The Unitary Micro-Immunotherapy Medicine Interferon-γ (4 CH) Displays Similar Immunostimulatory and Immunomodulatory Effects than Those of Biologically Active Human Interferon-γ on Various Cell Types.

As a cytokine, gamma-interferon (IFN-γ) is considered a key player in the fine-tuned orchestration of immune responses. The extreme cellular sensitivity to cytokines is attested by the fact that very few of these bioactive molecules per cell are enough to trigger cellular functions. These findings can, at least partially, explain how/why homeopathically-prepared cytokines, and especially micro-immunotherapy (MI) medicines, are able to drive cellular responses. We focused our fundamental research on a unitary MI preparation of IFN-γ, specifically employed at 4 CH, manufactured and impregnated onto sucrose-lactose pillules as all other MI medicines. We assessed the IFN-γ concentration in the medium after dilution of the IFN-γ (4 CH)-bearing pillules and we evaluated in vitro drug responses in a wide range of immune cells, and in endothelial cells. Our results showed that IFN-γ (4 CH) stimulated the proliferation, the activation and the phagocytic capabilities of primary immune cells, as well as modulated their cytokine-secretion and immunity-related markers' expression in a trend that is quite comparable with the well-recognized biological effects induced by IFN-γ. Altogether, these data provide novel and additional evidences on MI medicines, and specifically when active substances are prepared at 4 CH, thus suggesting the need for more investigations.

A review of machines and devices to potentize homeopathic medicines.

BACKGROUND: Potentization, consisting of serial dilution and succussion, is a key step in the manufacture of homeopathic medicines. Originally prescribed as a manual process, several attempts at mechanization have been published, patented and even commercialised in order to remove the human element and introduce reproducibility without drudgery. Various machines have been used over the years to prepare homeopathic medicines. Although these machines follow the same principles, i.e. energetically mixing the medicines and diluting them significantly, their mode of operation is different from each other. METHODS: This review paper surveys the main methods of preparation of homeopathic medicines. The main machines discussed are: Boericke's potentizer, Tyler Kent's instrument, John Alphonse's machine and the fluxion potentizer, which were used in the past, as well as more recent potentizers like arm-and-weight instruments, the K-Tronic potentizer and Quinn's machine. We review the construction and operating principle of each of these machines, along with their advantages and limitations. A scheme for relative performance assessment of these machines is proposed based on the parameters mechanical efficiency, physico-chemical efficiency, turbulence generation, energy dissipation, and accuracy of dilution. RESULTS: Quinn's machine and the arm-and-weight potentizer perform well for generating turbulence due to high impaction forces, while John Alphonse's machine is much more accurate in diluting the homeopathic medicines at every step. CONCLUSIONS: Both the commercial potentizers, Quinn's machine and the K-Tronic potentizer, are completely automated and therefore reduce the manual labour and variation in succussive forces during each step, which may produce uniformity in physico-chemical changes within the resulting homeopathic medicines.

Could the study of cavitation luminescence be useful in high dilution research?

Cavitation in agitated liquids has been discussed for over five decades as a phenomenon that could play a role in the appearance of structural changes in the solvent of potentised dilutions. However, its lack of specificity as well as the absence of experimental confirmation have so far confined the idea to theory. The light emission associated with cavitational bubble collapse can be used to detect and study cavitation in fluids. The phenomenon has been extensively studied when driven by ultrasound, where it is called sonoluminescence. Sonoluminescence spectra reflect extremely high temperature and pressure in the collapsing bubbles and are parameter sensitive. This article tries to examine whether, despite objections and difficulties, the detection or the study of cavitational luminescence in solutions during potentisation could be useful as a physical tool in high dilution research.

"Ultra High Dilution 1994" revisited 2015--the state of follow-up research.

BACKGROUND: The "Ultra High Dilution 1994" project was an endeavour to take stock of the findings and theories on homeopathic extreme dilutions that were under research at the time in areas of biology, biophysics, physics and medicine. The project finally materialized into an anthology assembling contributions of leading scientists in the field. Over the following two decades, it became widely quoted within the homeopathic community and also known in other research communities. The aim of the present project was to re-visit and review the 1994 studies from the perspective of 2015. METHOD: The original authors from 1994 or close laboratory colleagues were asked to contribute papers covering their research efforts and learnings in the period from 1994 up to 2015. These contributions were edited and cross-referenced, and a selection of further contributions was added. RESULTS: About a dozen contributions reported on follow-up experiments and studies, including further developments in theory. Only few of the models that had seemed promising in 1994 had not been followed up later. Most models presented in the original publication had meanwhile been submitted to intra-laboratory, multicentre or independent scrutiny. The results of the follow-up research seemed to have rewarded the efforts. Furthermore, contributions were provided on new models that had been inspired by the original ones or that may be candidates for further in-depth ultra high dilution (UHD) research. CONCLUSION: The project "Ultra High Dilution 1994 revisited 2015" is the latest output of what might be considered the "buena vista social club" of homeopathy research. However, it presents new developments and results of the older, established experimental models as well as a general survey of the state of UHD research.

Amphibians and ultra high diluted thyroxine--further experiments and re-analysis of data.

BACKGROUND: A model of thyroxine and metamorphosis of highland amphibians is frequently mentioned as an example of experiments on extremely diluted substances in discussions around 'homeopathy'. METHODS: The model was scrutinized by reanalysing the results of the initial researcher A and a second researcher B as well as of 5 external researchers C between 1990 and 2013. Rana temporaria larvae were taken from an alpine highland biotope. The test solution was thyroxine 10(-30) (T30x), tetra-iodo-thyronine sodium pentahydrate diluted with pure water in 26 steps of 1:10, being agitated after each step. Analogously prepared water (W30x) was used for control. Tadpoles were observed from the 2-legged to the 4-legged stage. Experiments were performed in different years, at different times of season, and their duration could vary. Frequencies of 4-legged animals, effect sizes and areas under the curves (AUCs) were calculated and regression analyses were performed to investigate possible correlations between year, season, duration etc. Experiments were in line with animal protection guidelines. RESULTS: The total set of data A + B + C as well as subsets A (initial researcher, N=286+293), B (second centre, 965 + 965) and C (5 external researchers, 690 + 690) showed an effect of extremely diluted agitated thyroxine reverse to that known of molecular thyroxin, i.e. test values were below control by 11.4% for A, 9.5% for B and 7.0% for C (p<0.001 for each of the subsets). The effect size (Cohen's d) was >0.8 (large) for both A and B and 0.74 (medium) for C. CONCLUSION: Although a perfect reproducibility was not obtained, this paradoxical phenomenon was generally consistent in different observations. Correlations were found between details of laboratory handling, as well as environment temperature, and the size of the results.

Immunological models in high dilution research following M Bastide.

In 1994, Madeleine Bastide described experimental models in immunology that were used during the 1980s to investigate high dilution effects on several biological systems. She classified the available papers in four categories: High dilutions of antigens; High dilutions of thymus, bursa and other hormones; High dilutions of cytokines; Immunopharmacological activity of silica. The studies about high dilutions of antigens were not continued after this period, but gave rise to a long process of a series of in vitro models on antigens and histamine dilutions, that led to the demonstration of the biological modulation effects of these preparations on basophil degranulation. During this process, a multi-centre study was performed, with a high degree of reproducibility among different independent laboratories. The studies about high diluted cytokines, thymulin and other hormones opened a new line of scientific investigation, about the regulatory properties of endogenous substances prepared according to homeopathic methods. The most frequently studied substance, thymulin, when administered to mice at 5cH potency, is able to improve the activity of phagocytes in different experimental situations, such as viral, bacterial and parasitic infections. The immunopharmacological activity of silica was demonstrated, at that time, as an in vivo illustration of the homeopathic 'similia principle'. More recently, studies on silica have assumed another focus: the putative role of silica as active contaminant present in high dilutions. This paper presents a follow-up summary on these items, considering the evolution of discoveries from 1994 to 2014.

Observations on entanglement, non-locality and ultra-high dilutions.

INTRODUCTION: Fundamental research into the scientific basis of the manufacture of ultra-high dilutions and their working in applications has evolved over the past twenty years since our last critical analysis of the field was published in 1994. New contenders from the realm of physics (entanglement, non-locality) have entered the scene. The vast majority within the community of the application of ultra-high dilutions are not physicists. This paper attempts to elucidate the concepts of entanglement, non-locality and their application in ultra-high dilution research (UHD). METHOD: A selected study on the activity of fundamental research into UHD is performed to gain insight into trends of development activity of fundamental research in this area. In an attempt to nurture further development of theoretical models in fundamental research in UHD, an attempt is made to made recent theoretical concepts more accessible to the larger community including practitioners, policy makers and beneficiaries of UHD. RESULTS: Fundamental research in UHD had a period of prolific activity and recognition at the turn of the millennium until about ten years ago. Since then, research output as well as its recognition receded sharply suggesting that a period of reflection and consolidation may be in progress. CONCLUSION: The study and the knowledge gained from more recent theoretical models in UHD and entanglement suggest that there may be some benefit in stocktaking of what we really know about the fundamental workings of UHD as well as identifying or developing models that include measurable predictors that go beyond metaphorical descriptors.

Replications of fundamental research models in ultra high dilutions 1994 and 2015--update on a bibliometric study.

INTRODUCTION: This paper focuses exclusively on experimental models with ultra high dilutions (i.e. beyond 10(-23)) that have been submitted to replication scrutiny. It updates previous surveys, considers suggestions made by the research community and compares the state of replication in 1994 with that in 2015. METHODS: Following literature research, biochemical, immunological, botanical, cell biological and zoological studies on ultra high dilutions (potencies) were included. Reports were grouped into initial studies, laboratory-internal, multicentre and external replications. Repetition could yield either comparable, or zero, or opposite results. The null-hypothesis was that test and control groups would not be distinguishable (zero effect). RESULTS: A total of 126 studies were found. From these, 28 were initial studies. When all 98 replicative studies were considered, 70.4% (i.e. 69) reported a result comparable to that of the initial study, 20.4% (20) zero effect and 9.2% (9) an opposite result. Both for the studies until 1994 and the studies 1995-2015 the null-hypothesis (dominance of zero results) should be rejected. Furthermore, the odds of finding a comparable result are generally higher than of finding an opposite result. Although this is true for all three types of replication studies, the fraction of comparable studies diminishes from laboratory-internal (total 82.9%) to multicentre (total 75%) to external (total 48.3%), while the fraction of opposite results was 4.9%, 10.7% and 13.8%. Furthermore, it became obvious that the probability of an external replication producing comparable results is bigger for models that had already been further scrutinized by the initial researchers. CONCLUSIONS: We found 28 experimental models which underwent replication. In total, 24 models were replicated with comparable results, 12 models with zero effect, and 6 models with opposite results. Five models were externally reproduced with comparable results. We encourage further replications of studies in order to learn more about the model systems used.

Wheat and ultra high diluted silver nitrate--further experiments and re-analysis of data.

BACKGROUND: Since 1926, an influence of a dilution of silver nitrate (24x) on the growth of coleoptiles of wheat seedlings was described. The aim of the study discussed here is the critical proof of the reliability of a test system which has been quoted as a basic model for the research on homoeopathy for decades. METHODS: Grains of winter wheat (Triticum aestivum) were observed under the influence of extremely diluted silver nitrate (10(-23)) prepared by stepwise dilution and agitation according to a protocol derived from homeopathy ('24x'). Analogously prepared water and/or inert water was used for control. Thirty experiments including 5000+5000 grains were performed by 5 researchers. RESULTS: Stalk lengths clearly indicate that development is enhanced by the probe silver nitrate 24x as compared to control. When the experiments 1989-1995 were pooled, means and SD for silver nitrate 24x-groups were 42.3±26.9 mm and for water control groups 34.7±22.2 mm. Verum stalk length was 21.9% bigger than control (100%) (p<0.01; d=0.31, i.e. small). For the experiments 1998-2014, means and SD were 73.7±21.7 mm and 60.5±20.9 mm. Verum stalk length was 21.7% bigger than control (100%) (p<0.01; d=0.62, i.e. medium). From the results one may hypothesize that the result is more marked in experiments showing an average mean of stalk length between ca. 50 and 90 mm in contrast to smaller or bigger mean lengths. CONCLUSION: The previous findings were confirmed by the results.

Wheat and ultra high diluted gibberellic acid--further experiments and re-analysis of data.

BACKGROUND: Following studies (a) on wheat seedlings and ultra high diluted silver nitrate, and (b) on amphibians and an ultra high diluted hormone, (c) a bio-assay on wheat and extremely diluted gibberellic acid was standardized. This assay was intended to combine the easy-to-handle aspect of (a) and biologically interesting aspects of (b). The purpose of the data analysis presented here was to investigate the influence of an extreme dilution of gibberellic acid on wheat stalk length and to determine the influence of external factors on the experimental outcome. METHODS: Grains of winter wheat (Triticum aestivum, Capo variety) were observed under the influence of extremely diluted gibberellic acid (10(-30)) prepared by stepwise dilution and agitation according to a protocol derived from homeopathy ('G30x'). Analogously prepared water was used for control ('W30x'). 16 experiments including 8000+8000 grains were performed by 9 researchers. RESULTS: Experiments that were performed between January and April showed inconsistent results, whereas most of the experiments performed between September and December showed shorter stalks in the G30x group. This was confirmed by correlation analysis (p<0.01). Thus winter/spring experiments and autumn experiments were analysed separately. When all 10 autumn experiments were pooled, mean stalk lengths (mm) were 48.3±21.4 for the verum group and 52.1±20.4 for control (mean±SD) at grain level (N=5000 per group) and ±5.3 and ±5.1 respectively at dish level. In other words, verum stalk length (92.67%) was 7.33% smaller than control stalk length (100%). The effect size is small when calculation is done on the basis of grains (d=0.18) but, due to the smaller SD at dish level, medium when done on the basis of dishes (d=0.73). The inhibiting effect was observed by 6 of the 6 researchers who performed the autumn experiments. CONCLUSION: The model may be useful for further research as there exists a theoretical justification due to previous studies with wheat and extremely diluted silver nitrate, as well as to previous studies with amphibians and diluted hormones, and its methods are well standardized. Data confirm the hypothesis that information can be stored in the test liquid, even at a dilution of the original substance beyond Avogadro's value; and that the wheat bio-assay is sensitive to such information.

Physiological effects of homeopathic medicines in closed phials--a critical evaluation.

BACKGROUND: Phials containing high dilutions of homeopathic remedies are commonly utilized in diagnostic tools in combination with electro acupuncture measuring electrical conductivity of the skin. The present research aimed to elucidate the interaction of the homeopathic remedy and the human organism. METHOD: The study protocol included the transition of a healthy subject to a diseased state utilizing a sub-harmful dose of a toxin. Subsequently, the subject was exposed to a homeopathic or placebo remedy in a closed glass phial and electrical conductivity was measured in open as well as in blind trials. A total of 229 data were collected from open trials and 750 data from blind trials. RESULT: The experimental data showed that homoeopathic remedies in closed glass phials may influence the electrical skin conductivity. The results of the blind trials differed considerably from those of the open trials. The percentage of correctly identified (verum and placebo) phials in the open trials (85%) was statistically different from those in the blind trials (65%). In both types of trials, however, the difference between the sealed phials containing the homeopathic medicine and the placebo was statistically significant. The results of measurements are directly linked to the way in which the tester holds the electrode in one hand and the subject's finger in the other, while putting pressure on the electrode. CONCLUSION: It was concluded that the tester's muscle tone is the intermediary in unconsciously selecting which phial influenced him in his specific contact with the subject.

Reporting Experiments in Homeopathic Basic Research (REHBaR).

INTRODUCTION: The aim of this study was to develop a criteria catalogue serving as a guideline for authors to improve quality of Reporting Experiments in Homeopathic Basic Research (REHBaR). Main focus was in the field of biochemical and biological experiments. So far, there was no guideline for scientists and authors available, unlike criteria catalogues common in clinical research. METHODS: A Delphi Process was conducted among experts who published experimental work within the last five years in this field. The process included a total of five rounds, three rounds of adjusting and phrasing plus two consensus conferences. RESULTS: A checklist of 23 items was achieved, augmented with detailed examples how to handle each item while compiling a publication. Background, objectives and possible hypotheses are necessary to be given in the part 'introduction'. The section 'materials and methods' is the most important part, where a detailed description of chosen controls, object of investigation, experimental setup, replication, parameters, intervention, allocation, blinding, and statistical methods is mandatory. In the 'results' section sufficient details on analysed data, descriptive as well as inferential are needed. Moreover, authors should discuss their results and interpret them in the context of current evidence. REHBaR was compiled for authors when preparing their manuscripts, and to be used by scientific journals in the reviewing process. CONCLUSIONS: Reporting experiments in basic research in homeopathy is an important issue to state the quality and validity of gained results. A guideline for REHBaR seemed to be the first step to come to a commitment what information is necessary to be given in a paper. More than that, the catalogue can serve as a statement what the standards in good basic research should be.

Gas nanobubbles and aqueous nanostructures: the crucial role of dynamization.

Nanobubbles (NBs) have been a subject of intensive research over the past decade. Their peculiar characteristics, including extremely low buoyancy, longevity, enhanced solubility of oxygen in water, zeta potentials and burst during collapse, have led to many applications in the industrial, biological and medical fields. NBs may form spontaneously from dissolved gas but the process is greatly enhanced by gas supersaturation and mechanical actions such as dynamization. Therefore, the formation of NBs during the preparation of homeopathic dilutions under atmospheric pressure cannot be ignored. I suggested in 2009 the involvement of NBs in nanometric superstructures revealed in high dilutions using NMR relaxation. These superstructures seemed to increase in size with dilution, well into the ultramolecular range (>12c). I report here new experiments that confirm the involvement of NBs and prove the crucial role of dynamization to create superstructures specific to the solute. A second dynamization was shown to enhance or regenerate these superstructures. I postulate that superstructures result from a nucleation process of NBs around the solute, with shells of highly organized water (with ions and silicates if any) which protect the solute against out-diffusion and behave as nucleation centres for further dilution steps. The sampling tip may play an active role by catching the superstructures and thus carry the encaged solute across the dilution range, possibly up to the ultramolecular range. The superstructures were not observed at low dilution, probably because of a destructuring of the solvent by the solute and/or of an inadequate gas/solute ratio.

Survey of immuno-allergological ultra high dilution research.

BACKGROUND: Experiments about basic research in Immuno-allergology reported by M. Bastide and B. Poitevin in Ultra High Dilution (1994) have been appraised from a 20 year perspective. The numerous experiments published mainly focus on immunological regulation, inflammatory process and basophil activation. They are analyzed according to one essential criterion: repeatability. METHODS: The commentary reflects the research details made available in a recently published literature review, also published in French. RESULTS: The regulatory effect of high dilution of bursin on immune response has been observed in multiple experiments but not reproduced by independent teams. The immunomodulating effect of Thymulin has been confirmed in mice. Rhus toxicodendron has an anti-inflammatory activity on different models, from mother tincture (TM) to very high dilutions. The homeopathic complex Canova activates macrophages in vitro and in vivo, induces lymphocyte proliferation, and reduces the size of tumors and mortality of sarcoma-bearing mice. Some homeopathic medicines used in clinical inflammation modulate in vitro the neutrophil activation, with variability in the protocols used and in the medicines tested. In allergology, high dilution histamine has an inhibitory effect on basophil activation in multicenter trials and with independent teams, either with methods implying a change in basophil staining or with flow cytometry. However, high dilution histamine had no effect in some well-conducted experiments. The inhibitory effect of Apis mellifica has not been studied with the flow cytometry method, as well as the activation of basophil by anti-IgE high dilution, published in Nature. CONCLUSIONS: Despite considerable research activity in immuno-allergology and a great increase in the number of publications, there is still not in this domain a "gold standard" trial in basic research in homeopathy. The most studied system, the inhibitory effect of histamine high dilutions on basophil activation, requires clarifications of various factors, including individual sensitivity. For scientific and epistemological reasons, the same work should be carried out for independent reproduction of the experiments conducted with anti-IgE and Apis mel high dilution, in complement of the new axes of research in immunoallergology developed since 20 years.

Electromagnetic and magnetic vector potential bio-information and water.

This work developed over the past 40 years starting from dielectric measurements on enzymes and the subsequent finding that the measurements were affected by electric, magnetic, electromagnetic fields and quantum fields. A request for help in the diagnosis and therapy of chemically sensitive patients who had become sensitive to their electromagnetic environment came in 1982. The same symptoms could be provoked by a chemical or a frequency challenge and this led to an appreciation of the synergy between chemical and frequency environmental sensitivities. Experimental cooperation with theoretical physicist Herbert Fröhlich FRS and others led to an understanding of the physics of coherent water in living systems and a mechanism for the memory of water for coherent frequencies. In a coherent system there are interacting frequencies proportionate to any velocity the system will support, in particular the velocity of light and the velocity of coherence diffusion. Thus, there can be biological interaction between the optical, microwave and ELF spectral regions. Frequency modulation of light scattered by bio-fields and its retention in recorded images is discussed. A 'nil-potent' frequency can erase a frequency signature and thence affect a biological system. Homeopathy is interpreted through the biological effects of coherent frequencies derived from the frequency signature of the 'Mother Tincture' and developed through dilution and succussion. A homeopathic potency has a frequency signature therefore it must be able to have a biological effect.

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.

The mathematics of dilution.

The major objection to homeopathic medicine is that the doses of medicine prescribed in some cases are too dilute for any active ingredient to be present. The medicines would hence be rendered inactive, necessitating novel explanations for the action. A further examination of dilution in the light of the Langmuir equation shows that homeopathic medicines may not be as dilute as a simplistic application of Avogadro's Principle suggests, due to surface effects.

Seasonal variation of the effect of extremely diluted agitated gibberellic acid (10e-30) on wheat stalk growth: a multiresearcher study.

The influence of a homeopathic high dilution of gibberellic acid on wheat growth was studied at different seasons of the year. Seedlings were allowed to develop under standardized conditions for 7 days; plants were harvested and stalk lengths were measured. The data obtained confirm previous findings, that ultrahigh diluted potentized gibberellic acid affects stalk growth. Furthermore, the outcome of the study suggests that experiments utilizing the bioassay presented should best be performed in autumn season. In winter and spring, respectively, no reliable effects were found.

Veterinary Biotypology: a review and new possibilities

The truths surrounding medical practices are seasonally challenged by innovative concepts that can aggregate changing procedures in many degrees. The Galtonian eugenics issues supported the pure-breed idea in dictatorial governments, and introduced mesological studies, turning possible to join genetic concepts to the physiology and psychology of the human organism. Following human medicine, more therapeutic models need to forthcoming in domestic animals. The companionship necessity and the highly responsive behavior have addressed the domestication of dogs and their relationship to owners, to an endpoint that both share the same pathologies. Thus, traditional human concepts of biotypology could be extended to companion animals. Grauvogl (1811-1877) proposeda simple biochemical correlation between physiological states and the miasmas of sick individuals (oxygenoid -syphilis, hydrogenoid -sycosis, carbo-nitrogenoid -psora). Antoine Nebel (1870-1954) correlated biochemical status with the musculoskeletal system and behavior as well. Leon Vannier (1880-1963) model, whose morphophysiological distortions and behavioral inconsistencies were explained by the carbon element and variations in its bonds with phosphorus or fluor radicals was another attempt to categorize and predefine physiology states. Following the advent of structural and functional identification of thyroid hormone in the 1940s, Henri Bernard described the neuro-morphofunctional plasticity of individuals guided by their predominant embryonic leaflet and consequent hormonal diseases. Methods:This work is a narrative review with the purpose of describing and discussingthe legacy ofbiotypology studies and their applicability in dog therapy, and proposinga new homeopathic approach in veterinary medicine based on the miasmas, also contributing to the scarcelyavailable literature. Results:Based on cellular exchanges and consequent metabolic rate, animals can be classified into psoric (no evidence of clinical signs, stable behavior, and adequate exonerative cellular processes); sycotic (cellular dysfunction with alterations in oxidative phosphorylation processes allowing accumulation of cellular toxins such as reactive oxygen and nitrogen species; clinically culminating in chronic inflammations in noble organs, and purulent discharges; unstable and polarized behavior) and syphilitic (whose cellular alterations have reached the molecular level, reducing protein expression and determining cellular toxicity and loss of function; indifferentbehavior). Generalities such as temperature influence, weight, thirst and feedingshall also be considered. Discussion and Conclusion: Thismodel could benefit stray animals, newly adopted or even from shelters, whose actual behavior is unknown, and the search for the Simillimum may be impaired.

Evaluation of the in vitrofungicidal activity of the dynamized essential oil of Aloysia polystachyabefore and after freezing

The method of preserving substances of natural origin should not only maintain the microbiological safety of the product but also the integrity of its therapeutic potential. Essential oils obtained from plants are complex mixtures of substances and it issuggested to keep them under refrigeration for better conservation. On the other hand, homeopathic mother tincture prepared from plant is kept at room temperature. Aim: This work aimed to evaluate if the freezing process changes the in vitro antifungal activity potential of the homeopathic preparation Aloysia polystachya1CH against Candida albicans. Methodology:The inoculum of C. albicansATCC 10231 was cultivated in culture médium Sabouroud (Himedia®), standardized on a spectrometer and distributed in a 96-well plate. Then, A. polystachya1CH was added to the wells, prepared accordingtothe Brazilian Homeopathic Pharmacopoeia (FHB, 3rd edition) from A. polystachya essencial oil. An aliquot of this homeopathic preparation was frozen and after 40 days it was submitted to the same methodology for evaluation of the antifungal activity. After incubation, the plates were read with triphenyltetrazolic (TTC) (Vetec®). Results and discussion: The results of the in vitroevaluation showed that the freezing process retained the antifungal activity of the dynamized essential oil of A. polystachya1CH against C. albicans. Conclusion: Under the conditions evaluated in this study, the freezing method presented as a viable method of conservation of dynamized plant material.