Effects of Homeopathic Preparations of Mercurius corrosivus on the Growth Rate of Moderately Mercury-Stressed Duckweed Lemna gibba L.

BACKGROUND: A bioassay with severely mercury-stressed duckweed (Lemna gibba L.) had revealed growth-inhibiting effects of homeopathically potentised mercury(II) chloride (Mercurius corrosivus, Merc-c.). We hypothesised that effects of potentised preparations are dependent on the stress level of the organisms used in the bioassay. The aim of the present investigation was to examine the response of duckweed to potentised Merc-c. at a lower stress level. METHODS: Duckweed was moderately stressed with 2.5 mg/L mercury(II) chloride for 48 hours. Afterwards plants grew in either Merc-c. (seven different potency levels, 24x-30x) or water controls (unsuccussed or succussed water) for 7 days. Growth rates of the frond (leaf) area were determined using a computerised image-analysis system for day 0-3 and 3-7. Three independent experiments with potentised Merc-c. and three systematic negative control experiments were performed. All experiments were randomised and blinded. RESULTS: Unsuccussed and succussed water did not significantly differ in their effects on duckweed growth rate. The systematic negative control experiments did not yield any significant effects, thus providing evidence for the stability of the experimental system. Data from the two control groups and the seven treatment groups (Merc-c. 24x-30x) were each pooled to increase statistical power. Duckweed growth rates for day 3-7 were enhanced (p < 0.05) after application of Merc-c. compared with the controls. Growth rates for day 0-3 were not influenced by the homeopathic preparations. CONCLUSIONS: Moderately mercury-stressed Lemna gibba L. yielded evidence of growth-enhancing specific effects of Merc-c. 24x-30x in the second observation period (day 3-7). This observation is complementary to previous experiments with severely mercury-stressed duckweed, in which a decrease in growth was observed in the first observation period (day 0-3). We hypothesise that the differing results are associated with the level of stress intensity (moderate vs. severe).

M1 homeopathic complex trigger effective responses against Leishmania (L) amazonensis in vivo and in vitro.

Leishmaniasis is a term referring to a range of clinical conditions caused by protozoan parasites of the genus Leishmania, Trypanosomatidae family, Kinetoplastida order that is transmitted by the bite of certain species of mosquitoes Phlebotominae subfamily. These parasites infect hosts wild and domestic mammals, considered as natural reservoirs and can also infect humans. Leishmania are obligate intramacrophage protozoa that have exclusively intracellular life style. This suggests that the amastigotes possess mechanisms to avoid killing by host cells. Cutaneous leishmaniasis, the most common form of the disease, causes ulcers on exposed parts of the body, leading to disfigurement, permanent scars, and stigma and in some cases disability. Many studies concluded that the cytokines profile and immune system of host have fundamental role in humans and animals natural self-healing. Conventional treatments are far from ideals and the search for new therapeutic alternatives is considered a strategic priority line of research by the World Health Organization. A promising approach in the field of basic research in homeopathy is the treatment of experimental infections with homeopathic drugs prepared from natural substances associations highly diluted, which comprise a combination of several different compounds considered as useful for a symptom or disease. Therefore, this study aimed to evaluate the effect of M1, a complex homeopathic product, in macrophage-Leishmania interaction in vitro and in vivo. It was used RAW cells lineage and BALB/c mice as a host for the promastigotes of L. amazonensis (WHOM/BR/75/Josefa). Several biochemical and morphological parameters were determined. Together, the harmonic results obtained in this study indicate that, in general, the highly diluted products trigger rapid and effective responses by living organisms, cells and mice, against Leishmania, by altering cytokines profile, by NO increasing (p<0.05), by decreasing parasitic load (p<0.001), and modifying classical maturation and biogenesis of parasitophorous vacuoles (p<0.001). M1 complex decreased endocytic index (p<0.001), and the % of infected macrophages (p<0.05), preventing the development of lesions (p<0.05) caused by L. amazonensis by increasing Th1 response (p<0.05). Therefore the M1complex can be a good candidate for a complementary therapy to conventional treatments, since all the parameters observed in vitro and in vivo improved. It could be an interesting clinical tool in association to a classical anti-parasitic treatment, maybe resulting in better quality of life to the patients, with less toxicity.

[Biological assay in quality control of animal medicines].

Animal medicine is a unique part of traditional Chinese medicine. They have strong effects, but their effective compounds are not entirely known. The efficiency and safety of animal medicines can't be effectively controlled by current quality assurance system and evaluation method, which has deeply influenced the development of animal medicines. Biological assay does not focus on efficacy of single component, but directly reflects the pharmacodynamics and safety of animal medicines by biological effect. With the development of biotechnology, many new technologies have emerged, such as biochip and high content analysis. Based on the related targets, pathways and key biochemical factors, the field of biological assay has been expanded. With advantages of pharmacology andoverall controllability, as well as the characteristics of in line with the quality control of Chinese Medicine, biological assay will become one of the important development directionsfor quality standardization of animal medicines.

The use of plant-based bioassays in homeopathic basic research.

OBJECTIVES: The objective was to evaluate homeopathic basic research studies that use plant-based bioassays. With this in view, a compilation was made of the findings of three systematic literature reviews covering plant-based bioassays in the three fields of healthy, abiotically, or biotically stressed plants. This compilation focused on investigations using advanced experimental methods and detailed descriptions, also with the aim of supporting the design of future experiments. METHODS: Publications included had to report on studies into the effects of homeopathic preparations on whole plants, seeds, plant parts and cells. Outcomes had to be measured by established procedures and statistically evaluated. A Manuscript Information Score (MIS) was applied using predefined criteria to identify publications with sufficient information for adequate interpretation (MIS ≥ 5). Additional evaluation focused on the use of adequate controls to investigate specific effects of homeopathic preparations, and on the use of systematic negative control (SNC) experiments to ensure the stability of the bioassay. Only a fraction of the studies reported here were performed with 'ultra high' dilutions, whereas other studies were performed with moderate or high dilutions. RESULTS: A total of 157 publications were identified, describing a total of 167 experimental studies. 84 studies included statistics and 48 had a MIS ≥ 5, thus allowing adequate interpretation. 29 studies had adequate controls to identify specific effects of homeopathic preparations, and reported significant effects of decimal and centesimal homeopathic potencies, including dilution levels beyond Avogadro's number. 10 studies reported use of SNC experiments, yielding evidence for the stability of the experimental set-up. CONCLUSION: Plant models appear to be a useful approach for investigating basic research questions relating to homeopathic preparations, but more independent replication trials are needed in order to verify the results found in single experiments. Adequate controls and SNC experiments should be implemented on a routine basis to exclude false-positive results.

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

BACKGROUND: Reproducibility of basic research investigations in homeopathy is challenging. This study investigated if formerly observed effects of homeopathically potentised gibberellic acid (GA3) on growth of duckweed (Lemna gibba L.) were reproducible. METHODS: Duckweed was grown in potencies (14x-30x) of GA3 and one time succussed and unsuccussed water controls. Outcome parameter area-related growth rate was determined by a computerised image analysis system. Three series including five independent blinded and randomised potency experiments (PE) each were carried out. System stability was controlled by three series of five systematic negative control (SNC) experiments. Gibbosity (a specific growth state of L. gibba) was investigated as possibly essential factor for reactivity of L. gibba towards potentised GA3 in one series of potency and SNC experiments, respectively. RESULTS: Only in the third series with gibbous L. gibba L. we observed a significant effect (p = 0.009, F-test) of the homeopathic treatment. However, growth rate increased in contrast to the former study, and most biologically active potency levels differed. Variability in PE was lower than in SNC experiments. The stability of the experimental system was verified by the SNC experiments. CONCLUSIONS: Gibbosity seems to be a necessary condition for reactivity of L. gibba to potentised GA3. Further still unknown conditions seem to govern effect direction and the pattern of active and inactive potency levels. When designing new reproducibility studies, the physiological state of the test organism must be considered. Variability might be an interesting parameter to investigate effects of homeopathic remedies in basic research.

[Investigation on pattern and methods of quality control for Chinese materia medica based on dao-di herbs and bioassay - bioassay for Coptis chinensis].

Establishment of bioassay methods is the technical issues to be faced with in the bioassay of Chinese materia medica. Taking the bioassay of Coptis chinensis Franch. as an example, the establishment process and application of the bioassay methods (including bio-potency and bio-activity fingerprint) were explained from the aspects of methodology, principle of selection, experimental design, method confirmation and data analysis. The common technologies were extracted and formed with the above aspects, so as to provide technical support for constructing pattern and method of the quality control for Chinese materia medica based on the dao-di herbs and bioassay.

Effects of homeopathic arsenicum album, nosode, and gibberellic acid preparations on the growth rate of arsenic-impaired duckweed (Lemna gibba L.).

This study evaluated the effects of homeopathically potentized Arsenicum album, nosode, and gibberellic acid in a bioassay with arsenic-stressed duckweed (Lemna gibba L.). The test substances were applied in nine potency levels (17x, 18x, 21x-24x, 28x, 30x, 33x) and compared with controls (unsuccussed and succussed water) regarding their influence on the plant's growth rate. Duckweed was stressed with arsenic(V) for 48 h. Afterwards, plants grew in either potentized substances or water controls for 6 days. Growth rates of frond (leaf) area and frond number were determined with a computerized image analysis system for different time intervals (days 0-2, 2-6, 0-6). Five independent experiments were evaluated for each test substance. Additionally, five water control experiments were analyzed to investigate the stability of the experimental setup (systematic negative control experiments). All experiments were randomized and blinded. The test system exhibited a low coefficient of variation (approximately equal to 1%). Unsuccussed and succussed water did not result in any significant differences in duckweed growth rate. Data from the control and treatment groups were pooled to increase statistical power. Growth rates for days 0-2 were not influenced by any homeopathic preparation. Growth rates for days 2-6 increased after application of potentized Arsenicum album regarding both frond area (p < 0.001) and frond number (p < 0.001), and by application of potentized nosode (frond area growth rate only, p < 0.01). Potencies of gibberellic acid did not influence duckweed growth rate. The systematic negative control experiments did not yield any significant effects. Thus, false-positive results can be excluded with high certainty. To conclude, the test system with L. gibba impaired by arsenic(V) was stable and reliable. It yielded evidence for specific effects of homeopathic Arsenicum album preparations and it will provide a valuable tool for future experiments that aim at revealing the mode of action of homeopathic preparations. It may also be useful to investigate the influence of external factors (e.g., heat, electromagnetic radiation) on the effects of homeopathic preparations.

Empirical investigation of preparations produced according to the European Pharmacopoeia monograph 1038.

According to the European Pharmacopoeia monograph 1038 (Praeparationes homoeopathicae), homeopathic preparations are produced by successive dilution and succussion steps. Dilution levels beyond Avogadro's limit, however, render specific effects implausible according to standard scientific knowledge. Accordingly, we were interested in a critical empirical investigation of preparations produced according to this monograph. Within a precursor study we developed a bioassay based on a fingerprint metabolomic analysis of Lepidium sativum seeds germinated in vitro in either homeopathic preparations or controls in a blinded and randomized assignment. Results of the precursor study were not consistent with the hypothesis that the effects of a Stannum metallicum 30x preparation are identical to placebo. In the present study we investigated the reproducibility of these effects after scrutinizing and optimizing experimental procedures. Ten independent experiments were performed in a blinded and randomized assignment in two independent laboratories. Additionally, 10 systematic negative water control experiments were performed in both laboratories to critically assess the stability of the experimental set-up. The effects of the Stannum metallicum 30x treatment were reproduced. The systematic negative control experiments did not yield false-positive results, indicating a stable experimental set-up. We thus repeatedly observed biological effects conflicting with the assumption that Stannum metallicum 30x is identical to placebo. We therefore wish to discuss whether these findings are to be considered a scientific anomaly or whether they might stimulate further investigations to clarify whether application of the European Pharmacopoeia monograph 1038 may result in pharmaceutical preparations with specific effects.

Development of a whole plant bioassay to test effects of potentized calcium carbonate in pillule formulation.

OBJECTIVES: From a pharmaceutical point of view, we see a need to develop stable preclinical test systems to identify and investigate effects of potentized remedies as used in Anthroposophic Medicine and Homeopathy. We evaluated a plant bioassay regarding its capacity to distinguish homeopathic remedies from placebo, applied as sucrose pillules. METHODS: Pea seed (Pisum sativum L) was soaked for 24 hours in water with dissolved homeopathic or placebo pillules, or in water only. Shoot length was measured 14 days after planting and treatment groups were compared by analysis of variance (ANOVA). The stability of the system was validated by systematic negative control experiments. RESULTS: The system is suitable to test a common application form - sucrose pillules - of a potentized preparation without influence of the pharmaceutical carrier substance. A screening of 13 potentized preparations revealed Calcium carbonicum to affect pea shoot growth (p < 0.05). Three independent series of main experiments were performed with potentized Calcium carbonicum to assess reproducibility. Meta-analysis of all data revealed significant effects of Calcium carbonicum 12c and 30c on pea shoot growth (p < 0.05), which were however dependent on the date of experiment and/or the experimental series. CONCLUSIONS: Potentized Calcium carbonicum, applied as sucrose pillules, influenced pea shoot growth in the assay investigated. However, due to the small effect size and due to the modulation of the effects by still unknown external factors, further optimization of this bioassay is necessary to be used in pharmaceutical quality control or in investigating the biological or pharmaceutical mode of action of potentized preparations.

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

OBJECTIVES: A bioassay with duckweed (Lemna gibba L.) was used to study the effects of homeopathic potencies on the plant'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<0.05, analysis of variance F-test) 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.

Production and measurement of exophthalmos-producing factor in guinea pigs.

Although the extent of proptosis in exophthalmic Graves' disease has been measured directly and shown to correlate with serum content of a bioassayable exophthalmus-producing factor (EPS;1), a comparable relationship in an experimental model has not been reported. Progressive exophthalmos, measured from photographs and expressed as a ratio of intercorneal distance to intersupraorbital ridge distance, was produced in male guinea pigs when thyroid status was altered either by surgical thyroidectomy supplemented with 131-I treatment or by the administration of 6-propyl-2 thiouracil (0.1% in chow). In both groups, at time of sacrifice, serum content of EPF estimated by a modified goldfish bioassay using a known exophthalmogeric TSH preparation (Ambinon, Organon-Oss) as standard was positively correlated (r equals 0.804) with the terminal degree of exophthalmos. Daily replacement therapy with T4 (15mug/kg body wt) failed to alter significantly the exophthalmos which developed, even when replacement was initiated prior to the alterations of thyroid gland function; this observation tends to eliminate thyroid hormone deficiency per se as the causal event in exophthalmos. T4 treatment did, however, reverse or prevent the rises in serum TSH levels (McKenzie bioassay) thus dissociating TSH activity from EPF activity in the guinea pig. Treatment of guinea pigs with synthetic TRH (0.5, 1.0 OR 10 mug/kg body wt) for 21 days failed to produce demonstrable exophthalmos or assayable EPF levels although plasma TSH was significantly elevated.

An integrated approach to the pharmacological evaluation of traditional materia medica.

A triple-tiered working model is projected for the pharmacological evaluation of crude drugs prescribed in traditional medical practices. Of the proposed component tests and procedures, the majority have had their usefulness already validated in natural product pharmacological research. First-level investigations (herbalist consultations/professional attestations, hippocratic screening and literature surveys) provide first-hand pharmacological information about traditional materia medica. Second-level investigation comprising bioassay-modelled tests (brine shrimp toxicity test, opiate receptor-binding studies, fertilized sea urchin egg test, platelet adenyl cyclase/phosphodiesterase assays, hexobarbital-induced sleeping time test, antimicrobial assays, isolated guinea pig ileum test and pharmacodynamic screening) are designed to extend the mode(s) of drug action suggested by the first-level procedures. Based on data from Level 1 and Level 2 findings, third-level experiments are specific tests tailored to confirm the pharmacodynamic/kinetic properties and clinical efficacy of traditional drugs. Level 2 and Level 3 tests may be combined in monitoring fractionations and subsequent isolation of unique crude drug constituents with potential application in conventional therapeutics.

[A hypotensive analog of platelet-activating factor (PAF) in pheretima].

Experiments showed that intravenous administration of pheretima decoction elicited pronounced decrease of hypotension in rats. Pretreatment with CV6209, a specific PAF antagonist, significantly inhibited the hypotensive activity of pheretima. Furthermore, an analog of PAF was separated from the total lipids of pheretima and its content was measured to be around 90-130 ng/g.

Development of a test system for homeopathic preparations using impaired duckweed (Lemna gibba L.).

OBJECTIVES: A bioassay with arsenic-stressed duckweed (Lemna gibba L.) was developed to study potentially regulative effects of homeopathic preparations. We compared potentized substances (nine different potency levels between 17 x and 33 x ) with two controls (unsuccussed and succussed water) regarding their influence on number- and area-related growth rate and color of fronds (leaves). Screening included 11 potentized substances: Arsenicum album, gibberellic acid, nosode, arsenic(V), phosphorus, Conchae, Acidum picrinicum, Argentum nitricum, Crotalus horridus, Hepar sulfuris, and Mercurius vivus naturalis. DESIGN: Duckweed was stressed with arsenic(V) for 48 hours. Afterwards, plants grew in either potentized substances or water controls for 6 days. Growth rate and color of fronds were determined with a computerized image analysis system for different time intervals (days 0-2, 2-6, 0-6). A systematic negative control experiment with unsuccussed water was used to investigate the stability of the bioassay. All experiments were randomized and blinded. RESULTS: Arsenicum album and nosode potencies increased frond number-related growth rate compared to controls (succussed water controls or pooled water controls [succussed and unsuccussed], p < 0.05, t test). Regarding color classification, no effects were observed. CONCLUSIONS: The experimental setup with L. gibba stressed by arsenic(V) provides a valuable tool to investigate regulative effects of potentized substances. In order to verify the effects of Arsenicum album and nosode potencies, further independent replication experiments are necessary.

Homeopathic drug standardization through biological evaluations: An untrodden avenue

There is a dearth of chemico-analytical or instrumental methods for standardization and quality control of higher dilutions of homeopathic drugs. Aim: This review highlights the challenges in standardization of anti-inflammatory homeopathic drugs and suggests a battery of biological assays for their standardization. Methods: We retrieved a total 57 scientific reports from the experimental studies and scientific reviews published between January 1999 and June 2014 related to anti-inflammatory homeopathic drugs and their high dilutions. These comprised of 18 reports on preclinical evaluation, 15 on source materials, 9 on isolated constituents and 15 studies on in-vitro experiments. Few recent citations which supported the initial studies were added later during the compilation of the manuscript. Conclusion: Standardization and quality control of homeopathic mother tinctures and high dilutions warrants an urgent attention. As biological activities are observed to be attributed to the high dilutions which are practically devoid of active ingredients, their standardization may be done through the suggested battery of biological investigations. It is suggested that the current methods of standardization of homeopathic drugs need to be upgraded to include sensitive, reproducible and relevant biological assays so that the end users are assured of the quality, efficacy, and safety of homeopathic dilutions...

Homeopathic drug standardization through biological evaluations: An untrodden avenue

Background: There is a dearth of chemico-analytical or instrumental methods for standardization and quality control of higher dilutions of homeopathic drugs. Aim: This review highlights the challenges in standardization of anti-inflammatory homeopathic drugs and suggests a battery of biological assays for their standardization. Methods: We retrieved a total 57 scientific reports from the experimental studies and scientific reviews published between January 1999 and June 2014 related to anti-inflammatory homeopathic drugs and their high dilutions. These comprised of 18 reports on preclinical evaluation, 15 on source materials, 9 on isolated constituents and 15 studies on in-vitro experiments. Few recent citations which supported the initial studies were added later during the compilation of the manuscript. Conclusion: Standardization and quality control of homeopathic mother tinctures and high dilutions warrants an urgent attention. As biological activities are observed to be attributed to the high dilutions which are practically devoid of active ingredients, their standardization may be done through the suggested battery of biological investigations. It is suggested that the current methods of standardization of homeopathic drugs need to be upgraded to include sensitive, reproducible and relevant biological assays so that the end users are assured of the quality, efficacy, and safety of homeopathic dilutions. (AU)

Lowland amphibians - recalculation of data on effects of diluted thyroxine / Anfíbios de terras baixas - recálculo dos dados sobre os efeitos de tiroxina diluída / Anfibios de tierras bajas - re-cálculo de datos sobre los efectos de tiroxina diluida

Our previous paper described methodological problems and a generally acceptable pooling method for metamorphosis experiments and application of that method to the results of multicentre experiments performed over the course of two decades (1990 - 2010) on highland amphibians (Rana temporaria) treated with a homeopathically prepared high dilution of thyroxine (?30x?). Differences between treatment groups thus calculated were in line with those obtained with other pooling methods: Thyroxine 30x does slow down metamorphosis in highland amphibians. This follow up paper provides a broader background on metamorphosis physiology and describes application of the pooling method to experiments with Rana temporaria from lowland biotopes both with a moderate dilution of thyroxine (?8x?) and with 30x. Analogously prepared water was used for control (water 8x or 30x). Development was, again as above, monitored by documenting the number of animals that had entered the 4-legged stage. Experiments were carried out between 1990 and 2000 by different researchers independently and in blind. As it is well known, metamorphosis can be speeded up by thyroxine 10-8 mol/l; interestingly, thyroxine 8x may produce a reverse, i.e. inhibiting effect (p < 0.01). In contrast to the inhibiting effect of thyroxine 30x on highland larvae (see above), 2-legged lowland larvae did not react to thyroxine 30x (p > 0.05). However, an inhibiting effect on lowland larvae was found when animals were treated from the spawn stage on (p < 0.01).

Nosso artículo anterior descreve problemas metodológicos e um método geralmente aceitável de agrupamento em experimentos sobre metamorfose e a aplicação deste método aos resultados de experimentos multicêntricos realizados no curso de duas décadas (1990 ? 2010) em anfíbios de terras altas (Rana temporaria) tratados com uma alta diluição de tiroxina preparada homeopaticamente (?30x?). As diferenças calculadas entre os grupos de tratamento concordam com as obtidas através de outros métodos de agrupamento: Tiroxina 30x de fato faz mais lenta a metamorfose nos anfíbios de terras altas. Este artigo posterior oferece um panorama mais amplo sobre a fisiologia da metamorfose e descreve a aplicação do método de agrupamento a experimentos com Rana temporaria de biótopos de terras baixas com diluição moderada (?8x?) e 30x de tiroxina. Foi utilizado como controle água preparada analogamente (água 8x e 30x). Novamente, o desenvolvimento foi monitorado documentando o número de animais que entraram no estágio de 4 patas. Os experimentos foram realizados entre 1990 e 2000 por diversos pesquisadores independentemente e em cego. Como é sabido, a metamorfose pode ser acelerada por tiroxina 10-8 mol/l; de maneira interessante, tiroxina 8x pode produzir o efeito inverso, ou seja, inibidor (p < 0.01). Contrariamente ao efeito inibidor de tiroxina 30x nas larvas de terras altas (vide acima), as larvas de 2 patas de terras baixas não responderam a tiroxina 30x (p > 0.05). No entanto, foi achado um efeito inibidor sobre as larvas de terras baixas quando os animais foram tratados a partir da desova (p < 0.01).

Nuestro artículo anterior describe problemas metodológicos y un método aceptable en general de agrupamiento en experimentos sobre metamorfosis y la aplicación de este método a los resultados de experimentos multicéntricos realizados durante dos décadas (1990 ? 2010) en anfibios de tierras altas (Rana temporaria) tratados con una alta dilución de tiroxina preparada homeopáticamente (?30x?). Las diferencias calculadas entre los grupos de tratamiento están de acuerdo con las obtenidas mediante otros métodos de agrupamiento: Tiroxina 30x fehacientemente enlentece la metamorfosis en anfibios de tierras altas. Este artículo posterior ofrece un panorama más amplio de la fisiología de la metamorfosis y describe la aplicación del método de agrupamiento en experimentos con Rana temporaria de biótopos de tierras bajas con dilución moderada (?8x?) y 30x de tiroxina. Se utilizó como control agua preparada análogamente (agua 8x y 30x). Nuevamente, el desarrollo fue monitoreado documentando el número de animales que entraron en el estadio de 4 patas. Os experimentos fueron realizados entre 1990 e 2000 por diversos investigadores independientemente y en ciego. Como se sabe, la metamorfosis puede ser acelerada por tiroxina 10-8 mol/l; de modo interesante, tiroxina 8x puede producir el efecto inverso, o sea, inhibidor (p < 0.01). Contrariamente al efecto inhibidor de tiroxina 30x en larvas de tierras altas (ver antes), las larvas de 2 patas de tierras bajas no respondieron a tiroxina 30x (p > 0.05). Sin embargo, se encontró efecto inhibidor en larvas de tierras bajas cuando los animales fueron tratados desde la freza (p < 0.01).

Lowland amphibians - recalculation of data on effects of diluted thyroxine / Anfíbios de terras baixas - recálculo dos dados sobre os efeitos de tiroxina diluída / Anfibios de tierras bajas - re-cálculo de datos sobre los efectos de tiroxina diluida

Our previous paper described methodological problems and a generally acceptable pooling method for metamorphosis experiments and application of that method to the results of multicentre experiments performed over the course of two decades (1990 - 2010) on highland amphibians (Rana temporaria) treated with a homeopathically prepared high dilution of thyroxine (?30x?). Differences between treatment groups thus calculated were in line with those obtained with other pooling methods: Thyroxine 30x does slow down metamorphosis in highland amphibians. This follow up paper provides a broader background on metamorphosis physiology and describes application of the pooling method to experiments with Rana temporaria from lowland biotopes both with a moderate dilution of thyroxine (?8x?) and with 30x. Analogously prepared water was used for control (water 8x or 30x). Development was, again as above, monitored by documenting the number of animals that had entered the 4-legged stage. Experiments were carried out between 1990 and 2000 by different researchers independently and in blind. As it is well known, metamorphosis can be speeded up by thyroxine 10-8 mol/l; interestingly, thyroxine 8x may produce a reverse, i.e. inhibiting effect (p < 0.01). In contrast to the inhibiting effect of thyroxine 30x on highland larvae (see above), 2-legged lowland larvae did not react to thyroxine 30x (p > 0.05). However, an inhibiting effect on lowland larvae was found when animals were treated from the spawn stage on (p < 0.01).(AU)

Nosso artículo anterior descreve problemas metodológicos e um método geralmente aceitável de agrupamento em experimentos sobre metamorfose e a aplicação deste método aos resultados de experimentos multicêntricos realizados no curso de duas décadas (1990 ? 2010) em anfíbios de terras altas (Rana temporaria) tratados com uma alta diluição de tiroxina preparada homeopaticamente (?30x?). As diferenças calculadas entre os grupos de tratamento concordam com as obtidas através de outros métodos de agrupamento: Tiroxina 30x de fato faz mais lenta a metamorfose nos anfíbios de terras altas. Este artigo posterior oferece um panorama mais amplo sobre a fisiologia da metamorfose e descreve a aplicação do método de agrupamento a experimentos com Rana temporaria de biótopos de terras baixas com diluição moderada (?8x?) e 30x de tiroxina. Foi utilizado como controle água preparada analogamente (água 8x e 30x). Novamente, o desenvolvimento foi monitorado documentando o número de animais que entraram no estágio de 4 patas. Os experimentos foram realizados entre 1990 e 2000 por diversos pesquisadores independentemente e em cego. Como é sabido, a metamorfose pode ser acelerada por tiroxina 10-8 mol/l; de maneira interessante, tiroxina 8x pode produzir o efeito inverso, ou seja, inibidor (p < 0.01). Contrariamente ao efeito inibidor de tiroxina 30x nas larvas de terras altas (vide acima), as larvas de 2 patas de terras baixas não responderam a tiroxina 30x (p > 0.05). No entanto, foi achado um efeito inibidor sobre as larvas de terras baixas quando os animais foram tratados a partir da desova (p < 0.01).(AU)

Nuestro artículo anterior describe problemas metodológicos y un método aceptable en general de agrupamiento en experimentos sobre metamorfosis y la aplicación de este método a los resultados de experimentos multicéntricos realizados durante dos décadas (1990 ? 2010) en anfibios de tierras altas (Rana temporaria) tratados con una alta dilución de tiroxina preparada homeopáticamente (?30x?). Las diferencias calculadas entre los grupos de tratamiento están de acuerdo con las obtenidas mediante otros métodos de agrupamiento: Tiroxina 30x fehacientemente enlentece la metamorfosis en anfibios de tierras altas. Este artículo posterior ofrece un panorama más amplio de la fisiología de la metamorfosis y describe la aplicación del método de agrupamiento en experimentos con Rana temporaria de biótopos de tierras bajas con dilución moderada (?8x?) y 30x de tiroxina. Se utilizó como control agua preparada análogamente (agua 8x y 30x). Nuevamente, el desarrollo fue monitoreado documentando el número de animales que entraron en el estadio de 4 patas. Os experimentos fueron realizados entre 1990 e 2000 por diversos investigadores independientemente y en ciego. Como se sabe, la metamorfosis puede ser acelerada por tiroxina 10-8 mol/l; de modo interesante, tiroxina 8x puede producir el efecto inverso, o sea, inhibidor (p < 0.01). Contrariamente al efecto inhibidor de tiroxina 30x en larvas de tierras altas (ver antes), las larvas de 2 patas de tierras bajas no respondieron a tiroxina 30x (p > 0.05). Sin embargo, se encontró efecto inhibidor en larvas de tierras bajas cuando los animales fueron tratados desde la freza (p < 0.01).(AU)

A new approach to the memory of water.

We investigated whether water has a 'memory' for succussion compared to unsuccussed controls. The method is based on a bioassay using dinoflagellates. The duration of memory measured by this method is at least 10 min, and may be longer. The effect may be based upon solitons. A hypothesis about the mode of action of homeopathy may be experimentally investigated by this method.

Search for an endogenous ligand for the opiate receptor.

It was investigated whether there is an endogenous factor in the brain which binds to the opiate receptor in neural tissue. Extracts from rat brain were processed in different ways; fractions were assayed for ability to inhibit the receptor binding of dihydromorphine. There was no evidence for high-molecular weight substances or lipid soluble substances with such ability. On the other hand, processing of an acid water extract of brain except the cerebellum (which was negative) yielded an active fraction with receptor blocking activity. This fraction was heat stable, polyionic and had an apparent molecular weight of 1000-1 200 dalton. These and other characteristics indicate that it might well be a peptide. The factor inhibited binding to the opiate receptor in synaptic plasma membranes of rat brain and to the receptor of the guinea-pig ileum although it was less effective on the latter, particularly after long-term incubation. The interaction between the factor and dihydromorphine was reversible and apparently competitive.