The PlantLIBRA project: how we intend to innovate the science of botanicals

The main aim of the EC-financed R&D project PlantLIBRA (PLANT food supplements: Levels of Intake, Benefit and Risk Assessment) is to foster the safe use of food supplements containing plants or botanical preparations, by enabling science-based decision making by regulators and stakeholders. To make informed decisions, competent authorities and industry need more accessible and quality-assured information, as well as better tools (e.g., databases) and procedures for safety and benefit assessments, supported by broadly accepted methodologies. Consequently, PlantLIBRA is working to develop, validate and disseminate data and methodologies for risk and benefit assessment of plant food supplements, and to implement sustainable international cooperation. International cooperation will help ensure the quality of botanicals imported in the EU. Moreover, the project will provide data on intake by conducting a harmonized consumption survey. Existing composition and safety data will be collated into a meta-database. New analytical data and methods will be investigated and validated. The consortium is working closely with competent authorities and stakeholders

Simple surface functionalization strategy for immunosensing detection of aflatoxin B1

This paper describes the strategy of functionalization of gold electrodes surfaces in aim to obtain a new impedimetric immunosensor for the detection of aflatoxin B1, a common toxic food contaminant. The immunosensor elaboration is based on immobilization of anti-aflatoxin antibody on gold electrodes modified with a cross-linked film of bovine serum albumin, by a four-step protocol. The immunosensor is based on a simple design and requires small volumes of toxic aflatoxin solution. All the steps of the immunosensor elaboration and the immunochemical reaction between aflatoxin and antibody were followed using atomic force microscopy (AFM) and electrochemical impedance spectroscopy (EIS). The resistance to charge transfer (Rct) was the most sensitive parameter to changes induced to the interfacial properties of the immunosensor by the incubation with aflatoxin and varied linearly with aflatoxin concentration in the range 1-20 ng/mL. The immunosensor was applied for the detection of aflatoxin in spiked plant extracts with good recovery factors

The synaptic blocker botulinum toxin A decreases the density and complexity of oligodendrocyte precursor cells in the adult mouse hippocampus

Oligodendrocyte progenitor cells (OPCs) are responsible for generating oligodendrocytes, the myelinating cells of the CNS. Life-long myelination is promoted by neuronal activity and is essential for neural network plasticity and learning. OPCs are known to contact synapses and it is proposed that neuronal synaptic activity in turn regulates their behavior. To examine this in the adult, we performed unilateral injection of the synaptic blocker botulinum neurotoxin A (BoNT/A) into the hippocampus of adult mice. We confirm BoNT/A cleaves SNAP-25 in the CA1 are of the hippocampus, which has been proven to block neurotransmission. Notably, BoNT/A significantly decreased OPC density and caused their shrinkage, as determined by immunolabeling for the OPC marker NG2. Furthermore, BoNT/A resulted in an overall decrease in the number of OPC processes, as well as a decrease in their lengths and branching frequency. These data indicate that synaptic activity is important for maintaining adult OPC numbers and cellular integrity, which is relevant to pathophysiological scenarios characterized by dysregulation of synaptic activity, such as age-related cognitive decline, Multiple Sclerosis and Alzheimer's disease

Ochratoxin A Detection on Antibody- Immobilized on BSA-Functionalized Gold Electrodes

Ochratoxin A (OTA)-a toxin produced by Aspergillus carbonarius, Aspergillus ochraceus, and Penicillium verrucosum-is one of the most-abundant food-contaminating mycotoxins. To avoid the risk of OTA consumption for humans and animals, the rapid detection and quantitation of OTA level in different commodities are of great importance. In this work, an impedimetric immunosensor for ochratoxin A (OTA) detection, a common toxic botanical contaminant, was developed via the immobilization of anti-OTA antibody on bovine serum albumin modified gold electrodes. A four-step reaction protocol was tested to modify the gold electrode and obtain the sensing substrate. All the steps of the immunosensor elaboration and also the immunochemical reaction between surface-bound antibody and ochratoxin A were analyzed using cyclic voltammetry and electrochemical impedance spectroscopy. Modification of the impedance due to the specific antigen-antibody reaction at immunosensor surface, was used in order to detect ochratoxin A. Linear proportionality of the charge transfer resistance to the concentration of OTA allows ochratoxin A detection in the range of 2.5-100 ng/mL

Phenolic profile and biological activity of table grapes (Vitis vinifera L.)

Table grapes are largely produced and consumed in the Mediterranean area. Furthermore, in the last years, the commercial interest in table grapes and other non-alcoholic grape products has notably increased worldwide. In addition to the nutritional aspects, polyphenol compounds in fresh grapes could exert positive effects on human health. The aim of this study was the characterization of the phenolic pattern of sixteen grape varieties and the evaluation of the associated antioxidant capacity and anti-inflammatory activity at gastric level. The methods used were: 1) Folin-Cocalteau\u2019s assay for the quantification of total polyphenol content; 2) High-Performance Liquid Chromatography (HPLC) coupled with Diode-Array Detector (DAD) to perform the quantitative analysis of grapes phenolic fraction 3) DPPH (1,1,-diphenil-2-picrylhydrazyl) spectrophotometric assay for the assessment of radical scavenging activity; 4) assessment of IL-8 release from human gastric epithelial cells to evaluate the anti-inflammatory activity of grape extracts. Some grapes, including seedless varieties, showed that the phenolic pattern was highly correlated with the biological activities, and, in particular, with peel and seeds portion. These results suggest that selected grape varieties could represent, also for consumers who do not drink wine, a source of healthy compounds potentially able to counteract oxidative stress and gastric inflammation

Duplication of clostridial binding domains for enhanced macromolecular delivery into neurons

Neurological diseases constitute a quarter of global disease burden and are expected to rise worldwide with the ageing of human populations. There is an increasing need to develop new molecular systems which can deliver drugs specifically into neurons, non-dividing cells meant to last a human lifetime. Neuronal drug delivery must rely on agents which can recognise neurons with high specificity and affinity. Here we used a recently introduced ‘stapling’ system to prepare macromolecules carrying duplicated binding domains from the clostridial family of neurotoxins. We engineered individual parts of clostridial neurotoxins separately and combined them using a strong alpha-helical bundle. We show that combining two identical binding domains of tetanus and botulinum type D neurotoxins, in a sterically defined way by protein stapling, allows enhanced intracellular delivery of molecules into neurons. We also engineered a botulinum neurotoxin type C variant with a duplicated binding domain which increased enzymatic delivery compared to the native type C toxin. We conclude that duplication of the binding parts of tetanus or botulinum neurotoxins will allow production of high avidity agents which could deliver imaging reagents and large therapeutic enzymes into neurons with superior efficiency

Optimization of electrochemical detection of L-Ascorbic acid from Plant Food Supplements using screen printed transducers

The importance of Ascorbic acid (vitamin C) in several biological processes, and during the last years as active compounds used in several pesticides (L-Ascorbic acid is used as a fungicide on glasshouse tomato, potato, and field and glasshouse flower bulbs) is a challenge for groups of researchers to develop new, rapid and sensitive method for its detection. Methods for the detection of Ascorbic acid (AA) were developed and optimised using differential pulse voltammetry (DPV) analysis with modified carbon-printed electrodes with cobalt phthalocyanine. There was studied detection of Ascorbic acid in hydrochloric acid 2% as supporting media. Results showed that the method developed for Ascorbic acid analyses can be used to assay different samples as teas and plant food supplements containing different botanicals

Beneficial effects of non-alcoholic grape-derived products on human health: a literature review

Vine is widely cultivated due to the economic value of wine and other grape derivatives. The grape berry is characterized by the presence of a wide variety of flavonoids, which have been investigated for their health promoting properties. Several epidemiological studies have shown that a moderate consumption of wine is associated with a J-shaped effect on some risk factors for chronic diseases. On the other hand, the wine market has shown a decreasing trend due to the frequent abuse of alcoholic beverages also by young people, as denounced by WHO. Accordingly, the scientific research in the field of non-alcoholic grape products has been further stimulated. The aim of this paper was a preliminary collection of data on human studies supporting the beneficial properties of unfermented grape products. The most convincing positive effects, observed in humans, consisted in the reduction of risk factors for cardiovascular diseases, such as hypertension and oxidative stress. Other human trials have been published in the area of: immune system, diabetes, cognitive functions, oral health, and cancer. Generally speaking, the findings listed in this review support the use of nonalcoholic grape derivatives, as a source of beneficial compounds for the human diet, even though further studies are necessar

Raisins in human health : a review

In the last years, the scientific research in the field of non-alcoholic grape products has increased significantly. Raisins are often evaluated negatively from the nutritional point of view, mainly for their high sugar content. On the other hand, some in vitro and in vivo studies have suggested that raisins could have healthy effects due to their positive phytochemical profile. The aim of this work was the collection of scientific studies performed in humans to assess critically the health-promoting effects of raisins, as a part of the normal/Mediterranean diet. In most cases, the beneficial effects of raisins have been assessed in intervention studies focused on cardiovascular area, diabetes and oral health, where a decrease in postprandial glycemia and insulinemia both in diabetic and healthy subjects has been observed. The positive effects were generally evident after a short-term consumption of about 70 g/die of raisins in comparison to a similar quantity of snacks or glucose solution. Surprisingly, some positive findings were shown in oral health. On these bases several findings support the suitability of raisins as a source of healthy compounds for human diet, but limits in the data published till now clearly support the need of new specifically designed trials

Plant Food Supplements with Anti-Inflammatory Properties: A Systematic Review (II)

The aim of this systematic review is to summarize the evidence for or against the efficacy of plant food supplements (PFS) for coping inflammatory conditions by considering epidemiological and human intervention studies. The review considers six botanical species commonly used as food supplements/medicinals: Urtica dioica L., Symphytum officinalis L., Calendula officinalis L., Curcuma longa L., Boswellia serrata Roxb., and Harpagophytum procumbens L. The search retrieved 579 publications. By removing the duplicates and applying the inclusion/exclusion criteria, the final number of papers was 47. No epidemiological data were found. The bibliographic search found no paper regarding the anti-inflammatory effects of Calendula officinalis L. and Symphytum officinalis L. by oral use. In spite of the long-term traditional use for inflammatory disorders, Curcuma longa L. and Harpagophytum procumbens L. warrant further investigation, whereas the efficacy of Urtica dioica L, even if the available data on hard endpoints are promising, requires other trials. Boswellia serrata Roxb. was found to be the most promising, since it shows the best efficacy for the treatment of pain/inflammatory conditions. In conclusion, it is advisable to conduct further studies with more homogeneous population and larger number of subjects by avoiding the heterogeneity of the herbal preparations considered