Use of bacaba peel for the development of hydroelectrolytic beverages and their consumer acceptance

Abstract The hydroelectrolytic beverages segment has been expading its market and introducing new flavors in order to meet the demand for new products. However, experimental studies find concerns about the chemical compositions of these drinks. The aim of this study was to develop a drink without synthetic coloring or flavoring, with functional attributes based on the bacaba (Oenocarpus bacaba Mart.) peel extract. Two hydroelectrolytic drinks were developed, one hypotonic and the other isotonic, containing 0.5 and 1.0% of bacaba peel extract. Physicochemical characterization, determination of total phenolic compounds, anthocyanins, and antioxidant capacity were perfomed, in addition to color evaluation, as well as sensory analysis by means of preference tests. The developed formulations showed potential antioxidant activity and natural red coloring due to the phenolic compounds and anthocyanins present in the beverages. The sensory evaluation indicated positive acceptance by the tasters regarding the addition of the bacaba peel extract to the beverage formulations. The developed formulations demonstrated that the use of the bacaba peel is a viable option for the production of sports drinks, acting as a natural dye and offering health benefits due to its bioactive compounds.

Essential Oil from the Stem Bark of Casuarina equisetifolia Exerts Anti-inflammatory and Anti-nociceptive Activities in Rats

Abstract Herein the chemical constituents and the anti-pain properties of the essential oil from the stem bark of Casuarina equisetifolia L. (Casuarinaceae) grown in Nigeria were evaluated. The essential oil was obtained by hydrodistillation method in an all glass Clevenger-type apparatus, and characterized by gas chromatography (GC-FID) and gas chromatography-mass spectrometry (GC-MS). The hot plate method was used to determine the anti-nociceptive property whereas the anti-inflammatory activity was evaluated by carrageenan-induced and formalin experimental models. The pale-yellow essential oil was obtained in yield of 0.21% (v/w), calculated on a dry weight basis. The main constituents of the essential oil were methyl salicylate (30.4%), a-zingiberene (15.5%), (E)-anethole (9.5%), b-bisabolene (8.6%), b- sesquiphellandrene (6.9%), and ar-curcumene (6.2%). In the anti-nociceptive study, the rate of inhibition increases as the doses of essential oil increases with optimum activity at the 30th and 60th min for all tested doses. The essential oil displayed anti-nociceptive activity independently of reaction time at the highest tested dose (200 mg/kg). The essential oil of C. equisetifolia moderately reduced pain responses in early and late phases of the formalin test. The oil inhibited the paw licking in the neurogenic phase (60-63%) compared to the late phase of the formalin test. The carrageenan- induced oedema model revealed the suppression of inflammatory mediators within the 1st - 3rd h. Thus, C. equisetifolia essential oil displayed both anti-nociceptive and anti-inflammatory activities independent of the dose tested. The anti-inflammatory and anti-nociceptive activities of C. equisetifolia essential oil are herein reported for the first time

Evaluation of Antioxidative Mechanisms In Vitro and Triterpenes Composition of Extracts from Silver Birch (Betula pendula Roth) and Black Birch (Betula obscura Kotula) Barks by FT-IR and HPLC-PDA.

Silver birch, Betula pendula Roth, is one of the most common trees in Europe. Due to its content of many biologically active substances, it has long been used in medicine and cosmetics, unlike the rare black birch, Betula obscura Kotula. The aim of the study was therefore to compare the antioxidant properties of extracts from the inner and outer bark layers of both birch trees towards the L929 line treated with acetaldehyde. Based on the lactate dehydrogenase test and the MTT test, 10 and 25% concentrations of extracts were selected for the antioxidant evaluation. All extracts at tested concentrations reduced the production of hydrogen peroxide, superoxide anion radical, and 25% extract decreased malonic aldehyde formation in acetaldehyde-treated cells. The chemical composition of bark extracts was accessed by IR and HPLC-PDA methods and surprisingly, revealed a high content of betulin and lupeol in the inner bark extract of B. obscura. Furthermore, IR analysis revealed differences in the chemical composition of the outer bark between black and silver birch extracts, indicating that black birch may be a valuable source of numerous biologically active substances. Further experiments are required to evaluate their potential against neuroinflammation, cancer, viral infections, as well as their usefulness in cosmetology.

Variation in the Phenolic Composition of Cork Stoppers from Different Geographical Origins.

Cork stopper granulates from five geographical origins from Portugal and six from Spain were analyzed regarding polyphenol composition by HPLC-DAD/ESI-MS and geographical discrimination studied by near-infrared spectroscopy (NIRS). The phenolic composition of the eleven origins ranged from 30 to 52 mg/g cork granulates, with vescavaloninic acid, castalagin, sanguisorbic acid dilactone, vescalagin, castavaloninic acid, dehydrated tergallic-C-Glc, and ellagic acid being the major compounds. NIRS revealed to be a powerful tool to discriminate origins and predict the concentration of polyphenols. However, polyphenols do not fully explain the discrimination of geographical origins. Variability in the polyphenol composition of cork stoppers is not significantly influenced by geographical location but probably may be more related to the plant genetics, tree age, and phytosanitary and edaphoclimatic conditions.

Evidence of traditionality of Brazilian medicinal plants: The case studies of Stryphnodendron adstringens (Mart.) Coville (barbatimão) barks and Copaifera spp. (copaíba) oleoresin in wound healing.

ETHNOPHARMACOLOGICAL RELEVANCE: The World Health Organization (WHO) recognizes the potential of plants used in secular traditional medicine and considers this an important source of evidence to assess their effectiveness and safety. Brazil is rich in biodiversity and traditional uses based on the Amerindian culture. However, many processes started with the arrival of the Portuguese in the year 1500. The successive economic cycles, for example, led to destruction of native vegetation and an intense cultural erosion. As a consequence, the information about the use of plants in the past centuries are dispersed and without interpretation. In this study a methodology to evidence the traditionality of Brazilian plants was demonstrated using data about barbatimão barks (Stryphnodendron adstringens (Mart.) Coville - Fabaceae) and Copaiba oleoresin (Copaifera spp. - Fabaceae) in wound healing, was established. MATERIAL AND METHODS: Data about use of the plants were recovered from bibliography published between 1576 and 2011. The books (101) were classified using weights, considering the date of publication and the source of Information. Older books that describe primary information received weight 10, while books written more recently and with secondary information received weight 0.4. A score for each category of medicinal use was calculated based on the books weights and the frequency of citation. A review about the current use of both plants was also performed from ethnobotanical studies published in journals. RESULTS AND DISCUSSION: The traditional secular use of barks of barbatimão and oleoresin of copaiba to treat wounds was confirmed based on the historic bibliographic research. The most frequent use of barbatimão in a timeline of 500 years of Brazil's history, was as astringent, whereas for copaíba was as healing of skin and mucosal lesions. The continuous and current use of these plants to treat wounds, confirmed by recent ethnobotanical studies, is an indicative of the resilience of these remedies and their effectiveness. CONCLUSION: The use of preparations containing barbatimão barks and copaiba oleoresin can be considered effective in the treatment of wounds. Nonetheless, it is necessary to improve the quality of the formulas as established by WHO.

Magnolia Officinalis Bark Extract Induces Depolarization of Pacemaker Potentials Through M2 and M3 Muscarinic Receptors in Cultured Murine Small Intestine Interstitial Cells of Cajal.

BACKGROUND: Magnolia officinalis Rehder and EH Wilson (M. officinalis) are traditional Chinese medicines widely used for gastrointestinal (GI) tract motility disorder in Asian countries. We investigated the effects of an ethanol extract of M. officinalis (MOE) on the pacemaker potentials of cultured interstitial cells of Cajal (ICCs) in vitro and its effects on GI motor functions in vivo. METHODS: We isolated ICCs from small intestines, and the whole-cell patch-clamp configuration was used to record the pacemaker potentials in cultured ICCs in vitro. Both gastric emptying (GE) and intestinal transit rates (ITRs) were investigated in normal and GI motility dysfunction (GMD) mice models in vivo. RESULTS: MOE depolarized ICC pacemaker potentials dose-dependently. Pretreatment with methoctramine (a muscarinic M2 receptor antagonist) and 4-DAMP (a muscarinic M3 receptor antagonist) inhibited the effects of MOE on the pacemaker potential relative to treatment with MOE alone. In addition, MOE depolarized pacemaker potentials after pretreatment with Y25130 (a 5-HT3 receptor antagonist), GR113808 (a 5-HT4 receptor antagonist) or SB269970 (a 5-HT7 receptor antagonist). However, pretreatment with RS39604 (a 5-HT4 receptor antagonist) blocked MOE-induced pacemaker potential depolarizations. Intracellular GDPßS inhibited MOE-induced pacemaker potential depolarization, as did pretreatment with Ca2+ free solution or thapsigargin. In normal mice, the GE and ITR values were significantly and dose-dependently increased by MOE. In loperamide-and cisplatin-induced GE delay models, MOE administration reversed the GE deficits. The ITRs of the GMD mice were significantly reduced relative to those of normal mice, which were significantly and dose-dependently reversed by MOE. CONCLUSION: These results suggest that MOE dose-dependently depolarizes ICCs pacemaker potentials through M2 and M3 receptors via internal and external Ca2+ regulation through G protein pathways in vitro. Moreover, MOE increased GE and ITRs in vivo in normal and GMD mouse models. Taken together, the results of this study show that MOE have the potential for development as a gastroprokinetic agent in GI motility function.

Castanea sativa Mill. Bark Extract Protects U-373 MG Cells and Rat Brain Slices Against Ischemia and Reperfusion Injury.

Ischemic brain injury is one of the most important causes of death worldwide. The use of one-drug-multi-target agents based on natural compounds is a promising therapeutic option for cerebral ischemia due to their pleiotropic properties. This study assessed the neuroprotective properties of Castanea sativa Mill. bark extract (ENC) in human astrocytoma U-373 MG cells subjected to oxygen-glucose deprivation and reperfusion and rat cortical slices subjected to ischemia-like conditions or treated with glutamate or hydrogen peroxide. Neuroprotective effects were determined by assessing cells or slices viability (MTT assay), ROS formation (DCFH-DA assay), apoptosis (sub G0/G1 peak), nuclear fragmentation and chromatin condensation (DAPI staining) as well as changes in lysosomes and mitochondria morphology (Acridine Orange and Rhodamine123 staining, respectively). ENC treatment before injury on U-373 MG cells (5-50 µg/ml) and cortical slices (50-100 µg/ml) provided neuroprotection, while lower or higher concentrations (100 µg/ml U-373 MG cells, 200 µg/ml brain slices) were ineffective. ENC addition during reperfusion or after the injury was not found to be effective. The results suggest that ENC might hold potential as preventive neuroprotective agent, and indicate the importance of further studies exploring its mechanism of action. J. Cell. Biochem. 118: 839-850, 2017. © 2016 Wiley Periodicals, Inc.

Comparative Analysis of DNA Barcoding and HPLC Fingerprint to Trace Species of Phellodendri Cortex, an Important Traditional Chinese Medicine from Multiple Sources.

Phellodendri Cortex is derived from the dried barks of Phellodendron genus species, has been extensively used in traditional Chinese medicine. The cortex is divided into two odorless crude drugs Guanhuangbo and Huangbo. Historically, it has been difficult to distinguish their identities due to a lack of identification methods. This study was executed to confirm the identity and to ensure the species traceability of Phellodendri Cortex. In the current study, analysis is based on the internal transcribed spacer (ITS) and psbA-trnH intergenic spacer (psbA-trnH) barcodes and HPLC fingerprint was carried out to guarantee the species traceability of Guanhuangbo and Huangbo. DNA barcoding data successfully identified the three plants of the Phellodendron genus species by ITS+psbA-trnH, with the ability to distinguish the species origin of Huangbo. Moreover, the psbA-trnH data distinguished Guanhuangbo and Huangbo except to trace species. The HPLC fingerprint data showed that Guanhuangbo was clearly different from Huangbo, but there was no difference between the two origins of Huangbo. Additionally, the result of hierarchical clustering analysis, based on chlorogenic acid, phellodendrine, magnoflorine, jatrorrhizine, palmatine and berberine, was consistent with the HPLC fingerprint analysis. These results show that DNA barcoding and HPLC fingerprint can discriminate Guanhuangbo and Huangbo. However, DNA barcoding is more powerful than HPLC fingerprint for species traceability in the identification of related species that are genetically similar. DNA barcoding is a useful scientific tool to accurately confirm the identities of medicinal materials from multiple sources.

Anxiolytic-like effect of Citrus limon (L. ) Burm f. essential oil inhalation on mice / Efeito ansiolítico da inalação do óleo essencial de Citrus limon (L. ) Burm f. em camundongos.

ABSTRACT Experimental in vivo study aimed to characterize the anxiolytic-like effect of the Citrus limon fruit peel’s essential oil (CLEO) in animal models of anxiety, besides evaluating the viability J774.A1 cells in vitro through the MTT reduction method at the concentrations of 10 and 100 µg/mL. The anxiolytic behavior was evaluated in Swiss mice (n = 8) using the methodology of Elevated Plus-Maze (EPM) and Open-Field (OF). CLEO was tested by inhalation at the doses of 100, 200, and 400 µL, and as control, animals were subjected to inhalation of the vehicle (saline solution 0.9% + Tween80®) and intraperitoneal administration of diazepam (1.5 mg/kg). In the cell viability assay, it was observed that none of the concentrations showed cytotoxicity. OF test showed significant anxiolytic activity at all tested doses of OECL, compared to the control group, without changing the motor performance of the animals. Corroborating OF data, the EPM test confirmed anxiolytic activity in at least two doses of the tested oil (200 and 400 µL), justified by the number of entries and increase in the percentage of time in the open arms. The data analysis of this study evidenced that inhalation of OECL was able to induce an anxiolytic behavior in mice; however, further studies are required to ensure its safe use by the population.

RESUMO O estudo experimental in vivo objetivou caracterizar o efeito ansiolítico do óleo essencial das cascas do fruto de Citrus limon (OECL) em modelos animais de ansiedade, além de avaliar in vitro a viabilidade de células J774.A1 através do método de redução do MTT nas concentrações de 10 e 100 µg/mL. O comportamento ansiolítico foi avaliado em camundongos Swiss (n = 8), utilizando a metodologia do Labirinto em Cruz Elevado (LCE) e do Campo Aberto (CA). O OECL foi testado por inalação nas doses de 100, 200 e 400 µL, e como controle, os grupos foram submetidos à inalação do veículo (Solução fisiológica 0,9% + Tween80®) e administração intraperitoneal de diazepam (1,5 mg/kg). No ensaio de viabilidade celular, observou-se que nenhuma das concentrações apresentou citotoxicidade. O teste de CA demonstrou atividade ansiolítica significativa em todas as doses testadas do OECL, comparada ao grupo controle, sem alterar a performance motora dos animais. Corroborando os dados do CA, o teste do LCE confirmou a atividade ansiolítica em pelo menos duas das doses do óleo testadas (200 e 400 µL), justificada pelo número de entradas e aumento na percentagem de tempo nos braços abertos. A análise dos dados deste estudo evidenciou que a inalação do OECL foi capaz de induzir um comportamento ansiolítico em camundongos; entretanto, outros estudos são necessários para garantir o seu uso seguro pela população.

Morphoanatomical characterization and chemical study of the internal portion of the stem bark of Sambucus australis Cham. & Schltdl / Caracterização morfoanatômica e estudo químico da porção interna da casca do caule de Sambucus australis Cham. & Schltdl.

ABSTRACT The internal part of the stem bark of this species is used to produce a homemade ointment in some regions of the state of Rio Grande do Sul. The purpose of this study is to characterize the morphoanatomy and identify the compounds present in the internal part of the stem bark of S. australis through chemical and histochemical methods. In addition, the best extraction conditions for the sample were determined. It was possible to quantify the rutin and total phenolic compounds, as well as define the Soxhlet method with an 80% hydroethanolic solution as the best method for extracting these compounds from the bark of the species. The portion of the stem bark that is popularly used could also be determined. Based on the results, new studies will be performed in order to identify other characteristics of the species and the possible reasons that sustain its traditional use.

RESUMO Sambucus australis Cham. & Schtdl. (Adoxaceae) é uma espécie arbórea nativa do sul do Brazil, conhecida como “sabugueiro”. A parte interna da casca do caule dessa espécie é utilizada na produção de uma pomada caseira em algumas regiões do estado do Rio Grande do Sul. O objetivo desse estudo é caracterizar a morfoanatomia e identificar compostos presentes na parte interna da casca do caule de S. australis por métodos químicos e histoquímicos. Ademais, realizou-se avaliações para determinar as melhores condições extrativas para a amostra. Foi possível quantificar rutina e compostos fenólicos totais, além de determinar o método Soxhlet com solvente hidroetanólico 80% como o melhor método para extrair esses compostos da casca da espécie. Ainda foi possível determinar a porção da casca do caule utilizada popularmente. Com base nos resultados, novas investigações serão realizadas a fim de determinar mais características da espécie e as possíveis razões que corroboram o uso tradicional.

Recipientes e substratos na produção de mudas e no cultivo hidropônico de tomilho (Thymus vulgaris L. ) / Containers and substrates on seedling production and hydroponic cultivation of thyme (Thymus vulgaris L. )

Bandejas com diferente número de células (128, 200 e 288) e dois substratos (fibra de casca de coco e casca de pinus) foram avaliados para a produção de mudas de tomilho e sobre o desempenho das plantas em sistema hidropônico. Para ambos os experimentos, o delineamento foi em blocos ao acaso. As mudas provenientes de estaquia foram avaliadas aos 30 dias quanto ao comprimento e massa seca de raízes. Aos 40 dias após o transplante das mudas, as plantas cultivadas em sistema hidropônico foram avaliadas quanto à massa fresca da parte aérea, massa seca da parte aérea e massa seca das raízes. A maior massa seca de raízes foi obtida para as mudas cultivadas em substrato à base de casca de fibra de coco. As mudas com maior comprimento de raízes foram obtidas nas bandejas com 128 e 200 células. As plantas de tomilho provenientes de mudas produzidas em bandejas de 128 células apresentaram maior massa fresca da parte aérea, massa seca da parte aérea e massa seca das raízes.

Trays with different number of cells (128, 200 and 288) and two substrates (coir and pine bark-based) were studied for thyme seedling production and plant development in hydroponic system. In both experiments, experimental design was in randomized blocks. Seedlings from cuttings were evaluated at 30 days as to root length and dry matter. At 40 days after transplanting, plants grown in hydroponic system were evaluated for shoot fresh matter, shoot dry matter and root dry matter. The highest root dry matter was detected in seedlings grown in coir-based substrate. Seedlings that presented the greatest root length were obtained by using trays with 128 and 200 cells. Thyme plants from seedlings produced in trays with 128 cells had higher shoot fresh matter, shoot dry matter and root dry matter.

Isolation and characterization of EST-SSR markers in Schima mertensiana (Theaceae) using pyrosequencing technology.

PREMISE OF THE STUDY: Microsatellite or simple sequence repeat (SSR) markers from expressed sequence tags (ESTs) in Schima mertensiana, an evergreen tree that is endemic to the Bonin Islands, were developed to facilitate evaluation of the species's population genetic structure. METHODS AND RESULTS: Using 149831 ESTs derived by pyrosequencing S. mertensiana complementary DNA (cDNA), 683 EST-SSRs were developed, 48 of which were tested for amplification. Thirty of the 48 showed clear amplification and detected polymorphism in sampled populations, with mean expected heterozygosity at the amplified loci ranging from 0.045 to 0.874 (average 0.545). CONCLUSIONS: The markers developed in this study can be used for future studies of the genetic structure of S. mertensiana and related species.

Rapid development of microsatellite markers for Pandanus boninensis (Pandanaceae) by pyrosequencing technology.

PREMISE OF THE STUDY: To facilitate rapid development of microsatellite or simple sequence repeat (SSR) markers, an expressed sequence tags (EST) database was constructed for Pandanus boninensis, an evergreen tree endemic to the Bonin Islands, using pyrosequencing technology. METHODS AND RESULTS: We designed primers for 340 EST-SSRs identified from 109620 pyrosequencing reads, 48 of which were tested for PCR amplification. Thirty-four primers provided clear amplification, and 26 of those 34 displayed clear polymorphic patterns in sampled populations, with mean expected heterozygosity at the amplified loci ranging from 0.022 to 0.742 (average 0.262). CONCLUSIONS: The developed markers are promising tools for future genetic studies of P. boninensis and related species.