Antibacterial and Antibiofilm Activity of Different Species of Fabiana sp. Extract Obtained via Maceration and Ultrasound-Assisted Extraction against Staphylococcus epidermidis.

Staphylococcus epidermidis is an opportunistic pathogen that, under certain conditions, can induce aggravated infectious processes, mainly in immunosuppressed patients. Moreover, S. epidermidis is one of the leading causes of medical device- and implant-associated infections and is also recognized as a canonical biofilm producer. Fabiana punensis, F. densa and F. patagonica are three medicinal plants that grow in arid environments in Argentina (Altoandina, Puna, Prepuna and Monte regions). In this work, we studied the antimicrobial activity of alcoholic extracts of these plant species obtained via maceration (M) and ultrasound-assisted extraction (UAE) against S. epidermidis. In addition, the antibiofilm activity of the F. densa extract was also evaluated. It was found that the extracts obtained via M did not present differences with those obtained via UAE regarding the chemical profile. F. densa showed the lowest minimum inhibitory concentration (MIC) value (75 µg GAE/mL). At concentrations higher than the MIC, the extract induced the release of cellular constituents. At the concentration of 1/8× MIC, the extract inhibited biofilm formation by 78%, reducing metabolic activity by 67%. On the other hand, it presented a low percentage of preformed biofilm removal. In all assays, gallic acid (GA) has been used as a reference antimicrobial compound. Finally, it was shown via microscopy visualization that the extract reduces adhesion to hydrophobic and hydrophilic surfaces. Thus, F. densa extracts could potentially be used for the antibiotic treatment of infections produced by S. epidermidis or as an inhibitor agent of production biofilm, avoiding infections caused by medical devices.

Gochnatia glutinosa (D.Don) D.Don ex Hook. & Arn.: A plant with medicinal value against inflammatory disorders and infections.

Gochnatia glutinosa is a shrub that grown in the Argentinean semiarid region (Monte region) used in the ancestral medicine as an antiseptic and anti-inflammatory agent. This study was aimed to examine the morpho-anatomical characteristics of G. glutinosa aerial parts, identify the chemical composition of traditionally used preparations to assess its pharmacobotanical characterization and evaluate its activity as antiseptic and anti-inflammatory to give scientific support to its traditional uses. G. glutinosa morpho-anatomical description was performed following standard histological techniques. Tincture and infusion of its aerial parts were prepared and were subjected to phytochemical analysis. Xanthine oxidase (XOD) and lipoxygenase (LOX) inhibition experiments, as well as ABTS•+, superoxide radical, and hydrogen peroxide scavenging activity, were carried out. The growth inhibition of methicillin-resistant Staphylococcus aureus (MRSA) strains was also determined. The morpho-anatomical traits of G. glutinosa leaves and stems were reported for the first time. The medicinal preparations exhibited a large amount of phenolic chemicals mainly flavonoids such as rhamnetin, arcapillin, rhamnacin, hesperetin, isorhamnetin, centaureidin, europetin 7-O-mehylmyricetin, cirsiliol, sakuranetin, genkwanin and eupatorine and also phenolic acids and diterpenoid derivatives. Both preparations had free radical scavenging activity and were able to reduce both XOD and LOX activity, indicating their anti-inflammatory properties. Besides, tincture was effective against all MRSA strains (MIC values ranging from 60 to 240 g DW/mL). The results obtained in this work scientifically support the medicinal popular use of G. glutinosa as an antiseptic and anti-inflammatory. The identification of bioactive compounds and their morpho-anatomical description contribute to the quality control of this medicinal plant from Argentine Calchaquí Valley.

Nicotiana tabacum Leaf Waste: Morphological Characterization and Chemical-Functional Analysis of Extracts Obtained from Powder Leaves by Using Green Solvents.

Tobacco cultivation and industrialization are characterized by the production of trillions of pre-harvest and post-harvest waste biomasses each year with the resulting negative effects on the environment. The leaves of blunt, pre-harvest waste, could be further used to obtain bioactive metabolites, i.e., polyphenols and alkaloids, for its potential cosmetic use. This study was conducted to obtain bio-compounds from pre-harvest tobacco leaf waste (var. Virginia) by applying conventional and green solvents (NaDES). Leaves and ground leaf waste were characterized based on their microscopic features. Conventional solvents, such as water, acetone, ethanol, and non-conventional solvents, such as Natural Deep Eutectic Solvents (NaDES), i.e., sucrose:lactic acid (LAS), frutose:glucose:sucrose (FGS), lactic acid:sucrose:water (SALA), choline chloride:urea (CU), and citric acid: propylene glycol (CAP) were used for bioactive extraction from tobacco waste powder. CU, FGS, and acetone/ethanol had similar behavior for the best extraction of alkaloids (6.37-11.23 mg ACE/g tobacco powder). LAS, FGS, SALA, and CU were more effective in phenolic compound extraction than conventional solvents (18.13-21.98 mg AGE/g tobacco powder). Because of this, LAS and SALA could be used to obtain phenolic-enriched extracts with lower alkaloid content rather than CU and FGS. Extracts of the powder obtained with conventional solvent or CU showed a high level of sugars (47 mg/g tobacco powder) The ABTS antioxidant capacity of tobacco leaf powder was higher in the extracts obtained with CU, FGS, and acetone (SC50 1.6-5 µg GAE/mL) while H2O2 scavenging activity was better in the extracts obtained with LAS, CAP and SALA (SC50 3.8-8.7 µg GAE/mL). Due to the biocompatibility of the NaDES with the components of tobacco leaf waste, the opportunity to apply these extracts directly in antioxidant formulations, such as cosmetics, phytotherapic, and other formulations of topic use seems promising. Furthermore, NaDES constituents, i.e., urea and organic acid can also have beneficial effects on the skin.

A Flavonoid-rich Zuccagnia punctata Extract Prevents High Fat Diet-induced Normal Weight Obesity in a Rabbit Model.

Oral administration of rich in flavonoids hydroalcoholic extract from Zuccagnia punctata (ZpE) improves lipid profile and prevents vascular dysfunction in hypercholesterolemic rabbits. This study aimed to evaluate the ability of ZpE to prevent metabolic and vascular alterations induced by high fat diet (HFD) on a metabolically obese and normal weight rabbit model. The major components of ZpE were analyzed by HPLC method. Rabbits were separated into six groups: 1-fed on standard chow (CD); 2-fed on HFD; 3, 4, 5- fed on HFD and orally administrated 2.5 mg, 5 mg or 10 mg GAE/day of ZpE, respectively (ZpE- HFD); 6- fed on HFD and orally administered 30 mg orlistat/day (Or-HFD). All diets were administrated by 6 weeks. The major compounds of ZpE identified were chalcones: 2',4'-dihydroxy-3'-methoxychalcone and 2',4'-dihydroxychalcone. Oral treatment with ZpE 5 mg GAE/day as well as orlistat prevented the HFD-induced increase of triglycerides, fasting glucose, intraperitoneal glucose test, white cells, and TyG index. Acetylcholine relaxation was reduced in arteries from HFD group and oral administration of ZpE reached this response to CD values. Contractile response to angiotensin II was lower in arteries from rabbits fed on HFD treated with ZpE 5 and 10 mg GAE/day than those of untreated rabbits. Moreover, ZpE could inhibit the activity of pancreatic lipase in vitro and in vivo. In conclusion the ZpE may prevent normal weight obesity by inhibiting the pancreatic lipase. Thus, the use of ZpE as a natural product in the prevention of metabolic syndrome and endothelial dysfunction is very promising.

Potential use of native fruits waste from Argentina as nonconventional sources of cosmetic ingredients.

BACKGROUND: Collagenase, hyaluronidase, elastase, and tyrosinase enzymes are overexpressed and overactive in the skin aging process and hydrolyze the components of the dermal extracellular matrix (ECM) of the skin; these enzymes produce the clinical framework of aging, which includes skin dryness, hyperpigmentation, wrinkles, and inelasticity. AIMS: The aim of this study was to explore the potential use of waste from two Argentine native fruits, namely Ziziphus mistol, and red and orange varieties of Solanum betaceum, as sources of bioactive compounds. METHODS: Phenolic enriched extracts (PEE) from waste of Z. mistol and S. betaceum were obtained, and their total contents of phenolics and flavonoids were evaluated. The bioactive properties of the extracts were analyzed by measuring their antioxidant capacity and the inhibitory activity on collagenase, hyaluronidase, elastase, and tyrosinase enzymes. RESULTS: The increased ability to inhibit the collagenase was demonstrated by the PEE of Z. mistol seeds and peel, while the enzyme elastase was mostly inhibited by extracts of S. betaceum skin. Z. mistol seed extract was the most active to inhibit hyaluronidase, reaching 96% inhibition at a concentration of 100 µg GAE/mL. The most active extracts to inhibit the tyrosinase enzyme were obtained from the peel of two varieties of chilto fruits, orange and red, and the mistol seed. CONCLUSIONS: The results obtained suggest that Z. mistol and S. betaceum waste may be considered as a source of bioactive phenolics. Here, Argentine native fruits waste is presented as a most promising alternative in cosmetic products, with future uses such as hydrogels, creams, or lotions.

Evaluation of the antibacterial synergism of two plant extracts belonging to Bignoniaceae family and development of a topical formulation

Abstract Fridericia caudigera and Cuspidaria convoluta (Bignoniaceae) species, which grow in the northwest of Argentina, have shown antibacterial effect against strains isolated from skin infections, and each one displayed synergism with commercial antibiotics. The aims of this work were to evaluate the antibacterial activity and toxicity of the combination of these two plant species, and to design a stable gel for topical use including the blend of extracts. The combination of extracts was evaluated for synergistic effects (chequerboard assay), genotoxicity (Ames test) and cytotoxicity (Artemia salina test). A gel was subsequently formulated with the combination of extracts using carboxymethylcellulose as a polymer. The following physico- chemical characteristics of the gel formulation: pH, viscosity, spreadability and total phenol content, as well as resistance to severe temperature changes, biological activity (diffusion in agar), in vitro permeation (Franz cells) and primary dermal irritation (Draize test) were analyzed. The combination of extracts showed a synergistic effect on pathogenic bacteria and was not toxic in the in vitro tests. The gel was stable and retained the antimicrobial activity of the original extracts. The formulation proposed in this work could constitute an alternative for primary skin infections since it proved to be safe for topical administration.

Flower beverages of native medicinal plants from Argentina (Acacia caven, Geoffroea decorticans and Larrea divaricata) as antioxidant and anti-inflammatory.

ETHNOPHARMACOLOGICAL RELEVANCE: Herbal beverages have been used as a natural part of the medicinal and food culture in northwestern Argentina. The flower beverages (infusion or decoction) of Acacia caven, Geoffroea decorticans and Larrea divaricata, three native species from arid and semiarid regions of Argentina are widely used as anti-inflammatory and anti-rheumatic by several local communities. AIM OF THE STUDY: The aim of this study was to analyze the phytochemical composition of some Argentine flower beverage and to validate its traditional use as an antioxidant and anti-inflammatory agent. MATERIALS AND METHODS: Phenolic profiles from all flower infusions and decoctions were analyzed by both spectrophotometric analysis and HPLC-DAD. ABTS•+; the scavenging activity of both hydrogen peroxide and hydroxyl radical was determined and finally, their ability to inhibit pro-inflammatory enzymes, such as xanthine oxidase (XOD), and lipoxygenase (LOX) was also assessed. RESULTS: The flower beverages of all assayed species showed a high level of phenolic compounds with similar chromatographic patterns in both infusions and decoctions of each plant species, the major components of which have been identified. The flower beverages, especially G. decorticans infusion and decoctions, displayed an important antioxidant activity (SC50 values between 18.14 and 47 µg/mL) through different mechanisms; all of them were able to inhibit the XOD enzyme activity and, consequently, the formation of uric acid and reactive oxygen species, the primary cause of arthritis-related diseases. The most active beverages as XOD inhibitor were G. decorticans flower infusion and decoctions (IC50 values of 20 and 35 µg/mL, respectively). Pro-inflammatory enzymes, such as LOX, were also inhibited by infusions and decoctions of G. decorticans, L. cuneifolia and A caven flowers, lessening inflammation mediators in all beverages. CONCLUSIONS: The present work validates the traditional medicinal use of flower beverages from Argentina as an anti-rheumatic and anti-inflammatory agent as it has been used for hundreds of years in several pathologies associated with oxidative stress.

Propolis from the Monte Region in Argentina: A Potential Phytotherapic and Food Functional Ingredient.

The aim of this review is to provide overall information on Argentine propolis and to shed light on its potential, especially the one from the Monte region so as to support future research in the field. Around 1999, the Argentine propolis began to be chemically and functionally characterized to give it greater added value. Because Argentina has a wide plant biodiversity, it is expected that its propolis will have various botanical origins, and consequently, a different chemical composition. To date, five types have been defined. Based on their functionality, several products have been developed for use in human and veterinary medicine and in animal and human food. Because the Argentine propolis with the greatest potential is that of the Monte eco-region, this review will describe the findings of the last 20 years on this propolis, its botanical source (Zuccagnia punctata Cav.), its chemical composition, and a description of markers of chemical quality (chalcones) and functionality. Propolis can regulate the activity of various pro-inflammatory enzymes and carbohydrate and lipid metabolism enzymes, as well as remove reactive oxygen and nitrogen species. Consequently, it can modulate metabolic syndrome and could be used as a functional ingredient in food. Furthermore, hydroalcoholic extracts can act against human and animal pathogenic bacteria and human yeast, and mycelial pathogenic fungi. The ability to stop the growth of post-harvest pathogenic bacteria and fungi was also demonstrated. For this reason, Argentine propolis are natural products capable of protecting crops and increasing the lifespan of harvested fruit and vegetables. Several reports indicate the potential of Argentine propolis to be used in innovative products to improve health, food preservation, and packaging. However, there is still much to learn about these natural products to make a wholesome use of them.

Potential use of medicinal plants from Argentinean highland as agent anti-photoaging.

BACKGROUND: The overexposure to sun ultraviolet (UV) radiation produce photoaging by effect of free radicals on lipid, protein, and nucleic acid or by direct activation of responsible enzymes of cleavage of extracellular matrix components (EMC). AIMS: To develop new anti-photoaging agents, the anti-aging activity of four "jarillas" standardized extracts from Argentina (Zuccagnia punctata Cav.,Larrea divaricata Cav.,Larrea cuneifolia Cav., and Larrea nitida Cav.) were examined. METHODS: The effect on the activity and expression of enzymes related to photoaging (collagenase, hyaluronidase, elastase, tyrosinase) and the antioxidant capacity of four "jarillas" extracts were examined. RESULTS: The results suggest that "jarillas" extracts could protect EMC by inhibition of skin aging-related enzymes and suppression of the expression of metalloproteinases (MMP-1) in human skin fibroblasts from photoinduced damage. Modulation of MMP-1 expression induced by UV radiation in human skin cells could be associated, at least in part, with the ROS scavenging capacity of "jarilla" extracts. The extract of Z punctata was more active than Larrea species extracts in all the trials. The activity of Z punctata and Larrea extracts could be ascribed to chalcones and lignans, major constituents of "jarillas," respectively. CONCLUSIONS: The results obtained would indicate the potential of these adapted species to live in arid zones of Argentina, exposed to high ultraviolet radiation in production of cosmetic products and justify the promotion of their sustainable use and the development of crops.

Antigenotoxic, antiproliferative and antimetastatic properties of a combination of native medicinal plants from Argentina.

ETHNOPHARMACOLOGICAL RELEVANCE: Jarilla is the common name of an appreciated group of native plants from the semi-arid region in Argentina (Larrea cuneifolia Cav., Larrea divaricata Cav. and Zuccagnia punctata Cav.) that have been historically consumed to heal respiratory, musculoskeletal and skin ailments, as well as recommended for weakness/tiredness, hypertension, diabetes and cancer treatment. It was previously reported that some biological properties could be improved when these plants are used jointly. Infusions of a defined mixture, composed by three Jarilla species, L. cuneifolia: L. divaricata: Z. punctata (0.5:0.25:0.25) (HM2) showed synergistic and additive effect on antioxidant activity even after passing through the gastro-duodenal tract. AIM OF THE STUDY: The main purpose of this work was to evaluate antigenotoxic, antitumor, and anti-metastatic properties of the Jarilla species that grow in the Northwest of Argentina and a herbal combination of them. MATERIAL AND METHODS: Infusions of Jarilla mixture (HM2), and of each single plant species were prepared. Phenolic profiles of infusions were analyzed by HPLC-ESI-MS/MS and two relevant chemical markers were quantified. The antigenotoxic activity was evaluated by using the Ames test and the Cytokinesis-Block Micronucleus (CBMN) assay against direct mutagens. Evaluations of both cytotoxicity and antiproliferative effects were conducted on tumor and non-tumor cell lines. Both in vivo tumoral growth and metastasis inhibition were evaluated by using a carcinoma model on Balb/c mice. RESULTS: HM2 mix could suppress genetic and chromosome mutations induced by 4-nitro-o-phenylendiamine (4-NPD) and doxorubicin. Herbal mixture and single plant infusions showed cytotoxic effect against mammary, uterus, and brain tumoral cells without a selective action vs normal human cell line. HM2 mix was able to reduce mammary tumor mass on the Balb/c mice model and showed a significant reduction in the number of metastatic nodules in the lungs. CONCLUSIONS: Our results suggest that the combinations of three Jarilla species from northwest Argentina would be a promising alternative to treat or slow down the development of chronic diseases, such as cancer.

Flourensia fiebrigii S.F. blake: A medicinal plant from the Argentinean highlands with potential use as anti-rheumatic and anti-inflammatory.

ETHNOPHARMACOLOGICAL RELEVANCE: Flourensia fiebrigii is a plant used in traditional medicine in the Argentine Calchaquí Valley as purgative, expectorant, anti-rheumatic and anti-inflammatory. AIM OF THE STUDY: The aim of this study was to analyze the macroscopic and microscopic characteristics of F. fiebrigii leaf and stem, the phytochemical composition of leaves ethanolic extracts and to validate its traditional use as anti-rheumatic and anti-inflammatory. MATERIALS AND METHODS: The macroscopic and microscopic description of F. fiebrigii leaf and stem was carried out. Two extracts (immersions and tinctures) from leaves were obtained. The phytochemical analysis and UHPLC-OT-MS metabolome fingerprinting of both extracts were performed. The anti-rheumatic and anti-inflammatory activities of both extracts were determined using enzymatic inhibition assays of xanthine-oxidase (XOD), secretory phospholipase A2 (sPLA2) and lipoxygenase (LOX). RESULTS: The macroscopic and micrographic characters of F. fiebrigii were described to allow the botanical characterization of the plant species. The leaves extracts showed a high level of phenolic compounds with similar chromatographic patterns. Forty-five compounds were identified based on UHPLC-OT-MS including several sesquiterpenes, chalcones, flavonoids, isoflavonoids, a lignan and phenylpropanoids phenolic acids that have been identified for the first time in this plant species. F. fiebrigii extracts were able to inhibit the XOD activity and, consequently, the formation of uric acid and reactive oxygen species, primary cause of diseases, such as gouty arthritis (IC50 values of 1.10-2.12 µg/mL). Pro-inflammatory enzymes like sPLA2 and LOX were also inhibited by F. fiebrigii extracts (IC50 values of 22.00-2.20 µg/mL) decreasing the production of inflammation mediators. CONCLUSIONS: The present work validates the traditional medicinal use of F. fiebrigii as anti-rheumatic and anti-inflammatory through the use of enzymatic assays. The presence of several chemical compounds with demonstrated anti-rheumatic and anti-inflammatory properties also supports the bioactivity of the F. fiebrigii.

Antifungal, anti-inflammatory and antioxidant activity of bi-herbal mixtures with medicinal plants from Argentinean highlands.

ETHNOPHARMACOLOGICAL RELEVANCE: Argentinean medicinal plants such as Larrea divaricata Cav., Larrea cuneifolia Cav., Larrea nitida Cav., Zuccagnia punctata Cav. and Tetraglochin andina Ciald. are used alone and in combination in traditional medicine by inhabitants from northwestern Argentina to solve mycosis, vaginal infections, gastrointestinal, respiratory, and inflammatory processes. AIM OF THE STUDY: To assess the effect of interactions between hydroalcoholic extracts of these five species of medicinal plants against yeast strains isolated from vaginal infections, select the most active mixtures and evaluate anti-inflammatory and antioxidant activities. MATERIAL AND METHODS: Synergy between the plant extracts was studied by using a broth microdilution assay by means of the checkerboard method against Saccharomyces cerevisiae, Candida albicans, and non albicans strains. The inhibitory effect on lipoxygenase and the antioxidant capacity in cell-free and cell systems were studied. The chemical profile was evaluated by qualitative and quantitative screening, whereas chemical markers were quantified by HPLC-DAD. RESULTS: A synergistic antifungal effect was observed in some binary combinations. Z. punctata/L. divaricata, Z. punctata/L. cuneifolia, and Z. punctata/L. nitida were the most active mixtures. Nordihydroguaiaretic acid and 2',4'-dihydroxychalcone, two antifungal compounds, present in these extracts, were identified and quantified by HPLC-DAD. Both single extracts and bi-herbal mixtures showed antioxidant activity (in cellular and in cell-free systems) and were active on pro-inflammatory enzymes (LOX). CONCLUSIONS: Our results indicated that the most active combinations of these species extracts could be useful in the treatment of vaginal infectious diseases caused by Saccharomyces cerevisiae and Candida spp. strains and in associated oxidative and inflammatory processes, supporting its traditional use. In addition, the results highlighted the phyto-therapeutic potential of total phytochemical compounds present in these medicinal plants.

Integral use of Argentinean Solanum betaceum red fruits as functional food ingredient to prevent metabolic syndrome: effect of in vitro simulated gastroduodenal digestion.

The "red chilto" (Solanum betaceum Cav) is a native fruit from the Yungas forest of Argentina. Red chilto is a neglected and underutilized native species (NUS). The objective of this work was to evaluate the potentiality of pulp, seed and skin from "red chilto" as a functional food ingredient to add value to a native resource of Argentine Yungas to promote sustainable integral use of it. The powders have low carbohydrate and sodium content and are a source of vitamin C, phenolic acids (rosmarinic acid and caffeoylquinic acid), anthocyanins, condensed tannins, carotenoids, potassium, and fiber. The phenolics of chilto powders showed, before and after simulated gastroduodenal digestion, antioxidant activity (ABTS•+; H2O2; O2 •; HO•) and were able to inhibit enzymes related to metabolic syndrome, such as α-glucosidase, α-amylase and lipase. Chilto powder showed hypoglycemic effect by increasing glucose adsorption, decreasing glucose diffusion rate and by promoting glucose transport across the cell membrane. These results suggest the potential of Argentinean "red chilto" fruits as functional food ingredients or dietary supplements to prevent metabolic syndrome principally by its antioxidant, hypoglycemic and hypolipemic effects.

Antifungal activity of phytotherapeutic preparation of Baccharis species from argentine Puna against clinically relevant fungi.

ETHNOPHARMACOLOGICAL RELEVANCE: B. boliviensis and B. tola are used in traditional medicine in the Argentine Puna to treat skin and soft tissue infections and inflammatory processes in humans and animals. AIM OF THE STUDY: To assess the potential of phytotherapeutic preparations of Baccharis species as antifungal agents against clinically relevant fungi and to determine the chemical composition of the extracts. MATERIAL AND METHODS: Phytotherapeutic preparations of B. boliviensis and B. tola collected in Argentine Puna were evaluated as an antifungal agent against clinically relevant fungi (yeast, non-dermatophytes, and dermatophytes) isolated of patients from a local Hospital, and reference strains, using macrodilution and microdilution assays. The bioactivity was supported by UHPLC-OT-MS metabolome fingerprinting. RESULTS: The results revealed that the plant preparations were active against most of evaluated fungal strains; B. boliviensis was more active than B. tola. Dermatophyte fungi strains were the most sensitive isolates. The phytotherapeutic preparation showed Minimal Inhibitory Concentration (MIC) values between 25 and 400 µg GAE/mL and Minimum Fungicidal Concentration (MFC) values between 50 and 400 µg GAE/mL. Regarding the phytochemical analysis, total phenolic and total flavonoid contents of hydroalcoholic preparation of B. boliviensis were greater than those of the B. tola extract. Both Baccharis species showed similar chromatographic patterns, fifty-two compounds were identified based on UHPLC-OT-MS including several terpenoids, flavonoids and phenolic acids that have been identified in this two endemic South American Baccharis species for the first time. Several identified compounds present antifungal properties, the presence of these compounds support the bioactivity of the Baccharis extracts. CONCLUSIONS: In this work the traditional use of both Baccharis species as an antimicrobial against commercial products resistant fungal strains was validate, principally against dermatophytes fungi such as T. rubrum, T. mentagrophytes, M. canis, and M. gypseum. These results indicate that the hydroalcoholic preparations could be used for the treatment of fungal infectious.

The use of jarilla native plants in a Diaguita-Calchaquí indigenous community from northwestern Argentina: An ethnobotanical, phytochemical and biological approach.

ETHNOPHARMACOLOGICAL RELEVANCE: In northwestern Argentina inhabit several ancient indigenous communities with diverse cultural and historical background. Geographical isolation has contributed to the prevalence of a native plant-based folk medicine; "jarilla" species are medicinal plants widely used in local communities for the treatment of mycosis, respiratory, gastrointestinal and rheumatic disorders. THE AIM OF THE STUDY: To assemble the traditional knowledge acquired through years with scientific data concerning to phytochemistry, antioxidant and anti-inflammatory potential of three "jarillas" species. MATERIAL AND METHODS: Ethnobotanical data of three "jarillas", Zuccagnia punctata (Zp), Larrea cuneifolia (Lc), and Larrea divaricata (Ld), were explored by interviewing native people from Indigenous Community of Amaicha del Valle, Tucumán. Phenolic profiles from each infusion were analyzed by HPLC-ESI-MS/MS. Antioxidant activity was determined by superoxide anion and hydrogen peroxide scavenging capacity, lipoperoxidation inhibition, and ferrous iron chelating activity. It was also assessed their ability to inhibit pro-inflammatory enzymes, such as xanthine oxidase, lipoxygenase, and hyaluronidase. RESULTS: Ethnobotanical interviews showed that local people use "jarillas" mainly as infusions and baths. It was reported different categories of uses, such as medicinal (10 curative applications), to religious purposes, tinctorial, as construction material and as fuel. From infusions prepared, the MS and MS/MS data allowed the identification of 27 compounds from Z. punctata, and 11 from both Larrea sp. The infusions showed an important antioxidant activity through different mechanisms, highlighting Zp and Lc in free radical scavenging capacity and Ld on lipid peroxidation inhibition and iron binding. They were also capable of inhibit xanthine oxidase and lipoxygenase enzymes, being Lc the most active one. CONCLUSIONS: This research work provides novel information concerning to several categories of traditional uses of "jarilla" species in a Diaguita-Calchaquí community and focus attention to infusions from a phytochemical and biological approach.

Argentinean Puna Plants with In Vitro Antioxidant and Anti-Inflammatory Activities as a Potential Nutraceutical.

Parastrephia lucida (Compositae), Tessaria absinthioides (Compositae), and Ephedra multiflora (Ephedraceae), three plant species from the Argentinean Puna (3600 m.a.s.l.) were selected for their anti-inflammatory and antioxidant properties to prepare mixtures to evaluate their use as nutraceuticals. Seven binary and ternary herbal mixtures made of ethanol 20% extracts of the selected plant species were prepared (Mixtures A to G), and they were analyzed for their effect on proinflammatory enzymes and their antioxidant activity in two cellular systems and in cell free systems. Toxicity tests were also carried out, and they were analyzed by high-performance liquid chromatography with a diode-array detector (HPLC-DAD) to quantify chemical markers. Mix A (equal parts of the three selected plant species) showed an important inhibitory capacity of different proinflammatory enzymes. Its potency on COX-2 was also higher than that of ibuprofen. Mix A and Mix G (P. lucida and T. absinthioides 1:1) showed a high antioxidant capacity in cellular and in cell-free systems. Toxicity assays further demonstrated their safety. This work shows the potential use of herbal mixtures made of medicinal plant species from the Argentinean Puna as nutraceutical or dietary supplements with antioxidant and anti-inflammatory activities. PRACTICAL APPLICATION: P. lucida, T. absinthioides, and E. multiflora are three plant species that are commonly used by Argentinean Puna inhabitants with medicinal purposes. Their proven safety, their antioxidant activity as well as their capacity to inhibit different proinflammatory enzymes make them attractive candidates to be used in combination as part of a dietary supplement aimed to prevent or palliate gastrointestinal and systemic inflammatory diseases. The use of native plant species as an alternative to more common and commercial plant species would have a positive impact on local communities' economies.

Oral administration of Zuccagnia punctata extract improves lipid profile, reduces oxidative stress and prevents vascular dysfunction in hypercholesterolemic rabbits.

BACKGROUND: The consumption of flavonoids has been shown to prevent cardiovascular diseases including atherosclerosis. In this sense, in a recent in vitro study we demonstrated that a rich in flavonoids extract from Zuccagnia punctata has beneficial effects on vascular function in aorta from hypercholesterolemic rabbits. PURPOSE: The aim of this study was to evaluate the ability of a hydroalcoholic extract from Z.puncata (ZpE) to prevent alterations induced by high cholesterol diet in rabbits. METHODS: The major components of the ZpE, flavonoids, were analyzed by using a validated reversed phase HPLC method. Rabbits were separated in five groups: fed standard chow (CD); CD orally administrated 2.5 mg, 5 mg or 10 mg GAE/day ZpE (ZpE- CD); fed 1% cholesterol-enriched chow (HD); HD orally administrated 2.5 mg GAE/day ZpE (ZpE-HD); HD orally administrated 2.5 mg rosuvastatin/day (Ro-HD). All diets were administrated by 6 weeks. Body weights (BW), mean blood pressure (MAP), heart rate (HR), visceral abdominal fat (VAF), organ weight (heart, kidney, liver) and vascular morphology were determined. Total cholesterol (TC), triglycerides (TG), fasting glucose (FG), aspartate amino transferase (AST), alanine amino transferase (ALT), bilirubin, creatinine, thiobarbituric acids reactive substances (TBARS) and glutathione reduced/oxidized index were measured in serum. Abdominal aorta was excised and vascular function was assessed by acetylcholine and sodium nitroprusiate relaxation and contractile response to norepinephrine and angiotensin II. RESULTS: The major compounds of ZpE identified were chalcones: 2',4'-dihydroxy-3'-methoxychalcone and 2',4'-dihydroxychalcone. Oral treatment with ZpE reduced MAP, TC, TG, TBARS, aortic intima/media ratio and increased glutathione reduced/oxidized index in HD rabbits. No differences were found in AST, ALT, bilirubin or creatinine. Acetylcholine relaxation was normalized and contractile response to norepinephrine and angiotensin II was reduced in ZpE-HD. CONCLUSION: Oral administration of ZpE as natural product in the prevention of cardiovascular disease related with hypercholesterolemia and endothelial dysfunction is very promising.

Electrosprayed chitosan microcapsules as delivery vehicles for vaginal phytoformulations.

The design of novel delivery systems to treat vaginal fungal infections is a topic of high interest. Chitosan, being itself antimicrobial and having good mucoadhesive properties, is an excellent candidate as a delivery matrix for active compounds. In this work, chitosan microcapsules containing dry extracts of Argentinean medicinal plants with proved biological properties (Larrea divaricata, L. cuneifolia, L. nitida, Zuccagnia punctata and Tetraglochin andina) were developed through electrospraying and compared with conventionally used tablets containing the same extracts. Total phenolics, loading efficacy, physical properties, morphology and particle size, molecular organization, water sorption capacity, release of bioactive compounds and biological properties were assessed. The encapsulation process or the inclusion in tablets did not degrade the bioactive compounds of the extracts. The release of phenolic compounds from chitosan microcapsules was faster than from tablets. The fingerprint of released phenolic compounds from microcapsules and tablets was similar to that from the dry extracts and the antioxidant and antifungal capacity remained unchanged. The FT-IR analysis suggested interactions between the chitosan and the extracts, which explained why the microcapsules kept the integrity in slightly acidic media. Increased solubility of the extracts when incorporated in the microcapsules was seen in simulated vaginal fluid, potentially increasing the bioavailability of bioactive compounds in the vaginal environment. This work highlights the potential of the chitosan-based delivery systems for phytomedicines with antifungal and antioxidant activity to be used in vulvovaginal candidiasis.

Effect of Wine Wastes Extracts on the Viability and Biofilm Formation of Pseudomonas aeruginosa and Staphylococcus aureus Strains.

In this work, we intended to inhibit the biofilm synthesis and the metabolism of Gram-positive and Gram-negative bacteria using two highly available wastes (stem and marc) obtained after the manufacturing of Torrontes wine at Cafayate, Argentina. Wine wastes contain a significant amount of bioactive compounds, mainly phenolic compounds, which makes them a potential source of compounds with beneficial properties to human health, as they could inhibit the virulence of pathogenic bacteria or protect the tissue against oxidative stress. Marc and stem extracts of Torrontes wine were evaluated for their ability to inhibit the metabolism and biofilm production of Pseudomonas aeruginosa and Staphylococcus aureus strains. The phytochemical composition and antioxidant activity of these extracts were also determined. The methanol and ethyl acetate extracts, which contained the highest amount of total polyphenolic, exhibited the highest scavenging capacity of ABTS and nitric oxide and the strongest Fe3+ reducing power and exhibited the highest level of inhibition of the biofilm formation and of the metabolic activity in bacterial biofilm. We also noticed a positive correlation between phenolic compounds content, the antioxidant activity, and the anti-biofilm capacity of the winemaking wastes. These results display the potentiality of wine wastes to prevent or reduce the formation of biofilm. Moreover, their abundance makes them an attractive and affordable source of antibiofilm and antioxidant agents.

Morphoanatomical and histochemical characterization of Larrea species from Northwestern of Argentina

Abstract Larrea divaricata Cav., L. cuneifolia Cav. and L. nitida Cav., Zygophyllaceae, are evergreen xerophytic shrubs occurring in Northwestern Argentina used in traditional medicine. The aim of this work was to characterize the morphology, anatomy and histochemistry of the vegetative organs of three Larrea species by light and scanning electron microscopy in order to provide supporting data for their correct identification and to determine the site of synthesis and accumulation of its main active compounds. The shape, number and percentage of coalescence of leaflets, presence or absence of mucrones and rachis and the shape of the stipules represented the main botanical differences between the studied Larrea species. Anatomically three species presented amphystomatic leaves, with thick resinous slightly striated cuticle with resinous deposits, polygonal epidermal cells with straight anticlinal walls, ciclocytic, brachy-paracytic and paracytic stomatal types, non-glandular trichomes and isolateral mesophyll. The position and abundance of the sclerenchyma at the mid vein and petiole transection allows the differentiation of the three species, been more abundant in L. cuneifolia. Secondary phloem and parenchyma cells presented abundant calcium oxalate druses and solitary rhomboidal crystals. Epidermal cells and cuticle layer of leaflets and stipules of the three species presented amber resin deposits and content which stained positively for polysaccharides, phenolic compounds, flavonoids and tannins, while mesophyll palisade cells showed small refracting droplets stained positively for lipophilic substances.