Antioxidant and Antiproliferative Activities of Phenolic Extracts of Eriobotrya japonica (Thunb.) Lindl. Fruits and Leaves.

Increasing interest in new sources of secondary metabolites as biologically active substances has resulted in an advanced study of many plant species. Loquat (Eriobotrya japonica (Thunb.) Lindl. = Rhaphiolepis bibas (Lour.) Galasso & Banfi, Rosaceae family), an evergreen, subtropical fruit tree, native to China and Japan, but cultivated in southern countries of Europe, is a species commonly used in folk medicine and may be an excellent source of bioactive compounds. Therefore, the aim of the present study was to evaluate the profile of the phenolic constituents of E. japonica fruits and leaves originating from Tuscany (Italy), as well as their in vitro antioxidant and chemopreventive activities on human cancer cell lines breast adenocarcinoma (MCF-7), colon adenocarcinoma (Caco-2 and HT-29), and glioblastoma (U87MG). Results revealed that the extract of leaves displayed higher antioxidant and anticancer potential than the fruit extract and contained 25 individual phenolic compounds that have been characterized and quantified by the UPLC-PDA-MS method. The antiproliferative activity was correlated with the content of polyphenolic compounds indicating that both fruits and leaves are a good source of antioxidants and may be exploited as nutraceuticals enriching food or as components for the cosmetic/pharmaceutical industry.

Chemical Profiling, Bioactive Properties, and Anticancer and Antimicrobial Potential of Juglans regia L. Leaves.

The aim of this study was to assess the biological potential of the polyphenolic fraction isolated from J. regia leaves, collected in the Subcarpathian region (Poland). The phenolic profile was determined using the UPLC-PDA-MS/MS method. Biological activity was determined by evaluating the antioxidant, anticancer, antibacterial, and antifungal effects. Prior to this study, the purified polyphenolic fraction was not been tested in this regard. A total of 40 phenolic compounds (104.28 mg/g dw) were identified, with quercetin 3-O-glucoside and quercetin pentosides dominating. The preparation was characterized by a high ability to chelate iron ions and capture O2•- and OH• radicals (reaching IC50 values of 388.61, 67.78 and 193.29 µg/mL, respectively). As for the anticancer activity, among the six tested cell lines, the preparation reduced the viability of the DLD-1, Caco-2, and MCF-7 lines the most, while in the antibacterial activity, among the seven tested strains, the highest susceptibility has been demonstrated against K. pneumoniae, S. pyogenes, and S. aureus. Depending on the needs, such a preparation can be widely used in the design of functional food and/or the cosmetics industry.

Walnut Male Flowers (Juglans regia L.) as a Functional Addition to Wheat Bread.

The aim of this study is to assess the effect of adding dried and ground walnut male flowers to wheat flour on selected physicochemical and antioxidant properties of the obtained wheat bread. The control sample was bread without the addition of walnut male flowers. In the tests, the addition of flowers was used in the amounts of 0.5%, 1.0%, 1.5%, 2.0% and 2.5% of the weight of the flour used in the control sample. Bread dough was prepared according to the direct method with the use of yeast. The parameters of the baking process of the control bread and of the test bread were calculated. The selected parameters determining the quality of the obtained bread, including the loaf volume, the specific volume, the crumb porosity, the crumb texture and the color were measured. The antioxidant potential and total polyphenol content were tested both in the dough and in the bread. The test results were analyzed statistically. It was found that the partial replacement of wheat flour with walnut male flowers resulted in a significant reduction in the loaves' volume and in the darkening of both the color of the dough and the crumb. The value of the L* parameter decreased with the increasing addition of walnut flowers. The lowest values of this parameter were found to be 53.87 (crust) and 39.94 (crumb) in the sample with 2.5% addition. The average volume of the loaves ranged from 565 cm3 (0.5%) to 675 cm3 (control). The use of the additive in the amount of 2.5% caused significant changes in the values of most of the examined parameters of the bread crumb texture. The addition of walnut male flowers to the flour had a significant effect on the antioxidant potential and the total polyphenol content of the tested doughs and breads. In the case of bread, the total polyphenol content value ranged from 96.90 mg GAE/g dw to 208.08 mg GAE/g dw. Similarly, the antioxidant potential increased with increasing walnut male flower supplementation. The antioxidant potential determined by the ABTS and FRAP methods ranged from 1.42 mmol TE/g dw to 2.02 mmol TE/g dw and from 0.08 mmol TE/g dw to 0.33 mmol TE/g dw, respectively. The obtained research results prove the application potential of walnut flowers in the design of food, with beneficial pro-health properties, and are an example of the use of plant by-products in the process of food enrichment.

Phytochemical Screening and Bioactive Properties of Juglans regia L. Pollen.

Pollen is one of the major by-products of the walnut tree, yet it is poorly investigated. Thus, the aim of this study was to investigate the total phenolics, flavonoids, antioxidant, anticancer potential, and polyphenol profile of pollen obtained from male Juglans regia flowers. A total of 24 phenolic compounds were identified in Juglans regia pollen and all of them were reported for the first time for this raw material. The content of polyphenols was 408.03 mg/100 g dry weight (dw) and the most abundant components were quercetin 3-O-sophoroside and 4',5,7-trihydroxy-3,6-dimethoxyflavone-7-O-beta-D-glucoside. The concentration of these compounds, as well as the total content of polyphenols and flavonoids, strongly determined the antioxidant and cytotoxic activity of Juglans regia pollen. Antioxidant action using the ABTS and CUPRAC methods had the values of 3.35 and 0.32 mmol TE/g dw, respectively. In turn, in the tests of chelating ability of ferrous ion, O2•- and OH- radical scavenging activity, of which the results were expressed as IC50, the values were equal to 335.01, 459.31, and 92.89 µg/mL, respectively. Among the six cancer cell lines, the strongest effect was demonstrated for Caco-2 (140.65 µg/mL) and MCF-7 (140.98 µg/mL) cells. The results provide valuable and previously unpublished data on the polyphenol composition and biological potential of Juglans regia pollen.

Phenolic Profile and Antioxidant, Antibacterial, and Antiproliferative Activity of Juglans regia L. Male Flowers.

Juglans regia L., walnut, is a large, long-living tree, cultivated in temperate climates around the world. It is highly appreciated for its nutritional kernels and high-quality timber. Its barks, leaves, and husk are used as dyes and in folk medicine as herbal remedies for several diseases. From a biological and chemical standpoint, relatively little is known about the male flowers of the tree. Therefore, the aim of the study was to evaluate the phenolic profile as well as in vitro antioxidant, antimicrobial, and antiproliferative activity of male Juglans regia L. flowers. Phenolic content was determined by UPLC/PDA/MS/MS analyses; antioxidant activity was assessed by five different methods; antimicrobial activity was evaluated against the six most common pathogenic strains of Gram-positive and Gram-negative bacteria, and antiproliferative properties were assessed against six cell lines. Most of the analyses carried out in this study were performed for the first time for this raw material. J. regia flower extract was characterized by a strong ability to scavenge DPPH˙ free radicals, hydroxyl radicals, and chelating metal ions. Among the examined bacterial strains and neoplastic lines, the strongest antimicrobial activity was shown against S. aureus, L. monocytogenes, and B. cereus, and cytotoxic activity against breast cancer, glioblastoma, and astrocytoma cells. Male J. regia flowers have also been found to be a rich source of phenolic compounds. The content of polyphenols in the extract was 4369.73 mg/100 g d.w., and 24 compounds from the group of flavonoids, phenolic acids, and juglunosides were identified. Additionally, a strong correlation between the content of polyphenols and the antioxidant capacity and cytotoxic activity was observed. This is why the tested J. regia flowers are an excellent source of effective natural antioxidant, antibacterial, and chemopreventive compounds that have potential to be used in the pharmaceutical or food industries.

Identification of a Potent and Selective 5-HT1A Receptor Agonist with In Vitro and In Vivo Antinociceptive Activity.

Opioids are the gold standard drugs for the treatment of acute and chronic severe pain, although their serious side effects constitute a big limitation. In the search for new and safer drugs, 5-HT1AR agonists are emerging as potential candidates in pain relief therapy. In this work, we evaluated the affinity and activity of enantiomers of the two newly synthesized, potent 5-HT1AR agonists N-[(2,2-diphenyl-1,3-dioxolan-4-yl)methyl]-2-[2-(pyridin-4-yl)phenoxy]ethan-1-ammonium hydrogenoxalate (rac-1) and N-((2,2-diphenyl-1,3-dioxolan-4-yl)methyl)-2-(2-(1-methyl-1H-imidazol-5-yl)phenoxy)ethan-1-ammonium hydrogenoxalate (rac-2) in vitro and in vivo. The role of chirality in the interaction with 5-HT1AR was evaluated by molecular docking. The activity of the rac-1 was tested in mouse models of acute pain (hot plate) and severe tonic nociceptive stimulation (intraplantar formalin test). Rac-1 was active in the formalin test with a reduction in paw licking in both phases at 10 mg/kg, and its effect was abolished by the selective 5-HT1AR antagonist, WAY-100635. The eutomer (S)-1, but not the racemate, was active during the hot plate test at 10 and 20 mg/kg, and this effect was abolished by 30 min treatment with WAY-100635 at 30 min. Similarly to 8-OH-DPAT, (S)-1 evoked a slow outward current and depressed spontaneous glutamatergic transmission in superficial dorsal horn neurons, more effectively than rac-1. The eutomer (S)-1 showed promising developability properties, such as high selectivity over 5-HT subtypes, no interaction with the µ receptors, and low hepato- and cardiotoxicity. Therefore, (S)-1 may represent a potential candidate for the treatment of acute and chronic pain without having the adverse effects that are commonly associated with the classic opioid drugs.

G protein-coupled receptor binding and pharmacological evaluation of indole-derived thiourea compounds.

Four 2-(1H-indol-3-yl)ethylthiourea derivatives were prepared by condensation of 2-(1H-indol-3-yl)ethanamine with the corresponding aryl/alkylisothiocyanates in a medium-polarity solvent. Their structures were confirmed by spectral techniques, and the molecular structure of 3 was determined by X-ray crystal analysis. For all derivatives, the binding affinities at the 5-HT2A and 5-HT2C receptors, as well as their functional activities at the 5-HT1A and D2 receptors, were determined. The arylthioureas 1 and 4 were the most active at the 5-HT1A receptor, showing, at the same time, significant selectivity over the studied 5-HT2 and D2 receptor subtypes. The compounds were tested for their pharmacological activities within the central nervous system in relevant mouse models. The involvement of the serotonergic system in the activity of 1 and 4 was indicated. The antinociceptive action of 4 was linked to its anti-inflammatory activity.

"Green preservatives": combating fungi in the food and feed industry by applying antifungal lactic acid bacteria.

Fungal food spoilage plays a pivotal role in the deterioration of food and feed systems and some of them are also able to produce toxic compounds for humans and animals. The mycotoxins produced by fungi can cause serious health hazards, including cancerogenic, immunotoxic, teratogenic, neurotoxic, nephrotoxic and hepatotoxic effects, and Kashin-Beck disease. In addition to this, fungal spoilage/pathogens are causing losses of marketable quality and hygiene of foodstuffs, resulting in major economic problem throughout the world. Nowadays, food spoilage can be prevented using physical and chemical methods, but no efficient strategy has been proposed so far to reduce the microbial growth ensuring public health. Therefore, lactic acid bacteria (LAB) can play an important role as natural preservatives. The protection of food products using LAB is mainly due to the production of antifungal compounds such as carboxylic acids, fatty acids, ethanol, carbon dioxide, hydrogen peroxide, and bacteriocins. In addition to this, LAB can also positively contribute to the flavor, texture, and nutritional value of food products. This review mainly focuses on the use of LAB for food preservation given their extensive industrial application in a wide range of foods and feeds. The attention points out the several industrial patents concerning the use of antifungal LAB as biocontrol agent against spoilage organisms in different fermented foods and feeds.

Lactobacillus amylovorus DSM 19280 as a novel food-grade antifungal agent for bakery products.

Mould spoilage is the main cause of substantial economic loss in bakery industry and might also cause public health problems due to the production of mycotoxins. The reduction of mould growth in bakery products is thus of crucial importance and there is great interest to develop safe and efficient strategies for this purpose. In this study Lactobacillus amylovorus DSM19280 has been shown to produce a wide spectrum of antifungal compounds active against common bread spoilage fungi. Among the indicator moulds, Aspergillus fumigatus and Fusarium culmorum were the most sensitive organisms. Several antifungal compounds were found to be present in synthetic medium inoculated with L. amylovorus DSM19280 strain, some of them being reported here for the first time. Wheat doughs fermented with L. amylovorus DSM19280 had good rheological properties and the breads thereof were of high quality as shown by rheofermentometer and texture analyser measurements. The results were compared with those obtained with a control non-antifungal L. amylovorus DSM20531(T) strain, a non-acidified and a chemically acidified dough. The quality of sourdough and bread fermented with L. amylovorus DSM 19280 was comparable to that obtained by using L. amylovorus DSM20531 (T). Additionally, breads were evaluated for the ability to retard the growth of Fusarium culmorum FST 4.05, Aspergillus niger FST4.21, Penicillium expansum FST 4.22, Penicillium roqueforti FST 4.11 and fungal flora from the bakery environment. The biological preservation of bread with L. amylovorus DSM 19280 was also compared to the most commonly used antifungal agent Calcium propionate. Breads containing sourdough fermented with L. amylovorus DSM 19280 were more effective in extending the shelf life of bread than the calcium propionate.

Chemical composition and antimicrobial activity of Clausena indica (Dalz) Oliv. (Rutaceae) essential oil from Vietnam.

The essential oil obtained from the branches and leaves of Clausena indica (Dalz) Oliv. (Rutaceae) has been analyzed by GC/MS. Fifty-three components of the essential oil, representing 96.9% of the total amount, were identified. The main constituents were myristicin (35.3%), terpinolene (16.7%), and delta-3-carene (11.3%). The essential oil was screened for antimicrobial activity, showing positive results against some bacterial strains. Moreover, a comparison between constituents of C. indica essential oil with the ones reported in the literature for other Clausena ssp. is pointed out and some chemotaxonomic considerations have been identified.

Quali-quantitative analyses of Flavonoids of Morus nigra L. and Morus alba L. (Moraceae) fruits.

Morus nigra L., belonging to the Moraceae family, is a decidious tree widely cultivated in Europe and West Asia. It has a long history of medicinal use in Chinese medicine, as a remedy for many kinds of diseases. The aim of the present study was to evaluate the profile of the phenolic constituents of M. nigra fruits and to compare their content with the fruits of another species of Morus, Morus alba, which is also very well known in folklore medicine. The fruits of black and white mulberries have been studied, and five compounds from the methanol extract have been identified by means of HPLC/PDA/ESI-MS. Four compounds (quercetin 3- O-glucoside, quercetin 3- O-rutinoside, kaempferol 3- O-rutinoside, and 5- O-caffeoylquinic acid) have been isolated by use of Sephadex LH-20 column chromatography and HPLC and characterized by means of NMR and ESI-MS. Furthermore, HPLC/PDA/ESI-MS analysis of the red pigment of M. nigra fruits revealed the presence of four anthocyanins recognized as cyanidin 3- O-glucoside, cyanidin 3- O-rutinoside, pelargonidin 3- O-glucoside, and pelargonidin 3- O-rutinoside. All of the compounds were quantified.

Phenolics of Arbutus unedo L. (Ericaceae) fruits: identification of anthocyanins and gallic acid derivatives.

Arbutus unedo L., the strawberry tree (Ericaceae family), is an evergreen shrub or small tree, typical of the Mediterranean fringe and climate. The aim of the present study was to evaluate the profile of the phenolic constituents of A. unedo fruits. Seven compounds were purified by Sephadex LH-20 column chromatography of the MeOH extract followed by HPLC and were characterized as arbutin, beta-D-glucogalline, gallic acid 4-O-beta-D-glucopyranoside, 3-O-galloylquinic acid, 5-O-galloylquinic acid, 3-O-galloylshikimic acid, and 5-O-galloylshikimic acid, by means of NMR and ESI-MS analyses. Moreover, LC-PDA-MS analysis of the red pigment of A. unedo fruits revealed the presence of three anthocyanins recognized as cyanidin 3-O-beta-D-galactopyranoside, delphinidin 3-O-beta-D-glucopyranoside, and cyanidin 3-O-beta-D-arabinopyranoside. These pigments were also quantified.