Antiviral and Cytotoxic Activities of Ilex aquifolium Silver Queen in the Context of Chemical Profiling of Two Ilex Species.

The leaves of Ilex paraguariensis (known as Yerba mate), used as a popular beverage, are a very well-recognized plant material with various biological activities, including analeptic (because of caffeine), anti-obesity (phenolics, saponins), antimicrobial, and antiviral (phenolics, saponins). Here, the chemical compositions of the leaves of two European Ilex species (× meserveae and aquifolium) with three varieties each were investigated. The terpenoid, saponin, and polyphenolic fractions were submitted for LC-MS or GC-MS analysis against a standard Mate leaf. In addition, the aroma profiles of all the species were analysed using HS-SPME-Arrow prior to GC-MS analysis. All fractions were subjected to antiviral and cytotoxic assays. We found 86 compounds in all accessions, with limonene, linalool, and p-cymene being predominant. There were minor similarities between the volatile compositions of the European and South American species. We found ursolic and oleanolic acid to be the main compounds in the terpenoid fraction. Mono-caffeoylquinic acids and di-caffeoylquinic acids were the main constituents of the polar fractions. About 180 compounds from the saponin group were tentatively identified, of which 9 and 3 were selected as distinctive markers for I. meserveae and I. aquifolium, respectively. Based on chemical screening, I. aquifolium Silver Queen was chosen as the source of terpenoid and saponin fractions and polyphenol extracts. The most substantial inhibition of cancer cell growth was observed with saponin in the case of the MCF7 (human breast cancer) cell line, while for LoVo and L929 cell lines (human colorectal cancer and reference mouse fibroblasts), it was slightly weaker. These results should be analysed further as a promising chemoprevention of colorectal and gastrointestinal cancers. Saponin and polyphenolic extracts exhibited similar activities against HSV-1 and HAdV-5, with 4-log reduction in virus titres. This study focuses our attention on a field of potential antiviral formulations derived from European holly.

Variation in Flavonoid Compounds, Volatiles and Yield Related Traits in Different Iranian Rosa damascena Mill. Cultivars Based on SPME Arrow and LC-MS/MS.

Damask rose (Rosa damascena Mill.) is an aromatic industrial plant with different applications. Selection of cultivars with high-value metabolites such as flavonoids-with acceptable yields-can lead to elite cultivars for mass propagation in various industries. A field experiment was carried out in a randomized complete block design (RCBD) to evaluate metabolites and some yield-related morphological data. In the present investigation, for the first time 13 flavonoid components of nine Iranian damask rose cultivars were compared using LC-MS/MS. As a result, 13 flavonoids were identified, most of which were reported for the first time in rose petals. Phloridzin (72.59-375.92 mg/100 g dw), diosmetin (82.48-153.16 mg/100 g dw) and biochanin A (0-1066.89 mg/100 g dw) were the most abundant, followed by trans-chalcone (0-106.29 mg/100 g dw) and diosmin (41.55-84.57 mg/100 g dw). Levels of naringenin also ranged from 3.77 in B111 to 54.70 mg/100 g dw in C294, while luteolin varied from 4.37 in B111 to 28.87 mg/100 g dw in C294. The SPME Arrow technique also was applied to determine the real aroma of the studied cultivars. Phenethyl alcohol was the most abundant compound, in the range of 69.28 to 77.58%. The highest citronellol/geraniol (C/G) was observed in D234 (4.52%) and D237 (4.30%), while the lowest amount belonged to A104 (1.28%). Rose oxide, as the most crucial factor for odor, ranged from 0.06% in D237 to 0.15% in D211. Based on cluster and principal component analysis (PCA), D234 cultivar can be suggested as a promising cultivar with high yield, high C/G content and high rose oxide, while D234 and C294 were the most valuable cultivars in terms of flavonoids with high yield. Finally, these cultivars can be introduced for further breeding programs and industrial cultivation.

Effect of Drying Methods on Chemical and Sensory Properties of Cannabis sativa Leaves.

Hemp is used as a source of fiber, oil and bioactive substances including volatile and cannabinoid-containing substances. This paper presents, for the first time, results on the evaluation of drying methods (convective, vacuum-microwave and combined convective pre-drying and vacuum-microwave finishing drying) of hemp leaves on the qualitative and quantitative changes in secondary metabolites, including essential oils, cannabinoids and sterols. A ranking and descriptive test of hemp leaves was also performed. Drying kinetics was presented using three models, including logarithmic, Midilli and modified Page. The SPME-Arrow technique was used to determine 41 volatile compounds, of which caryophyllene, ß-myrcene and α-humulene were dominant in dried and fresh leaves. Regarding the essential oils obtained, 64 were identified, with caryophyllene, humulene epoxide II and limonene being the dominant ones. For preserving the highest amount of oils, the best method was the convective pre-drying followed by vacuum-microwave finishing drying (CD60-VMD) combined method, where the retention of volatile compounds was 36.08%, whereas the CD70 and 240-VMD methods resulted in the highest loss of 83%. The predominant cannabinoids in fresh hemp leaves were CBDA 6.05 and CBD 2.19 mg g-1. Drying caused no change in the cannabinoid profile of the plant material. ß-Sitosterol, campesterol and lupeol were dominant in the phytosterol and triterpene fractions. No changes in either quality or quantity were observed in any of the variants found.

Chemical Investigations in Kelussia odoratissima Mozaff. Leaves Based on Comprehensive Analytical Methods: LC-MS, SPME, and GC-MS Analyses.

Kelussia odoratissima Mozaff. is a species of Apiaceae endemic to the Zagros Mountains in Iran. In the present investigation, for the first time, the polyphenolic compounds and flavonoids of its leaves were determined by liquid chromatography-mass spectrometry (LC-MS). As a result, p-coumaric acid, ferulic acid, caffeic acid, chlorogenic acid, acetyl phloroglucinol, vanillic acid, m-coumaric acid, and 4-methylsiringol were determined as the main phenolic compounds, while 3-hydroxyflavone, flavone, quercetin, rutin, neohesperidin, polydatin, and diosmin were the main flavonoid components, of which chlorogenic acid (303.08 µL/gDW), neohesperidin (38.37 µL/gDw), and diosmin (28.62 µL/gDW) were the most abundant. Solid-phase microextraction (SPME) was also used to determine the chemical compounds. Based on SPME, (Z)-undec-6-en-2-one (17.48%) and (Z)-butylidenephthalide (4.348%) were the major components. Based on GC-MS analyses, (Z)-ligustilide was the main compound; however, some new compounds were also determined, including 3-ethylisobenzofuran-1 (3H)-one, (E)-ligugustilide, and E-n-butylidene phthalide. Also, for the first time, we have identified EOs ethyl and isobutyl phthalides on the basis of the obtained EI-MS spectra. Finally, the fragmentation of phthalides is also discussed in this research.

Natural Appetite Control: Consumer Perception of Food-Based Appetite Regulating Aromas.

According to the WHO, the number of overweight people (BMI ≥ 25) and obese people (BMI ≥ 30) is constantly growing. On the other hand, the number of elderly people (≥60 years old) in 2020 reached 1.4 billion worldwide. Both mentioned groups demonstrate their individual and characteristic appetite disorders. In light of the side effects of appetite stimulating drugs, which interfere with diabetics, hypertension and thrombosis medicines or diet supplements with doubtful effectiveness in reducing appetite, new and natural alternatives are highly demanded. Therefore, the present study focusses on the search for natural food aromas, which may have potential for appetite regulation. A survey was carried out among consumers with excess body weight (BMI ≥ 25) and the elderly (≥60 years old). Food products and meals pointed out by the survey participants were subjected to volatile analysis by HS-SPME Arrow followed by GC-MS. As a result, a group of volatiles and their odor characteristic were determined for appetite stimulation or reduction, which may suggest that the actual composition of food aroma is more significant than the character of the aroma. Those results may be a basis for designing appetite regulating agents, in which the mechanism of action will be based only on olfaction activity.

In Vitro Evaluation of Antiprotozoal Properties, Cytotoxicity Effect and Anticancer Activity of New Essential-Oil Based Phytoncide Mixtures.

Protozoa, in both humans and animals, are one of the leading causes of disease. International programmes introduced in many countries have helped reduce the incidence of disease. However, it has recently become increasingly difficult to achieve the goals set for the coming years. One of the main reasons for this, as with other pathogenic organisms, such as bacteria and fungi, is the increasing resistance to current methods of treating and preventing infection. Therefore, new therapies with high efficacy are needed. In the present study, the novel mixtures of essential oils (EOs), clove, garlic, Ceylon cinnamon, and rosemary with organic acids (acetic, propionic, lactic) and metal ions (Cu, Mn, Zn) were tested against five selected model protozoa (Euglena gracilis, Gregarina blattarum, Amoeba proteus, Paramecium caudatum, Pentatrichomonas hominis). The cytotoxicity and potential anticancer activity of the obtained combinations were tested on the human fibroblasts (NHDF) and human cancer cell lines (A549, MCF7, LoVo, HT29). All of the mixtures showed very good antiprotozoal properties. The most efficient were the combination of clove and rosemary essential oils, mixtures of acids, and Mn ions. The LD50 values were in the range of 0.001-0.006% and the LD100 values were 0.002-0.008%. All of the tested mixtures did not show cytotoxicity against normal cells, but did show growth inhibition against cancer cell lines. The most cytotoxic against cancer cells were combinations with cinnamon essential oil. Nevertheless, the proposed combinations containing essential oils, organic acids, and metal ions have high antiprotozoal activity, with low toxicity to healthy human cells.

Chemical investigation on Salvia officinalis L. Affected by multiple drying techniques - The comprehensive analytical approach (HS-SPME, GC-MS, LC-MS/MS, GC-O and NMR).

In light of large scale production of Salvia officinalis L. and its complex storage and delivery chain, the efficient preservation process is required. At this moment, the most popular preservation method is drying, therefore a comprehensive experiment to evaluate the influence of multiple drying techniques on sage was conducted. Convective drying at 40, 50 and 60 °C, vacuum-microwave drying with powers 240, 360 and 480 W and combined drying consisting of convective pre-drying at 50 °C followed by vacuum-microwave finish-drying with power 360 W were applied. To evaluate the effect of particular procedures chemical analyses were performed, namely evaluation of changes in bioactive volatile constituents, odour-active compounds and various groups of non-volatile bioactive constituents of sage. The obtained results shown, that to receive the product with expected quality, it is necessary to identify the designation of the product before treatment, since particular groups of phytochemicals react differently during drying.

Biochemical and Molecular Investigation of the Effect of Saponins and Terpenoids Derived from Leaves of Ilex aquifolium on Lipid Metabolism of Obese Zucker Rats.

Ilex paraguariensis, the holly tree, is a plant with recognized biological properties, whose aqueous infusions are known as "Yerba mate", that regulate lipid metabolism, reduce obesity, and improve brain stimulation. In the present study, the effect of standardized saponin and terpenoid fractions of a European taxon, Ilex aquifolium, on blood biochemical parameters in a rat model of metabolic disorder, (fa/fa) Zucker, are presented. The profiles of the volatile fractions of two species and six European varieties of Ilex were investigated. After selecting the best variety, the saponin and terpenoid fractions were isolated and standardized, and animals were fed 10 mg kg−1 b.w. for 8 weeks. A statistically significant decrease in liver adiposity was observed, confirmed by histology and quantitative identification (gas chromatography−mass spectrometry analyses of hepatic lipids. RT-qPCR analysis of gene expression in the aorta revealed that the administration of the terpenoid fraction downregulated LOX-1, suggesting a reduction in atherosclerotic stimuli. In addition, a statistically significant reduction (p < 0.05) in PPARγ for the saponin fraction was observed in the liver. The expression of the ACAT-1 gene in the liver, responsible for the formation of cholesterol esters, increased significantly in the group receiving the terpenoid fraction compared to the control, which was also confirmed by the analysis of individual blood biochemical parameters. The opposite effect was observed for saponins. Taking the above into account, it is shown for the first time that Ilex aquifolium can be a source of compounds that positively influence lipid metabolism.

Systematic Investigation of the Effects of Seven Plant Extracts on the Physiological Parameters, Yield, and Nutritional Quality of Radish (Raphanus sativus var. sativus).

The modern agricultural sector faces the challenge of addressing the needs of the fast-growing global population. This process should be both high-yielding and sustainable, without creating risks for the environment and human health. Therefore, natural products are gaining attention in the production of safe and nutritious food. In a systematic effort to develop affordable and effective biostimulants, we examined the impact of botanical extracts on the growth and physiological parameters of radish plants under field conditions. Ultrasound-assisted extraction, mechanical homogenization, and water were used for the production of potential plant-based biostimulants. Foliar applications of the bio-products, developed and used in our study, have led to an increase in the examined parameters (total yield, dry weight, photosynthetic pigments, vitamin C, nitrates, and micro- and macroelements). A decrease in the total phenolic compounds content was also noted, as well as a varied impact on the steam volatile compounds, fatty acids, sterol, and glucosinolates composition. The most beneficial effects on radish, in terms of physiological and biochemical properties, were found in groups treated with extracts based on the common dandelion, valerian, and giant goldenrod. This innovative approach presented in our study could provide a valuable tool for sustainable horticultural production.

Effect of Botanical Extracts on the Growth and Nutritional Quality of Field-Grown White Head Cabbage (Brassica oleracea var. capitata).

Nutraceuticals and functional foods are gaining more attention amongst consumers interested in nutritious food. The consumption of foodstuffs with a high content of phytochemicals has been proven to provide various health benefits. The application of biostimulants is a potential strategy to fortify cultivated plants with beneficial bioactive compounds. Nevertheless, it has not yet been established whether the proposed higher plants (St. John's wort, giant goldenrod, common dandelion, red clover, nettle, and valerian) are appropriate for the production of potential bio-products enhancing the nutritional value of white cabbage. Therefore, this research examines the impact of botanical extracts on the growth and nutritional quality of cabbage grown under field conditions. Two extraction methods were used for the production of water-based bio-products, namely: ultrasound-assisted extraction and mechanical homogenisation. Bio-products were applied as foliar sprays to evaluate their impact on total yield, dry weight, photosynthetic pigments, polyphenols, antioxidant activity, vitamin C, nitrates, micro- and macroelements, volatile compounds, fatty acids, sterols, and sugars. Botanical extracts showed different effects on the examined parameters. The best results in terms of physiological and biochemical properties of cabbage were obtained for extracts from common dandelion, valerian, nettle, and giant goldenrod. When enriched with nutrients, vegetables can constitute a valuable component of functional food.

Activity of Liquid and Volatile Fractions of Essential Oils against Biofilm Formed by Selected Reference Strains on Polystyrene and Hydroxyapatite Surfaces.

Biofilms are surface-attached, structured microbial communities displaying higher tolerance to antimicrobial agents in comparison to planktonic cells. An estimated 80% of all infections are thought to be biofilm-related. The drying pipeline of new antibiotics efficient against biofilm-forming pathogens urges the search for alternative routes of treatment. Essential Oils (EOs), extracted from medicinally important plants, are a reservoir of bioactive compounds that may serve as a foothold in investigating novel antibiofilm compounds. The aim of this study was to compare antimicrobial activity of liquid and volatile fractions of tested EOs against biofilm-forming pathogens using different techniques. In this research, we tested five EOs, extracted from Syzygium aromaticum L., Boswelia serrata Roxb., Juniperus virginiana L., Pelargonium graveolens L. and Melaleuca alternifolia Cheel., against planktonic and biofilm forms of five selected reference strains, namely Staphylococcus aureus, Enterococcus faecalis, Klebsiella pneumoniae, Pseudomonas aeruginosa, Escherichia coli, and Candida albicans. To obtain cohesive results, we applied four various methodological approaches: to assess the activity of the liquid fraction of EOs, disc diffusion and the microdilution method were applied; to test EOs' volatile fraction, the AntiBioVol assay and modified Antibiofilm Dressing Activity Measurement (A.D.A.M.) were used. The molecular composition and dynamics of antimicrobial substances released from specific EOs was measured using Gas Chromatography-Mass Spectrometry (GC-MS). The antimicrobial potency of EO's volatile fraction against biofilm formed by tested strains differed from that of the liquid fraction and was related to the molecular weight of volatile compounds. The liquid fraction of CW-EO and volatile fraction of F-EO acted in the strongest manner against biofilm of C. albicans. The addition of 0.5% Tween 20 to liquid phase, enhanced activity of G-EO against E. coli and K. pneumoniae biofilm. EO activity depended on the microbial species it was applied against and the chosen assessment methodology. While all tested EOs have shown a certain level of antimicrobial and antibiofilm effect, our results indicate that the choice of EO to be applied against a specific biofilm-forming pathogen requires careful consideration with regard to the above-listed aspects. Nevertheless, the results presented in this research contribute to the growing body of evidence indicating the beneficial effects of EOs, which may be applied to fight biofilm-forming pathogens.

Biological Potential and Chemical Profile of European Varieties of Ilex.

Plants of the genus Ilex are widespread throughout the world, with its best-known representative being Ilex paraguraiensis from South America. The European species Ilex aquifolium shows similarities in its terpenoid, sugar and phenolic acid profiles. Using aqueous extracts of Ilex aquifolium as a supplement in Wistar rats showed that, despite the lack of caffeine, it had strong hypocholesterolemic effects. In addition, a reduction in oxidative lipid degradation and a decrease in hepatic steatosis in histopathological studies were observed. The results of this study suggest that extracts from the European species Ilex aquifolium may have potential as an alternative treatment for hyperlipidemia.

Field-Scale Evaluation of Botanical Extracts Effect on the Yield, Chemical Composition and Antioxidant Activity of Celeriac (Apium graveolens L. Var. rapaceum).

The use of higher plants for the production of plant growth biostimulants is receiving increased attention among scientists, farmers, investors, consumers and regulators. The aim of the present study was to examine the possibility of converting plants commonly occurring in Europe (St. John's wort, giant goldenrod, common dandelion, red clover, nettle, valerian) into valuable and easy to use bio-products. The biostimulating activity of botanical extracts and their effect on the chemical composition of celeriac were identified. Plant-based extracts, obtained by ultrasound-assisted extraction and mechanical homogenisation, were tested in field trials. It was found that the obtained formulations increased the total yield of leaves rosettes and roots, the dry weight of leaves rosettes and roots, the content of chlorophyll a + b and carotenoids, the greenness index of leaves, the content of vitamin C in leaves and roots. They mostly decreased the content of polyphenols and antioxidant activities in leaves but increased them in roots and conversely affected the nitrates content. Extracts showed a varied impact on the content of micro and macroelements, as well as the composition of volatile compounds and fatty acids in the celeriac biomass. Due to the modulatory properties of the tested products, they may be used successfully in sustainable horticulture.

Volatile Composition and Sensory Properties as Quality Attributes of Fresh and Dried Hemp Flowers (Cannabis sativa L.).

Flowers of hemp (Cannabis sativa L.) are widely used in cosmetics, food, and in the pharmaceutical industry. The drying process plays a key role in retention of aroma and also in the quality of products. Seven variants of hemp flower drying, including convection drying (CD), vacuum-microwave drying (VMD), and combined drying consisting of convective pre-drying followed by vacuum-microwave finishing drying (CPD-VMFD) were checked in this study. For each process, we applied the two-term model. Dried material was submitted to color and chromatographical assessments. Analyses of obtained essential oil showed the presence of 93 volatile compounds, predominantly ß-myrcene, limonene, and ß-(E)-caryophyllene, as well as α-humulene. Application of 240 W during VMD and 50 °C during CD gave the highest retention of aroma compounds, amounting to 85 and 76%, respectively, but with huge color changes. Additionally, sensory analysis proved that drying with a microwave power of 240 W provides a product most similar to fresh material.

Metabolism, Ketosis Treatment and Milk Production after Using Glycerol in Dairy Cows: A Review.

The aim of this paper is to review and systematize the current state of knowledge on glycol metabolism in cattle. Glycerol, derived from biodiesel production, must be purified in order to be a useful product for feeding livestock. The use of glycerol in the feeding of ruminants can be justified for several reasons: (i) it is a source of energy in the ration, (ii) it is a glucogenic precursor, and (iii) it may have an effect on milk composition. The high energy value of glycerol provides the opportunity to use this raw material as a partial grain substitute in cattle feed rations. Dietary supplementation of glycerol is associated with increased propionate, butyrate, valerate, and isovalerate concentrations in the rumen. Glycerol can be used at up to 10%-15% of the dietary dry matter (DM) and is well-established as a treatment for ketosis in cows. Glycerol increases plasma glucose and may reduce non-esterified fatty acids and ß-hydroxybutyrate levels. The use of glycerol does not have a clear effect on DM intake, milk yield, or milk composition. However, some authors have reported an increase in milk yield after glycerol supplementation associated with decreased milk fat concentration. It is also possible that the concentration in the milk of odd-chain fatty acids and cis-9, trans-11 conjugated linoleic acid may increase after glycerol application.

Polish Propolis-Chemical Composition and Biological Effects in Tongue Cancer Cells and Macrophages.

The purpose of this study was to compare the chemical composition and biological properties of Polish propolis. Ethanol, ethanol-hexane, hexane and hexane-ethanol extracts of propolis from three different regions of Poland were prepared. On the basis of the evaluation of their chemical composition as well as the extraction yield and free radical scavenging activity, the ethanol and hexane-ethanol extractions were proposed as the most effective methods. Subsequently, the biological properties of the extracts were evaluated to investigate the selectivity of an anticancer effect on tongue cancer cells in comparison to normal gingival fibroblasts. The obtained products demonstrated anticancer activity against tongue cancer cells. Additionally, when the lowest extract concentration (100 µg/mL) was applied, they were not cytotoxic to gingival fibroblasts. Finally, a possible anti-inflammatory potential of the prepared products was revealed, as reduced mitochondrial activity and proliferation of macrophages exposed to the extracts were observed. The results obtained indicate a potential of Polish propolis as a natural product with cancer-selective toxicity and anti-inflammatory effect. However, further studies are still needed to thoroughly explain the molecular mechanisms of its action and to obtain the promising health benefits of this versatile natural product.