Biomolecules and Natural Medicine Preparations: Analysis of New Sources of Bioactive Compounds from Ribes and Rubus spp. buds

It is well known that plants are important sources for the preparation of natural remedies as they contain many biologically active compounds. In particular, polyphenols, terpenic compounds, organic acids, and vitamins are the most widely occurring groups of phytochemicals. Some endemic species may be used for the production of herbal preparations containing phytochemicals with significant bioactivity, as antioxidant activity and anti-inflammatory capacities, and health benefits. Blackberry sprouts and blackcurrant buds are known to contain appreciable levels of bioactive compounds, including flavonols, phenolic acids, monoterpenes, vitamin C, and catechins, with several clinical effects. The aim of this research was to perform an analytical study of blackcurrant and blackberry bud-preparations, in order to identify and quantify the main biomarkers, obtaining a specific phytochemical fingerprint to evaluate the single botanical class contribution to total phytocomplex and relative bioactivity, using a High Performance Liquid Chromatograph−Diode Array Detector; the same analyses were performed both on the University laboratory and commercial preparations. Different chromatographic methods were used to determine concentrations of biomolecules in the preparations, allowing for quantification of statistically significant differences in their bioactive compound content both in the case of Ribes nigrum and Rubus cultivated varieties at different harvest stages. In blackcurrant bud-extracts the most important class was organic acids (50.98%) followed by monoterpenes (14.05%), while in blackberry preparations the main bioactive classes were catechins (50.06%) and organic acids (27.34%). Chemical, pharmaceutical and agronomic-environmental knowledge could be important for obtaining label certifications for the valorization of specific genotypes, with high clinical and pharmaceutical value: this study allowed to develop an effective tool for the natural preparation quality control and bioactivity evaluation through the chemical fingerprinting of bud preparations

Proximal and distal control for ligand binding in neuroglobin: role of the CD loop and evidence for His64 gating

Neuroglobin (Ngb) is predominantly expressed in neurons of the central and peripheral nervous systems and it clearly seems to be involved in neuroprotection. Engineering Ngb to observe structural and dynamic alterations associated with perturbation in ligand binding might reveal important structural determinants, and could shed light on key features related to its mechanism of action. Our results highlight the relevance of the CD loop and of Phe106 as distal and proximal controls involved in ligand binding in murine neuroglobin. We observed the effects of individual and combined mutations of the CD loop and Phe106 that conferred to Ngb higher CO binding velocities, which we correlate with the following structural observations: the mutant F106A shows, upon CO binding, a reduced heme sliding hindrance, with the heme present in a peculiar double conformation, whereas in the CD loop mutant "Gly-loop", the original network of interactions between the loop and the heme was abolished, enhancing binding via facilitated gating out of the distal His64. Finally, the double mutant, combining both mutations, showed a synergistic effect on CO binding rates. Resonance Raman spectroscopy and MD simulations support our findings on structural dynamics and heme interactions in wild type and mutated Ngbs

Medicinl plants, chemical composition and quality: may blackcurrant buds and blackberry sprouts be a new polyphenol source for herbal preparations?

It is well known that plants are important sources for the preparation of natural remedies as they contain many biologically active compounds: in particular, phenolic compounds are one of the most widely occurring groups of phytochemicals. Some endemic species may be used for the production of herbal preparations containing phytochemicals with significant antioxidant capacities and health benefits: blackberry sprouts and blackcurrant buds are known to contain appreciable levels of phenolic compounds, including flavonols, phenolic acids and catechins. The aim of this research was to perform an analytical study of blackcurrant and blackberry bud-preparations, in order to identify the main bioactive polyphenolic compounds, to study the total polyphenolic content and to obtain a specific profile of the main polyphenols contained in these products using a High Performance Liquid Chromatograph - Diode Array Detector; the same analyses were performed both on the University lab preparations and on commercial preparations.Differentchromatographic methodswere usedto determine concentrations of phytochemical compounds in the preparations, allowing to quantifystatistically significant differencesin their polyphenolic content both in the case of Ribes nigrum and Rubus ulmifolius. The assessment of chemical composition and bioactivities of the plant-derived products could help in find out new sources of natural antioxidants and other health-promoting compounds: only with the deep knowledge of the bioactive composition of plant preparations it will be possible to develop a new generation of standardized, effect-optimized, mono- and multi-extract preparations

On the use of Life Cycle Assessment to improve agronomists’ knowledge and skills toward sustainable agricultural systems

Purpose. In agricultural and forestry sciences higher education, environmental sustainability is most often taught through the discussion of examples of green agricultural practices, such as precision farming, and more rarely by taking a more general point of departure in environmental assessment methods, such as Life Cycle Assessment (LCA). Nevertheless, we think that teaching LCA in the agronomists’ curriculum might significantly contribute to enhance students’ systemic perspective on agricultural sustainability. The purpose of this paper is to highlight which additional knowledge and skills may be given to agronomists thorough the teaching of LCA.Design/Methodology/Approach. We designed two short courses focused on LCA to be followed by students at the Bachelor´s degree in Agronomy (University of Turin, Italy) and at the Master´s degree in Sustainability of Agro-food Networks (UNESCO Chair for Sustainable Development, Turin, Italy). After the courses, students filled in a questionnaire about their opinions on the usefulness and value taken from the short courses. Findings. From students’ answers in the questionnaire and their comments during both teaching sessions, it was possible to point out four key aspects acquired by students during the courses: (I) Complexity of agricultural systems. Application of LCA requires to describe the energy flows and material cycles of the system under study and to decide the allocation of environmental impacts to specific phases of the production. (II) Systemic view of the farms. The need to identify boundaries between technical and natural systems for impact assessment highlights the strong interconnection between the two of them. (III) The problem of efficiency. The application of LCA may highlight that productions that are efficient from an agronomic point of view may not be as efficient from an environmental point of view. (IV) Conceptions about sustainable agriculture. During the group work, students were asked to highlight (if possible) the paradigm of sustainability of the authors of the scientific papers and to discuss it. This way, they were able to reflect on the complexity of the concept on environmental sustainability.Practical Implications. Teaching LCA in an interactive course, agronomists discussed pivotal concepts for environmental sustainability, such as system thinking, the problem of efficiency as well as conceptions about sustainable agriculture. All of these aspects reflect positively on the professional life of the agronomists, even if they will not apply any environmental impact methods in their future careers.Originality/Value. This paper describes a pioneer research in which LCA is used as a pure educational tool for understanding the environmental efficiency of agricultural systems, but also founding concepts of environmental sustainability in the agricultural sector

Subcellular localization of the five members of the human steroid 5α-reductase family

In humans the steroid 5a-reductase (SRD5A) family comprises five integral membrane enzymes that carry out reduction of a double bond in lipidic substrates: D4-3-keto steroids, polyprenol and trans-enoyl CoA. The best-characterized reaction is the conversion of testosterone into the more potent dihydrotestosterone carried out by SRD5A1-2. Some controversy exists on their possible nuclear or endoplasmic reticulum localization. We report the cloning and transient expression in HeLa cells of the five members of the human steroid 5a-reductase family as both N- and Cterminus green fluorescent protein tagged protein constructs. Following the intrinsic fluorescence of the tag, we have determined that the subcellular localization of these enzymes is in the endoplasmic reticulum, upon expression in HeLa cells. The presence of the tag at either end of the polypeptide chain can affect protein expression and, in the case of trans enoyl-CoA reductase, it induces the formation of protein aggregates