Grapevine woody tissues accumulate stilbenoids following bud burst.

MAIN CONCLUSION: After bud burst, a transcriptional reprogramming of the shikimate and phenylpropanoid pathways occurs in grapevine canes resulting in the accumulation of stilbenoids like resveratrol and viniferin. Stilbenoids are phenylpropanoid compounds with important biological properties and biotechnological applications that are synthesized in grapevine in response to different stresses. Although they are found in woody tissues, such as canes and buds, their biosynthesis and accumulation have been essentially described in berries. We have previously shown that transcripts encoding secondary metabolism enzymes accumulate in grapevine canes following the transition from dormancy (E-L 1) to bud burst (E-L 4) suggesting that secondary metabolites may accumulate in grapevine canes during this transition. In the present study, using UPLC-MS we demonstrate the accumulation of important metabolites such as ferulic acid and the stilbenoids E-resveratrol, E-piceatannol and E-ε-viniferin. Stilbenoids accumulation correlated with the increased expression of several stilbene synthase genes and of VviMYB14, encoding a transcription factor that regulates stilbene biosynthesis. In addition, a general stimulation of the plastidial shikimate pathway was observed. Taken together, results show that important secondary metabolites accumulate in the woody canes during bud burst. These findings may aid biotechnological approaches aimed at extracting biologically active phenolic compounds, including stilbenoids, from grapevine woody tissues.

Opuntia ficus-indica Fruit: A Systematic Review of Its Phytochemicals and Pharmacological Activities.

The use of Opuntia ficus-indica fruits in the agro-food sector is increasing for a multiplicity of players. This renewed interest is, in part, due to its organoleptic characteristics, nutritional value and health benefits. Furthermore, industries from different sectors intend to make use of its vast array of metabolites to be used in different fields. This trend represents an economic growth opportunity for several partners who could find new opportunities exploring non-conventional fruits, and such is the case for Opuntia ficus-indica. O. ficus-indica originates from Mexico, belongs to the Cactaceae family and is commonly known as opuntia, prickly pear or cactus pear. The species produces flowers, cladodes and fruits that are consumed either in raw or in processed products. Recent publications described that consumption of the fruit improves human health, exhibiting antioxidant activity and other relevant pharmacological activities through enzymatic and non-enzymatic mechanisms. Thus, we provide a systematic, scientific and rational review for researchers, consumers and other relevant stakeholders regarding the chemical composition and biological activities of O. ficus-indica fruits.

Monoclonal antibody based immunoassay: An alternative way for aquatic environmental selenium detection.

Environmental concerns about human health encouraged increasing methodological interest in selenium (Se), which is an essential non-metal trace element and varies within a narrow concentration range between essential and toxic. In this study, two types of long-armed Se haptens (Se-hapten-lc-NHS) were synthesized for the first time using active ester formalization. In producing monoclonal antibodies (mAbs), the derivatization of haptenized Se at para- (meta-) and ortho-sites showed different properties. Finally, a mAb derived from hybridoma 5A52 was confirmed to be capable of establishing an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA). There was a successful quantitative determination of Se4+ with a detection range of 17 to 207 pmol mL-1 and a limit of detection of approximately 3.9 pmol mL-1. The mAb was found to be remarkably sensitive and specific, with no evidence of cross-reactivity with other ions. The assay was validated for four kinds of Se forms in water samples and showed satisfactory recoveries between 80 % and 108 %, with coefficients of variation of 2.1 %-11 %. The method proposed in our study offers a useful protocol for the rapid screening of Se and provides an alternative solution for the analysis of Se in aquatic environments.

Hypericum perforatum and Its Potential Antiplatelet Effect.

BACKGROUND: Hypericum perforatum (HP) is currently one of the most consumed medicinal plants in the world. In traditional Chinese medicine, the herb hypericum (Guan Ye Lian Qiao) belongs to the group of plants that clarify heat. It is also used to treat various types of infection and inflammation. In contrast to the extensive literature on the antidepressant effects of HP, little is known about its action on platelets. The main objective of this work was to investigate the possible relevance of HP to platelet function. METHODS: We characterized the profile of platelet activation in the presence of HP extracts through an evaluation of molecular markers by flow cytometry: mobilization of intracellular Ca++ and expression of platelet receptors such as activated GPIIbIIIa and P-selectin (CD62). RESULTS: The results indicated a possible inhibitory effect of HP on the platelet activation response, which could be explained by the effect on intracellular calcium mobilization and the expression of activated GPIIbIIIa receptors. Despite of the limitations of an in vitro study, our results provide evidence of the possible mechanisms of action of HP. CONCLUSIONS: Further studies are needed to elucidate the effect of HP on hemostasis, but it may be recognized as a substance with antiplatelet properties.

Protection against Paraquat-Induced Oxidative Stress by Curcuma longa Extract-Loaded Polymeric Nanoparticles in Zebrafish Embryos.

The link between oxidative stress and environmental factors plays an important role in chronic degenerative diseases; therefore, exogenous antioxidants could be an effective alternative to combat disease progression and/or most significant symptoms. Curcuma longa L. (CL), commonly known as turmeric, is mostly composed of curcumin, a multivalent molecule described as having antioxidant, anti-inflammatory and neuroprotective properties. Poor chemical stability and low oral bioavailability and, consequently, poor absorption, rapid metabolism, and limited tissue distribution are major restrictions to its applicability. The advent of nanotechnology, by combining nanosacale with multi-functionality and bioavailability improvement, offers an opportunity to overcome these limitations. Therefore, in this work, poly-Ɛ-caprolactone (PCL) nanoparticles were developed to incorporate the methanolic extract of CL, and their bioactivity was assessed in comparison to free or encapsulated curcumin. Their toxicity was evaluated using zebrafish embryos by applying the Fish Embryo Acute Toxicity test, following recommended OECD guidelines. The protective effect against paraquat-induced oxidative damage of CL extract, free or encapsulated in PCL nanoparticles, was evaluated. This herbicide is known to cause oxidative damage and greatly affect neuromotor functions. The overall results indicate that CL-loaded PCL nanoparticles have an interesting protective capacity against paraquat-induced damage, particularly in neuromuscular development that goes well beyond that of CL extract itself and other known antioxidants.

How Does Hedonic Aroma Impact Long-Term Anxiety, Depression, and Quality of Life in Women with Breast Cancer? A Cross-Lagged Panel Model Analysis.

Depression and anxiety are common symptoms during and after adjuvant chemotherapy treatment for breast cancer (BC), with implications on quality of life (QoL). The present study evaluates the temporal relationship between anxiety, depression, and QoL (primary outcomes), as well as the impact of hedonic aroma (essential oils) on this relationship. This is a secondary analysis of a previously reported randomized controlled trial, with two groups: an experimental group (n = 56), who were subjected to the inhalation of a self-selected essential oil during chemotherapy, and a control group (n = 56), who were only subjected to the standard treatment. The hedonic aroma intervention occurred in the second (T1), third (T2), and fourth (T3) chemotherapy sessions, three weeks apart from each other. The follow-up (T4) assessments took place three months after the end of the treatment. Cross-lagged panel models were estimated in the path analysis framework, using structural equation modeling methodology. Regarding the control group, the cross-lagged panel model showed that anxiety at T1 predicted anxiety at T3, which in turn predicted both QoL and depression at T4. In the experimental group, hedonic aroma intervention was associated with stability of anxiety and QoL over time from T1 to T3, with no longitudinal prediction at T4. For women undergoing standard chemotherapy treatment, anxiety was the main longitudinal precursor to depression and QoL three months after chemotherapy. Thus, essential oils could complement chemotherapy treatment for early-stage BC as a way to improve long-term emotional and QoL-related adjustment.

Development of quantum dot-linked immunosorbent assay (QLISA) and ELISA for the detection of sunset yellow in foods and beverages.

Sunset yellow (SY) is widely used as food colorant. Excess and illegal use of SY could pose potential health risk. Quantum dots have been successfully used in biological research due to the high photoluminescence and high resistance to photobleaching. To analyze SY efficiently, quantum dot-linked immunosorbent assay (QLISA) and enzyme-linked immunosorbent assay (ic-ELISA) were developed on the basis of generated monoclonal antibody. A carboxyl group was introduced to SY and coupled with carrier proteins to synthesize artificial antigen. Under the optimal conditions, inhibitory concentrations (IC50) of SY were 1.9 ng/mL (ic-ELISA) and 3.4 ng/mL (QLISA); the limits of detection (LODs) were 0.2 ng/mL (ic-ELISA) and 1.0 ng/mL (QLISA), respectively. Cross-reactivities of the mAb toward eight kinds of analogues were<0.01%. The recovery rates in spiked foods and beverages were 75.6%∼120.1% (ic-ELISA) and 74.0%∼114.1% (QLISA), respectively.

Quercetin-biapigenin nanoparticles are effective to penetrate the blood-brain barrier.

Search for efficient therapeutic agents for central nervous system (CNS) disorders has been extensive. Nevertheless, blood-brain barrier (BBB) is an obstacle that prevents the majority of compounds to act in these diseases. It is, thus, of extreme relevance the BBB overcome, in order to deliver a drugs therapeutically active concentration to the action site, with the least losses and interaction with other organs, tissues, or cells. The present study aimed to investigate the potential protective effect of quercetin-biapigenin encapsulated into poly(Ɛ-polycaprolactone) (PCL) nanoparticles against t-BOOH-induced oxidative stress in several brain cell lines, as well as evaluate the permeability of those active molecules through an in vitro BBB model. The three cell lines under study (BV-2, hcmec/D3, and U87) presented different reactions to t-BOOH. In general, quercetin-biapigenin PCL-loaded nanoparticles were able to minimize compound toxicity they convey, regardless the cell line. Quercetin-biapigenin PCL-loaded nanoparticles (Papp of approximately 80 × 10-6 cm/s) revealed to be more permeable than free compounds (Papp of approximately 50 × 10-6 cm/s). As of our knowledge, this is the first report of quercetin-biapigenin PCL-loaded nanoparticle activity in brain cells. It is also the first determining its permeability through BBB, as an effective nanocarrier for brain delivery.

In vivo anti-inflammatory and antinociceptive effects, and in vitro antioxidant, antiglycant and anti-neuroinflammatory actions of Syzygium malaccense.

Syzygium malaccense is popularly used to treat inflammation and pain-related ailments. The species was assessed regarding its antioxidant, antiglycant, anti-inflammatory, including anti-neuroinflammatory, and antinociceptive activities. Different models were employed to measure S. malaccense extract (ESM) antioxidant activity. The antiglycant activity was determined using the glucose-induced protein glycation model. LPS-induced neuroinflammation on murine BV-2 microglial cell line was used for anti-neuroinflammatory activity evaluation. The croton oil-induced ear edema test was accomplished to evaluate the in vivo anti-inflammatory activity. Acetic acid-induced writhing together with formalin-induced paw licking assays were performed to evaluate the antinociceptive potential. Finally, the chemical characterization was accomplished by a UHPLC-MS analysis. ESM presented relevant antioxidant and antiglycant activity. NO production by BV-2 cells was reduced, indicating the relevant neuroprotective activity. ESM significantly decreased the mice ear edema induced by croton oil and the nociceptive stimulus induced by acetic acid and formalin by central and peripheral mechanisms. The flavonoids myricitrin, myricetin and quercetin were identified and, as far as we know, the alkaloid reserpine was reported in the species for the first time. The antioxidant and antiglycant potential of ESM, may be related to the in vivo anti-inflammatory and antinociceptive effects, and to the in vitro neuroinflammation inhibition.

Transcriptional responses of Hypericum perforatum cells to Agrobacterium tumefaciens and differential gene expression in dark glands.

Hypericum perforatum L. (St. John's wort) is a well-known medicinal plant that possesses secondary metabolites with beneficial pharmacological properties. However, improvement in the production of secondary metabolites via genetic manipulation is a challenging task as H. perforatum remains recalcitrant to Agrobacterium tumefaciens-mediated transformation. Here, the transcripts of key genes involved in several plant defence responses (secondary metabolites, RNA silencing, reactive oxygen species (ROS) and specific defence genes) were investigated in H. perforatum suspension cells inoculated with A. tumefaciens by quantitative real-time PCR. Results indicated that key genes from the xanthone, hypericin and melatonin biosynthesis pathways, the ROS-detoxification enzyme HpAOX, as well as the defence genes Hyp-1 and HpPGIP, were all upregulated to rapidly respond to A. tumefaciens elicitation in H. perforatum. By contrast, expression levels of genes involved in hyperforin and flavonoid biosynthesis pathways were markedly downregulated upon A. tumefaciens elicitation. In addition, we compared the expression patterns of key genes in H. perforatum leaf tissues with and without dark glands, a major site of secondary metabolite production. Overall, we provide evidence for the upregulation of several phenylpropanoid pathway genes in response to elicitation by Agrobacterium, suggesting that production of secondary metabolites could modulate H. perforatum recalcitrance to A. tumefaciens-mediated transformation.

A sensitive sandwich ELISA using a modified biotin-streptavidin amplified system for histamine detection in fish, prawn and crab.

Histamine poisoning from seafood is a significant public health and safety concern. To detect histamine sensitively and accurately, a novel competitive sandwich immunoassay using a modified biotin-streptavidin system coupling with polylysine was developed. Using this strategy, a sandwich ELISA with an IC50 value of 112.8 ng mL-1 and a broad linear range of 11.7-1500 ng mL-1 with a correlation coefficient of 0.9942 was validated. Without any sample derivatization procedure, the recovery of histamine ranged from 80.19% to 108.3% with a coefficient of variation of 1.43-11.7% in tuna, prawn and crab. The sandwich ELISA had a detectionlimit of 5.86 ng mL-1, which was 15-fold lower than an indirect competitive ELISA (ic-ELISA). This simple, sensitive and accurate method can be applied to detect histamine in routine seafood samples.

Neurotherapeutic effect of Hyptis spp. leaf extracts in Caenorhabditis elegans models of tauopathy and polyglutamine disease: Role of the glutathione redox cycle.

Hyptis suaveolens (HS), Hyptis pectinata (HP) and Hyptis marrubioides (HM) are plants used in folk medicine for treatment of several diseases. Here, we tested the in vivo antioxidant and neuroprotective potential of methanolic extracts from these plants, containing several rosmarinic acid derivatives and isoquercetin. In C. elegans, HS, HP and HM leaf extracts enhanced the antioxidant responses through the induction of specific antioxidant enzymes and demonstrated neurotherapeutic potential in transgenic models of genetically determined human neurodegenerative diseases - Frontotemporal dementia with parkinsonism linked to chromosome 17 and Machado-Joseph disease. Chronic treatment of disease models with HS, HP and HM leaf extracts improved the animals' motor function and increased their tolerance to an oxidative insult. The restorative effect of HM extract in motor performance of both disease models required the presence of glutathione reductase (gsr-1), an enzyme that assures the glutathione redox cycle, highlighting the role of this pathway and unveiling a common candidate therapeutic target for these diseases. Our findings strengthen the relevance of plant-derived bioactive compound discovery for neurodegenerative disorders that remain without effective treatment.

Data on the effects of Hyptis spp. and Lycium spp. plant extracts in C. elegans models of genetically determined neurodegenerative diseases.

Here, we present the data on the biological effects of Hyptis spp. and Lycium spp. plant extracts in Caenorhabditis elegans (C. elegans) models of neurodegenerative diseases, which is related to the work presented in the article "Neurotherapeutic effect of Hyptis spp. leaf extracts in C. elegans models of tauopathy and polyglutamine disease: role of the glutathione redox cycle" [1]. This dataset was generated to define non-toxic concentrations of these plant extracts and to assess their impact on the motor phenotype and oxidative stress resistance of transgenic C. elegans models of two genetically defined neurodegenerative diseases: Machado-Joseph disease and Frontotemporal dementia with Parkinsonism associated to the chromosome 17. The impact of the plant extracts on toxicity was assessed using the food-clearance assay, absorbance being measured daily for seven days at 595 nm to quantify Escherichia coli (E. coli) strain OP50 bacteria consumption. Worm length and motor behaviour, including spontaneous and stimulated movement, were analysed using videos acquired with an Olympus SZX7 stereomicroscope with an integrated camera (Olympus SC30) and processed using the Image J® software and the Wrmtrck plugin. The resistance to oxidative stress induced by 240 µM juglone was assessed by determining the percentage of live animals after 1 hour of exposure.

Development of ELISA and chemiluminescence enzyme immunoassay for quantification of histamine in drug products and food samples.

Histamine (HA) is a biogenic amine associated with allergies and food poisoning. It is an important indicator of food freshness and quality. In recent years, a series of medical negligence cases have been reported to be related to the intravenous injection of antibiotics produced via fermentation with fish peptone due to HA contamination. To detect HA efficiently, mouse monoclonal antibody was developed. An enzyme-linked immunosorbent assay (ELISA) and a chemiluminescence enzyme immunoassay (CLEIA) were developed and compared with conventional HPLC analysis. Both immunoassays showed low cross-reactivity, low 50% inhibitive concentration (IC50; 1.2 µg/mL and 1.1 µg/mL), low limits of detection (LODs, IC10; 89.0 ng/mL and 73.4 ng/mL), and appreciable recoveries in spiked foods and drugs (from 73.4 to 131.0% and from 77.0 to 119.0%, espectively), demonstrating that the developed methods are sensitive, specific, fast, and reliable for HA detection in complicated real samples. Graphical abstract.

Development, characterization, antioxidant and hepatoprotective properties of poly(Ɛ-caprolactone) nanoparticles loaded with a neuroprotective fraction of Hypericum perforatum.

Quercetin and biapigenin are antioxidant and neuroprotective compounds present in Hypericum perforatum, with potential application in neurodegenerative diseases. Quercetin has shown to have excellent antioxidant activities. Biapigenin possesses a distinctive mechanism of action, preventing the onset of calcium deregulation and mitochondrial dysfunction. The main aim of this study was to establish a new delivery system encapsulating H. perforatum neuroprotective fraction, isolated from it,containing quercetin and biapigenin into poly(Ɛ-caprolactone) (PCL) nanoparticles. It also aimed to study its hepatoprotective potential. Different formulation parameters were optimized, resulting in a stable formulation of polymer:compounds ratio 1:0.1 with a mean particle size of 185nm, zeta potential around -20mV and association efficiency close to 100%. Compounds in vitro release, under physiological conditions, showed an initial burst followed by a sustained release. The antioxidant functional properties of these compounds were not altered by encapsulation. PCL-loaded nanoparticles protected HepG2 cells from intrinsic compound toxicity at high concentrations. Depending on the incubation regimen, quercetin-biapigenin PCL-loaded nanoparticles or free compounds were more effective in protecting HepG2 cells against tert-butylhydroperoxide-induced toxicity. This is the first report of the encapsulation of a quercetin-biapigenin mixture in a polymeric matrix, specifically, in PCL, with synergic anti-oxidant and hepatoprotective effects.

Differential phenolic production in leaves of Vitis vinifera cv. Alvarinho affected with esca disease.

Esca is a destructive disease of complex etiology affecting grapevines worldwide. A major constraint to the study and control of esca is that the disease is not diagnosed until external leaf and/or fruit symptoms are visible; however external symptoms usually appear several years after infection onset. We studied the phenolic content of V. vinifera cv. Alvarinho leaves using high performance liquid chromatography-diode array detection-mass spectrometry (HPLC-DAD-MS)/LC-MS. Leaves from affected cordons with and without visible symptoms (diseased and apparently healthy leaves, respectively) and leaves from asymptomatic cordons (healthy leaves) were analyzed. Application of principal components analysis (PCA) to HPLC data showed a clear separation between diseased, apparently healthy, and healthy leaves, with the apparently healthy leaves clustered in a medial position. Several compounds were highly correlated with diseased leaves indicating a differential phenolic production due to esca disease in V. vinifera cv. Alvarinho leaves. Total phenolic production was shown to significantly increase in diseased leaves, compared to healthy leaves, with apparently healthy leaves containing a medial amount. Trans-caffeoyltartaric acid, trans-coumaroyl-tartaric acid, quercetin-3-O-glucoside, quercetin-3-O-galactoside, kaempferol-3-glucoside and myricetin were identified among the compounds associated with disease and their content shown to change similarly to total phenolic production. This study shows that it is possible to discriminate between diseased, healthy and apparently healthy leaves by applying PCA to HPLC data.

Neuroprotective Activity of Hypericum perforatum and Its Major Components.

Hypericum perforatum is a perennial plant, with worldwide distribution, commonly known as St. John's wort. It has been used for centuries in traditional medicine for the treatment of several disorders, such as minor burns, anxiety, and mild to moderate depression. In the past years, its antidepressant properties have been extensively studied. Despite that, other H. perforatum biological activities, as its neuroprotective properties have also been evaluated. The present review aims to provide a comprehensive summary of the main biologically active compounds of H. perforatum, as for its chemistry, pharmacological activities, drug interactions and adverse reactions and gather scattered information about its neuroprotective abilities. As for this, it has been demonstrated that H. perforatum extracts and several of its major molecular components have the ability to protect against toxic insults, either directly, through neuroprotective mechanisms, or indirectly, through is antioxidant properties. H. perforatum has therefore the potential to become an effective neuroprotective therapeutic agent, despite further studies that need to be carried out.

Antimicrobial activity of cream incorporated with silver nanoparticles biosynthesized from Withania somnifera.

We report on the antimicrobial activity of a cream formulation of silver nanoparticles (AgNPs), biosynthesized using Withania somnifera extract. Aqueous extracts of leaves promoted efficient green synthesis of AgNPs compared to fruits and root extracts of W. somnifera. Biosynthesized AgNPs were characterized for their size and shape by physical-chemical techniques such as UV-visible spectroscopy, laser Doppler anemometry, transmission electron microscopy, scanning electron microscopy, atomic force microscopy, X-ray diffraction, and X-ray energy dispersive spectroscopy. After confirming the antimicrobial potential of AgNPs, they were incorporated into a cream. Cream formulations of AgNPs and AgNO3 were prepared and compared for their antimicrobial activity against human pathogens (Staphylococcus aureus, Pseudomonas aeruginosa, Proteus vulgaris, Escherichia coli, and Candida albicans) and a plant pathogen (Agrobacterium tumefaciens). Our results show that AgNP creams possess significantly higher antimicrobial activity against the tested organisms.

Impact of an educational hands-on project on the antimicrobial, antitumor and anti-inflammatory properties of plants on Portuguese students' awareness, knowledge, and competences.

Promoting environmental and health education is crucial to allow students to make conscious decisions based on scientific criteria. The study is based on the outcomes of an Educational Project implemented with Portuguese students and consisted of several activities, exploring pre-existent Scientific Gardens at the School, aiming to investigate the antibacterial, antitumor and anti-inflammatory properties of plant extracts, with posterior incorporation in soaps and creams. A logo and a webpage were also created. The effectiveness of the project was assessed via the application of a questionnaire (pre- and post-test) and observations of the participants in terms of engagement and interaction with all individuals involved in the project. This project increased the knowledge about autochthonous plants and the potential medical properties of the corresponding plant extracts and increased the awareness about the correct design of scientific experiments and the importance of the use of experimental models of disease. The students regarded their experiences as exciting and valuable and believed that the project helped to improve their understanding and increase their interest in these subjects and in science in general. This study emphasizes the importance of raising students' awareness on the valorization of autochthonous plants and exploitation of their medicinal properties.