Phytochemical Profiles and Antimicrobial Activity of Selected Populus spp. Bud Extracts.

Buds of poplar trees (Populus species) are often covered with sticky, usually polyphenol-rich, exudates. Moreover, accessible data showed that some Populus bud extracts may be excellent antibacterial agents, especially against Gram-positive bacteria. Due to the fragmentary nature of the data found, we conducted a systematic screening study. The antimicrobial activity of two extract types (semi-polar-ethanolic and polar-ethanolic-water (50/50; V/V)) from 27 bud samples of different poplar taxons were compared. Antimicrobial assays were performed against Gram-positive (five strains) and Gram-negative (six strains) bacteria as well as fungi (three strains) and covered the determination of minimal inhibitory, bactericidal, and fungicidal concentrations. The composition of extracts was later investigated by ultra-high-performance liquid chromatography coupled with ultraviolet detection (UHPLC-DAD) and with electrospray-quadrupole-time-of-flight tandem mass spectrometry (UHPLC-ESI-qTOF-MS). As a result, most of the extracts exhibited good (MIC ≤ 62.5 µg/mL) or moderate (62.5 < MIC ≤ 500 µg/mL) activity against Gram-positives and Helicobacter pylori, as well as fungi. The most active were ethanolic extracts from P. trichocarpa, P. trichocarpa clone 'Robusta', and P. tacamahaca × P. trichocarpa. The strongest activity was observed for P. tacamahaca × P. trichocarpa. Antibacterial activity was supposedly connected with the abundant presence of flavonoids (pinobanksin, pinobanksin 3-acetate, chrysin, pinocembrin, galangin, isosakuranetin dihydrochalcone, pinocembrin dihydrochalcone, and 2',6'-dihydroxy-4'-methoxydihydrochalcone), hydroxycinnamic acids monoesters (p-methoxycinnamic acid cinnamyl ester, caffeic acid phenethylate and different isomers of prenyl esters), and some minor components (balsacones).

Polyphenol Profile of Cistus × incanus L. and Its Relevance to Antioxidant Effect and α-Glucosidase Inhibition.

The European Food Safety Authority recommends C. incanus as a natural source of antioxidants. Its activity is essentially determined by polyphenols, although specific compounds are not finally indicated. The available plant material comes from different subspecies and locations, which can lead to differences in chemical composition and potency. For this reason, we conducted a detailed analysis of the polyphenol content and antioxidant activity of 52 different C. incanus teas from Turkey, Albania, Greece, and unspecified regions. We focused special attention on ellagitannins, which have not been properly determined so far. Besides oxidative stress, hyperglycemia is an essential component of cardiometabolic diseases. Therefore, in subsequent experiments, we evaluated the ability of C. incanus extracts and individual polyphenols to inhibit α-glucosidase. Using statistical methods, we analyzed how differences in chemical composition affect activity. The results showed that C. incanus is a rich source of ellagitannins (2.5-19%), which dominate among polyphenols (5.5-23%). Turkish-origin products had higher ellagitannin content and a greater antioxidant effect (FRAP, ABTS) than Albanian and Greek products. In contrast, the flavonoid and phenolic acid contents and DPPH values were at similar levels in all products. An in-depth analysis of their composition indicated that all groups of polyphenols are involved in the antioxidant effect, but a significant contribution can be attributed to ellagitannins and flavonoids. C. incanus extracts showed a high capacity to inhibit α-glucosidase activity (IC50 125-145 µg/mL). Ellagitannins were the most effective inhibitors (IC50 0.7-1.1 µM), with a potency exceeding acarbose (3.3 mM). In conclusion, C. incanus, due to the presence of ellagitannins and flavonoids, exhibits powerful antioxidant and α-glucosidase inhibitory effects.

Fragaria × ananassa cv. Senga Sengana Leaf: An Agricultural Waste with Antiglycation Potential and High Content of Ellagitannins, Flavonols, and 2-Pyrone-4,6-dicarboxylic Acid.

Strawberry leaves are considered a valuable waste material; so far, mainly due to their antioxidant properties. Since the annual production of this crop is high, our study aimed to thoroughly examine the chemical composition and antidiabetes-related bioactivity of Fragaria × ananassa leaf of its popular and productive cultivar Senga Sengana. Leaves from three different seasons, collected after fruiting, were extensively analyzed (UHPLC-qTOF-MS/MS, HPLC-DAD). Some individual components were isolated and quantified, including specific flavonol diglycosides (e.g., 3-O-[ß-xylosyl(1‴→2″)]-ß-glucuronosides). The separated quercetin glycosides were tested in an antiglycation assay, and their methylglyoxal uptake capacity was measured. In addition, the biodegradable polyester precursor 2-pyrone-4,6-dicarboxylic acid (PDC) was confirmed at relatively high levels, providing further opportunity for strawberry leaf utilization. We want to bring to the attention of the food, pharmaceutical, and cosmetic industries the Senga Sengana strawberry leaf as a new botanical raw material. It is rich in PDC, ellagitannins, and flavonols-potent glycation inhibitors.

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.

Quantification of Salicylates and Flavonoids in Poplar Bark and Leaves Based on IR, NIR, and Raman Spectra.

Poplar bark and leaves can be an attractive source of salicylates and other biologically active compounds used in medicine. However, the biochemical variability of poplar material requires a standardization prior to processing. The official analytical protocols used in the pharmaceutical industry rely on the extraction of active compounds, which makes their determination long and costly. An analysis of plant materials in their native state can be performed using vibrational spectroscopy. This paper presents for the first time a comparison of diffuse reflectance in the near- and mid-infrared regions, attenuated total reflection, and Raman spectroscopy used for the simultaneous determination of salicylates and flavonoids in poplar bark and leaves. Based on 185 spectra of various poplar species and hybrid powdered samples, partial least squares regression models, characterized by the relative standard errors of prediction in the 4.5-9.9% range for both calibration and validation sets, were developed. These models allow for fast and precise quantification of the studied active compounds in poplar bark and leaves without any chemical sample treatment.

Sanguinarine-Chelerythrine Fraction of Coptis chinensis Exerts Anti-inflammatory Activity in Carrageenan Paw Oedema Test in Rats and Reveals Reduced Gastrotoxicity.

Inflammatory diseases are a common therapeutic problem and nonsteroidal anti-inflammatory drugs are not deprived of side effects, of which ulcerogenic activity is one of the most frequent. The aim of the study was to evaluate the anti-inflammatory activity of the sanguinarine-chelerythrine (SC) fraction of Coptis chinensis and its influence on the integrity of gastric mucosa. The study was conducted on sixty male rats randomly divided into six experimental groups: two control groups (a negative control group CON and a positive control group CAR); three groups receiving an investigational fraction of C. chinensis (1, 5, 10 mg/kg i.g.) named SC1, SC5, and SC10, respectively; and a group receiving indomethacin (IND) (10 mg/kg i.g.) as a reference drug. In all animals, the carrageenan-induced paw oedema was measured; PGE2 release, TNFα production, and MMP-9 concentration in inflamed tissue were determined. Additionally, the macroscopic and microscopic damage of gastric mucosa was evaluated. Administration of SC dose-dependently inhibited the second phase of carrageenan rat paw oedema and PGE2 release, decreased the production of TNFα, and reduced the concentration of MMP-9, and the efficacy of the highest dose was comparable to the effect of IND. Contrary to IND, no gastrotoxic activity of SC was detected. The investigated sanguinarine-chelerythrine fraction of C. chinensis seems to be a promising candidate for further research on new anti-inflammatory and analgesic drugs characterized with a safer gastric profile compared to existing NSAIDs.

An in-depth look into a well-known herbal drug: Fingerprinting, isolation, identification, and content estimation of saponins in different Strophanthus seeds.

Seeds of Strophanthus species are known as a source of rapid-acting cardenolides. These water-soluble glycosides are listed as the sole critical constituents of this raw herbal drug. A non-standard cardioprotective medication with ouabain-containing oral remedies has become popular in Europe as a result of the withdrawal of corresponding registered drugs from the market. However, the bioequivalence of pure ouabain solutions, tinctures, and home-made extracts from Strophanthus seeds is unknown. Thus, this study aimed to update the information on the composition of Strophanthus seeds used for this purpose. The distribution of two main saponins and about 90 previously unreported compounds, tentatively identified as saponins in eleven Strophanthus species, was systematically evaluated by ultra-high-performance liquid chromatography mass spectrometry (UHPLC-MS) and -MS/MS. Seeds of S. gratus were selected to isolate the dominant unreported triterpenoids, bidesmosides of echinocystic and oleanolic acid. Their structures were established by HRMS, MS/MS, as well as by NMR techniques. The total saponin content, estimated by UHPLC-MS, was up to 1%. The detected saponins could influence the peroral bioavailability of hardly absorbable Strophanthus cardenolides and exhibit their own activity. This finding may be relevant when Strophanthus preparations (containing both saponins and cardiac glycosides) are used, particularly when homemade preparations are administered.

Composition and Antimicrobial Activity of Ilex Leaves Water Extracts.

Plants from the Ilex genus are known for properties such as antimicrobial and anti-inflammatory activity, can act as antiobesity agents and thus can be helpful in medicine. Some holly species, such as Ilex paraguariensis (widely known in the form of popular beverage: yerba mate), have been investigated, while others have been partially researched or remain unknown. Therefore, we performed qualitative and quantitative phytochemical analyses and screened antimicrobial properties of lesser-studied species (I. aquifolium L., I. aquifolium 'Argentea Marginata' and I. × meserveae 'Blue Angel'). I. paraguariensis was used as a standard species for comparison purposes. Investigations were performed on water extracts due to their expected activity and composition. Antimicrobial research included evaluating minimal inhibitory, bactericidal (Staphylococcus aureus and Escherichia coli) and fungicidal concentration (Candida albicans, Alternaria alternata, Fusarium oxysporum, and Aspergillus niger) of extracts. The influence of the extracts on the production, eradication, and viability of bacterial biofilms was also analysed. It was established that Ilex paraguariensis possesses the richest profile of hydroxycinnamic acids derivatives in terms of component concentration and diversity. Ilex spp., especially I. × meserveae, contain a slightly higher amount of flavonoids and more different flavonoid derivatives than I. paraguariensis. However, the strongest antibacterial activity was shown by I. aquifolium L. and its cultivar 'Argentea Marginata' in terms of minimal inhibitory, bactericidal and fungicidal concentration, and biofilm assays. Extracts from both species significantly reduced the biofilm viability of S. aureus as well, which may be of use in the production of multicomponent lavaseptics, antiseptics, diuretics (supporting urinary tract infection therapy) and, due to their action on fungi, additives to growth media for specific fungi. The significant content of saponins enables Ilex extracts to be used as natural emulsifiers, for example, in cosmetics. Moreover, relatively high chlorogenic acid and rutin content may suggest use of Ilex spp. to treat obesity, digestive problems, in chemoprevention, and as preservatives in the food industry.

The real Theriac - panacea, poisonous drug or quackery?

ETHNOPHARMACOLOGICAL RELEVANCE: Theriac is considered the most popular cure-all multi-ingredient medicine and has been used for more than two millennia. It has also been used as one of the most important anti-epidemic drugs up to the 19th c., treated as an emergency medicine in case of e.g. bubonic plague. AIM OF THE STUDY: Until now, no reliable information regarding the pharmacological effect of the treacle was available, including its possible toxic or narcotic properties. In order to change the state of knowledge in this matter we have selected the Theriac recipe that had been actually used for producing the treacle in 1630, which was confirmed by the official municipal documents of the time. METHODS: The recipe was written in Latin, with the use of pre-Linnean nomenclature and then apothecary common names, which required translation into the modern scientific language in order to get reliable pharmacological conclusions. The information from historical sources has been compiled with the pharmacological data concerning the most potent compounds, which for the first time made it possible to calculate the amounts of active compounds in the doses taken by then patients. RESULTS: Only two species included in Theriac can be harmful in humans: poppy and sea squill, but in both cases the calculated quantity of morphine and cardiac glycosides, respectively, were below toxic level. There are no indications, both from the historical and pharmacological point of view, for Theriac being toxic or narcotic in patients, when used as prescribed. CONCLUSIONS: As for now, the most probable is that the treacle owed its postulated efficacy in the main indications to the placebo effect. Still, the results should be further confirmed by reconstructing the actual Theriac and subjecting it to modern tests and analyses.

The Effect of Ilex × meserveae S. Y. Hu Extract and Its Fractions on Renal Morphology in Rats Fed with Normal and High-Cholesterol Diet.

Therapeutic properties of Ilex species are widely used in natural medicine. Ilex × meserveae may become a potential substitute for Ilex paraguariensis (Yerba Mate). As a part of the preliminary safety verification of this European Ilex hybrid vs. Yerba Mate, an eight-week study concerning the impact of regular administration of leaves of both species on kidneys was conducted. The standard water infusion and three dominant fractions of Ilex × meserveae leaves' constituents (polyphenols, saponins and less polar terpenoids) were separately tried on 96 male Wistar rats divided into 8-member groups. Animals were divided into two basic nutritional groups: the first one was rats fed standard feed and the second on was rats fed with high-cholesterol diet (20 g of cholesterol per kg of standard feed). Postmortem morphometric evaluation of stained kidney samples concerned the filtration barrier elements, which are crucial in proper diuresis. The results showed that saponins present in the hydroalcoholic dry extract (administered in a dose of 10 mg/kg of body weight/day) as well as in water infusions (1:20) from Ilex × meserveae and Ilex paraguariensis do not demonstrate nephrotoxicity but conversely, have a protective role on kidney status in animals fed with a normal diet and in a high-cholesterol diet.

Isolation and structural determination of flavan-3-ol derivatives from the Polypodium vulgare L. rhizomes water extract.

Polypodium vulgare L. (Polypodiaceae) is a fern used in traditional Polish medicine as an expectorant to treat cough and pertussis. Additionally, it was used as a diuretic in renal diseases, especially in chronic nephritis and pyelonephritis. In our study, a water extract was prepared from the rhizome of common polypody and subsequently fractionated on a resin column. As a result, the mixture of flavan-3-ol derivatives was obtained after the column elution with 60% methanol. Further purification by various chromatographic techniques led us to the isolation of (+)-afzelechin (1), a new previously not reported (+)-afzelechin-7-O-α-l-arabinofuranoside (2), and three other monomer flavan-3-ol glycosides: (+)-afzelechin-7-O-ß-d-apiofuranoside (3), (+)-catechin-7-O-α-l-arabinofuranoside (4) and (+)-catechin-7-O-ß-d-apiofuranoside (5). Structures of the compounds were established by HR-ESI-MS, 1D and 2D NMR spectroscopy. The HSQC and HMBC NMR techniques were used in the structure elucidation of the position of sugar attachment.

In Search of High-Yielding and Single-Compound-Yielding Plants: New Sources of Pharmaceutically Important Saponins from the Primulaceae Family.

So far, only a few primrose species have been analyzed regarding their saponin composition and content. Moreover, the roots of only two of them are defined by the European Union (EU) Pharmacopoeia monograph and commercially utilized by the pharmaceutical industry. Thus, this study intended to find some new sources of main triterpene saponins from Primulae radix, namely primulasaponins I and II together with the closely related sakurasosaponin. Using isolated standards, UHPLC-ESI-HRMS served to assess over 155 Primulaceae members qualitatively and quantitatively. Nine examples of plants accumulating over 5% of primulasaponin I in their roots were found. Among them, in one case, it was found as the almost sole secondary metabolite with the concentration of 15-20% (Primula grandis L.). A reasonable content of primulasaponin II was found to be typical for Primula vulgaris Huds. and P. megaseifolia Boiss. & Bal. The sakurasosaponin level was found in seven species to exceed 5%. The finding of new, single and rich sources of the abovementioned biomolecules among species that were never analyzed phytochemically is important for future research and economic benefit. The chemotaxonomic significance of the occurrence of these three saponins in Primulaceae is discussed.

Treatment of an oronasal fistula in a patient on bisphosphonate therapy: A case study.

Bisphophonates (BPs) are a group of drugs used in treating bone diseases, which may lead to the development of the osteonecrosis of the jaw (ONJ). The negative impact of BPs on angiogenesis is among the causes of ONJ. The specific mechanisms of complications are unknown. What is taken into consideration is the trauma background, which, in combination with the implemented BP treatment, can induce bone necrosis. One of the possible consequences of necrotic change progression is the development of an oronasal fistula. Treatment generally requires a surgical intervention.The paper describes the course of treatment of an oronasal fistula in a patient with BP osteitis, currently using an upper denture. The fistula arose a year after the removal of a protruding sequestrum in the region of the hard palate. An attempt was made to treat the fistula by the mobilization of soft tissues from the palate and the bilayered closure of the fistula with the use of a pedicled connective tissue graft on the greater palatine artery, along with a Tinti-Parma-Benfenati (TPB) flap. The patient was subjected to appropriate post-procedural measures. Regular follow-ups did not reveal any abnormalities in the course of healing.The use of the abovementioned procedure proved to be an effective method of treatment of an oronasal fistula. The use of a pedicled connective tissue graft for the closure of the oronasal fistula caused by BP therapy had a significant effect on the treatment outcome.

Analysis of Polyphenolic Composition of a Herbal Medicinal Product-Peppermint Tincture.

The pharmacological activity of peppermint leaf (Menthae piperitae folium) for medical use is mainly attributed to the presence of essential oil, which, according to the European Pharmacopoeia (Ph. Eur.), should constitute not less than 12 mL/kg of raw material. The content of polyphenols in peppermint-based preparations, except peppermint leaf dry extract, has not yet been considered as an essential parameter in the pharmacopeial assessment of peppermint quality. This study concerns the evaluation of the presence of representatives of polyphenolic compounds in 23 commercial peppermint tinctures (ethanolic extracts) purchased in pharmacies in Poland. The non-volatile polyphenolic fraction was investigated, and the presence of flavonoids and phenolic acids was quantified. High performance liquid chromatography coupled with a diode-array detector (HPLC-DAD) and an electrospray ionization mass spectrometer (U(H)PLC-ESI-MS) were used in the experiment. The study showed that eriocitrin, luteolin-7-O-rutinoside, and rosmarinic acid were the main polyphenolic components of the peppermint tinctures, as previously reported for peppermint leaf. Despite this, the research shows the extremely diverse content of the mentioned compounds in analyzed commercial medicinal products. In light of these results, it seems that the pharmacopeial assessment for the peppermint leaf (Ph. Eur.) and peppermint tincture (Polish Pharmacopoeia (FP)) requires correction and supplementation.

Lanostane-Type Saponins from Vitaliana primuliflora.

The phytochemistry of the genera Androsace, Cortusa, Soldanella, and Vitaliana, belonging to the Primulaceae family is not well studied so far. Hence, in this paper, we present the results of UHPLC-MS/MS analysis of several primrose family members as well as isolation and structure determination of two new saponins from Vitaliana primuliflora subsp. praetutiana. These two nor-triterpenoid saponins were characterized as (23S)-17α,23-epoxy-29-hydroxy-3ß-[(O-ß-d-glucopyranosyl-(1→2)-O-α-l-rhamnopyranosyl-(1→2)-O-ß-d-glucopyranosyl-(1→2)-O-α-l-arabinopyranosyl-(1→6)-ß-d-glucopyranosyl)oxy]-27-nor-lanost-8-en-25-one and (23S)-17α,23-epoxy-29-hydroxy-3ß-[(O-α-l-rhamnopyranosyl-(1→2)-O-ß-d-glucopyranosyl-(1→2)-O-α-l-arabinopyranosyl-(1→6)-ß-d-glucopyranosyl)oxy]-27-nor-lanost-8-en-25-one, respectively. Their structures were determined by high resolution mass spectrometry (HRMS), tandem mass spectrometry (MS/MS), one- and two-dimensional nuclear magnetic resonance spectroscopy (1D-, and 2D-NMR) analyses. So far, the 27-nor-lanostane monodesmosides were rarely found in dicotyledon plants. Therefore their presence in Vitaliana and also in Androsace species belonging to the Aretia section is unique and reported here for the first time. Additionally, eleven other saponins were determined by HRMS and MS/MS spectra. The isolated lanostane saponins can be considered as chemotaxonomic markers of the family Primulaceae.

Nitrogen compounds in Phacelia tanacetifolia Benth. honey: First time report on occurrence of (-)-5-epi-lithospermoside, uridine, adenine and xanthine in honey.

Lacy phacelia (Phacelia tanacetifolia Borkh.) honey composition was screened by UHPLC-DAD-QqTOF-MS. The targeted analysis revealed 6 major nitrogen compounds including aromatic amino acids (tyrosine, phenylalanine), purine derivatives (adenine, xanthine), nucleoside (uridine) and rare non-cyanogenic cyanoglucoside, (-)-5-epi-lithospermoside ((2Z)-2-[(4R,5R,6S)-4,5-dihydroxy-6-(ß-d-glucopyranosyl)oxycyclohex-2-en-1-ylidene]acetonitrile). Their identity was confirmed by different analytical tools: HRMS, co-chromatography with standard compound or comprehensive NMR experiments. All the compounds, except amino acids, were reported and determined in honey for the first time. The amount of the compounds was quantified in 16 unifloral phacelia samples: adenine (18.45 ±â€¯4.63 mg/kg), xanthine (10.53 ±â€¯2.98 mg/kg), uridine (42.84 ±â€¯9.26 mg/kg), tyrosine (14.66 ±â€¯10.22 mg/kg), (-)-5-epi-lithospermoside (70.61 ±â€¯31.37 mg/kg) and phenylalanine (20.41 ±â€¯11.99 mg/kg). The (-)-5-epi-lithospermoside content is significantly correlated with P. tanacetifolia pollen percentage (R2 = 0.5612, p < 0.001) and it is proposed as a potential marker of botanical origin for phacelia honey.

Triterpenoid Acids as Important Antiproliferative Constituents of European Elderberry Fruits.

In Europe, both the fruits and flowers of Sambucus nigra L. have been used against cold, as well as laxative, diaphoretic, and diuretic remedies. There are also a number of commercially available food products that contain elderberry juice, puréed or dried elderberries. Recent comprehensive literature data on pharmacology and chemistry of Sambuci fructus have encouraged us to screen extracts with different polarities from this plant material against cancer cell lines. The cytotoxic activity of the ethyl acetate and aqueous acetone extracts from elderberries as well as detected triterpenoids on human colon adenocarcinoma cell line (LoVo) and human breast cancer cell line (MCF-7) was investigated by sulforhodamine B assay. Moreover, cell migration assay was conducted for triterpenoid fraction and pure compounds. Aqueous acetone extract possessed much lower IC50 value in cancer cell lines compared to ethyl acetate extract. The latter manifested high cytotoxicity against studied cell lines, suggesting that nonpolar compounds are responsible for the cytotoxic activity. Indeed, the phytochemical analysis revealed that ursolic and oleanolic acids are the main triterpenoids in the mentioned extract of which ursolic acid showed the highest activity with IC50 values of 10.7 µg/mL on MCF-7 and 7.7 µg/mL on LoVo cells.

In vitro evaluation of the antioxidant and antimicrobial activity of DIMBOA [2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one].

The aim of this study was to evaluate the in vitro antioxidant and antimicrobial properties of the natural cyclic hydroxamic acid: 2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one (DIMBOA). Antioxidant activity of the isolated DIMBOA was examined using DPPH, FRAP and ABTS tests. It was found that DIMBOA exhibits a potent free-radical scavenging activity and a weaker iron (III) ions reducing activity. Antimicrobial activity against selected G(+), G(-) bacterial strains and against yeasts-like reference strains of fungi was investigated using disk-diffusion method. It has been shown that DIMBOA possess growth inhibitory properties against many strains of studied bacteria and fungi, such as Staphylococcus aureus, Escherichia coli as well as against Saccharomyces cerevisiae.

ß-Aescin at subinhibitory concentration (sub-MIC) enhances susceptibility of Candida glabrata clinical isolates to nystatin.

Aescin (escin) derived from the seeds of horse chestnut (Aesculus hippocastanum L.) is a natural mixture of triterpene saponins exhibiting a wide variety of pharmacological properties, including antiinflammatory, analgesic, and antipyretic activities. However, data concerning antifungal activities of these compounds are limited. This study aims to evaluate the in vitro antifungal susceptibility of Candida glabrata clinical isolates to α-aescin sodium, ß-aescin crystalline and ß-aescin sodium using the disk diffusion (DD) and broth microdilution (BMD) methods. Moreover, the influence of subinhibitory concentration (0.5×MIC) of ß-aescins on the nystatin MIC was also studied. In general, the results obtained by the DD assay correlated well with those obtained by the BMD method. Both ß-aescins effectively inhibited the growth of all 24 strains tested. The minimum inhibitory concentration (MIC) values ranging from 8 to 32 µg/ml for ß-aescin crystalline, whereas those of ß-aescin sodium were slightly lower and ranged from 4 to 16 µg/ml. In contrast, α-aescin sodium was found to be completely ineffective against the strains studied. MIC values of nystatin were reduced 2-16-fold and 2-4-fold in the presence of subinhibitory concentration of ß-aescin crystalline and ß-aescin sodium, respectively. Results of the present study may suggest the additive interaction between ß-aescin and nystatin.

Determination of ursolic and oleanolic acid in Sambuci fructus.

Elderberries are used in the preparation of pie, jelly, punch, wine, or liqueur, as well as in many herbal remedies and food supplements. Elderberry products may provide diaphoretic, diuretic, antioxidant, and immunostimulant activities that offer protection against cold and flu. Herein, we report for the first time the qualitative and quantitative evaluation of two isomeric triterpenoids isolated from Sambuci fructus. The analysis revealed that ursolic acid and oleanolic acid are present in Sambuci fructus. The average concentration of ursolic acid was ca. three times higher than the concentration of oleanolic acid. The triterpenoids were detected and quantified using chromatographic methods such as TLC and HPLC. Spectroscopic techniques, including HR-MS and 2D-NMR, allowed unequivocal structure determination.