The use of ultra-diluted preparations method in agriculture was introduced with agro-homeopathy, which allows to influence biological processes of plants by either accelerating or delaying growth. Moreover, it can contribute to the control of plagues and diseases, directly promoting an increase of the yield and an improvement of product qualitative traits. Since the pioneering works of Kolisko on wheat germination [1] and Junker on growth of microorganisms (paramecium, yeast, fungi) [2], in the last 30 years work has flourished from independent researchers from worldwide (Americas, Europe and Australasia). The international research works on agro-homeopathy can be conceptually divided in two main groups: effects of ultra-diluted preparations on crop growth and applicability for crop disease control. The first type of investigations usually are carried out on both healthy organisms for determining the growth stimulation of treatments and on abiotically stressed plants (i.e. heavy metal over-exposition, salt excess, water and nutrients deficiency) for determining the re-growth stimulation of ultra-dilutions [3,4]. The second type of investigations are usually performed on artificially diseased organisms (i.e. fungal and viral pathogens or nematode infection), which may react more markedly to homeopathic treatments than healthy ones [5]. Unfortunately, on the basis of the extensive critical review of published papers, there is little firm evidence to support the reliability of the reported results. Except for a limited number of publications, the most common drawbacks of agro-homeopathy researches are the poor experimental methodology and the inadequate statistical analysis. Moreover, since there is no agricultural homeopathic pharmacopoeia, much work is required to find suitable remedies, potencies and dose levels. Considering the criticism on the practical applicability of ultra-diluted preparations, in order to be accepted as a valid part of agricultural practices a well structured research strategy for agro-homeopathy is needed. This is often hampered by methodological problems as well as by the general underinvestment on the academic and nonacademic research structures. Fundamental researches based on collaborative approaches (i.e. ring tests on selected crop models) and on common experimental protocols (i.e. statistical robustness) are the keys for determining the worldwide acceptability of agro-homeopathy as a sustainable agro-technique. Statement of conflict of interest Authors declare there is no conflict of interest. Statement of financial support Authors declare that this study received no funding. Bibliography 1. Kolisko L. Physiologischer und physikalischer Nachweis der Wirksamkeit kleinster Entitäten. 1923; Stuttgart, Verlag Der Kommende Tag AG. 2. Junker H.. Die Wirkung extremer Potenzverdünnungen auf Organismen. Pflugers Arch ges Phys 1928; 219B, 5/6, 647-672. 3. Jäger T, Scherr C, Shah D, Majewsky V, Betti L, Trebbi G, Bonamin L, Simões-Wüst AP, Wolf U, Simon M, Heusser P, Baumgartner S. Use of homeopathic preparations in experimental studies with abiotically stressed plants. Homeopathy 2011; 100: 275-287 4. Majewsky V, Heuwieser, Shah D, Scherr C, Jaeger Tim, Betti L, Trebbi G, Bonamin L, Klocke P, Baumpartner S. Use of homeopathic preparations in experimental studies with healthy plants. Homeopathy 2009; 98: 228-243. 5. Betti L, Trebbi G, Majewsky V, Scherr C, Shah-Rossi D, Jäger T, Baumgartner S. Use of homeopathic preparations in phytopathological models and in field trials: a critical review. Homeopathy 2009; 98: 244-266.

The important functions of the endothelium and the relationship between cardiovascular risk factors and endothelial dysfunction suggest the primary role of this tissue as a target for dietary strategies aimed at the prevention from related diseases. Cereals are key component of a healthy and balanced diet, and the presence of non-specific lipid-transfer protein 2 (nsLTP2) in wheat represents an added value to contribute to maintain the functionality of the vascular endothelium and consequently of the cardiovascular system. Indeed, nsLTP2 downregulates the expression of the main cell adhesion molecules induced by a pro-inflammatory cytokine and, meanwhile, upregulates heme oxigenase-1, exerting a cytoprotective/anti-inflammatory activity. Therefore, nsLTP2 might represent a food-derived tool to protect the vascular system against several pathological conditions.

Modern agriculture must meet the changing needs of society both in terms of food quality and with regards to the environmental issues related to primary production (e.g., biodiversity, habitats, the quality of water and air, and climate change). In this context, win-win solutions at the farm level are required, with more sustainable food production, as well as, a reduction in air, water, and soil pollution, and loss of biodiversity and natural resources. To address these issues, a new emerging discipline called “systemic agro-homeopathy” is currently being developed. This approach considers plants and their interactions with the environment as a unified agroecosystem, called the “holon” and it is based on the use of substances at ultrahigh dilutions that are safe for farmers and have no ecological side effects. This report describes the principles and the application of this approach.

Introduction “Agro-homeopathy” plays an important role in sustainable agriculture providing cost-saving and residue-free plant treatments for the improvement of yield and the management of diseases and pests [1]. However, one of the main difficulties in “agro-homeopathy” is the exact choice of the right remedy and dilution, which requires the performance of many time-consuming trials. The droplet evaporation method could constitute a screening method for evaluating the efficacy of extremely low doses and high dilutions on plants. Droplet patterns prepared out of wheat seed leakages show differences in their complexity in relation to the seed vigour [2]. Here we show that this structure-forming capacity of seed leakages prepared in water or in ultra-high diluted (UHD) solutions may give a reliable picture of the treatment’s stimulating or inhibiting influence on plant vitality. Materials and Methods Part of the seeds was stressed with 0.1% As2O3 (As) aqueous solution [3]. The droplet patterns were prepared out of four samples: non-stressed (ns) seeds in water, ns-seeds in As 45x, stressed (s) seeds in water, s-seeds in As 45x. The experiment was performed in a 3 day repetition, 3 replicates per day, and 5 droplets per each replicate. The resulting patterns were evaluated by means of the ImageJ software [4] for their local connected fractal dimension (LCFD) values. The plant vigour of the four samples was evaluated by means of in vitro wheat growth [5, 6]. Results The mean LCFD of patterns deriving from s-seeds was significantly lower (1.4) vs. ns-seeds (1.7). Moreover, for both ns- and s-seeds, the LCFD was significantly higher with treatment As 45x vs. water (1.8 vs. 1.7 for ns-seeds, and 1.5 vs. 1.2 for s-seeds). The results of the growth test showed a significant vigour decrease caused by As stress (shoot length 19.7mm) vs. ns-seeds (22.4mm); a significant stimulating effect following As 45x treatment was observed for both s-seeds (25.1mm) and ns-seeds (31.4). Discussion The droplet patterns seem to be a sensitive tool for visualizing the effectiveness of UHD on seeds. The experiment showed that the pattern complexity increased with the application of the treatment for both ns- and s-seeds. The growth test provided a further verification of these results. Conclusions Further confirmations are required, but even as it is now the droplet evaporation method may be considered a promising tool for “agro-homeopathic” screening tests. Authors declare that there is no conflict of interest and that this study received no funding. References Betti L, Trebbi G, Majewsky V, Scherr C, Shah-Rossi D, Jaeger T, Baumgartner S. Use of homeopathic preparations in phytopathological models and in field trials: a critical review. Homeopathy 2009; 98: 244-266. Kokornaczyk MO, Dinelli G, Marotti I, Benedettelli S, Nani D, Betti L. Self-organized crystallization patterns from evaporating droplets of Common wheat grain leakages as a potential tool for quality analysis. TheScientificWorldJ. 2011; 11: 1712-1725. Brizzi M, Elia V, Trebbi G, Nani D, Peruzzi M, Betti L. The efficacy of ultra molecular aqueous dilutions on a wheat germination model as a function of heat and aging-time. e-CAM 2011; Article ID 696298: 11p. doi: 10.1093/ecam/nep217. Collins TJ. ImmageJ for microscopy. BioTechniques. 2007; 43: 25-30. Betti L, Brizzi M, Nani D, Peruzzi M. Effect of high dilutions of Arsenicum album on wheat seedlings from seed poisoned with the same substance. Br. Homeopath. J. 1997; 86: 86-89. Brizzi M, Lazzarato L, Nani D, Borghini F, Peruzzi M, Betti L. A biostatistical insight into the As2O3 high dilution effects on the rate and variability of wheat seedling growth. Forsch. Komplementmed. 2005; 12: 277-283.