Wild and commercial samples of Achillea millefolium L.: proximate composition and individual compounds obtained by chromatography

Medicinal plants have been used since ancient times and emerge nowadays as alternative to synthetic products, due to their richness in bioactive compounds. In a society that requires new and safer products, due to the growing concern with health and nutrition, medicinal plants are now being used not only in traditional medicine but also in a number of food and pharmaceutical products [1]. Achillea millefolium L., belongs to Asteraceae family and it is commonly known as yarrow, very common in mountain meadows, pathways, crop fields and homegardens. Widespread across Europe, it’s infusion, decoction and alcoholic extract are widely used as an herbal remedy to treat digestive problems, diabetes, hepato-biliary diseases and amenorrhea, showing also antitumor, antimicrobial, anti-inflammatory and antioxidant properties [2,3]. In the present work, commercial and wild samples of A. millefoilum were characterized regarding the proximate composition and individual compounds namely, free sugars, organic acids, fatty acids and tocopherols, determined by chromatographic techniques coupled to different detectors (HPLC-RI, HPLC-DAD, GC-FID e HPLC-fluorescence, respectively). Carbohydrates, followed by proteins, were the major macronutrients in both samples. Commercial yarrow gave higher content of fat (and saturated fatty acids, mainly palmitic acid C16:0), proteins, ash, energetic value and total sugars (including fructose, glucose, sucrose and trehalose). Wild sample revealed higher levels of carbohydrates; it also showed raffinose (not detected in the commercial sample), polysaturated fatty acids (mainly linoleic acid, C18:2n-6) and organic acids (including malic, oxalic and quinic acids). Regarding tocopherols, both samples showed similar profile, although the wild sample gave higher levels of total tocopherols; γ-Tocopherol was the most abundant isoform; δ-Tocopherol was not found in the samples. Data obtained are clear evidence that traditional medicinal plants can be used not only in household products but also in pharmaceutical and food industry as a source of new and safer bioactive compounds

Chemical characterization of wild and commercial samples of Achillea millefolium L.

As plantas medicinais têm vindo a ser usadas desde tempos ancestrais e surgem hoje em dia como uma alternativa aos produtos sintéticos, devido à sua riqueza em compostos bioativos. Achillea millefolium L. pertence à família das Asteraceae e é vulgarmente conhecida como milefólio ou milfolhada, sendo muito comum em prados, caminhos, campos de cultivo e quintais. No presente trabalho, foram caracterizadas amostras comerciais e silvestres de A. millefolium em termos de composição nutricional e perfil em açúcares livres, ácidos orgânicos, ácidos gordos e tocoferóis, determinados por técnicas cromatográficas acopladas a diferentes detectores (HPLC -RI, HPLC -PDA, GC -FID e HPLC -fluorescência, respetivamente). Os hidratos de carbono, seguidos das proteínas, foram os macronutrientes maioritários em ambas as amostras. A amostra comercial mostrou um teor mais elevado de gordura, proteínas, cinzas, valor energético e açúcares totais A amostra silvestre revelou maior conteúdo em hidratos de carbono; também revelou a presença de rafinose, ácidos gordos polinsaturados e ácidos orgânicos. Relativamente aos tocoferóis, ambas as amostras revelaram um perfil muito semelhante, apesar da amostra silvestre ter mostrado uma maior concentração em tocoferóis totais. Os resultados obtidos são uma prova clara que as plantas usadas na medicina tradicional podem ter aplicabilidade não só em produtos caseiros mas também na indústria alimentar e farmacêutica como fonte de compostos bioativos.Medicinal plants have been used since ancient times and emerge nowadays as an alternative for synthetic products, due to their richness in bioactive compounds. Achillea millefolium L., commonly known as yarrow, belongs to the Asteraceae family being found in meadows, pathways, crop fields and homegardens. In the present work, commercial and wild samples of A. millefolium were characterized in terms of nutritional composition and free sugars, organic acids, fatty acids and tocopherols profile, determined by chromatographic techniques coupled to different detectors (HPLC -RI, HPLC -PDA, GC -FID and HPLC -fluorescence, respectively). Carbohydrates, followed by proteins, were the majority macronutrients found in both samples. The commercial sample showed a higher content of fat, proteins, ash, energy and total sugars. The wild sample revealed a higher content in carbohydrates, also showing the presence of raffinose, polyunsaturated fatty acids and organic acids. Regarding tocopherols, both samples showed a very similar profile, although the wild sample exhibited a higher content in total tocopherols. The results obtained highlight the fact that the plants used in traditional medicine may have applicability not only in homemade products but also in food and pharmaceutical industry, as a source of bioactive compounds.À Fundação para a Ciência e a Tecnologia (CIMO -PEst-OE/AGR/UI0690/2014; REQIMTE- -PEst -C/EQB/LA0006/2014), M.I. Dias (SFRH/ BD/84485/2012), R.C. Alves (SFRH/BPD/68883/2010) e L. Barros (SFRH/BPD/107855/2015).info:eu-repo/semantics/publishedVersio

A comparative study of bioactive properties of wild and commercial Achillea millefolium L.

Achillea millefolium L., commonly known as yarrow, is a medicinal plant with high bioactive value. Its infusion, decoction and alcoholic extract are widely used in Europe to treat digestive and intestinal problems, but also due to their antitumor, antimicrobial, anti-inflammatory and antioxidant properties [1]. In the present work, methanolic extract, infusion and decoction of wild and commercial yarrow were studied for their antioxidant properties and antitumor potential, evaluated by free radicals scavenging activity, reducing power and lipid peroxidation inhibition and by estimation of the growth inhibitory activity in human tumor cell lines, respectively. Overall, cultivated yarrow showed the highest antioxidant activity, presenting the lowest EC50 values. Decoctions of both samples revealed the highest DPPH scavenging activity (0.25 and 0.20 mg/ml, respectively), 13-carotene bleaching inhibition {0.18 and 0.22 mg/ml) and TBARS inhibition (0.04 and 0.08 mg/ml), while infusions presented the highest reducing power (0.12 and 0.13 mg/ml). The samples showed a higher lipid peroxidation inhibition, but a lower DPPH scavenging activity than methanolic extract of A. millefolium from Turkey (892.67 and 45.60 11g/ml, respectively) [2]. The infusion of wild yarrow showed the highest potential against breast (MCF-7; 8 150=17.04 1-Jg/ml) and hepatocellular (HepG2; 37.60 1-Jg/ml) carcinoma cell lines, while the methanolic extract of commercial yarrow was most potent against non-small cell lung (NCI H460; 26.64 IJg/ml), colon (HCT-15, 13.90 1-Jg/ml) and cervical (Hela, 19.68 IJg/ml) carcinoma cell lines. The results obtained with the decoction and infusion of cultivated yarrow against MCF-7 line are consistent with the ones reported with ethanolic extract of A. millefolium from Iran (8150=64 .078 1-Jg/ml) [3].This is a groundbreaking study on the comparison of different extracts of A. millefolium, showing that medicinal plants can be used not only in traditional medicine but also as a source of bioactive products, namely antioxidants and antitumorals.Foundation for Science and Technology (FCT, Portugal) for financial support to the research centre CIMO (PEst-OE/AGR/UI0690/2011 ) and REQUIMTE (PEst-C/EQB/LA0006/2011 ). M.l. Dias, L. Barros and R.C. Alves also thank to FCT, POPH-OREN and FSE for their grants (SFRH/BD/84485/2012, SFRH/BPD/4609/2008 and SFRH/BPD/68883/201 0, respectively)

Comparative analysis on parasite and host bioactive properties - a Cytinus hypocistis (L.) L. case study

Cytinus hypocistis (L.) L. is a rootless, stemless, and leafless holoparasite with a vegetative body reduced to an endophytic system that only grows inside the host [1,2]. Although to date, most studies on plant parasitism were focused on nutrient transfer from host to the parasite and the influence of parasites on host plants, a growing number of studies have documented the transfer of non-nutrient molecules. The transference of phytohormones, secondary metabolites, RNAs, and proteins suggests that hosts may significantly impact parasite physiology and ecology [3].The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to CIMO (UIDB/00690/2020). A. R. Silva is grateful to FCT and FSE for her Doctoral Grant (SFRH/BD/145834/2019) and L. Barros for her contract through the institutional scientific employment program.info:eu-repo/semantics/publishedVersio

Antioxidant properties of flowers and vegetative parts of wild Taraxacum sect. Ruderalia

The species of the genus Taraxacum are known as dendalion (due to the shape of the leaves) and are commonly found in the Northern Hemisphere, in inhabiting fields and roadsides with warmer temperatures. Dendalion infusion and decoction are used in traditional medicine to treat kidney disease, dyspepsia, arthritic and rheumatic complaints, skin problems like eczema and even diabetes mellitus. Besides the pharmaceutical uses, the whole plant is included in many food products [1]. One of the most commonly consumed species is T. officinale; however, traditional collectors have sometimes difficulties in distinguish different Taraxacum species. In the present study, two samples of wild Taraxacum sect. Rudera/ia collected in Bragan9a, flowers and vegetative parts, were studied for their antioxidant activity. Free radicals (DPPH- 2,2-diphenyl-1- picrylhydrazyl) scavenging activity, reducing power and lipid peroxidation inhibition were evaluated on the methanolic extract, infusion and decoction of dandelion samples. The results of the antioxidant activity of the flowers and vegetative parts were very similar. Decoctions of both samples showed the highest DPPH scavenging activity (EC50= 0.42 and 0.12 mg/ml for flowers and vegetative parts, respectively). The decoction of vegetative parts also showed the highest reducing power (EC50= 0.16 mg/ml). The decoction of flowers and the infusion of vegetative parts showed very similar results for 13-carotene bleaching inhibition (EC50= 0.40 and 0.46 mg/ml, respectively) .The methanolic extract of vegetative parts showed the highest activity in TSARS (thiobarbituric acid reactive substances) assay (EC50= 0.13 mg/ml), although the infusion revealed also a low EC50 value for the same assay (0.16 mg/ml). As far as we know, there are no previously studies on the comparison of the antioxidant activity of different extracts and parts of this species of dendalion. More studies will be conducted to evaluate the activity against human tumour cell lines and to characterize the bioactive compounds present in the different extracts.FCT for financial support to the research centre CIMO (PEst-OE/AGR/UI0690/2011) and REOUIMTE (PEst-C/EOB/LA0006/2011 ). M.l. Dias, L. Barros and R.C. Alves also thank to FCT, POPH-OREN and FSE for their grants (SFRH/80/84485/2012, SFRH/BPD/4609/2008 and SFRH/BPD/68883/201 0, respectively)

Nutritional and antioxidant contributions of Laurus nobilis L. leaves: would be more suitable a wild or a cultivated sample?

Medicinal and aromatic plants are used since ancient times in folk medicine and traditional food, but also in novel pharmaceutical preparations. The controversy lies in the use of cultivated and/or wild plants presenting both advantages and disadvantages in biological, ecological but also economic terms. Herein, cultivated and wild samples of Laurus nobilis L. were chemically characterized regarding nutritional value, free sugars, organic acids, fatty acids and tocopherols. Furthermore, the antioxidant activity (scavenging activity, reducing power and lipid peroxidation inhibition) and individual phenolic profile of L. nobilis extracts and infusions were evaluated. Data showed that the wild sample gave higher nutritional contribution related to a higher content of proteins, free sugars, organic acids, PUFA and tocopherols. It also gave better PUFA/SFA and n − 6/n − 3 ratios. Regarding antioxidant activity and phenolic compounds, it was the cultivated sample (mostly the infusion) that showed the highest values. The present study supports the arguments defending the use of wild and cultivated medicinal and aromatic plants as both present very interesting features, whether nutritional or antioxidant, that can be an assessed by their consumption. In vitro culture could be applied to L. nobilis as a production methodology that allows combination of the benefits of wild and cultivated samples.Fundação para a Ciência e a Tecnologia (FCT, Portugal) for financial support to CIMO (strategic project PEst-OE/AGR/UI0690/2011) and REQIMTE (PEst-C/EQB/LA0006/2011). M.I. Dias, L. Barros and R.C. Alves also thank to FCT, POPH-QREN and FSE for their grants (SFRH/BD/84485/2012, SFRH/BPD/4609/2008 and SFRH/BPD/68883/2010, respectively)

Biodegradable drug-eluting stents: Targeting urothelial tumors of upper urinary tract

INTRODUCTION & OBJECTIVES: Urothelial tumors of upper urinary tract are ranked among the most common types of cancers worldwide. The current standard therapy to prevent recurrence is intravesical Bacillus Calmetteâ Guerin (BCG) immunotherapy, but it presents several disadvantages such as BCG failure and intolerance. Another way is to use chemotherapy, which is generally better tolerated that BCG. In this case, drugs such as epirubicin, doxorubicin, paclitaxel and gemcitabine are used. Nevertheless, intravesical chemotherapy only prevents recurrence in the short-term. These failings can be partially attributed to the short residence time and low bioavailability of the drug within the upper urinary tract and the cancer cells, resulting in a need for frequent drug instillation. To avoid these problems, biodegradable ureteral stents impregnated by supercritical fluid CO2 (SCF) with each of the four anti-cancer drugs were produced. MATERIAL & METHODS: Four formulations with different concentrations of gelatin and alginate and crosslink agent were tested and bismuth was added to confer radiopaque properties to the stent. The preliminary in vivo validation studies in female domestic pigs was conducted at the University of Minho, Braga, after formal approval by the institutionâ s review board and in accordance with its internal ethical protocol for animal experiments. Paclitaxel, epirubicin, doxorubicin and gemcitabine were impregnated in the stents and the release kinetics was measured in artificial urine solution (AUS) for 9 days by UV spectroscopy in a microplate reader. The anti-tumoral effect of the developed stents in transitional cell carcinoma (TCC) and HUVEC primary cells, used as control, was evaluated. RESULTS: The in vivo validation of this second-generation of ureteral stents performed was herein demonstrated. Biodegradable ureteral stents were placed in the ureters of a female pigs, following the normal surgical procedure. The animals remained asymptomatic, with normal urine flow. The in vitro release study in AUS of the stent impregnated showed a higher release in the first 72h for the four anti-cancer drugs impregnated after this time the plateau was achieved and the stent degraded after 9 days. The direct and indirect contact of the anti-cancer biodegradable stents with the TCC and HUVEC cell lines confirm the anti-tumor effect of the stents impregnated with the four anti-cancer drugs, reducing around 75% of the viability of the TCC cell line after 72h and no killing effect in the HUVEC cells. CONCLUSIONS: The use of biodegradable ureteral stent in urology clinical practice not only reduce the stent-related symptoms but also open new treatment therapyâ s, like in urothelial tumors of upper urinary tract. Furthermore, we have demonstrated the clinical validation in vivo pig model. This study has thus shown the killing efficacy of the anti-cancer drug eluting biodegradable stents in vitro for the TCC cell line, with no toxicity observed in the control, non-cancerous cells.The direct and indirect contact of the anti-cancer biodegradable stents with the TCC and HUVEC cell lines confirm the anti-tumor effect of the stents impregnated with the four anti-cancer drugs, reducing around 75% of the viability of the TCC cell line after 72h and no killing effect in the HUVEC cells. This study has thus shown the killing efficacy of the anti-cancer drug eluting biodegradable stents in vitro for the TCC cell line, with no toxicity observed in the control, non-cancerous cells

Polymer electrolytes for electrochromic devices through solvent casting and sol-gel routes

Ionically conductive membranes of gelatin and d-PCL(530)/siloxane doped with cyano-based ionic liquids (ILs) were prepared through solvent casting and sol-gel methods, respectively. The membranes were characterized in terms of ionic conductivity, thermal behavior, morphology, and structure. All samples, except the d-PCL(530)/siloxane matrix, exhibited a predominantly amorphous morphology. The samples prepared through solvent casting and sol-gel displayed a minimum thermal stability of 170 and 230 ºC, respectively. The ionic conductivity varied accordingly with the type, quantity, and length of the alkyl chain of the cation of the ILs. The sample with the highest ionic conductivity was gelatin0.5[C2mim][N(CN)2] with 2.40 x 10-3 S. cm-1 at 25 ºC and 1.68 x 10-2 S. cm-1 at 95 ºC. The good results of ionic conductivity encouraged the assembly and characterization of prototypes of electrochromic devices (ECDs). The best results were obtained with glass/ITO/WO3/gelatin1[C2mim][SCN]/CeO2-TiO2/ITO/glass configuration that showed a fast color switching time (~ 15 s) and a good open circuit memory (~ 4 hours). The ECD changed its color from pale blue to transparent, and its charge density decreased from -17.53 to - 2.71 mC. cm-2 during 640 color/bleaching cycles.This work was supported by Fundação para a Ciência e a Tecnologia (FCT) in the framework of the Research unit GREEN-it "Bioresources for Sustainability" (UID/Multi/04551/2013), Laboratório Associado para a Química Verde – Tecnologias e Processos Limpos-LAQV (UID/QUI/50006/2013), grant SRFH/BD/90366/2012 (R.L.) and a contract under Investigador FCT 2012 program (J.M.S.S.E.). It was also co-financed by FEDER through the COMPETE Program and PT2020 Partnership Agreement (POCI-01-0145-FEDER – 007265). M.M. Silva acknowledges CNPq (PVE grant 406617/2013-9) for the mobility grant provided by this institution. A. Pawlicka and R.C. Sabadini acknowledge CNPq (grant 305029/2013-4 and 152252/2016-9), and F.C. Sentanin acknowledges CAPES (grant PNPD20131739- 33002045017P6).info:eu-repo/semantics/publishedVersio

Targeting urothelial tumors of upper urinary tract with drug-eluting stents impregnated by supercritical fluids

Urothelial tumors of upper urinary tract are ranked among the most common types of cancers worldwide and it has been considered as one of the more expensive to treat due of its long-term propensity of recurrence. The current standard therapy to prevent recurrence is intravesical Bacillus Calmette–Guerin (BCG) immunotherapy, but it presents several disadvantages such as BCG failure and intolerance. Another way is to use chemotherapy, that has been reported to be generally better tolerated that BCG. In this case, drugs such as epirubicin, doxorubicin, paclitaxel and gemcitabine are used. Nevertheless, intravesical chemotherapy only prevents recurrence in the short-term[1], [2]. These failings can be partially attributed to the short residence time and low penetration of the drug within the upper urinary tract and the cancer cells, resulting in a need for frequent drug instillation [3]. To avoid these problems, biodegradable ureteral stents impregnated by supercritical fluid CO2 (SCF) with each of the four anti-cancer drugs were produced (figure 1). Four types of drug-eluting biodegradable stents were studied, impregnated with paclitaxel, epirubicin, doxorubicin and gemcitabine. The release kinetics of the impregnated drugs from the anti-cancer drug-eluting stents was measured in artificial urine solution (AUS) for 9 days. The in vitro drugs release from the impregnated biodegradable ureteral stents was analyzed using a microplate reader. The in vitro release study in AUS showed a higher release in the first 72h for the four anti-cancer drugs impregnated after this time the plateau was achieved and the stent degrades after 9 days. Regarding the amount of impregnated drugs by SCF the gemcitabine showed higher amount (109 μg) and the lower amount was obtained for paclitaxel (67 ng). The diffusion coefficient and the impregnation yield were calculated. The anti-tumoral effect of the developed stents in transitional cell carcinoma (TCC) - T24 cell lines was evaluated. T24 cell line was exposed to graded concentrations (0.01 to 2000 ng/ml) of the four drugs for both 4 and 72 hours to determine the sensitivities to each drug (IC50). Toxicity as a result of both direct and indirect contact of the cell lines with the different material conditions of biodegradable stent were studied. The four anti-cancer drugs showed a concentrationdependent inhibitory effect on the T24 and HUVEC cell lines with IC50’s for paclitaxel of 7.30ng and 501.50ng, respectively. The T24 cell line shows to be more sensitive than HUVEC cell line for all the anti-cancer drugs tested. The direct and indirect contact of the anti-cancer biodegradable stents with the T24 and HUVEC cell lines confirm the anti-tumor effect of the stents impregnated with the four anti-cancer drugs, reducing around 75% of the viability of the T24 cell line after 72h and no killing effect in the HUVEC cells. Finally, this study has shown the killing efficacy of the anti-cancer drug eluting biodegradable stents in vitro for the T24 cell lines, with no toxicity observed in the control, non-cancerous cells.Luso­- American Foundation's Grant for Internships in the University of California, Berkeley, 2015/CON5/CAN8; FCT PhD Grant (SFRH/BD/97203/2013); European Union's Seventh Framework Programme (FP7/2007­2013) under grant agreement n° REGPOT­CT2012­316331­ POLARIS; Project “Novel smart and biomimetic materials for innovative regenerative medicine approaches (Ref.: RL1 ­ ABMR ­ NORTE­01­0124­FEDER­000016)” cofinanced by North Portugal Regional Operational Programme (ON.2 – O Novo Norte), under the National Strategic Reference Framework (NSRF), through the European Regional Development Fund (ERDF

Water and carbon dioxide: green solvents for the extraction of collagen/gelatin from marine sponges

"Publication Date (Web): December 23, 2014"Marine sponges are extremely rich in natural products and are considered a promising biological resource. The major objective of this work is to couple a green extraction process with a natural origin raw material to obtain sponge origin collagen/gelatin for biomedical applications. Marine sponge collagen has unique physicochemical properties, but its application is hindered by the lack of availability due to inefficient extraction methodologies. Traditional extraction methods are time consuming as they involve several operating steps and large amounts of solvents. In this work, we propose a new extraction methodology under mild operating conditions in which water is acidified with carbon dioxide (CO2) to promote the extraction of collagen/gelatin from different marine sponge species. An extraction yield of approximately 50% of collagen/gelatin was achieved. The results of Fourier transformed infrared spectroscopy (FTIR), circular dichroism (CD), and differential scanning calorimetry (DSC) spectra suggest a mixture of collagen/gelatin with high purity, and the analysis of the amino acid composition has shown similarities with collagen from other marine sources. Additionally, in vitro cytotoxicity studies did not demonstrate any toxicity effects for three of the extracts.The authors are grateful for financial support of FCT through Grant EXP/QEQ:EPS/0745/2012, SWIMS (Subcritical Water Isolation of compounds from Marine Sponges). The funding from the European Union Seventh Framework Programme (FP7/2007-2013) under grant agreement numbers REGPOT-CT2012-316331-POLARIS and KBBE-2010-266033 (project SPECIAL), as well as from ERDF under the project "Novel smart and biomimetic materials for innovative regenerative medicine approaches" RLI-ABMR-NORTE-01-0124-FEDER-000016), cofinanced by North Portugal Regional Operational Programme (ON.2,O Novo Norte), under the National Strategic Reference Framework (NSRF) are also gratefully ackowledged. The authors are also truly thankfull to Prof. Micha flan (Tel Aviv University, Israel), Dr. Ronald Osinga (Porifarma, The Netherlands), Dr. Antonio Sara and Dr. Martina Milanese (Studio Associato GAIA, Italy), and Dr. Joana Xavier (University of Azores) for the kind offer of marine sponges samples