines (CCL2, CCL5, CXCL8 and CXCL9) were evaluated in PF and culture supernatants generated by unstimulated and CD3/CD28/IL-2-stimulated CD4+ T cells cultured in the presence of PF. The effect of PF on the generation of Treg and Th17 cells in CD4+ T cell cultures, as well as the natural cytotoxic activity of peripheral blood mononuclear cells, was also investigated. Concentrations of IL-6, IL-10, CCL2, CXCL8 and CXCL9 were significantly upregulated in the PF from women with endometriosis when compared to control women, whereas concentrations of other cytokines and chemokines were unaffected. The culturing of unstimulated and CD3/CD28/IL-2-stimulated CD4+ T cells in the presence of endometriotic PF resulted in the downregulation of their IL-2, IFN-γ, IL-17A and TNF production as compared to culture medium alone. On the other side, endometriotic PF significantly stimulated the production of IL-4 and IL-10. Endometriotic PF also stimulated the release of CCL2 and CXCL8, whereas the production of CCL5 and CXCL9 was downregulated. Endometriotic PF stimulated the generation of Treg cells and had an inhibitory effect on the generation of Th17 cells in cultures of CD4+ T cells. It also inhibited the NK cell cytotoxic activity of the peripheral blood lymphocytes. These results strongly imply that the PF from patients with endometriosis has immunoregulatory/immunosuppressive activity and shifts the Th1/Th2 cytokine balance toward the Th2 response, which may account for deviation of local and systemic immune responses. However, a similar trend, albeit not a statistically significant one, was also observed in case of PF from women without endometriosis, thus suggesting that peritoneal milieu may in general display some immunoregulatory/immunosuppressive properties. It should be stressed, however, that our present observations were made on a relatively small number of PF samples and further studies are needed to reveal possible mechanism(s) responsible for this phenomenon.

The Safety Assessment of Toxic Metals in Commonly Used Herbs, Spices, Tea, and Coffee in Poland.

The presented study was aimed at the determination of the level of contamination with heavy metals (Cd, Pb, As, and Hg) in 240 samples of plant materials, i.e., herbal raw materials, spices, tea, and coffee. Moreover, a probabilistic risk assessment (noncarcinogenic and carcinogenic risks) was estimated by models including target hazard quotient (THQ) and cancer risk (CR). The samples were subjected to microwave mineralisation with the use of HNO3 (65%), while the determination of the content of the elements was performed with the use of inductively coupled plasma mass spectrometer (ICP-MS) and a mercury analyser. The element which was characterised by the highest level of accumulation in the analysed samples was lead (from 0.010 to 5.680 mg/kg). Among the heavy metals under analysis, the lowest concentration was noted in the case of mercury (from 0.005 to 0.030 mg/kg). A notably higher level of contamination with heavy metals was noted in the analysed samples of herbs and spices (0.005-5.680 mg/kg), compared to samples of tea and coffee (0.005-0.791 mg/kg). According to the guidelines of the World Health Organisation (WHO) concerning the limits of contamination of samples of herbal raw materials with heavy metals, lead levels exceeding the limits were only noted in 24 samples of herbs (18%). In all of the analysed samples of spices, tea, and coffee, no instances of exceeded limits were noted for any of the analysed heavy metals. The values of TTHQmax (in relation to the consumption of the analysed products) were as follows: up to 4.23 × 10-2 for spices, up to 2.51 × 10-1 for herbs, up to 4.03 × 10-2 for China tea, and up to 1.25 × 10-1 for roasted coffee beans. As the value of THQ ≤1, there is no probability of the appearance of undesirable effects related to the consumption of the analysed group of raw materials and products of plant origin. The CR value for As (max. value) was 1.29 × 10-5, which is lower than the maximum acceptable level of 1 × 10-4 suggested by United States Environmental Protection Agency (USEPA).

Optimization of Glycerol-Water Extraction of Selected Bioactive Compounds from Peppermint and Common Nettle.

Current trends in the industry indicate that extraction solvents should conform with the ideals of so-called "green chemistry". Therefore, the objective of the presented study was to optimize the conditions for the extraction of polyphenols, flavonoids and chlorophyll from peppermint leaves (Mentha × piperita L., Lamiaceae) and from common nettle leaves (Urtica dioica L., Urticaceae) via green chemistry. The obtained experimental results were subjected to modelling by means of the multiple regression method, while the optimization of the system was addressed via the application of the desirability function. As a result of the use of glycerol-water systems for the extraction of the tested active compounds from mint leaves and nettle leaves, extracts with higher concentrations of polyphenols, flavonoids and chlorophyll were most often obtained, when compared with the use of classical solvents such as water and ethanol. In this work, we demonstrate that the extraction temperature has significant influence on the concentration of the determined components in the extracts. To obtain the highest values of the analysed parameters, leaves of peppermint should be extracted with glycerol-water mixture at the proportions of 30.5:69.5 at a temperature of 50 °C, while the optimal conditions for the extraction of leaves of common nettle were the glycerol-water proportions of 12.5:87.5 and extraction temperature of 20 °C. Comparing the average percentage differences between the highest values of the analysed parameters obtained in the experiment and the approximated values for various temperatures with the level of desirability, one can note a high correlation that, in the analysed examples, amounted to 0.9681. The study showed that glycerol can be an alternative solvent in the extraction of polyphenols, flavonoids and chlorophyll, replacing, e.g., ethanol-which, for various reasons, cannot always be used.

Myorelaxant Effect of the Dysphania ambrosioides Essential Oil on Sus scrofa domesticus Coronary Artery and Its Toxicity in the Drosophila melanogaster Model.

PURPOSE: Alternative methods for the use of animals in research have gained increasing importance, due to assessments evaluating the real need for their use and the development of legislation that regulates the subject. The principle of the 3R's (replacement, reduction and refinement) has been an important reference, such that in vitro, ex vivo and cord replacement methods have achieved a prominent place in research. METHODS: Therefore, due to successful results from studies developed with these methods, the present study aimed to evaluate the myorelaxant effect of the Dysphania ambrosioides essential oil (EODa) using a Sus scrofa domesticus coronary artery model, and the toxicity of both the Dysphania ambrosioides essential oil and its major constituent, α-terpinene, against Drosophila melanogaster in toxicity and negative geotaxis assays. RESULTS: The EODa relaxed the smooth muscle of swine coronary arteries precontracted with K+ and 5-HT in assays using Sus scrofa domesticus coronary arteries. The toxicity results presented LC50 values of 1.546 mg/mL and 2.282 mg/mL for the EODa and α-terpinene, respectively, thus showing the EODa and α-terpinene presented toxicity to these dipterans, with the EODa being more toxic. CONCLUSIONS: Moreover, the results reveal the possibility of using the EODa in vascular disease studies since it promoted the relaxation of the Sus scrofa domesticus coronary smooth muscle.

Pentosidine, advanced glycation end product, in acute ischaemic stroke patients with and without atrial rhythm disturbances.

Atrial fibrillation (AF) and atherosclerotic disease are independent risk factors for acute ischaemic stroke (AIS). The optimal biological marker which could allow differentiation between AF and non-AF AIS patients is still not available. AIM OF THE STUDY: Aim of the present study was to investigate the role of pentosidine as a potential biological marker for AF in an AIS patient group. MATERIALS AND METHODS: Sixty-three acute ischaemic hemispheric stroke patients were recruited and divided into two groups according to the presumed underlying mechanism: with or without atrial rhythm disorders. Ten healthy volunteers were a reference group for serum level of pentosidine. Carotid artery ultrasound was performed, and common carotid artery stiffness and intima-media thickness were measured. Serum levels of pentosidine and selected routine biochemical risk factors for atherosclerosis (cholesterol and its lipoprotein fractions, homocysteine) were examined. RESULTS: A higher serum level of pentosidine was observed in patients without atrial fibrillation (1,509 ± 485.13pmol/ml); a statistically significant difference was observed compared to the reference group (1,041.52 ± 411.17pmol/ml; p = 0.01), but not the AF patients (1,438.19 ± 495.97pmol/ml; p = 0.59). No significant difference in the non-AF group compared to the AF group for carotid intima-media thickness (IMT)/stiffness and pentosidine serum level was recorded. CONCLUSIONS AND CLINICAL IMPLICATIONS: A higher serum level of pentosidine was observed in AIS patients without atrial fibrillation compared to the healthy volunteers. According to the results of the present study, no difference between these patients in the selected risk factors of atherosclerosis were observed. Further studies are needed to identify a reliable marker of AF that would bring added value to the standard diagnostic workup after acute ischaemic stroke.