Series of experiments performed with the droplet evaporation method on low potencies

Homeopathic preparations in low potencies, containing still measurable quantities of the starting substance, constitute a unique research field in homeopathic basic research. Here a series of experiments is presented carried out by means of the droplet evaporation method (DEM), investigating the specificity of the method, and presumed effects of the succussion procedure applied in the production of homeopathic preparations. Methods:DEM analysis consisted in the evaporation of droplets of the potencies perse placed on microscope slides. Resulting patterns were photographed. Images were evaluated by means of ImageJ software, by measuring grey level distribution, texture, and fractality. The experimentation consisted of four series: (i) screening (1x­6x potencies from 19 substances), (ii) differentiation experiments (2x­6x potencies of Echinacea, Baptisia, Luffa, and Spongia), (iii) differentiation between succussed (100 or 10 times) and unsuccussed samples (Echinacea 2x, Baptisia 3x, Baptisia 4x, Luffa 4x, and Spongia 6x). (iv) investigation of the influence upon the patterns of single compounds present in a remedy complex. The experimental set-up stability was examined by systematic positive control experiments. Results:(i) Homeopathic preparations of mineral origin showed the greatest form variety, whereas those of vegetal origin created fractal patterns in the potency range 2x­4x. (ii) Differentiation of potencies of different origin at the same dilution level was possible from 2x to 4x. (iii) In all potency levels, succussed (100 and 10 times) and unsuccussed variants could be significantly differentiated. Significant differences between all variants were found in some cases in potency levels 4x and higher. In general, application of succussion reduced size, homogeneity, and complexity of the DEM patterns. (iv) Patterns of a remedy complex Luffa 4x -Mercurius bijodatum 9x showed a clear predominance of the Luffa 4x; however also the second component, present in a much lower concentration, influenced significantly the pattern of the remedy complex as also differed significantly from the pattern of succussed water control. Conclusions:The results suggest that DEM is a suitable tool for scientific investigation of homeopathic preparations in the low potency range. DEM might be applied to assess further research questions, such different potentization procedures (vessel shape, overhead volume, material), storing time, and difference between batches.

Formation of Self-organized Structures in Drying Droplets: a Promising Analytical Tool for Homeopathy Basic Research

The droplet evaporation method (DEM) is based on the evaporation-induced pattern formation in droplets and is applied mainly for medical diagnosis[1].Here, we present aseries of experiments performed by our team showing DEMs potential also forhomeopathy basic research, in particular, for the investigation of(i) low potencies, (ii) low potency complexes (physical model), and (iii) the action of high potencies (plant-based model).Methods:(i) DEM differentiated significantly between Luffa, Baptisia, Echinacea, and Spongiauntil 4x[2]. Furthermore, the patterns varied in function of the numberof succussion strokes (0, 10, or 100) applied during potentization[3]. The performance of chaotic succussions vs. laminar flow vs. slight mixing during the potentization of Viscum album quercus3x influenced the DEM patterns; the chaotic succussions reduced, whereas laminar flow enhanced the patterns complexity vs. the unsuccussed control.(ii) The addition of Mercurius bijodatus9x to Luffa4x changed significantly the DEM patterns, even if the material quantity present in the 9x potency lied far beyond that of ultrapure water.(iii) Leakages obtained by placing healthy or arsenic-damaged wheat-seeds into Arsenicum album45x orheat-damaged intoZincum metallicum30c vs. water created significantly different DEM structures [4, 5]. Results:The damaged seeds put into the potency created structures characterized by a higher complexity than those obtained from damaged seeds put into control water. Furthermore, the potency action seemed to increase with rising numbers ofsuccussion strokes applied during potentization,ascould be shown by means of DEM patterns and germination rate using the same wheat-seed model[6].In all our studies, the pattern evaluation was computerized (texture and fractal analysis performed by means of ImageJ) or based on deep-learning algorithms and the robustness of the experimental system was checked by means of systematic control experiments.Conclusion:DEM together with other similarmethods has also been reviewed by our team for what concerns theapplication in homeopathy basic research[7].

Physicochemical and UV/VIS measurements of UHD solutionsof water, potentized water,and different potencies of organic substance

Conventional science regards the study of UHD (highly homeopathically potentized) solutions as pseudo-science. However, an increasing number of rigorous scientific investigations demonstrate differences in physicochemical and physical characteristics of such solutions. Strictly chemically regarded, they correspond to highly distilled water. Our research team developed a system of physicochemical and UV spectrographic measurements, whereby the differences may be consistently confirmed with high statistical significance.Methods:For measurement of the physicochemical parameters,we usedpH,electrical conductivity,and oxidation-reduction potential (ORP)sensors.For UV/VIS spectroscopymeasurements,we useda Macherey-Nagelspectrophotometer.For UHD research, we used two batches of substances: Russian (R) and Brazilian (B). In R, distilled water (W) was used for dilutions and potencies as follows: potentized water (W cH9), potentized dilution of antibodies to interferon-gamma (Abs IFNγ cH9), the same original substance in the form of a mixture of potencies (Abs IFNγ cH12, cH30, cH50, shortly Abs IFNγ mix). Furthermore, we prepared higher potencies of the substances (supplementary potentiation) in a specially prepared solution and measured their characteristics. In B, the solution was used for further dilutions and potencies as follows: potentized water (W cH1) and Glyphosate potencies (Gly cH6 => cH8, cH30 => cH 32, and cH200 => cH 202).For direct or post-hoc analysis, we used Wilcoxon signed-rank test, two-tailed.Results:UV-VIS spectroscopy (R): measurements of received liquids potentized for further cH1 show statistically significant differences between all substances, except between water W and W cH9 at 260 nm. Significant differences (p-values) were as follows: Abs IFNγ mixvs. W= 0.007; Abs IFNγ mixvs.W cH9=0.008; Abs IFNγ cH9vs. W=0.044; Abs IFNγ cH9vs.W cH9= 0.026; Abs IFNγ mixvs.W cH9= 0.007; W vs.W cH9= 0.506.Physicochemical measurements:R: measurementsdemonstrated statistical difference only in pH (Abs IFNγ mix towards all others). Significant differences (p-values) were as follows: Abs IFNγ mixvs. W=0.022; Abs IFNγ mixvs.W cH9=0.005; Abs IFNγ mixvs.Abs IFNγ cH9=0.025. After supplementarypotentiation, we obtained a more conspicuous picture with many statistical differences in conductivity and ORP, ranging from p= 0.001 to 0.046.A difference between water and potentized water has also been demonstrated.B: the measurements demonstrated statistical differences mainly in pH between Gly cH8 and the rest and between Gly cH202 and W cH1.Significant differences (p-values) were as follows:Gly cH8vs. Gly cH32 =0.027; Gly cH8vs. Gly cH202 = 0.011; Gly cH8vs. W cH1= 0.014; Gly cH202vs. W cH1= 0.034.Conclusion:UV/VIS at wavelength 260 nm Abs IFNγ mix discloses a pattern similar to exclusion zone (EZ)water at 270 nm.By additional potentiation and with physicochemical measurements, we obtained higher statistical differences than in the original dilutions.In contrast, UV/VIS spectroscopy showed more conspicuous results without additional potentiation. However, the very act of succussion becomes very distinct

Free energy perturbation (FEP)-guided scaffold hopping

Scaffold hopping refers to computer-aided screening for active compounds with different structures against the same receptor to enrich privileged scaffolds, which is a topic of high interest in organic and medicinal chemistry. However, most approaches cannot efficiently predict the potency level of candidates after scaffold hopping. Herein, we identified potent PDE5 inhibitors with a novel scaffold via a free energy perturbation (FEP)-guided scaffold-hopping strategy, and FEP shows great advantages to precisely predict the theoretical binding potencies ΔG FEP between ligands and their target, which were more consistent with the experimental binding potencies ΔG EXP (the mean absolute deviations | Δ G FEP - Δ G EXP |  < 2 kcal/mol) than those ΔG MM-PBSA or ΔG MM-GBSA predicted by the MM-PBSA or MM-GBSA method. Lead L12 had an IC50 of 8.7 nmol/L and exhibited a different binding pattern in its crystal structure with PDE5 from the famous starting drug tadalafil. Our work provides the first report via the FEP-guided scaffold hopping strategy for potent inhibitor discovery with a novel scaffold, implying that it will have a variety of future applications in rational molecular design and drug discovery.

Structure-based discovery of orally efficient inhibitors via unique interactions with H-pocket of PDE8 for the treatment of vascular dementia

Our previous study demonstrated that phosphodiesterase 8 (PDE8) could work as a potential target for vascular dementia (VaD) using a chemical probe 3a. However, compound 3a is a chiral compound which was obtained by chiral resolution on HPLC, restricting its usage in clinic. Herein, a series of non-chiral 9-benzyl-2-chloro-adenine derivatives were discovered as novel PDE8 inhibitors. Lead 15 exhibited potent inhibitory activity against PDE8A (IC50 = 11 nmol/L), high selectivity over other PDEs, and remarkable drug-like properties (worthy to mention is that its bioavailability was up to 100%). Oral administration of 15 significantly improved the cAMP level of the right brain and exhibited dose-dependent effects on cognitive improvement in a VaD mouse model. Notably, the X-ray crystal structure of the PDE8A-15 complex showed that the potent affinity and high selectivity of 15 might come from the distinctive interactions with H-pocket including T-shaped π-π interactions with Phe785 as well as a unique H-bond network, which have never been observed in other PDE-inhibitor complex before, providing new strategies for the further rational design of novel selective inhibitors against PDE8.

Free water molecules and hydrogen bonding form the basis of variation in homeopathic potencies as revealed by vibrational spectroscopy

Using Fourier Transform Infrared spectroscopy (FTIR) we have demonstrated that homeopathic potencies of Natrum mur, Cantharis, Nux vomica and Sulphur show differences with respect to the number of free water molecules and strength of hydrogen bonding. The purpose of the present study is to confirm this phenomenon in three potencies of two more drugs Calcarea carb and Silicea. Design: The potencies used for each of the two drugs were 30cH, 200cH and 1000cH. The control was 90% ethanol as also the potentized drugs. The control, as well as the potencies, were diluted with distilled water to reduce the level of ethanol to 0.03 molar fraction in each of them. FTIR spectra of all the potentized drugs, control and sterile distilled water (reference water) were taken in the wave number region of 4000-2800 cm-1. The full width at half maximum (fwhm) of OH band was measured for each spectrum. The width was divided into two in the middle. The difference spectrum (absorbance of drug solution - absorbance of reference water) for each potency and the control was obtained after normalization of the spectrum at 3410 cm-1. One difference spectrum so obtained for a potency was subtracted from another to find out if there is a difference between two different potencies. Results: The half width half maximum (hwhm) in both the high and low-frequency sides of the OH band is far less narrow in potencies than in the control as compared to that in water. The difference spectra for different potencies show different levels of fall in intensity at the wave number region of dip at 3630 cm-1...

Reação histológica de exposições pulpares em dentes de ratos à aplicação do laser de baixa intensidade somente ou em associação ao capeamento com hidróxido de cálcio / Histological reaction in exposed pulp in teeth of mice to the application of the low power laser only ot in association to the dental pulp capping with calcium hydroxide

O presente estudo avaliou a reação histológica do efeito do laser de baixa intensidade diodo semicondutor de arseneto de gálio e alumínio (GaAlAs, com comprimento de onda de 785 nm, infravermelho, com potência de emissão de 50 mW, densidade de energia de 3 J/cm2 , em emissão contínua, por meio de ponta especial de fibra óptica, por um período de 2 segundos) em exposições pulpares induzidas em dentes de ratos. Foram utilizados 60 ratos, dos quais cada animal ofereceu dois dentes (totalizando 120), que foram distribuídos em 5 grupos experimentais: grupo I (controle), exposição capeada com hidróxido de cálcio; grupo II, laser imediato + hidróxido de cálcio; grupo III, laser imediato + hidróxido de cálcio + laser após 24 horas; grupo IV, laser imediato + hidróxido de cálcio + laser após 24 e 48 horas; e grupo V, somente a aplicação do laser. Decorridos os períodos experimentais de sete e 30 dias, os animais foram mortos, as peças removidas e preparadas para análise histológica. De acordo com a proposta e as condições específicas deste trabalho, os resultados nos permitem concluir que as associações laser imediato + hidróxido de cálcio e laser imediato + hidróxido de cálcio + laser 24 horas, foram os grupos que apresentaram os melhores resultados tanto aos sete dias (efeito antiinflamatório), quanto aos trinta dias (efeito no reparo tecidual

The present study evaluated the histological reaction of the low power laser effect, diode laser semiconductor of gallium aluminum arsenide (GaAlAs, with wavelength of 785 nm, infra-red, potency of emission of 50 mW, energy density of 3 J/cm2 , with continuous emission, through special tip of optic fiber, for a period of two seconds) in exposed pulp induced in teeth of mice. Sixty mice were used, of which each animal offered two teeth (totaling 120), that were distributed in 5 experimental groups: group I (control group) exposed capping with calcium hydroxide; group II, immediate laser + calcium hydroxide; group III, immediate laser + calcium hydroxide + laser after 24 hours; group IV, immediate laser + calcium hydroxide + laser after 24 and 48 hours; and group V, only laser application. After the experimental periods of seven and thirty days, the animals were killed, the pieces were removed and prepared for histological analysis. In agreement with the proposal and the specific conditions of this study, the results allow us conclude that the associations of immediate laser + calcium hydroxide and immediate laser + calcium hydroxide + laser after 24 hours, were the groups that presented the best results in the periods of seven days (anti-inflammatory effect), and thirty days (effect in the tissue repair