Advanced Extraction Techniques Combined with Natural Deep Eutectic Solvents for Extracting Phenolic Compounds from Pomegranate (Punica granatum L.) Peels.

Pomegranate (Punica granatum L.) peel is a potential source of bioactive phenolic compounds such as ellagic acid and α- and ß-punicalagin. This work explores the efficiency of natural deep eutectic solvents combined with ultrasound-assisted extraction (UAE) and pressurized liquid extraction (PLE) for their extraction. Five NaDESs were evaluated by employing UAE (25 °C, for 50 min) to determine their total phenolic content (Folin-Ciocalteu assay) and ellagic acid and α- and ß-punicalagin contents (high-performance liquid chromatography (HPLC-DAD)). The NaDES composed of choline chloride (ChCl) and glycerol (Gly) (1:2, molar ratio) was the most efficient in the UAE when compared with the rest of the NaDESs and water extracts. Therefore, ChCl:Gly was further evaluated using PLE at different temperatures (40, 80, 120 and 160 °C). The PLE-NaDES extract obtained at 80 °C for 20 min at 1500 psi exhibited the highest contents of ellagic acid and α- and ß-punicalagin compared to the rest of the temperatures and PLE-water extracts obtained under the same extraction conditions. Combining UAE or PLE with a NaDES emerges as a sustainable alternative for extracting ellagic acid and α- and ß-punicalagin from pomegranate peel.

Ellagic Acid: A Review on Its Natural Sources, Chemical Stability, and Therapeutic Potential.

Ellagic acid (EA) is a bioactive polyphenolic compound naturally occurring as secondary metabolite in many plant taxa. EA content is considerable in pomegranate (Punica granatum L.) and in wood and bark of some tree species. Structurally, EA is a dilactone of hexahydroxydiphenic acid (HHDP), a dimeric gallic acid derivative, produced mainly by hydrolysis of ellagitannins, a widely distributed group of secondary metabolites. EA is attracting attention due to its antioxidant, anti-inflammatory, antimutagenic, and antiproliferative properties. EA displayed pharmacological effects in various in vitro and in vivo model systems. Furthermore, EA has also been well documented for its antiallergic, antiatherosclerotic, cardioprotective, hepatoprotective, nephroprotective, and neuroprotective properties. This review reports on the health-promoting effects of EA, along with possible mechanisms of its action in maintaining the health status, by summarizing the literature related to the therapeutic potential of this polyphenolic in the treatment of several human diseases.

Jabuticaba residues (Myrciaria jaboticaba (Vell.) Berg) are rich sources of valuable compounds with bioactive properties.

Jabuticaba (Myrciaria jaboticaba (Vell.) Berg) is a Brazilian berry, very appreciated for in natura consumption. However, its epicarp is not normally consumed due to its stiffness and astringent taste, and in manufacture of products from jabuticaba fruit, it is responsible for the generation of large amounts of residues. The exploration of by-products is becoming important for the obtainment of valuable bioactive compounds for food and pharmaceutical industries. In this context, jabuticaba epicarp was studied regarding its chemical composition, namely in terms of phenolic compounds, tocopherols, and organic acids, and its bioactive properties, such as antioxidant, anti-proliferate, anti-inflammatory, and antimicrobial activities. A total of sixteen phenolic compounds, four tocopherols and six organic acids were identified in jabuticaba epicarp. Regarding bioactive properties, it showed high antioxidant activity, also presenting moderate anti-inflammatory, anti-proliferative, and antimicrobial activities. The extract did not present hepatotoxicity, confirming the possibility of its applications without toxicity issues.

Hydrolysable Tannins and Biological Activities of Meriania hernandoi and Meriania nobilis (Melastomataceae).

A bio-guided study of leaf extracts allowed the isolation of two new macrobicyclic hydrolysable tannins, namely merianin A (1) and merianin B (2), and oct-1-en-3-yl ß-xylopyranosyl-(1"-6')-ß-glucopyranoside (3) from Meriania hernandoi, in addition to 11 known compounds reported for the first time in the Meriania genus. The structures were elucidated by spectroscopic analyses including one- and two-dimensional NMR techniques and mass spectrometry. The bioactivities of the compounds were determined by measuring the DPPH radical scavenging activity and by carrying out antioxidant power assays (FRAP), etiolated wheat coleoptile assays and phytotoxicity assays on the standard target species Lycopersicum esculentum W. (tomato). Compounds 1 and 2 exhibited the best free radical scavenging activities, with FRS50 values of 2.0 and 1.9 µM, respectively.

Ellagitannins: Bioavailability, Purification and Biotechnological Degradation.

The ellagitannins are a group of phenolic compounds with biological activities. Ellagic acid is the product obtained from hydrolysis of ellagitannins. Information related to the biosynthesis of ellagitannins still been scarce and confused. The ellagitannins are obtained from plants and their purification process implies mainly the use of chromatographic techniques. The ellagitannin acyl hydrolase (EAH) also known as ellagitannase is an enzyme capable of hydrolyzing the ester bonds of ellagitannins and the consequent releasing of ellagic acid. Information about the EAH is not clear because the enzyme had showed different activities due to the low purity or complexity of substrates and there is no available information about the biochemical, physicochemical and molecular characteristics of EAH. The present review describes information related to the sources, biosynthesis and the purification of ellagitannins and a current assessment on the production of ellagitannase.

Cytotoxic and Chemopreventive Effects of Gemin D Against Different Mutagens Using In Vitro and In Vivo Assays.

BACKGROUND: Gemin D (GD) is an ellagitannin found in several plant species rich in phenolic compounds. Its many beneficial properties include antioxidant and antitumoral. OBJECTIVE: The present study assessed the genotoxicity, cytotoxicity, antigenotoxicity, and anticytotoxicity of GD by in vitro and in vivo assays. METHOD: The Ames mutagenicity assay in Salmonella typhimurium, Micronucleus and Comet tests in mice were used to evaluate the biological activities mentioned above. To assess the GD's protective effects against DNA damage induced by different mutagens we performed co-, pre- and/or post-treatment in these assays. RESULTS: There was no genotoxic effect of GD via Ames and Micronucleus tests, but in the Comet assay the highest dose induced DNA damage. This same highest dose presented a significant cytotoxicity in mice. In the antigenotoxicity, GD protected DNA against the action of 4-nitroquinoline-1-oxide and sodium azide by Ames test, and also against the harmful action of cyclophosphamide in pre- and co-treatment by Micronucleus and Comet tests, but it did not protect DNA in post-treatment. Regarding to anticytotoxicity, GD provoked an anticytotoxic effect only during pre-treatment. CONCLUSION: Therefore, GD showed relevant antigenotoxic, anticytotoxic and cytotoxic effects, which indicate that it may be a probable candidate for chemoprevention or for the development of new cancer therapies.

The Protective Potential of Phyllanthus niruri and Corilagin on Gastric Lesions Induced in Rodents by Different Harmful Agents.

The gastroprotective effect of the methanol extract of Phyllantus niruri and its main constituent, corilagin, were studied in vivo. The extract (50, 125, or 250 mg/kg, p. o.) inhibited ethanol-induced lesions in rats by 43 % (p < 0.001), 69 % (p < 0.001), and 99 % (p < 0.001), respectively. It also inhibited the formation of indomethacin-induced gastric ulcers in rats by 80 % (p < 0.01), 89 % (p < 0.01), and 97 % (p < 0.01). A decrease in lipid hydroperoxide levels (p < 0.01) and in myeloperoxidase activity (p < 0.05) evidenced a reduction of oxidative damage and neutrophil infiltration in gastric tissues from ulcerated mice using ethanol/HCl. Potent in vitro free radical scavenger activity (IC50 = 0.07) using the DPPH assay was observed. In contrast, the extract (250 mg/kg, i. d.) did not show antisecretory activity in pylorus-ligated rats, and also failed to inhibit the H+,K+-ATPase activity in vitro. However, in pylorus-ligated rats, the extract (50, 125, and 250 mg/kg, i. d.) increased adhered mucus content by 22 % (p < 0.05), 28 % (p < 0.01), and 38 % (p < 0.01), respectively. The involvement of prostaglandins, nonprotein endogenous sulfhydryl compounds, α2-receptors, and endogenous nitric oxide in the gastroprotection elicited by the extract was also evaluated. Finally, corilagin reduced the lesion area of ethanol-induced gastric ulcers in mice by 88 % (30 mg/kg, p. o.; p < 0.001). Based on these results, it has been concluded that P. niruri methanol extract possesses gastroprotective activity by different and complementary pathways, which together promote an improvement in gastric cytoprotection. The presence of corilagin may partially explain the effectiveness of the extract against gastric damage.

Identification of Ellagitannins and Flavonoids from Eugenia brasilienses Lam. (Grumixama) by HPLC-ESI-MS/MS.

The grumixama (Eugenia brasiliensis Lam.), also known as Brazilian cherry, is a fruit native to Brazil. This study identified the flavonoids in the flesh and seeds and ellagitannin in the flesh of purple and yellow varieties. The physicochemical characteristics and antioxidant capacity of these fruits were also evaluated. Anthocyanins and flavonols were found in high levels in the flesh of purple (32-180 mg 100 g(-1) FW) and yellow grumixama (13-41 mg 100 g(-1) FW), respectively. The major flavonoids identified were cyanidin 3-glucoside and quercetin aglycone. Furthermore, ellagitannins were found in high levels in the flesh of purple (82-243 mg ellagic acid equiv 100 g(-1) FW) and yellow grumixama (92 mg ellagic acid equiv 100 g(-1) FW) and seeds (2220-2905 mg ellagic acid equiv 100 g(-1) FW). The ellagitannin profiles of both varieties were first characterized in which pedunculagin isomers, strictinin isomers, and ellagic acid galloyl hexoside were the major ellagitannins identified. In summary, both varieties of the grumixama fruit as well as the seeds could be good sources of bioactive compounds, mainly ellagitannins.

Hydrolyzable tannins from hydroalcoholic extract from Poincianella pluviosa stem bark and its wound-healing properties: phytochemical investigations and influence on in vitro cell physiology of human keratinocytes and dermal fibroblasts.

Extracts from Poincianella pluviosa stem bark are used in traditional medicine of South America for its wound healing properties. For validation of this traditional use and for rationalizing a potential pharmaceutical development towards standardized preparations bioassay-guided fractionation of EtOH-water (1:1v/v) extract (crude extract, CE) of P. pluviosa bark was performed. HaCaT keratinocytes cell line and human primary dermal fibroblasts (pNHDF) were used as in vitro systems. Significant stimulation of mitochondrial activity was found for CE on both cell types, which caused a strong increase of cell proliferation of keratinocytes. Fractionation of CE over Sephadex LH20 revealed two inactive fractions (FA and FB) and an active fraction FC, which was further fractionated by MPLC into 4 subfractions. Subfraction FC1 increased mitochondrial activity and proliferation of keratinocytes and dermal fibroblasts in a dose dependent manner (10 to 100 µg/mL) and did not show necrotic cytotoxicity on keratinocytes (LDH release assay). FC1 was investigated by ESI-MS/MS and solid-state (13)C NMR which confirmed the presence of various polyphenols and hydrolyzable tannins. MS studies suggest the presence of pyrogallol (1), gallic acid (2), gallic acid methyl ester (3), ellagic acid (4), corilagin (5), 1,4,6-tri-O-galloyl-glucose (6), tellimagrandin I (7), 1,2,3,6-tetra-O-galloyl-glucose (8), mallotinic acid (9), tellimagrandin II (10), 1,2,3,4,6-penta-O-galloyl-glucose (11), geraniin (12), and mallotusinic acid (13).

Antifungal activity of pomegranate peel extract and isolated compound punicalagin against dermatophytes.

BACKGROUND: Dermatophyte species infect the epidermis and appendages, often with serious social and health-economic consequences. The hydroalcoholic extract of pomegranate fruit peel showed activity against the dermatophyte fungi Trichophyton mentagrophytes, T. rubrum, Microsporum canis and M. gypseum. METHODS: Hydroalcoholic extract was prepared with pomegranate peels. This crude extract was fractionated and submitted to liquid-liquid partition, resulting in an active fraction which was fractionated in a Sephadex LH-20 column, followed by a Lobar column. The structure of the active compound was established with the use of spectroscopic methods. RESULTS: The crude extract of pomegranate fruit peel showed activity against the dermatophytes Trichophyton mentagrophytes, T. rubrum, Microsporum canis, and M. gypseum, with MICs values of 125 µg/ml and 250 µg/ml, respectively for each genus. Punicalagin was isolated and identified by spectroscopic analysis. The crude extract and punicalagin showed activity against the conidial and hyphal stages of the fungi. The cytotoxicity assay showed selectivity for fungal cells than for mammalian cells. CONCLUSIONS: These results indicated that the crude extract and punicalagin had a greater antifungal activity against T. rubrum, indicating that the pomegranate is a good target for study to obtain a new antidermatophyte medicine.

Quantification of polyphenols and evaluation of antimicrobial, analgesic and anti-inflammatory activities of aqueous and acetone-water extracts of Libidibia ferrea, Parapiptadenia rigida and Psidium guajava.

ETHNOPHARMACOLOGICAL RELEVANCE: Vast numbers of plant species from northeastern Brazil have not yet been phytochemically or biologically evaluated. AIM OF THE STUDY: The goal of this work was to obtain, characterize and show the antimicrobial, analgesic and anti-inflammatory activities of aqueous and acetone-water extracts of Libidibia ferrea, Parapiptadenia rigida and Psidium guajava. MATERIALS AND METHODS: The plant material (100g) was dried, and the crude extracts were obtained by using turbo-extraction (10%; w/v) with water or acetone:water (7:3, v/v) as the extraction solvent. High-performance liquid chromatography (HPLC) methods were used to screen the crude extracts for hydrolysable tannins (gallic acid) and condensed tannins (catechins). The antibacterial activity was evaluated by agar-diffusion and microdilution methods against Gram-positive strains (Staphylococcus aureus ATCC 25923, Staphylococcus epidermidis INCQS 00016, Enterococcus faecalis ATCC 29212 and a clinical isolate of methicillin-resistant Staphylococcus aureus) as well as Gram-negative strains (Escherichia coli ATCC 25922, Salmonella enteritidis INCQS 00258, Shigella flexneri and Klebsiella pneumoniae). To evaluate the anti-inflammatory activity, a leukocyte migration model was used. Analgesic activity was determined by the hot plate test and the acetic acid-induced abdominal writhing test. Data were analyzed by analysis of variance (ANOVA) at a significance level of 5%. RESULTS: Parapiptadenia rigida presented the highest amount of total polyphenols (35.82 ± 0.20%), while the greatest catechin content was found in the acetone-water extract of Psidium guajava (EAWPg; 1.04 µg/g). The largest amounts of catechins were found in the aqueous extract of Libidibia ferrea (EALf; 1.07 µg/g) and the acetone-water extract of Parapiptadenia rigida (EAWPr; 1.0 µg/g). All extracts showed activity against Gram-positive bacteria. The aqueous and acetone-water extracts of Psidium guajava showed the greatest inhibition zones in the agar diffusion tests. In the evaluation of the minimum inhibitory concentration (MIC), the most susceptible Gram-positive bacterium was Staphylococcus epidermidis and the most susceptible Gram-negative bacterium was Shigella flexneri. EAPg and EAWPg showed the greatest MIC values. All extracts were significant inhibitors of leukocyte migration (p<0.05). Using the writhing test, significant analgesic activity was found for EAPr (50 mg/kg), EAWPr (100 mg/kg and 200 mg/kg) and EAWPg (50 mg/kg) (p<0.05). CONCLUSIONS: Thus, the appropriate extraction procedure preserves the chemical components such as gallic acid and catechin, and showed antimicrobial, anti-inflammatory and analgesic properties.

Antioxidant and antiangiogenic activity of Astronium graveolens Jacq. leaves.

Angiogenesis is involved in many physiological and pathological conditions. Natural compounds with antioxidant activity have also been reported to possess potent antiangiogenic properties by regulating angiogenesis modulators such as vascular endothelial growth factor (VEGF). Based on this, we screened the antioxidant and antiangiogenic activities of Astronium graveolens leaf extracts by a DPPH test and a competitive enzyme-linked immunosorbent assay, respectively. MeOH extract expressed a significant free radical-scavenging activity (EC50 = 37.65 µg/mL) and it was able to inhibit the interaction between placental growth factor (PlGF) (placental growth factor), a VEGF family member, and its receptor Flt-1 by more than 50% at 1 mg/mL. 1,2,3,4,6-Penta-O-galloyl-d-glucopyranose, 6 is the most active compound of the extract. It exhibited a high potency in scavenging DPPH (EC50 = 2.16 µg/mL) and reduced by 58% the PlGF/Flt-1 interaction at a concentration of 50 µM. Moreover, the known compounds (1-6) have been isolated for the first time in A. graveolens.

Characterization of four Phyllanthus species using liquid chromatography coupled to tandem mass spectrometry.

This paper reports a comparison of four Phyllanthus species (P. amarus, P. stipulatus, P. niruri and P. tenellus), commonly known as stone breaker, by the characterization of the chemical profile of their aqueous extracts. Such characterization was carried out using liquid chromatography coupled to ion trap tandem mass spectrometry (LC-IT-MS(n)) under reversed-phase gradient elution mode. The results of MS/MS and MS(3) on-line experiments, using the electrospray ionization source in the positive and negative mode, are extensively discussed. Furthermore, quercetin-3-O-ß-d-glucuronopyranoside was isolated in multimilligram scale from the aqueous extract of P. stipulatus and characterized by mass spectrometry and NMR. Although it is an unusual flavonol in natural products, LC-IT-MS(n) experiments showed it to be present also in P. amarus.

Atividade anti-helmíntica de plantas taniníferas do semiárido nordestino sobre nematóides gastrintestinais de pequenos ruminantes

O estudo de plantas taniníferas é uma alternativa que vem sendo pesquisada para o controle de nematóides gastrintestinais de pequenos ruminantes. Portanto, o objetivo desse estudo foi avaliar in vitro e in vivo a atividade a anti-helmíntica de plantas taniníferas do semiárido nordestino, como Anadenanthera colubrina (Vell.) Brenan (angico), Leucaena leucocephala (Lam.) de Wit (leucena), Mimosa tenuiflora (Willd.) Poir (jurema preta) e Myracrodruon urundeuva (Engl.) Fr. Allemão (aroeira), sobre nematóides gastrintestinais de pequenos ruminantes. Para avaliação in vitro foram preparados extratos acetona:água (70:30) das folhas e dos caules de cada planta. [...] Os resultados dos ensaios in vitro e in vivo foram submetidos a ANOVA e ao teste Kruskal-Wallis (p0,05). A análise histológica indicou que o número de eosinófilos e linfócitos foi semelhante nos três grupos (p>0,05). Apesar dos promissores resultados obtidos in vitro, as folhas e os caules de M. tenuiflora não apresentam efeito sobre o estabelecimento larvar de H. contortus em ovinos.

The study of tanniferous plants with anthelmintic activity is an alternative that has been studied for control of gastrointestinal nematodes of small ruminants. Thus the aim of this study was to evaluate in vitro and in vivo anthelmintic activity of plants tanniferous of northeastern semi-arid, asAnadenanthera colubrina(Vell.) Brenan ( angico), Leucaena leucocephala (Lam.) de Wit (leucena), Mimosa tenuiflora (Willd.) Poir (jurema preta) and Myracrodruon urundeuva (Engl.) Fr. Allemão (aroeira), on gastrointestinal nematodes ofsmall ruminants.In the evaluation in vitro acetone :ater (70:30) extracts were prepared from leaves and stems of each plant. [...] The results of in vitro and in vivo tests were submitted to ANOVA and Krusk al-Wallis test (p0.05) .The histological analysis indicated that the number of eosinophils and lymphocytes was similar in the three groups (p>0,05). Despite promising in vitro results, the leaves and stems of M. tenuiflora has no effect on larval establishment of H. contortus in sheep.

Antimicrobial activity of an Amazon medicinal plant (Chancapiedra) (Phyllanthus niruri L.) against Helicobacter pylori and lactic acid bacteria.

The potential of water extracts of the Amazon medicinal plant Chancapiedra (Phyllanthus niruri L.) from Ecuador and Peru for antimicrobial activity against Helicobacter pylori and different strains of lactic acid bacteria such as Lactobacillus acidophilus, Lactobacillus casei and Lactobacillus plantarum was investigated. H. pylori was inhibited by both water extracts in a dose dependent manner, whereas lactic acid bacterial growth was not affected. Both extracts contained ellagic acid and hydroxycinnamic acid derivatives and exhibited high free radical scavenging linked-antioxidant activities (89%). However, gallic acid was detected only in the Ecuadorian extract. Preliminary studies on the mode of action of Chancapiedra against H. pylori revealed that inhibition may not involve proline dehydrogenase-based oxidative phosphorylation inhibition associated with simple mono-phenolics and could involve ellagitannins or other non-phenolic compounds through a yet unknown mechanism. This study provides evidence about the potential of Chancapiedra for H. pylori inhibition without affecting beneficial lactic acid bacteria.

Characterization of gallotannins from Astronium species by flow injection analysis- electrospray ionization-ion trap-tandem mass spectrometry and matrix-assisted laser desorption/ionization time-of- flight mass spectrometry.

The species Astronium urundeuva (Allemao) Engl. and Astronium graveolens Jacq., which are used in Brazilian folk medicine to treat allergies, inflammation, diarrhea and ulcers, were investigated for their composition. The aim of this study was to define a rapid and reliable analytical approach, based on the flow-injection analysis-electrospray ionization-ion trap-tandem mass spectrometry (FIA-ESI-IT-MS-MS) and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-ToF-MS), to investigate the full range of hydrolyzable tannins present in the extracts of these Astronium species. The MALDI-ToF-MS analysis allowed us to ascertain the presence of hydrolysable tannins in both Astronium species as a series of gallotannins with degrees of polymerization of 7 to 13 galloyl units. Moreover, the analysis by FIA-ESI-IT-MS-MS, as well as confirming this result and chemically defining gallotannins as galloylglucose compounds, highlighted the presence of further classes of hydrolysable tannins, such as hexahydrodiphenoyl esters of glucose and some gallic acid derivatives, providing information about their structure by a careful study of their fragmentation patterns. Finally, the evaluation of the number of positional isomers of gallotannins occurring in both Astronium species was obtained by high-performance liquid chromatography-electrospray ionization-ion trap mass spectrometry (HPLC/ESI-IT-MS). This is the first mass spectrometric evidence relating to the existence of gallotannins in Astronium genus.

Antifungal effects of Ellagitannin isolated from leaves of Ocotea odorifera (Lauraceae).

Ocotea odorifera is a medicinal plant that is popularly known in Brazil as "canela-sassafrás" and is used to treat dermatosis. This study investigated the antifungal properties of O. odorifera. The methanol extract of O. odorifera was submitted to successive chromatographic separation and yielded Tellimagrandin II (TEL). Candida parapsilosis strain ATCC 22019 was used to determine the minimum inhibitory (MIC) and fungicidal concentrations, and to study the synergistic action with nystatin (NYS), amphotericin (AMP), and fluconazole (FLU). After treatment, the morphology of the yeast was analysed by scanning electron microscopy. Cytotoxicity was assessed in Vero cells, and genotoxicity by the micronucleus test. The TEL structure was proposed based on NMR and comparison with literature data and ESI-MSMS analysis. The compound showed potent inhibitory activity against C. parapsilosis, with a MIC of 1.6 µM. TEL acted synergistically with NYS, AMP, and FLU, and caused morphological alterations in the yeast cells. The methanolic extract showed low cytotoxicity in vitro and in vivo, and was not mutagenic in mice (P < 0.05). The use of O. odorifera in traditional medicine seems to have a valid basis, in view of the antifungal activity of TEL demonstrated in this study, and may contribute to potential drug development.

Potent antifungal activity of extracts and pure compound isolated from pomegranate peels and synergism with fluconazole against Candida albicans.

Activity-guided repeated fractionation of crude hydro alcoholic extract prepared from the fruit peel of Punica granatum on a silica-gel column yielded a compound that exhibited strong antifungal activity against Candida spp. Based on spectral analyses, the compound was identified as punicalagin. Punicalagin showed strong activity against Candida albicans and Candida parapsilosis, with MICs of 3.9 and 1.9 microg/ml, respectively. The combination of punicalagin and fluconazole showed a synergistic interaction. MIC for fluconazole decreased twofold when combined with the extract. The FIC index was 0.25. The synergism observed in disk-diffusion and checkerboard assays was confirmed in time-kill curves. The effect of punicalagin on the morphology and ultrastructure in treated yeast cells was examined by scanning and transmission electron microscopy. An irregular budding pattern and pseudohyphae were seen in treated yeasts. By transmission electron microscopy, treated cells showed a thickened cell wall, changes in the space between cell wall and the plasma membrane, vacuoles, and a reduction in cytoplasmic content. Since the punicalagin concentration effective in vitro is achievable in vivo, the combination of this agent with fluconazole represents an attractive prospect for the development of new management strategies for candidiasis, and should be investigated further in in vivo models.

Simultaneous removal of chromium and leather dye from simulated tannery effluent by photoelectrochemistry.

The feasibility of the photobleaching of a leather acid dye, acid red 151, simultaneously to degradation of anionic surfactant, Tamol, and reduction of Cr(VI) to the less toxic Cr(III) was investigated by photoelectrocatalytic oxidation. The best experimental conditions were found to be pH 2.0 and 0.1 mol L(-1) sodium sulfate when the nanoporous Ti/TiO2 photo anode was biased at +1.0 V and submitted to UV-irradiation. The photoelectrocatalytic oxidation promotes 100% discoloration, reducing around 98-100% of Cr(VI) and achieving an abatement of 95% of the original total organic carbon. The effect of pH, the applied potential, the Cr(VI) concentration and the complexation reaction between Cr(VI) and acid red dye were evaluated as to their effect on the kinetics of the reaction.

Induction of salivary polypeptides associated with parotid hypertrophy by gallotannins administered topically into the mouse mouth.

Isoproterenol-induced salivary polypeptides (IISP), a group of proline-rich proteins synthesized by mouse parotids, have been considered as markers for isoproterenol-induced parotid hypertrophy. Rodents fed diets containing high-tannin cereals (sorghum), also develop parotid hypertrophy. To test whether tannins are directly involved in provoking sialotrophic growth, we studied the effect of intraperitoneal and topical oral administrations of tannic acid (TA) on the induction of IISP polypeptides in endogamic mice (A/Snell). TA was characterized by HPLC chromatography and spectral analysis and shown to be composed solely of gallotannins, a complex family of glucose and gallic acid esters. IISP polypeptides were monitored in saliva by SDS-polyacrylamide gel electrophoresis during 36 h after ending TA stimulation. Single daily intraperitoneal administrations of TA for 3 consecutive days (0.033 mg/g bw/day), at variance of parallel administrations of isoproterenol (0.042 mg/g bw/day) failed to induce IISP polypeptides. However, repeated topical applications of TA into the mouse mouths (1.21 mg/g bw divided into three equal doses given at 4-h intervals within a single day) resulted in unequivocal induction of IISP polypeptides. That response was clearly intensified by increasing the stimulation frequency to eight equivalent doses given at 1.5-h intervals within a single day (corresponding to 3.23 mg/g bw) and even further by repeating this protocol for 3 days. Under these productive schemes of stimulations by TA, electrophoretic fractionation of parotid homogenates showed new polypeptide bands migrating in parallel to salivary IISP. These results suggest that topically administered gallotannins are effective inducers of trophic growth in mouse parotids.