Quality comparison of organic and conventional wheat by use of common and holistic methods of analysis

The market for organic food is strongly growing. There are many reasons why consumers buy products from organic agriculture. Some reasons are attributed to the product quality as well as to the assumption about the production process used in organic agriculture. Organic food stands for a sustainable and socially acceptable production. In addition, the desire for a healthy nutrition is important. So in summary consumers chose organic products because they trust in a number of attributes related to the product itself and it’s production process. During the last years, food scandals developed in conventional agriculture as well as in organic agriculture. Therefore the consumers lost some of their confidence in organic products. With a growing production the risk for more scandals might be rising. Thus emerged a need for methods, which can examine the quality of organically produced food (authenticity examination) and compare it with the quality of conventional products. In such comparison studies it is of particular importance to use a possibly wide range of analysis methods, because the quality differences due to the farming system can appear in crops in various forms – in the contents of singular compounds, as well as in the structural features, which only can be analyzed on the whole product (Kokornaczyk et al., 2008). Therefore, the common and holistic methods of analysis have been united in the current research in order to receive complementary information about the given samples. The aim of the research was to examine and compare the quality of the organically and conventionally grown grain samples. The samples derived from the Mediterranean Arable System Comparison Trial (MASCOT) in Italian Toscana, a long-term experiment launched in 2001 and carried out at the Interdepartmental Centre for Agri-environmental Research “E. Avanzi” (CIRAA) of the University of Pisa. The choice of the applied analysis methods was based on the founds available in literature. The organically and conventionally grown wheat often differs in total protein content, which depends on the availability of N and is also an important indicator of the technological quality of grain (Alföldi et al., 2006). The content of carotenoids, which belong to the secondary plant metabolites, was found to differ in organic and conventional crops because of the different growth conditions, like exposure to pests and diseases (Alföldi et al., 2006). Many studies were conducted on the antioxidants contents present in different wheat varieties (Zhou et al., 2004; Adom et al., 2003). But to our knowledge there is still limited literature on the contents of antioxidants in wheat samples deriving from differently managed farming systems. Studies indicate that holistic methods, such as biocrystallization, are especially suitable for authenticity tests of organic produce, hence a validation of these methods has been demanded (Siderer et al., 2005)

Influence of temperature and relative humidity on patterns formed in dried plasma and serum droplets

In the present methodological study, we investigated the influence of different evaporation conditions upon patterns formed in desiccated plasma and serum droplets; furthermore, we determined the potential of such patterns to distinguish between four donors. Our results show that the development of different pattern features strongly depended on relative humidity: lower relative humidity triggered the formation of cracks, whereas higher relative humidity favored the development of inner crystalline structures. Settings that allowed for the best donor differentiation and at the same time showed high stability of the experimental system were 24.5 ◦C /15% rH and 30.5 ◦C /45% rH for plasma and 30.5 ◦C /15% rH and 36.6 ◦C /45% rH for serum droplets. The results suggest that for the development of diagnostic tests based on pattern formation in evaporating plasma and serum droplets the choice of right evaporation conditions may influence the experimental system stability as also the test accuracy

Droplet evaporation method applied to test the efficacy of Zincum metallicum 30c on stressed and non-stressed wheat seeds

Background: The droplet evaporation method (DEM) has been recently proposed as a possible tool to test the efficacy of ultra high dilutions (UHDs) [1]. Here we applied the same methodology consisting in the evaporation of droplets from leakages obtained by placing wheat seeds in UHDs to test whether DEM patterns vary in function of the tested treatment: Zincum metallicum 30c (ZM), lactose 30c as dynamized control (L), and unsuccussed, undiluted water as negative control (C). Since our previous study [1] showed that there is a significant increase in the UHD action in the stressed model, with respect to the non-stressed one, also in the present experiment we tested ZM, L and C on both stressed (s-seeds) and non-stressed wheat seeds (ns-seeds). Aims: The aim of the experiment was to test whether treatments ZM, L, and C applied on ns- and s-seeds influence the DEM pattern characteristics. Materials and methods: Whole, undamaged wheat seeds (Triticum aestivum cv. Inallettabile, harvesting year 2010) were used for the experiment, following the experimental protocol described in [1]. The distinction between s- and ns-seeds is based upon different storing conditions from the harvest until the analysis day: the ns-seeds were kept in controlled conditions at 5°C and in the dark, whereas the s-seeds were kept in lab at varying temperature, humidity and light conditions. As far as treatment preparation is concerned, ZM and L were obtained from triturations 3c (received from the Federal University of Rio de Janeiro) by vertical mechanical succussions and following dilutions; negative control (C) was ultra-pure water from the same water batch used for dilutions. The experiment was repeated on 6 days within one month; each seed lot (ns- and s-seeds) was analyzed on 3 different days (s-seeds on 10th, 11th and 24th February 2014 and ns-seeds on 12th, 26th February and 3th March 2014), with three treatment-replicates for ZM and L and two for C, three repetitions for each treatment-replicate and 5 droplets per repetition (360 patterns for each seed lot). The complexity degree of polycrystalline structures obtained from the evaporating droplets was measured by means of the ImageJ software [2] by calculating their local connected fractal dimensions (LCFD). The data on moon phase, moon position and moon distance from Earth at the beginning of each evaporation process were collected from the online tool virtuelles Telescope [3]. All data were processed by a three-way ANOVA at a significance level of p ≤ 0.05. Correlations between the moon data and LCFD were evaluated by r Pearson coefficient. Results: The effect of the treatment on LCFD values of DEM patterns was significant only in the stressed model: ZM showed a crystallization inhibiting action vs. C on all 3 analysis days, whereas a significant difference between ZM and L could be observed only on first and third analysis day (Fig 1a). ANOVA analysis showed that the overall LCFD means for the s-seed lot differed significantly between each other: 1.33 (a) for C, 1.26 (b) for L, and 1.19 (c) for ZM. Moreover, the LCFD values of both seed lots showed a strong day factor influence. A possible explanation of this finding might be the influence of the moon (position, phase, and distance from Earth) on the crystallization process: strong correlation between the LCFD of the patterns and moon data were found (r values were from -0.72 to -0.97). Figure 1: Graphical representation of the mean local connected fractal dimension values (LCFD) of droplet evaporation patterns obtained from stressed and non-stressed wheat seeds following treatments with Zincum metallicum 30c (ZM), lactose 30c (L), and unsuccussed/undiluted water (C). Discussion: The present study confirms a pre-sensitization effect towards UHD action in stressed models: a significant inhibiting effect of ZM 30c was found in all experimentation day. Furthermore, the strong correlations observed between the LCFD values and moon data indicate that the complexity of polycrystalline structures from evaporating droplets of wheat seed leakages might be affected by tidal forces. Since DEM needs to be standardized, these correlations can be considered only simultaneous and not causal, however their strength gives good reasons for further studies. Conclusions: The results of the present pilot-study seem to encourage further DEM experiments on s-seeds following UHD treatments. For further confirmation of the inhibiting effect of ZM on s-seeds, germination tests should be planned. Finally, the performance of DEM experimentations during days and hours with equal tidal influence on gravity might be helpful for the reduction of the day factor. Keywords:Ultra high dilutions, droplet evaporation method, patterns, Zincum metallicu

Preliminary Study on Force-Like Effects between As45x, Water, and Wheat Seeds Performed by Means of the Droplet Evaporation Method

Background: The droplet evaporation method (DEM) is based on pattern formation in evaporating droplets. It has been shown that the local connected fractal dimension (LCFD; a complexity measure) of crystalline structures grown in evaporating droplets of wheat seed leakages depended upon whether the seeds were treated with an ultra-high dilution (UHD) or water control.1, 2 In homeopathy it is usually assumed, that UHD's need to come in direct (physical) contact with the organism to be treated to exhibit their action; however, recently, it has been demonstrated on a wheat seed germination model that UHD's may influence organisms also at a distance, i.e. without any physical contact. 3 In this case the UHD's effectiveness decreases with increasing distance. Here we test whether DEM might serve also as a tool to study such force-like influences. In a series of three DEM experiments (E1-3) we studied (i) force-like effects occurring between As2O3 45x treatment (As45x) and undiluted, unsuccussed, ultrapure water (W), (ii) As45x and wheat seeds, as also (iii) whether force-like effects may be shielded by means of aluminum foil, commonly applied for such purposes in homeopathic research

Multidimensional Outcome Parameters in a Cress Seedling-CuCl2 Crystallization Assay to Corroborate Specific Effects of Stannum metallicum 30x Compared to Lactose 30x.

BACKGROUND  Previously we developed a test system which yielded highly significant evidence for specific effects of a Stannum metallicum 30x preparation in a multi-center replication trial. This test system is based on cress seed germination in homeopathic or control samples, CuCl2 crystallization of the cress extract, and subsequent digital textural image analysis of the resulting crystallization patterns. OBJECTIVES  The current study aimed to investigate whether three novel outcome parameters could further corroborate and possibly characterize the specific effects of Stannum metallicum 30x. METHODS  To this end, (1) cress seedling length, (2) a second texture analysis parameter, entropy and (3) the local connected fractal dimension (LCFD) of crystallization patterns as a measure of complexity were considered. The stability of the experimental setup was monitored throughout the entire investigation with systematic negative control (SNC) experiments. RESULTS  Cress length and entropy revealed a time-modulated potency treatment effect, in the absence of a significant main treatment effect. This indicated that the effect of the potency treatment varied significantly across the different experimental days. LCFD yielded a highly significant potency treatment effect. In addition, a significant interaction of treatment with experimental day seems to indicate a modulation of this effect. No significant effects were observed in any of the evaluations of the SNC experiments, indicative of a stable experimental setup and a reliable and specific treatment effect. Neither significant nor strong correlations were found between the four parameters, indicating that they reflect different effects of Stannum metallicum 30x on the organism treated. CONCLUSION  This quadruple characterization of the biological effects of Stannum metallicum 30x provides an unprecedented opportunity for basic homeopathy research into, among others, the presumed specificity of homeopathic preparations

Vortex-like vs. turbulent mixing of a Viscum album preparation affects crystalline structures formed in dried droplets.

Various types of motion introduced into a solution can affect, among other factors, the alignment and positioning of molecules, the agglomeration of large molecules, oxidation processes, and the production of microparticles and microbubbles. We employed turbulent mixing vs. laminar flow induced by a vortex vs. diffusion-based mixing during the production of Viscum album Quercus L. 10-3 following the guidelines for manufacturing homeopathic preparations. The differently mixed preparation variants were analyzed using the droplet evaporation method. The crystalline structures formed in dried droplets were photographed and analyzed using computer-supported image analysis and deep learning. Computer-supported evaluation and deep learning revealed that the patterns of the variant succussed under turbulence are characterized by lower complexity, whereas those obtained from the vortex-mixed variant are characterized by greater complexity compared to the diffusion-based mixed control variant. The droplet evaporation method could provide a relatively inexpensive means of testing the effects of liquid flow and serve as an alternative to currently used methods

Deep learning applied to analyze patterns from evaporated droplets of Viscum album extracts.

This paper introduces a deep learning based methodology for analyzing the self-assembled, fractal-like structures formed in evaporated droplets. To this end, an extensive image database of such structures of the plant extract Viscum album Quercus [Formula: see text] was used, prepared by three different mixing procedures (turbulent, laminar, and diffusion based). The proposed pattern analysis approach is based on two stages: (1) automatic selection of patches that exhibit rich texture along the database; and (2) clustering of patches in accordance with prevalent texture by means of a Dense Convolutional Neural Network. The fractality of the patterns in each cluster is verified through Local Connected Fractal Dimension histograms. Experiments with Gray-Level Co-Occurrence matrices are performed to determine the benefit of the proposed approach in comparison with well established image analysis techniques. For the investigated plant extract, significant differences were found between the production modalities; whereas the patterns obtained by laminar flow showed the highest fractal structure, the patterns obtained by the application of turbulent mixture exhibited the lowest fractality. Our approach is the first to analyze, at the pure image level, the clustering properties of regions of interest within a database of evaporated droplets. This allows a greater description and differentiation of the patterns formed through different mixing procedures

Self-Organized Crystallization Patterns from Evaporating Droplets of Common Wheat Grain Leakages as a Potential Tool for Quality Analysis

We studied the evaporation-induced pattern formation in droplets of common wheat kernel leakages prepared out of ancient and modern wheat cultivars as a possible tool for wheat quality analysis. The experiments showed that the substances which passed into the water during the soaking of the kernels created crystalline structures with different degrees of complexity while the droplets were evaporating. The forms ranged from spots and simple structures with single ramifications, through dendrites, up to highly organized hexagonal shapes and fractal-like structures. The patterns were observed and photographed using dark field microscopy in small magnifications. The evaluation of the patterns was performed both visually and by means of the fractal dimension analysis. From the results, it can be inferred that the wheat cultivars differed in their pattern-forming capacities. Two of the analyzed wheat cultivars showed poor pattern formation, whereas another two created well-formed and complex patterns. Additionally, the wheat cultivars were analyzed for their vigor by means of the germination test and measurement of the electrical conductivity of the grain leakages. The results showed that the more vigorous cultivars also created more complex patterns, whereas the weaker cultivars created predominantly poor forms. This observation suggests a correlation between the wheat seed quality and droplet evaporation patterns

Scientific guidelines for preclinical research on potentised preparations manufactured according to current pharmacopoeias-the PrePoP guidelines.

OBJECTIVE Pharmacopoeias regulate the manufacture of potentised pharmaceutical preparations used in different branches of complementary and integrative medicine. The physicochemical properties and biological activity of these preparations are often investigated in preclinical research, yet no guidelines for experimental research currently exist in this area. The present PrePoP guidelines aim to provide recommendations to promote high-quality, statistically sound, and reproducible preclinical research on potentised preparations. METHODS Input was gathered from researchers nominated by the relevant scientific societies using a simplified Delphi consensus approach covering the most relevant aspects of basic research methodology in the field including appropriate controls, sample preparation and handling, and statistics. After three rounds of feedback, a consensus was finally reached on the most important aspects and considerations for conducting high-quality research on potentised preparations. RESULTS We present a series of recommendations on a range of topics including experimental controls, system stability, blinding and randomisation, environmental influences, and procedures for the preparation of potentised samples and controls, and we address some specific challenges of this research field. CONCLUSION This expert consensus process resulted in a robust set of methodological guidelines for research on potentised preparations and provides a valuable framework that will inform and improve the quality of subsequent research in this emerging field. PLEASE CITE THIS ARTICLE AS Tournier AL, Bonamin LV, Buchheim-Schmidt S, Cartwright S, Dombrowsky C, Doesburg P, Holandino C, Kokornaczyk MO, van de Kraats EB, López-Carvallo JA, Nandy P, Mazón-Suástegui JM, Mirzajani F, Poitevin B, Scherr C, Thieves K, Würtenberger S, Baumgartner S. Scientific guidelines for preclinical research on potentised preparations manufactured according to current pharmacopoeias-the PrePoP guidelines. J Integr Med. 2024; Epub ahead of print