Evaluation of Preclinical Assays to Investigate an Anthroposophic Pharmaceutical Process Applied to Mistletoe ( Viscum album

Extracts from European mistletoe (Viscum album L.) developed in anthroposophic medicine are based on specific pharmaceutical procedures to enhance remedy efficacy. One such anthroposophic pharmaceutical process was evaluated regarding effects on cancer cell toxicity in vitro and on colchicine tumor formation in Lepidium sativum. Anthroposophically processed Viscum album extract (APVAE) was produced by mixing winter and summer mistletoe extracts in the edge of a high-speed rotating disk and was compared with manually mixed Viscum album extract (VAE). The antiproliferative effect of VAE/APVAE was determined in five cell lines (NCI-H460, DU-145, HCC1143, MV3, and PA-TU-8902) by WST-1 assay in vitro; no difference was found between VAE and APVAE in any cell line tested (P>0.14). Incidence of colchicine tumor formation was assessed by measurement of the root/shoot-ratio of seedlings of Lepidium sativum treated with colchicine as well as VAE, APVAE, or water. Colchicine tumor formation decreased after application of VAE (−5.4% compared to water, P<0.001) and was even stronger by APVAE (−8.8% compared to water, P<0.001). The high-speed mistletoe extract mixing process investigated thus did not influence toxicity against cancer cells but seemed to sustain morphostasis and to enhance resistance against external noxious influences leading to phenomenological malformations

Test System Stability and Natural Variability of a Lemna Gibba L. Bioassay

BACKGROUND: In ecotoxicological and environmental studies Lemna spp. are used as test organisms due to their small size, rapid predominantly vegetative reproduction, easy handling and high sensitivity to various chemicals. However, there is not much information available concerning spatial and temporal stability of experimental set-ups used for Lemna bioassays, though this is essential for interpretation and reliability of results. We therefore investigated stability and natural variability of a Lemna gibba bioassay assessing area-related and frond number-related growth rates under controlled laboratory conditions over about one year. METHODOLOGY/PRINCIPAL FINDINGS: Lemna gibba L. was grown in beakers with Steinberg medium for one week. Area-related and frond number-related growth rates (r(area) and r(num)) were determined with a non-destructive image processing system. To assess inter-experimental stability, 35 independent experiments were performed with 10 beakers each in the course of one year. We observed changes in growth rates by a factor of two over time. These did not correlate well with temperature or relative humidity in the growth chamber. In order to assess intra-experimental stability, we analysed six systematic negative control experiments (nontoxicant tests) with 96 replicate beakers each. Evaluation showed that the chosen experimental set-up was stable and did not produce false positive results. The coefficient of variation was lower for r(area) (2.99%) than for r(num) (4.27%). CONCLUSIONS/SIGNIFICANCE: It is hypothesised that the variations in growth rates over time under controlled conditions are partly due to endogenic periodicities in Lemna gibba. The relevance of these variations for toxicity investigations should be investigated more closely. Area-related growth rate seems to be more precise as non-destructive calculation parameter than number-related growth rate. Furthermore, we propose two new validity criteria for Lemna gibba bioassays: variability of average specific and section-by-section segmented growth rate, complementary to average specific growth rate as the only validity criterion existing in guidelines for duckweed bioassays

Systems thinking : an approach for understanding 'eco-agri-food systems'

The TEEBAgriFood ‘Scientific and Economic Foundations’ report addresses the core theoretical issues and controversies underpinning the evaluation of the nexus between the agri-food sector, biodiversity and ecosystem services and externalities including human health impacts from agriculture on a global scale. It argues the need for a ‘systems thinking‘ approach, draws out issues related to health, nutrition, equity and livelihoods, presents a Framework for evaluation and describes how it can be applied, and identifies theories and pathways for transformational change

Systematic review of conceptual criticisms of homeopathy.

Homeopathy is the subject of frequent debates, especially in public media. This systematic review aims to give an overview of conceptual criticisms of homeopathy in the scientific literature. The literature search was conducted in four databases (EMBASE, PubMed, Web of Science, PhilPapers) on August 25, 2020. Included were peer reviewed articles in English or German criticising the basic concepts of homeopathy as main topic; excluded were articles criticising homeopathy primarily based on analysis of empirical clinical and/or preclinical data. The formal structure of publications included was evaluated regarding the recommended structure for scientific publications (IMRaD, acronym for 'Introduction', 'Methods', 'Results' and 'Discussion'). Arguments criticising the concepts of homeopathy were extracted and classified into groups. The literature search revealed 5139 articles, of which 15 articles (published between 1959 and 2020) met the inclusion criteria. These articles complied only partly with the IMRaD structure; just four articles considered with 8 or 9 IMRaD criteria the majority of the defined 11 IMRaD criteria. Extracted arguments against the concepts of homeopathy were classified into five groups: 'Conflict with current scientific principles and the foundations of modern medicine', 'Lack of a scientific basis', 'Arguments based on scientific theories', 'Ethical considerations and social consequences', 'Lack of empirical clinical evidence'. This classification is intended to provide a basis for future in-depth scientific analyses and discussions. Based on the number of articles found in the peer reviewed literature, it can be concluded that the on-going discussion about homeopathy in the public media is not reflected in a corresponding academic debate

Reproducibility of the effects of homeopathically potentised Argentum nitricum on the growth of Lemna gibba L. in a randomised and blinded bioassay

Background: A previous study reported a significant statistical interaction between experiment date and treatment effect of Argentum nitricum 14x–30x on the growth rate of duckweed (Lemna gibba L.). The aim of the present study was to investigate the stability of the test system and intra-laboratory reproducibility of the effects found. Methods: Duckweed was treated with A. nitricum potencies (14x–30x) as well as succussed and unsuccussed water controls. The outcome parameter area-related growth rate for day 0–7 was determined by a computerised image analysis system in two series of independent randomised and blinded experiments. Systematic negative control (SNC) experiments were carried out to investigate test system stability. Statistical analysis was performed with full two-way analysis of variance (ANOVA) and protected Fisher's Least Significant Difference (LSD) test. Results: In the first repetition series we found a significant treatment effect (p = 0.016), while in the second series no effect was observed. The negative control experiments showed that the experimental system was stable. An a posteriori subgroup analysis concerning gibbosity revealed the importance of this growth state of L. gibba for successful reproduction of the statistically significant interaction in the original study; flat: no interaction (p = 0.762); slight gibbosity: no interaction (p = 0.356); medium gibbosity: significant interaction (p = 0.031), high gibbosity: highly significant interaction (p = 0.005). Conclusions: With the original study design (disregarding gibbosity status of L. gibba) results of the original study could not be reproduced sensu stricto. We conclude that the growth state gibbosity is crucial for successful reproduction of the original study. Different physiological states of the test organisms used for bioassays for homeopathic basic research must carefully be considered

Effects of Potentised Substances on Growth Kinetics of Saccharomyces cerevisiae and Schizosaccharomyces pombe

Background: Homeopathic potencies are used as specific remedies in complementary medicine. Since the mode of action is unknown, the presumed specificity is discussed controversially. Objective: This study investigated the effects of potentised substances on two yeast species, Saccharomyces cerevisiae and Schizosaccharomyces pombe, in a stable and reliable test system with systematic negative controls. Materials and Methods: Yeast cells were cultivated in either potentised substances or water controls in microplates and their growth kinetics were measured photometrically. Water control runs were performed repeatedly to investigate the stability of the experimental set-up (systematic negative controls). Results: 4 out of 14 screened substances seem to have affected the growth curve parameters slope or yield. Out of these substances, azoxystrobin and phosphorus were chosen for 8 further replication experiments, which partly confirmed the results of the screening. On the average of all experiments, azoxystrobin affected the slope of the growth curve of Saccharomyces cerevisiae (p < 0.05), and phosphorus affected the slope of the growth curve of Schizosaccharomyces pombe (p < 0.05). No effects were seen in the water control runs. In addition, significant interactions between treatment with potentised substances and experiment number were observed in all experiments with potentised substances (p < 0.01), but not in the water control runs. Conclusions: Both yeast species reacted to certain potentised substances by changing their growth kinetics. However, the interactions found point to additional factors of still unknown nature, that modulate the effects of potentised substances. This stable test system with yeasts may be suitable for further studies regarding the efficacy of homeopathic potencies

Phase-Transition-Induced Pattern Formation Applied to Basic Research on Homeopathy: A Systematic Review.

BACKGROUND Methods based on phase-transition-induced pattern formation (PTPF) are increasingly used in medical research. Frequent application fields are medical diagnosis and basic research in homeopathy. Here, we present a systematic review of experimental studies concerning PTPF-based methods applied to homeopathy research. We also aimed at categorizing the PTPF methods included in this review. MATERIALS AND METHODS Experimental studies were collected from scientific databases (PubMed, Web of Science, Russian eLibrary) and from experts in the research field in question, following the PRISMA guidelines. The studies were rated according to pre-defined scientific criteria. RESULTS The review included 15 experimental studies. We identified seven different PTPF methods applied in 12 experimental models. Among these methods, phase-transition was triggered through evaporation, freezing, or solution, and in most cases led to the formation of crystals. First experimental studies concerning the application of PTPF methods in homeopathic research were performed in the first half of the 20th century; however, they were not continued in the following years. Only in the last decade, different research groups re-launched the idea, introducing new experimental approaches and computerized pattern evaluation techniques. The here-identified PTPF methods are for the first time proposed to be classified as one group of methods based on the same basic physical phenomenon. CONCLUSIONS Although the number of experimental studies in the area is still rather limited, the long tradition in the application of PTPF methods and the dynamics of the present developments point out the high potential of these methods and indicate that they might meet the demand for scientific methods to study potentized preparations

Do Potentised Substances have an Effect on Microorgisms?

Do Potentised Substances have an Effect on Microorgisms

S (2007) Duckweed (Lemna gibba L.) as test organism for homeopathic potencies

ABSTRACT Objectives: A bioassay with duckweed (Lemna gibba L.) was used to study the effects of homeopathic potencies on the plant&apos;s growth rate. Screening included 12 substances: argentum nitricum, copper sulfate, gibberellic acid, 3-indole acetic acid, kinetin, lactose, lemna minor, methyl jasmonate, metoxuron, phosphorus, potassium nitrate, and sulfur. Each substance was tested in the potency range 14x-30x. Controls were unsuccussed and succussed water. Design: In randomized and blinded experiments, duckweed was grown in either potentized substances or water controls over 7 days. Frond (leaf) growth was measured regularly with a computerized image analysis system and growth rates were calculated for different time intervals (day 0 -7, 0 -3, 3 -7). Additionally, a water control run with unsuccussed water as the only test substance was performed to determine the variability of the bioassay. Results: For the water control run, the between-group coefficient of variance for groups of five replicates was 0.87% for the frond area-related average specific growth rate r (area) compared to 1.60% for the frond number-related average specific growth rate r (num) . Thus, the former is the preferred parameter to be used. Of twelve tested substances, potentized argentum nitricum, phosphorus, and kinetin significantly (p Ͻ 0.05, analysis of variance Ftest) affected the main parameter: frond area-related average specific growth rate (day 0 -7). Segmented area growth rates (day 0 -3 or 3 -7) were affected by potentized argentum nitricum, gibberellic acid, lactose, and phosphorus. Conclusions: The described experimental set-up with L. gibba as test organism appears to be a promising new model system to investigate effects of potentized substances. Yet larger sets of replication experiments with selected test substances and systematic negative controls are necessary to verify the effects found. 93