scholarly journals Development of a biocrystallisation assay for examining effects of homeopathic preparations using cress seedlings

2021 ◽  
Vol 11 (40) ◽  
pp. 202-203
Author(s):  
Stephan Baumgartner ◽  
Paul Doesburg ◽  
Scherr Claudia ◽  
Jens-Otto Andersen
Keyword(s):  
Image Analysis ◽  
Outcome Measure ◽  
Growth Parameters ◽  
Germination Rate ◽  
Basic Research ◽  
Order Effect ◽  
Lepidium Sativum ◽  
Organic Additives ◽  
Plant Bioassays

Background: A major challenge of homeopathic basic research is to develop test systems that yield consistent results. Outcome of plant bioassays is usually based on growth parameters (e.g. germination rate, seedling length, leaf area). Aims: We aimed to evaluate the potential of a crystallisation method with additives (“biocrystallisation”) as complementary outcome measure. This method used is based on the crystallographic phenomenon that when crystallising watery solutions of dihydrate CuCl2 in the presence of organic additives (juices/extracts), reproducible dendritic crystal structures are observed. The resulting biocrystallograms can be evaluated visually and/or by computerized image analysis. Methods: Cress seeds (Lepidium sativum L.) germinated and grew in vitro in either Stannum met. 30x or water 30x. Per experiment, six coded (blinded) 30x preparations were applied in randomized order, representing three independent replicates of the two treatments. Seedlings grew for 96 hours in darkness and were subsequently processed into a watery extract. Biocrystallisation was performed on circular glass plates in 6-fold replication per treatment group, yielding 36 biocrystallograms per experiment. A total of 15 independent experiments were performed at two independent laboratories. Biocrystallograms were scanned and analysed by computerized texture image analysis, using 15 second-order parameters as outcome measure. 3-way-ANOVA with the independent parameters treatment (n=2), internal replicate (n=3), and number of experiment (n=15) was used to analyse the data. Results: All 15 texture analysis variables yielded significant or highly significant results for the homeopathic treatment. Two variables yielded differences between the internal replicates, most probably due to a processing order effect. There were only minor differences between the results of the two laboratories. Conclusions: The texture of biocrystallograms of homeopathically treated cress exhibited specific characteristics, differentiating water 30x and Stannum met. 30x. Thus, the biocrystallisation method seems to be a promising complementary outcome measure for plant bioassays investigating effects of homeopathic preparations.

scholarly journals Development of a Biocrystallisation Assay for Examining Effects of Homeopathic Preparations Using Cress Seedlings

2012 ◽  
Vol 2012 ◽  
pp. 1-14 ◽  
Author(s):  
S. Baumgartner ◽  
P. Doesburg ◽  
C. Scherr ◽  
J.-O. Andersen
Keyword(s):  
Image Analysis ◽  
Texture Analysis ◽  
Outcome Measure ◽  
Basic Research ◽  
Order Effect ◽  
Lepidium Sativum ◽  
Homeopathic Treatment ◽  
Processing Order ◽  
Plant Bioassays

A major challenge in basic research into homeopathic potentisation is to develop bioassays that yield consistent results. We evaluated the potential of a seedling-biocrystallisation method. Cress seeds (Lepidium sativumL.) germinated and grew for 4 daysin vitroinStannum metallicum30x or water 30x in blinded and randomized assignment. 15 experiments were performed at two laboratories. CuCl2-biocrystallisation of seedlings extracted in the homeopathic preparations was performed on circular glass plates. Resulting biocrystallograms were analysed by computerized textural image analysis. All texture analysis variables analysed yielded significant results for the homeopathic treatment; thus the texture of the biocrystallograms of homeopathically treated cress exhibited specific characteristics. Two texture analysis variables yielded differences between the internal replicates, most probably due to a processing order effect. There were only minor differences between the results of the two laboratories. The biocrystallisation method seems to be a promising complementary outcome measure for plant bioassays investigating effects of homeopathic preparations.

scholarly journals Examination of specific effects of different homeopathic preparations on cress seedlings with a CuCl2-biocrystallization assay

2021 ◽  
Vol 15 (4) ◽  
pp. 10-10
Author(s):  
Anezka Marie Sokol ◽  
Paul Doesburg ◽  
Claudia Scherr ◽  
Tim Jäger ◽  
Annekathrin Ãœcker ◽  
...  
Keyword(s):  
Image Analysis ◽  
Texture Analysis ◽  
Lepidium Sativum ◽  
Follow Up Study ◽  
Specific Effects ◽  
Glass Plates ◽  
Homeopathic Remedies ◽  
Lepidium Sativum L

The effect of six different homeopathic remedies on cress (Lepidium sativum L.) has been examined by CuCl2-biocrystallization. The method was based on the protocols of [1] and a follow up study (not yet published). Cress seeds germinated and grew for four days in vitro in a 30x potentization of Stannum metallicum, Arsenic Album, Mercurius metallicum, Sulphur, Silicea or Lactose (control) in a blinded and fully randomized assignment. Each remedy was prepared sterile and divided into 20 bottles used in 10 experiments. CuCl2-biocrystallization of seedlings extracted in the homeopathic preparations was performed on circular glass plates. Resulting biocrystallograms were analysed by digital textural image analysis. Texture analysis variables analysed yielded significant results between the control and some of the homeopathic treatments, as well as between a number of the different treatments. As the texture of the biocrystallograms of homeopathically treated cress exhibited effects depending on the remedies, the CuCl2-biocrystallization method will be further applied to a larger study investigating specific effects of different homeopathic preparations.

scholarly journals Germination and Protocorm Formation of Ophrys Sphegodes Mill. – In Vitro Protocol for a Rare Orchid Species

2018 ◽  
Vol 67 (3-4) ◽  
pp. 196-201
Author(s):  
Jovana Dulić ◽  
Mirjana Ljubojević ◽  
Ines Prlainović ◽  
Goran Barać ◽  
Tijana Narandžić ◽  
...  
Keyword(s):  
Seed Germination ◽  
Germination Rate ◽  
Casein Hydrolysate ◽  
Habitat Destruction ◽  
Ex Situ ◽  
Organic Additives ◽  
Orchid Species ◽  
Short Period ◽  
Protocorm Formation

Summary Ophrys sphegodes Mill. is a wild orchid species which is threatened and protected due to its pollination biology, small seed and habitat destruction. The aim of this study was to establish asymbiotic germination protocol for the purpose of ex situ conservation. Two basal media Knudson C (KC) and Malmgren (MM), supplemented with organic additives (peptone (PE), L-glutamin (A)e, folic acid, casein hydrolysate (CA)) added separately and control media KC--C and MM--C were used in the present research. All the nutrition media contained 2% sucrose, 7% agar and 1% activated carbon, while their pH was adjusted to 5.8 ± 0.02 before autoclaving at 121 ° C for 20 minutes. The seeds were examined under two illumination conditions, 0/24 light/dark (L/D) and 16/8 L/D. The presented results indicate a huge influence of illumination and nutrition media on the seed germination and protocorm formation. The seed germination was overall significantly more successful in dark conditions (0/24 L/D) than with lighting (16/8 L/D). Protocorm, rhizoids and shoot formation were achieved only on the seeds cultured on MM medium, while the KC medium caused only swelling of the embryo. Organic additives had positive influence on the germination rate. According to the obtained results, the best germination rate and seedling development were achieved on MM-PE media, cultured in dark. The presented procedure accelerates the germination period and can provide a large number of plants in a relatively short period of time so it can be used for conservation programs and mass production protocol.

scholarly journals The Screening of Potassium- and Phosphate-Solubilizing Actinobacteria and the Assessment of Their Ability to Promote Wheat Growth Parameters

2021 ◽  
Vol 9 (3) ◽  
pp. 470
Author(s):  
Kenza Boubekri ◽  
Abdoulaye Soumare ◽  
Ilham Mardad ◽  
Karim Lyamlouli ◽  
Mohamed Hafidi ◽  
...  
Keyword(s):  
Soil Fertility ◽  
Chemical Weathering ◽  
Growth Parameters ◽  
Germination Rate ◽  
Dry Weight ◽  
Shoot Dry Weight ◽  
Streptomyces Griseorubens ◽  
Phosphate Solubilizing ◽  
Nocardiopsis Alba

Soil fertility and plant nutrition require an adequate management of essential macronutrients such as potassium (K) and phosphorus (P), which are mandatory for plant development. Bioleaching of K and P bearing minerals improves their chemical weathering and increases the performance of the biofertilization strategies. In this study, in vitro and greenhouse experiments were carried out to investigate P and K solubilization traits of nine Actinobacteria (P13, P14, P15, P16, P17, P18, BC3, BC10, and BC11) under fertilization with rock phosphate (RP). K and P solubilization were evaluated on Alexandrov and NBRIP media containing mica and six RP samples, respectively. The actinobacterial strains were able to solubilize K in Alexandrov medium supplemented with RP. However, when soluble P was used instead of RP, only four strains of Actinobacteria (Streptomyces alboviridis P18–Streptomyces griseorubens BC3–Streptomyces griseorubens BC10 and Nocardiopsis alba BC11) solubilized K. The solubilization values of K ranged from 2.6 to 41.45 mg/L while those of P varied from 0.1 to 32 mg/L. Moreover, all strains were able to produce IAA, siderophore, HCN, and ammonia and significantly improved the germination rate and the vigor index of wheat. The pot experiments revealed that four strains (Streptomyces alboviridis P18, Streptomyces griseorubens BC3, Streptomyces griseorubens BC10, and Nocardiopsis alba BC11) significantly improved the growth parameters of wheat, namely root length (1.75–23.84%), root volume (41.57–71.46%), root dry weight (46.89–162.41%), shoot length (8.92–23.56%), and shoot dry weight (2.56–65.68%) compared to the uninoculated control. These findings showed that Streptomyces griseorubens BC10 and Nocardiopsis alba BC11 are promising candidates for the implementation of efficient biofertilization strategies to improve soil fertility and plant yield under rock P and rock K fertilization.

Interaction of Tomatoes (Solanum lycopersicum L.) Transformed by rapA1 Gene with Pseudomonas sp. 102 Bacteria Resistant to High Cadmium Concentrations as a Basis for Effective Symbiotic Phytoremediation System

2019 ◽  
pp. 38-48
Author(s):  
L.R. Khakimova ◽  
A.M. Lavina ◽  
L.R. Karimova ◽  
V.V. Fedyaev ◽  
An.Kh. Baymiev ◽  
...  
Keyword(s):  
Growth Parameters ◽  
Basic Research ◽  
Toxic Effects ◽  
Research Centre ◽  
Pseudomonas Sp ◽  
Tomato Plants ◽  
Solanum Lycopersicum L ◽  
Plant Growth Promoting ◽  
High Concentrations ◽  
The Government

A Pseudomonas sp. 102 strain, which is highly resistant to toxic effects of cadmium and has plant growth-promoting activity, can significantly increase growth parameters and biomass of tomato plants, including those observed under toxic effects of cadmium. The greatest positive effect was observed in plants transformed with the bacterial adhesin gene rapA1, the product of which is important for colonization of plant roots by bacteria. It was also shown that shoots of transgenic tomato plants accumulated the greatest amount of cadmium during inoculation with Pseudomonas sp. 102. The ability to extract high concentrations of cadmium and accumulate a large biomass under stress opens up prospects for the further use of associative interactions between tomato and Pseudomonas for phytoremediation. phytoremediation, cadmium, tomato, Pseudomonas, inoculation, agglutinins, This study was carried out using the equipment of the Biomika Centre for Collective Use of the Institute of Biochemistry and Genetics (Ufa Federal Research Centre, Russian Academy of Sciences) as part of the government task (project no. AAAA-A16-1160203500284). This study was supported by the Russian Foundation for Basic Research (project nos. 18-34-20004 and 18-34-00033) and 18-344-0033 mol_a_ved and 34-00033 mol_a).

scholarly journals In vitro Osmotic Stress Memory in Chrisanthemum hybridum: Structural and Physiological Responses

2017 ◽  
Vol 27 (2) ◽  
pp. 161-169 ◽  
Author(s):  
Lidiia Samarina ◽  
Valentina Malyarovskaya ◽  
Yulija Abilfazova ◽  
Natalia Platonova ◽  
Kristina Klemeshova ◽  
...  
Keyword(s):  
Osmotic Stress ◽  
Relative Water Content ◽  
Soluble Protein ◽  
Growth Parameters ◽  
Physiological Responses ◽  
Proline Content ◽  
Stress Exposure ◽  
Stress Memory ◽  
Relative Water

Structural and physiological responses of chrysanthemum to repeated osmotic stress were studied. Plants were cultured for 2 weeks (for each stress1 and stress 2) on half MS supplemented with mannitol 100 mM (Treatment I) and 200 mM (Treatment II). First stress inhibited growth parameters stronger than second stress in treatment I. In treatment II both stress events strongly inhibited growth parameters of micro‐shoots. Proline content exceeded control 6 ‐ 8 times after 1st stress, and 2 ‐ 5 times after the 2nd stress in treatments I and II, respectively. Soluble protein was accumulated in leaves during both stress exposures, and 2 ‐ 2.5 times exceeded control after the 2nd stress. Relative water content in both treatments increased after the 2nd stress exposure. In treatment II chlorophyll а and carotenoids contents were 8.78 and 4.62 mg/g comparing to control (4.21 and 2.25 mg/g, respectively) after the 1st stress. But after the 2nd stress there was no difference with control.Plant Tissue Cult. & Biotech. 27(2): 161-169, 2017 (December)

scholarly journals Machine learning-based classification of mitochondrial morphology in primary neurons and brain

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Garrett M. Fogo ◽  
Anthony R. Anzell ◽  
Kathleen J. Maheras ◽  
Sarita Raghunayakula ◽  
Joseph M. Wider ◽  
...  
Keyword(s):  
Image Analysis ◽  
Cortical Neurons ◽  
Mitochondrial Dynamics ◽  
Automated Image Analysis ◽  
Mitochondrial Morphology ◽  
Analysis Pipeline ◽  
Genetic Ablation ◽  
Block Face

AbstractThe mitochondrial network continually undergoes events of fission and fusion. Under physiologic conditions, the network is in equilibrium and is characterized by the presence of both elongated and punctate mitochondria. However, this balanced, homeostatic mitochondrial profile can change morphologic distribution in response to various stressors. Therefore, it is imperative to develop a method that robustly measures mitochondrial morphology with high accuracy. Here, we developed a semi-automated image analysis pipeline for the quantitation of mitochondrial morphology for both in vitro and in vivo applications. The image analysis pipeline was generated and validated utilizing images of primary cortical neurons from transgenic mice, allowing genetic ablation of key components of mitochondrial dynamics. This analysis pipeline was further extended to evaluate mitochondrial morphology in vivo through immunolabeling of brain sections as well as serial block-face scanning electron microscopy. These data demonstrate a highly specific and sensitive method that accurately classifies distinct physiological and pathological mitochondrial morphologies. Furthermore, this workflow employs the use of readily available, free open-source software designed for high throughput image processing, segmentation, and analysis that is customizable to various biological models.

Interaction of Neutrophils with Endothelial Cells, Fibroblasts and Their Extracellular Matrices: Microscopic and Computerised Analysis

1988 ◽  
Vol 16 (1) ◽  
pp. 48-53
Author(s):  
Marina Ziche ◽  
Lucia Morbidelli ◽  
Annalisa Rubino ◽  
Piero Dolara ◽  
Stefano Bianchi ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Image Analysis ◽  
Vascular Endothelial Cells ◽  
Extracellular Matrices ◽  
Polymorphonuclear Neutrophil ◽  
Initial Event ◽  
Computerised Image Analysis ◽  
Capillary Endothelial Cells ◽  
Vitro Model

Polymorphonuclear neutrophil (PMN) interaction with vascular endothelial cells is the initial event in the migration of neutrophils through blood vessel walls before reaching inflammation sites in tissues. The interaction between fibroblasts and endothelial cells and their extracellular matrices might be modulated by the activation of neutrophils that occurs at inflammatory reaction sites. We have used an in vitro model to study PMN function, measuring the adhesion of human PMNs to capillary endothelial cells and fibroblasts grown in culture and to their extracellular matrices. The interaction was measured in basal conditions and in the presence of the chemotactic effector, formyl-methionyl-leucyl-phenylalanine (FMLP at the concentration of 10 7M). Adhesion was expressed by the number of adherent PMNs/mm2 on a histological specimen. Moreover, we have adapted a program for image analysis to quantify neutrophil adhesion. Three times more PMNs adhered to matrices than to monolayers, and adherence could be increased by the presence of 10-7M FMLP, except in the case of fibroblast monolayers. We found a good correlation between microscopic observation and computerised image analysis measuring PMN adhesiveness to extracellular matrices.

scholarly journals Academic collaborative models fostering the translation of physiological in vitro systems from basic research into drug discovery

2021 ◽  
Author(s):  
Alessandra Silvestri ◽  
Francisca Vicente ◽  
María J. Vicent ◽  
Bahne Stechmann ◽  
Wolfgang Fecke
Keyword(s):  
Drug Discovery ◽  
Basic Research ◽  
Collaborative Models ◽  
In Vitro Systems

Neurotoxicity of Seven MEIC Chemicals Evaluated in Organotypic Cultures of Chick Embryonic Dorsal Root Ganglia

1997 ◽  
Vol 25 (3) ◽  
pp. 303-309
Author(s):  
Václav Mandys ◽  
Katerina Jirsová ◽  
Jirí Vrana
Keyword(s):  
Toxic Effect ◽  
Neurite Outgrowth ◽  
Dorsal Root Ganglia ◽  
Dorsal Root ◽  
Growth Parameters ◽  
In Vitro Cytotoxicity ◽  
Organotypic Cultures ◽  
Response Curves ◽  
Agar Culture

The neurotoxic effects of seven selected Multicenter Evaluation of In Vitro Cytotoxicity programme chemicals (methanol, ethanol, isopropanol, sodium chloride, potassium chloride, iron [II] sulphate and chloroform) were evaluated in organotypic cultures of chick embryonic dorsal root ganglia (DRG), maintained in a soft agar culture medium. Two growth parameters of neurite outgrowth from the ganglia — the mean radial length of neurites and the area of neurite outgrowth — were used to evaluate the toxicities of the chemicals. Dose-dependent decreases of both parameters were observed in all experiments. IC50 values (the concentration causing 50% inhibition of growth) were calculated from the dose-response curves established at three time-points during culture, i.e. 24, 48 and 72 hours. The lowest toxic effect was observed in cultures exposed to methanol (the IC50 ranging from 580mM to 1020mM). The highest toxic effect was observed in cultures exposed to iron (II) sulphate (the IC50 ranging from 1.2mM to 1.7mM). The results of other recent experiments suggest that organotypic cultures of DRG can be used during in vitro studies on target organ toxicity within the peripheral nervous system. Moreover, these cultures preserve the internal organisation of the tissue, maintain intercellular contacts, and thus reflect the in vitro situation, more precisely than other cell cultures.

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