A Comparative Analysis of Phytochemicals, Metal Ions, Volatile Metabolites in Heart Wood, Stem Bark and Leaves of Salix alba L. along with in Vitro Antioxidant, Antacid, Antimicrobial Activities for Sake of Environment Conservation by Substitution of Stem Bark With Leaf.

The genus of Salix is used in food, medicine and nutraceuticals, and standardized by using the single marker compound Salicin only. Stem bark is the official part used for the preparation of various drugs, nutraceuticals and food products, which may lead to overexploitation and damage of tree. There is need to search substitution of the stem bark with leaf of Salix alba L. (SA), which is yet not reported. Comparative phytochemicals viz. Salicin, Procyanidin B1 and Catechin were quantified in the various parts of SA viz. heart wood (SA-HW), stem bark (SA-SB) and leaves (SA-L) of Salix alba L.by using newly developed HPLC method. It was observed that SA-HW and SA-L contained far better amount of Salicin, Procyanidin B and Catechin as compared to SA-SB (SA-HW~SA-L≫SA-SB). Essential and toxic metal ions of all three parts were analysed using newly developed ICP-OES method, where SA-L were founded as a rich source of micronutrients and essential metal ions as compared to SA-SB and SA-HW. GC-MS analysis has shown the presence of fatty acids and volatile compounds. The observed TPC and TFC values for all three parts were ranged from 2.69 to 32.30 mg GAE/g of wt. and 37.57 to 220.76 mg QCE/g of wt. respectively. In DPPH assay the IC50 values of SA-SB, SA-HW, and SA-L were 1.09 (±0.02), 5.42 (±0.08), and 8.82 (±0.10) mg/mL, respectively. The order of antibacterial activities against E. coli, S. aureus, P. aeruginosa, and B. subtilis strains was SA-L>SA-HW>SA-SB with strong antibacterial activities against S. aureus, and B. subtilis strains. The antacid activities order was SA-L>SA-SB>SA-HW. The leaves of SA have shown significant source of nutrients, phytochemicals and medicinal properties than SA-HW and SA-SB. The leaves of SA may be considered as substitute of stem bark to save the environment or to avoid over exploitation, but after the complete pharmacological and toxicological studies.

Immunochemical Characterization of Salix alba (S. alba) Pollen Allergens and Evaluation of the Cross-Reactivity Pattern with Common Allergenic Pollen Grains.

BACKGROUND: Pollens, particularly tree and plant pollens, are one of the major causes of allergic respiratory diseases worldwide. Allergy to pollens of different species of Salix trees has been reported in various regions of the world. The most common type of Salix tree in Iran is white willow (Salix alba). OBJECTIVES: This study aimed to identify and determine the immunochemical characteristics of allergenic proteins in S. alba tree pollen extract using SDS-PAGE and IgE- immunoblotting methods. Moreover, the cross-reaction pattern of the specific IgE antibody of S. alba tree pollen proteins with pollen allergens of common allergenic trees, i.e., Populus nigra (P. nigra), Cupressus sempervirens (C. sempervirens), Pinus brutia (P. brutia) and Platanus orientalis (P. orientalis) in the region was investigated. METHODS: The reaction of allergenic proteins in S. alba pollen extract with specific IgE antibodies in patients' sera was investigated using SDS-PAGE and IgE-immunoblotting methods. The cross-reaction of specific IgE antibodies of the proteins present in S. alba pollen extract with pollen allergens of common allergenic trees in the region was investigated using ELISA and immunoblotting inhibition methods. In silico methods such as phylogenetic tree drawing and alignment of amino acid sequences were used to examine the evolutionary relationship and homology structure of common allergenic proteins (Panallergens) responsible for cross reactions. RESULTS: More than 11 protein bands binding to specific IgE antibodies in patients' sera with a molecular weight between 13 and 95 kDa were identified in the S. alba tree pollen extract. ELISA and immunoblotting inhibition results showed that P. nigra extract could inhibit the binding of IgE antibodies to S. alba pollen extract proteins to a greater extent than C. sempervirens, P. brutia, and P. orientalis tree extracts. In silico methods investigated the results of ELISA and immunoblotting inhibition methods. Moreover, a high structural homology and evolutionary relationship were observed between S. alba and P. nigra tree pollen panallergens. CONCLUSION: In this study, it was found that more than 80 % of the sensitive patients who were examined had specific IgE antibodies reacting with the approximately a 15 kDa-protein present in the S. alba pollen extract. Furthermore, the specific IgE-binding proteins found in the pollens of S. alba and P. nigra trees had relative structural homology, and it is likely that if recombinant forms are produced, they can be used for diagnostic and therapeutic purposes for both of the trees.

The effect of Salix alba L. bark extract on dark circles in vitro and in vivo.

OBJECTIVE: Dark circles in the infraorbital area are a common cosmetic concern among individuals because they exhibit fatigue and are undesirable across all ages. Of the dark circle etiologies, blood stasis by poor-vascular integrity can cause darkening of the lower eyelid skin, which might be alleviated by reduced endothelial permeability. In this study, we investigated the effects of Salix alba bark extract (SABE) on the synthesis of hyaluronic acid (HA) in fibroblasts and vascular integrity protection from inflammatory cytokine. We also performed a clinical trial investigating the effect of SABE on dark circles. METHODS: To confirm the effect of SABE on HA synthesis in human dermal fibroblasts (HDFs), we performed ELISA and real-time PCR. We investigated the interaction HDF-secreted substance with vascular integrity, and human dermal microvascular endothelial cells (HMEC-1) were treated with conditioned medium (CM) from HDF treated with or without SABE. Subsequently, we conducted a clinical study on 29 subjects by having them apply SABE containing cream for 8 weeks. RESULTS: Salix alba bark extract treatment increased HA synthesis and regulated HMW-HA-related gene expressions in HDF. CM from SABE-treated HDF alleviated endothelial permeability and led to improved vascular integrity in HMEC-1 cells. Treatment with the cream containing 2% SABE for 8 weeks improved the parameters measuring dark circles, skin microcirculation and elasticity. CONCLUSION: Our results showed that SABE could protect against dark circles in vitro, and that topical treatment of SABE improved the clinical indexes of dark circles in a clinical study. Therefore, SABE can be used as an active ingredient for improving dark circles.

OBJECTIF: Les cernes dans la région infra-orbitaire sont un problème cosmétique fréquent chez les patients, car elles témoignent de la fatigue et sont indésirables à tout âge. Parmi les étiologies de cerne, la stase sanguine due à une mauvaise intégrité vasculaire peut entraîner un assombrissement de la peau de la paupière inférieure qui peut être atténué par une réduction de la perméabilité endothéliale. Dans cette étude, nous avons étudié les effets de l'extrait d'écorce de Salix alba sur la synthèse de l'acide hyaluronique (AH) dans les fibroblastes, et la protection de l'intégrité vasculaire contre les cytokines inflammatoires. Nous avons également réalisé une étude clinique portant sur l'effet de l'extrait d'écorce de Salix alba sur les cernes. MÉTHODES: Pour confirmer l'effet de l'extrait d'écorce de Salix alba sur la synthèse de l'AH dans les fibroblastes dermiques humains (Human Dermal Fibroblasts, HDF), nous avons réalisé un test ELISA et un test PCR en temps réel. Nous avons étudié l'interaction entre la substance sécrétée par les HDF et l'intégrité vasculaire, et les cellules endothéliales microvasculaires dermiques humaines (Human Dermal Microvascular Endothelial Cells, HDMEC-1) ont été traitées avec un milieu conditionné pour les HDF traité avec ou sans extrait d'écorce de Salix alba. Par la suite, nous avons mené une étude clinique auprès de 29 sujets en leur demandant d'appliquer une crème à base d'extrait d'écorce de Salix alba pendant 8 semaines. RÉSULTATS: Le traitement par extrait d'écorce de Salix alba a augmenté la synthèse de l'AH et régulé les expressions géniques liées à l'acide hyaluronique à haut poids moléculaire dans les HDF. Les milieux conditionnés pour les HDF traités par extrait d'écorce de Salix alba ont atténué la perméabilité endothéliale et ont permis une amélioration de l'intégrité vasculaire des cellules HMEC-1. Le traitement avec la crème contenant 2% d'extrait d'écorce de Salix alba pendant 8 semaines a amélioré les paramètres de mesure des cernes, la microcirculation cutanée et l'élasticité. CONCLUSION: Nos résultats ont montré que l'extrait d'écorce de Salix alba pouvait protéger contre les cernes in vitro, et que le traitement topique par extrait d'écorce de Salix alba améliorait les indices cliniques des cernes dans une étude clinique. L'extrait d'écorce de Salix alba peut donc être utilisé comme principe actif pour améliorer les cernes.

Chemical Profile and Biological Effects of an Herbal Mixture for the Development of an Oil-in-Water Cream.

Three individual hydroalcoholic extracts derived from Hamamelis virginiana leaves, Krameria lappacea root, Salix alba bark, and the resulting herbal mixture (HM) were assessed for the phytochemical profile as well as for antibacterial and cytotoxic potential. The chemical composition of the individual extracts and of their mixture was analyzed by chromatographical (LC-MS) and spectrophotometrical methods. The antimicrobial properties were evaluated by using the agar-well diffusion and the broth microdilution assays, whereas the potential cytotoxicity was investigated on human keratinocyte cell line by MTT method and apoptosis test. The HM composition revealed important amounts of valuable polyphenolic compounds provided from the individual extracts, having synergistic biological effects. All tested extracts displayed in vitro antimicrobial properties, with a significantly higher efficacy noticed for the HM when tested against Staphylococcus aureus. Moreover, none of the tested extracts was responsible for in vitro cytotoxicity against the human keratinocytes in the selected concentration range. Furthermore, the HM was included in an oil-in-water cream for the nonpharmacological treatment of seborrheic dermatitis, developed and optimized by using a QbD approach. A D-optimal experimental plan with four factors that varied on two levels was used to investigate the effect of the quantitative variation of the formulation factors (emulsifier, co-emulsifier, thickening agent, oily phase ratio) on the characteristics of the cream in terms of firmness, consistency, adhesiveness, stringiness, spreadability, and viscosity. Based on the experimental results, an optimal formulation containing 2.5% emulsifier and 20% oily phase was prepared and analyzed. The obtained results showed appropriate quality characteristics of this novel cream, which may be used in the future to manage the associated symptoms of seborrheic dermatitis.

Antibacterial and antifungal activity of methanolic extracts of Salix alba L. against various disease causing pathogens

Abstract The present study was aimed to manifest the antibacterial and antifungal activity of methanolic extracts of Salix alba L. against seven Gram-positive and Gram-negative bacterial pathogens e.g. Streptococcus pyogenes, Staphylococcus aureus (1), S. aureus (2), Shigella sonnei, Escherichia coli (1), E. coli (2) and Neisseria gonorrhoeae and three fungal isolates from the air such as Aspergillus terreus, A. ornatus, and Rhizopus stolonifer. Two different serotypes of S. aureus and E. coli were used. The agar well-diffusion method results showed the dose-dependent response of plant extracts against bacterial and fungal strains while some organisms were found resistant e.g. E. coli (1), S. sonnei, A. terreus and R. stolonifer. The highest antibacterial activity was recorded at 17.000±1.732 mm from 100 mg/mL of leaves methanolic extracts against S. pyogenes while the activity of most of the pathogens decreased after 24 h of incubation. The highest antifungal activity was reported at 11.833±1.0 mm against A. ornatus at 50 mg/mL after 48 h of the incubation period. These experimental findings endorse the use of S. alba in ethnopharmacological formulations and suggest the use of methanolic extracts of the said plant to develop drugs to control the proliferation of resistant disease causing pathogenic microbes.

Resumo O presente estudo teve como objetivo manifestar a atividade antibacteriana e antifúngica de extratos metanólicos de Salix alba L. contra sete patógenos bacterianos Gram-positivos e Gram-negativos. Streptococcus pyogenes, Staphylococcus aureus (1), S. aureus (2), Shigella sonnei, Escherichia coli (1), E. coli (2) e Neisseria gonorrhoeae e três isolados de fungos do ar, como Aspergillus terreus, A. ornatus, e Rhizopus stolonifer. Dois sorotipos diferentes de S. aureus e E. coli foram usados. Os resultados do método de difusão em ágar mostraram a resposta dependente da dose de extratos de plantas contra cepas de bactérias e fungos, enquanto alguns organismos foram considerados resistentes, e.g. E. coli (1), S. sonnei, A. terreus e R. stolonifer. A maior atividade antibacteriana foi registrada em 17.000 ± 1.732 de 100 mg/mL de extratos metanólicos de folhas contra S. pyogenes, enquanto a atividade da maioria dos patógenos diminuiu após 24 h de incubação. A maior atividade antifúngica foi relatada em 11,833 ± 1,0 contra A. ornatus a 50 mg/mL após 48 h do período de incubação. Esses achados experimentais endossam o uso de S. alba em formulações etnofarmacológicas e sugerem o uso de extratos metanólicos da referida planta para o desenvolvimento de fármacos que controlem a proliferação de doenças resistentes que causam micróbios patogênicos.

Antibacterial and antifungal activity of methanolic extracts of Salix alba L. against various disease causing pathogens / Atividade antibacteriana e antifúngica de extratos metanólicos de Salix alba L. contra vários patógenos que causam doenças

Abstract The present study was aimed to manifest the antibacterial and antifungal activity of methanolic extracts of Salix alba L. against seven Gram-positive and Gram-negative bacterial pathogens e.g. Streptococcus pyogenes, Staphylococcus aureus (1), S. aureus (2), Shigella sonnei, Escherichia coli (1), E. coli (2) and Neisseria gonorrhoeae and three fungal isolates from the air such as Aspergillus terreus, A. ornatus, and Rhizopus stolonifer. Two different serotypes of S. aureus and E. coli were used. The agar well-diffusion method results showed the dose-dependent response of plant extracts against bacterial and fungal strains while some organisms were found resistant e.g. E. coli (1), S. sonnei, A. terreus and R. stolonifer. The highest antibacterial activity was recorded at 17.000±1.732 mm from 100 mg/mL of leaves methanolic extracts against S. pyogenes while the activity of most of the pathogens decreased after 24 h of incubation. The highest antifungal activity was reported at 11.833±1.0 mm against A. ornatus at 50 mg/mL after 48 h of the incubation period. These experimental findings endorse the use of S. alba in ethnopharmacological formulations and suggest the use of methanolic extracts of the said plant to develop drugs to control the proliferation of resistant disease causing pathogenic microbes.

Resumo O presente estudo teve como objetivo manifestar a atividade antibacteriana e antifúngica de extratos metanólicos de Salix alba L. contra sete patógenos bacterianos Gram-positivos e Gram-negativos. Streptococcus pyogenes, Staphylococcus aureus (1), S. aureus (2), Shigella sonnei, Escherichia coli (1), E. coli (2) e Neisseria gonorrhoeae e três isolados de fungos do ar, como Aspergillus terreus, A. ornatus, e Rhizopus stolonifer. Dois sorotipos diferentes de S. aureus e E. coli foram usados. Os resultados do método de difusão em ágar mostraram a resposta dependente da dose de extratos de plantas contra cepas de bactérias e fungos, enquanto alguns organismos foram considerados resistentes, e.g. E. coli (1), S. sonnei, A. terreus e R. stolonifer. A maior atividade antibacteriana foi registrada em 17.000 ± 1.732 de 100 mg/mL de extratos metanólicos de folhas contra S. pyogenes, enquanto a atividade da maioria dos patógenos diminuiu após 24 h de incubação. A maior atividade antifúngica foi relatada em 11,833 ± 1,0 contra A. ornatus a 50 mg/mL após 48 h do período de incubação. Esses achados experimentais endossam o uso de S. alba em formulações etnofarmacológicas e sugerem o uso de extratos metanólicos da referida planta para o desenvolvimento de fármacos que controlem a proliferação de doenças resistentes que causam micróbios patogênicos.

Influence of Dietary Supplementation with Boswellia serrata and Salix alba on Performance and Blood Biochemistry in Free-Range Leghorn Laying Hens.

This study was conducted to evaluate the safety and the beneficial effects of dietary supplementation with Boswellia serrata (Bs) and Salix alba (Sa) in Leghorn hens during the critical pre-laying and laying phases. A total of 120 pullets, 17 weeks of age, were assigned to two groups (Control­C; Treated­T, n = 60 each). For 12 weeks, the T group received a diet supplemented with 0.3% of dry extracts of Bs (5%) and Sa (5%). The study lasted 19 weeks. Productive performance, serum analytes, H/L ratio, IgA and anti-IBV antibodies were investigated. Water intake was significantly higher, while body and egg weight was significantly lower for the T group (p < 0.05). No other differences were detected in performance parameters, serum analytes, IgA and H/L ratio excluding t0, with a significantly (p < 0.05) higher H/R ratio and higher titers of anti-IBV antibody for the T group. Overall, the data obtained in this study show that the supplementation with Bs and Sa was safe and resulted in an increase in water consumption, a decrease in egg weight, and a sedative effect in the hens. In the future, it would be interesting to test this supplement in hens reared on intensive farms.

The Physicochemical and Antimicrobial Properties of Silver/Gold Nanoparticles Obtained by "Green Synthesis" from Willow Bark and Their Formulations as Potential Innovative Pharmaceutical Substances.

Green chemistry is a pharmaceutical industry tool, which, when implemented correctly, can lead to a minimization in resource consumption and waste. An aqueous extract of Salix alba L. was employed for the efficient and rapid synthesis of silver/gold particle nanostructures via an inexpensive, nontoxic and eco-friendly procedure. The nanoparticles were physicochemically characterized using ultraviolet-visible spectroscopy (UV-Vis), Fourier transform infrared spectroscopy (FT-IR), dynamic light scattering (DLS), X-ray diffraction (XRD) and scanning electron microscopy (SEM), with the best stability of up to one year in the solution obtained for silver nanoparticles without any chemical additives. A comparison of the antimicrobial effect of silver/gold nanoparticles and their formulations (hydrogels, ointments, aqueous solutions) showed that both metallic nanoparticles have antibacterial and antibiofilm effects, with silver-based hydrogels having particularly high antibiofilm efficiency. The highest antibacterial and antibiofilm efficacies were obtained against Pseudomonas aeruginosa when using silver nanoparticle hydrogels, with antibiofilm efficacies of over 75% registered. The hydrogels incorporating green nanoparticles displayed a 200% increased bacterial efficiency when compared to the controls and their components. All silver nanoparticle formulations were ecologically obtained by "green synthesis" and were shown to have an antimicrobial effect or potential as keratinocyte-acting pharmaceutical substances for ameliorating infectious psoriasis wounds.

Phytodepuration of Nitrate Contaminated Water Using Four Different Tree Species.

Water pollution by excessive amounts of nitrate (NO3-) has become a global issue. Technologies to clean up nitrate-contaminated water bodies include phytoremediation. In this context, this research aimed to evaluate four tree species (Salix alba L., Populus alba L., Corylus avellana L. and Sambucus nigra L.) to remediate nitrate-contaminated waters (100 and 300 mg L-1). Some physiological parameters showed that S. alba L. and P. alba L. increased particularly photosynthetic activity, chlorophyll content, dry weight, and transpired water, following the treatments with the above NO3- concentrations. Furthermore, these species were more efficient than the others studied in the phytodepuration of water contaminated by the two NO3- levels. In particular, within 15 days of treatment, S. alba L. and P. alba L. removed nitrate quantities ranging from 39 to 78%. Differently, C. avellana L. and S. nigra L. did not show particular responses regarding the physiological traits studied. Nonetheless, these species removed up to 30% of nitrate from water. In conclusion, these data provide exciting indications on the chance of using S. alba L. and P. alba L. to populate buffer strips to avoid NO3- environmental dispersion in agricultural areas.

Influence of Dietary Supplementation of Salix alba Bark on Performance, Oxidative Stress Parameters in Liver and Gut Microflora of Broilers.

The paper aimed to analyse the effect of dietary Salix alba L. bark powder (SAB) on broiler performance in terms of oxidative stress parameters in liver and gut microflora. One hundred and eighty Cobb 500 broiler chicks (14 days) were allotted to three homogeneous treatments (SAB 0%; SAB 0.025%; SAB 0.05%). The broilers were housed in an environmentally controlled space (10 replicates, six broilers/replicate). Compared to dietary control treatment (SAB 0%), the other treatments included 0.025% SAB (SAB 0.025%) and 0.05% SAB (SAB 0.05%). The results showed that SAB powder used in broiler diet had a high total phenolic content. Regarding the performance results, significant differences between experimental and control treatments were recorded only for average daily feed intake (35-42 days). The broilers fed with SAB powder had a significantly lower hepatic level of malondialdehide and glutathione, a higher total antioxidant capacity than those fed control treatment, and demonstrated a positive effect on the development of non-pathogenic bacteria (lactobacilli) but a decrease in the population of pathogenic ones (E. coli, staphylococci). Our findings suggested that dietary 0.05% SAB powder could be an effective solution to impede the oxidative stress in broiler liver and to improve gut microflora.

Evaluation of Salix alba, Juglans regia and Populus nigra as biomonitors of PTEs in the riparian soils of the Sava River.

A large number of human activities result in the release of potentially toxic elements (PTEs) into the environment, which could lead to the degradation of riparian areas. This study aimed to evaluate the potential of Salix alba, Juglans regia and Populus nigra for the biomonitoring of PTEs in the riparian soils of the Sava River. Levels of seven PTEs (As, Cd, Cr, Cu, Ni, Pb and Zn) were measured in the soils, roots and leaves of plants at selected sampling sites and evaluated according to bioaccumulation and translocation factors. The obtained results showed that in riparian soils, As, Cr, Cu, Ni and Zn were at levels considered to be critical for plants. The levels of As, Cd, Cr, Ni and Zn measured in roots of Salix alba and As, Cr, Ni and Zn in its leaves were toxic for plant tissue. Toxic levels of Cr were also measured in the roots of Juglans regia and As in its leaves, as well as As and Cr in the roots of Populus nigra, and Zn in its leaves. Bioconcentration and translocation factors showed that S. alba and P. nigra have potential for the phytoextraction of Zn and Cd, while J. regia has potential for the phytoextraction of As. In terms of phytostabilization potential, S. alba proved to be good for the phytostabilization of Cd and Cu, and J. regia for the phytostabilization of Cr, As, Ni and Pb, while P. nigra showed potential for the phytostabilization of Cr, Ni, Pb and Cu.

GC-MS analysis of the methanolic extracts of Smilax china and Salix alba and their antioxidant activity.

Smilax china L. (family Smilacaceae) and Salix alba L. (family Salicaceae) are plants that have been traditionally used to treat various ailments in Indian and Chinese medicine. A quantitative estimation of the methanolic extracts of these plants was performed by GC-MS analysis to obtain insight into its phytoconstituents responsible for therapeutic action. The antioxidant potential of the methanol extracts of Smilax china (MESC) and Salix alba (MESA) were assessed with DPPH by using a UV spectrophotometer at a wavelength of 517 nm. The prevailing compounds found in MESC were lactam sugars including 2,5-dimethyl-2,4-dihydroxy-3(2H)-furanon (1.40%), 1,5-anhydro-6-deoxyhexo-2,3-diulose (4.33%), and alpha-methyl-1-sorboside (1.80%); the two alkaloids found were 1,4-methane-4,4a,5,6,7,8,9,9a-octahydro-10,10-dimethyl cyclohepta[d] pyridazine (0.87%) and 1,3,7-trimethyl-2,6-dioxopurine(0.54%); terpenes included deltacadinene (0.39%), terpineol, (+)-cedrol (22.13%), 3-thujanol (0.77%), and 9,10-dehydro-cycloisolongifolene (0.34%); fatty acids included cis-vaccenic acid (4.98%) and telfairic acid (1.10%); esters included 1,2,3-propanetriol diacetate (7.56%), 7-hexadecenoic acid, methyl ester (1.77%), eicosanoic acid, and methyl ester (0.95%); and glycerol included 1,2,3-propanetriol (28.75%). The interesting compounds found in MESA were reducing sugars like D-allose (4.40%) and pyrogallol (10.48%), alkaloids like caffeine (63.49%), and esters like methyl octadecanoate (0.53%). Both fractions revealed considerable antioxidant activity. The reported existing phenolic compounds and terpenes are responsible for the antioxidant activity of the plant extracts.

Xenobiotics metabolization in Salix alba leaves uncovered by mass spectrometry imaging.

INTRODUCTION: Micropollutants are increasingly monitored as their presence in the environment is rising due to human activities, and they are potential threats to living organisms. OBJECTIVES: This study aimed at understanding the role of plants in xenobiotics removal from polluted environments by following xenobiotics metabolism in leaf tissues. METHODS: Different classes of micropollutants were investigated using liquid chromatography (LC) coupled to quadrupole-time of flight (Q-TOF) high resolution mass spectrometry (HRMS). The tissue localization of xenobiotics in the leaves of a spontaneous (not planted by humans) Salix alba growing near the water flux was further investigated using matrix-assisted laser desorption ionization (MALDI) mass spectrometry imaging (MSI). RESULTS: The LC-Q-TOF analysis revealed the distribution of micropollutants in three different compartments of a tertiary treatment wetland. When further investing the metabolic profile of S. alba leaves using MSI, different distribution patterns were observed in specific leaf tissues. Xenobiotic metabolites were predicted and could also be tentatively identified in S. alba leaves, shedding new light on the metabolic processes at play in leaves to manage xenobiotics uptake from a polluted environment. CONCLUSION: Using complementary metabolomics approaches, this study performed a large-scale exploration of micropollutants spreading in the environment at the exit of a tertiary treatment wetland. The use of MSI coupled with the prediction of xenobiotic metabolites yielded novel insights into plant metabolism during chronical exposure to low doses of a mixture of micropollutants.

Multiple pharmacological and toxicological investigations on Tanacetum parthenium and Salix alba extracts: Focus on potential application as anti-migraine agents.

Migraine is one of the most common neurological disorder, which has long been related to brain serotonin (5-HT) depletion and neuro-inflammation. Despite many treatment options are available, the frequent occurrence of unacceptable adverse effects further supports the research toward nutraceuticals and herbal preparations, among which Tanacetum parthenium and Salix alba showed promising anti-inflammatory and neuro-modulatory activities. The impact of extract treatment on astrocyte viability, spontaneous migration and apoptosis was evaluated. Anti-inflammatory/anti-oxidant effects were investigated on isolated rat cortexes exposed to a neurotoxic stimulus. The lactate dehydrogenase (LDH) release, nitrite levels and 5-HT turnover were evaluated, as well. A proteomic analysis was focused on specific neuronal proteins and a fingerprint analysis was carried out on selected phenolic compounds. Both extracts appeared able to exert in vitro anti-oxidant and anti-apoptotic effects. S. alba and T. parthenium extracts reduced LDH release, nitrite levels and 5-HT turnover induced by neurotoxic stimulus. The downregulation of selected proteins suggest a neurotoxicity, which could be ascribed to an elevated content of gallic acid in both S. alba and T. parthenium extracts. Concluding, both extracts exert neuroprotective effects, although the downregulation of key proteins involved in neuron physiology suggest caution in their use as food supplements.

Salix alba (white willow) medicinal plant presents genotoxic effects in human cultured leukocytes.

Salix alba (SA), commonly known as white willow, is a plant used in folk medicine for the treatment of chronic and acute inflammation, infection, pain, and fever. The phytochemical characterization of the bark extract of this plant indicated that its main component is salicin, a precursor of the anti-inflammatory agent acetylsalicylic acid. Considering the lack of studies evaluating the genetic toxicity and cytotoxic action of SA bark extract on human cells, as well as the chemical characterization of its major phenolic compounds, the present study was designed to (1) investigate the cytotoxic and genotoxic potential of SA bark extract on human peripheral leukocyte cells and human hepatoma cell line HepG2, and (2) characterize its major phenolic constituents. The phenolic compounds found were salicylic acid, salicin, salidroside, saligenin, tremulodin, salicoylsalicin, salicortin, and tremulacin. The results using trypan blue staining test showed viability decreases (viability less than 70%) for concentrations of SA extract equal and higher to 200 µg/ml. Low genotoxic activity (comet assay) was exhibited for 50 and 100 µg/ml SA extract in human leukocytes. SA did not exert a marked clastogenic/aneugenic effect on leukocytes and HepG2 human cells. Data suggest that the genotoxic effects of SA bark extract occur when it is not metabolized by liver enzymes.

Salix alba attenuated oxidative stress in the heart and kidney of hypercholesterolemic rabbits.

OBJECTIVE: Oxidative stress plays a critical role in the development of hypercholesterolemia-induced complications. This study evaluated the effects of aspirin and Salix alba hydroethanolic extract on oxidative stress in the heart and kidney of hypercholesterolemic rabbits. MATERIALS AND METHODS: The antioxidant activity, as well as total phenolic and salicin content of S. alba (Sa) extract were assessed by DPPH radical scavenging activity, Folin-Ciocalteu and HPLC methods, respectively. Animals were divided into two groups of control (fed with normal chow), and HD (fed with high cholesterol diet for 6 weeks). Then, hypercholesterolemic animals allocated to the following treatment groups: CHO (received HD), Sa extract (HD plus extract 60 and 120 mg/kg), and aspirin (HD plus aspirin 120 mg/kg) and received the treatments on a daily basis for 6 weeks. MDA, GSH, and nitrite concentrations as well as the activities of SOD and CAT were evaluated in cardiac and kidney tissues. RESULTS: The scavenging activity, total phenolic content and salicin were 19.1 µg/ml (IC50), 153.75 ± 3.6 mg of gallic acid/g, and 18.03 µg/mg, respectively. In comparison to CHO group, MDA levels were diminished in Sa and ASA groups but GSH levels were improved. NO metabolites increased in the heart of Sa 120 mg/kg group and in the kidney of all Sa and ASA treated groups. SOD activity increased only in the heart of Sa groups and in the kidney of Sa and ASA groups. CAT activity increased in the heart and kidney tissues of all Sa and ASA treated groups. CONCLUSION: The results showed S. alba extract improved redox homeostasis in heart and kidney tissues of hypercholesterolemic rabbits. The extract antioxidant property may be related to its phenolic content.

Usage Proposal of a common urban decorative tree (Salix alba L.) to monitor the dispersion of gaseous mercury: A case study from Turda (Romania).

Closure of chloralkali plants poses a risk of abandonment of important sources of gaseous mercury. In this work, an assessment has been made of the potential for pollution from one of these plants in the proximity of a densely populated town in central Romania. The work involved a comparison between two major types of monitoring survey: biomonitoring using leaves of a tree common in urban environments; and LUMEX-based gaseous mercury analysis. For biomonitoring, 21 samples from Salix alba L. trees were taken in Turda area. Atmospheric monitoring included two mobile surveys and one at a fixed location. The results from both monitoring systems show similarities in gaseous mercury dispersion patterns, with high mercury contents clearly related to the presence of the chloralkali plant. Particularly high levels were measured in the following situations: (i) in a 'smog' area related with thermal inversion and (ii) during dusk. Direct monitoring suffered from limitations in acquiring information, especially in a medium-long time range, but biomonitoring provided these data and is capable of covering studies on temporary trends or comparative assessments between European cities with contrasting gaseous mercury sources. The thermal speciation of mercury contents indicates that the whole fraction of mercury in leaves corresponds to organic mercury. This finding implies a non-reversible uptake process, which in turn ensures the applicability of this technique to biomonitor long-term exposure. As a conclusion, the assessment of gaseous mercury pollution based on biomonitoring using S. alba has proven to be a useful, reliable and cost-effective methodology.

Risk assessment of white willow (Salix alba) in food.

This Technical Report contains a description of the activities within the work programme of the EU-FORA Fellowship on the risk assessment of white willow in food. The bark of different varieties of willow has had a long history of medical use as a means to reduce fever and as a painkiller. Willow bark is also used in weight loss and sports performance food supplements. The labelling of these products usually does not mention any restrictions to the length of use. The recommended doses for foods differ, sometimes exceeding doses recommended for pharmaceuticals. A systematic literature review on adverse effects potentially resulting from oral exposure to white willow (Salix alba) was performed. The aim of the study was to assess the risk for humans when consuming white willow bark in food. The preliminary results show that despite the long history of use only very limited data on toxicity of white willow bark are available. However, anaphylactic reactions in people with a history of allergy to salicylates may occur. Some other adverse effects of salicylates are considered to be of low relevance for the long-time consumption of white willow bark, mainly due to relatively low concentrations of salicin and the presence of compounds with gastroprotective action. However, it seems that the content of heavy metals, mainly cadmium, should be further addressed in risk assessment of white willow bark in food.

Bioprofiling of Salicaceae bud extracts through high-performance thin-layer chromatography hyphenated to biochemical, microbiological and chemical detections.

The buds of poplars (Populus L.) and willows (Salix L.), both from the same family (Salicaceae Mirbel), are increasingly used in gemmotherapy and importantly contribute to the production of the physiologically active propolis by European bee Apis mellifera L. In order to study their phenolic profiles, polar extracts of buds from P. nigra L. were compared to those of P. alba L. and S. alba L. through high-performance thin-layer chromatography (HPTLC). Five chemotypical patterns were distinguished after derivatisation with the Natural Product reagent and confirmed by principal component analysis. The HPTLC analysis was directly hyphenated to various microbiological and biochemical assays as well as spectrometric techniques, directly linking to active molecules in the chromatograms. At a glance, polyvalent compounds were evident when all derivatisation and activity assays, to which HPTLC was hyphenated at ease, were combined together. In Populus buds, at least three antimicrobial compound zones were detected using Aliivibrio fischeri and Bacillus subtilis bioassays, and one phyto-œstrogen with the planar yeast œstrogen screen. In all samples, several inhibitors of acetyl- and butyrylcholinesterase and rabbit liver esterase were detected. Hyphenation to high resolution mass spectrometry supported the assignment of bioactive compounds, as shown for chrysin as selective cholinesterase inhibitor as well as caffeic acid and galangin as antimicrobials in P. nigra and P. alba. This fast and cost-efficient method can be appropriately extended and applied to the botanical origin determination and quality control of bud extracts and propolis samples.

Ethyl acetate Salix alba leaves extract-loaded chitosan-based hydrogel film for wound dressing applications.

High toxicity and multidrug resistance associated with various standard antimicrobial drugs have necessitated search for safer alternatives in plant-derived materials. In this study, we performed biological examination of chitosan-based hydrogel film loaded with ethyl acetate Salix alba leaves extract against 11 standard laboratory strains. FTIR showed regeneration of saccharide peak in CP1A at 1047 cm(-1) and increased in height of other peaks. DSC exothermic decomposition peaks at 112 °C, 175 °C and 251 °C reveal the effect of extract on hydrogel film. From FESEM images, three-dimensional cross-linking and extract easily seen in the globular form from the surface. MTT assay on HEK 293 cells showed that CP1A was non-toxic. Minimum inhibitory concentration ranges from 4000 µg/ml to 125 µg/ml. Enterococcus faecium, Candida glabrata and Candida tropicalis were the most resistant, while Salmonella typhi and Candida guilliermondii were the most susceptible micro-organisms.