Extreme dilutions of mercuric chloride produce longer binding interaction with ?-amylase as compared to blank solvents

Background:High and ultra-low doses of HgCl2modulate α-amylase starch interaction through two different binding sites of the enzyme. Mercurius corrosivus at ultra-high dilution (UHD) is reported to have a long duration of action for 30-60 days.Objectives:(i) To see whether the mother tincture and UHDs of mercuric chloride could directly act on α-amylase without intervention of its substrate starch. (ii) To see whether the MT and UHDs produce longer binding interaction with α-amylase than the control.Methods:Mercuric chloride MT (0.15 M) or Merc corθ and its three UHDs, 30, 200, 1000 cH called potencies were in 90% ethanol. Blank 90% ethanol and deionised and distilled (DD) water were used as controls. All ethanol based ligands were diluted with DD water to reduce EtOH content to 0.09%. Using an ITC instrument each ligand was injected once at 10 µl into15µM α-amylase at 250C and the reaction was observed for 33.3 min. Results:While Merccorθ produced endothermic reaction, all other ligands did exothermic reaction. All ligands sustained heat change for the entire period of observation (33.3 min). Conclusions:Merc corθ and potencies produced exactly the opposite reaction at the protein binding sites. Merc cor200 cH and 1000 cH produced highest heat change (6 micro cal/sec) during interaction with α-amylase.(AU)

High and ultra low concentrations of Mercuric chloride initiate their specific action on binding sites of invertase and modify its interaction with sucrose

Background: Mercuric chloride is known to inhibit the activity of enzymes. It is used in homeopathy at ultra low concentration (ULC) and is known as Mercurius corrosivus (Merc cor). ULCs of Merc cor are reported to promote enzyme activity. Objective: To see whether the mother tincture (θ) of Merc cor and its ULCs interact with an enzyme invertase at its binding sites and influence enzyme's action on its substrate sucrose. Methods: Merc cor θ (0.15 M HgCl2) was diluted with deionized and distilled (DD) water 1:100 and succussed 10 times to prepare Merc cor 1 cH or 1st potency. This potency was further diluted and succussed in 200 and 1000 steps to prepare 200cH and 1000cH potencies, respectively. Merc cor 200 cH and 1000cH were prepared in 90% ethanol. The two potencies and blank 90% EtOH were diluted with DD water 1:1000 to minimize ethanol content to a negligible amount 0.09%. The control was DD water (0.99g/M). The drugs, EtOH and water control were mixed separately with 0.037 mM invertase in DD water. Using an isothermal calorimetry (ITC) instrument the substrate sucrose (65mM) was injected at 2 µl every 2 min into 300 µl invertase solution 20 times at 25 0C. Molecular modeling study was done to predict possible binding sites and nature of binding between the enzyme and HgCl2, and between the enzyme and water. Potencies after dilution are virtually water. Fluorescence spectra of invertase (4µM) mixed with drug/control solutions were also obtained to see the effect of drugs on protein folding. Results: Thermodynamic parameters like binding constant (K), change in enthalpy(ΔH), entropy(ΔS) and Gibbs free energy(ΔG) showed marked variation in treatment effects on the enzyme. Molecular modeling study also shows variation in binding between invertase and HgCl2, and between invertase and water. Fluorescence spectra show variation in quenching related to different treatments. Conclusion: Merc cor mother tincture and its potencies interact at different binding sites of invertase and modify the enzyme's action on sucrose. So, potencies act as modulators of ligand, sucrose. Drug solutions induce conformational changes in the enzyme. (au)

Extremely dilution of strychnos Nux vomica mitigates alcohol-induced reduction in enthalpies associated with free water molecules in a fish brain

Introduction: Alcohol intoxication affects aquaporins in the glial cells of brain resulting in oedema. Nux vomica, a homeopathic drug of plant origin, is known to counteract alcohol effect. The objective of this present study is to find out the level of free water molecules in the brain of a teleost fish under ethanol intoxication. The second objective is to determine whether Nux vomica could restore the level of free water in the alcohol treated fish. Materials and methods: One group of fish was exposed to 456 mM ethanol for 30 min, another exposed first to a solution of Nux vomica 200c for 20 min and then to 456 mM ethanol for 30 min. The third group served as an untreated control. The mid brain of each fish was kept in an aluminium sample pan and its free water level was assessed by differential scanning calorimetry (DSC). Results: All alcohol treated fish showed significant reduction in the level of free water molecules as compared to the untreated control. Treatment with Nux vomica increased the level of free water in the brain significantly as compared to the untreated alcoholic group. Conclusion: Alcohol intoxication reduces free water molecules in the fish brain. Nux vom might have acted on aquaporins in the glial cells thereby increasing the level of free water in the brain. (AU)

Cantharis 200c counters heat stress in germinating seeds of Vigna unguiculata (L) Walp

Background Thermal stress during early imbibitional phase of germination causes disruption of redox-homeostasis by increasing accumulation of ROS Reactive Oxygen species (assessed in terms of hydrogen peroxide) and significant reduction of antioxidative defense (assessed in terms of catalase and peroxidase) in germinating tissues of cowpea (Vigna ungiculata). It also induces oxidative damage to newly assembled membrane system by aggravating membrane lipid peroxidation [measured in terms of thiobarbituric acid reactive substances (TBARS)]. Cantharis is a homeopathic remedy used for treating burn injuries in humans. The objective is to see whether potentized Cantharis could ameliorate heat stress in germinating seeds of cowpea. Methodology Seeds, imbibed in water overnight, were divided into four groups (n=50/group): Control I (Water 200c), Control II (Ethanol 200c), treated with Cantharis 200c and untreated and unstressed. Water soaked seeds were dipped in control/drug solution for 5 min and then washed. Control II and Cantharis 200c were diluted with water 1:1000 to minimize the ethanol effect. Except the fourth group, all other groups were subjected to heat stress (450C for 8 hours). All the groups were allowed to germinate for 5 days in germination chamber at 25 ± 20C. Groups 1 and 2 (Control I and II) served as systematic negative controls throughout the experiment. Ten independent replications were performed for each group in parallel. All experiments were randomized and blinded. Results The systematic negative controls (I, II) did not produce any significant effect. The results in terms of germination, growth, soluble sugar, protein, accumulation of reactive oxygen species and loss of membrane permeability clearly exhibit that Cantharis 200c could mitigate heat stress significantly (p<0.05) as compared to the controls I, II. Final germination percentage after 120 hrs was found to be 60%, 48%, 86%, 88%, for groups 1, 2, 3, 4, respectively. Total chlorophyll (Chl-a + Chl-b), carotenoids and total chl/carotenoid ratio were found to be highest in groups 3 and 4. Assessment of membrane lipid peroxidation in terms of TBARS and accumulation of reactive oxygen species (H2O2) showed minimum concentration in group 3 and 4. Conclusion Cantharis 200c can counteract the deleterious effects of heat stress on juvenile tissues and membranes and can be used effectively to counter heat stress in tropical countries especially during summer. (AU)

Raman spectroscopy shows difference in drugs at ultrahigh dilution prepared with stepwise mechanical agitation

OBJECTIVE: The present study aims at deciphering the nature of the water structure of two ultrahigh diluted (UHD) homeopathic drugs by Laser Raman Spectroscopy. METHOD: Two homeopathic drugs Sulphur and Natrum mur in three UHD 30cH, 200cH and 1000cH were selected for the study. Raman spectra of the drugs and their medium (90% ethanol) were obtained in the wave number region of 2600-3800 cm-1. The intensity ratio at vibration frequencies between 3200 and 3420 (R1) and that between 3620 and 3420 (R2) was calculated for each UHD as well as the control. RESULTS: Raman spectra shows differences in intensities in different UHDs and their control in the stretching vibrations of CH and OH groups. The three UHDs of each drug show an inverse relationship with respect to the R1 values. However, for R2 the relationship of UHD for each drug is positive. CONCLUSION: R1 provides information about the relative number of OH groups with strong and weak hydrogen bonds. R2 suggests the relative number of OH groups with broken and weak hydrogen bonds. Judged from R1 values the lower is the rank of UHD, the stronger is the H-bond of the OH groups. In the light of R2 values, the higher is the UHD rank the more abundant is the free OH groups. So, hydrogen bond strength and free OH groups together make an effective UHD rank relating to Sulphur and Natrum mur.

Raman spectroscopy reveals variation in free OH groups and hydrogen bond strength in ultrahigh dilutions

OBJECTIVE: To decipher the nature of water structure in two ultrahigh diluted (UHD) homeopathic drugs by Laser Raman Spectroscopy. METHOD: Two homeopathic drugs Calcarea carbonica (Calc.) and Sepia officinalis (Sep.) in 8cH, 202cH, and 1002cH and their diluent medium 90% ethanol in 8cH and 202cH were used in the present study. Laser Raman spectra of all the samples were obtained in the wave number region of 2400 – 4200 cm-1. The intensity ratio at vibration frequencies between 3200 and 3420 (R1) and that between 3620 and 3420 (R2) were calculated for each UHD of the samples. RESULTS: The spectra show a marked difference in intensities in the stretching vibrations of CH and OH groups of all the samples. R1 values for three UHDs of Calc. and Sep. show negative and positive relationships, respectively. In the case of R2 values, the relationship in three UHDs is 81002 for Calc., and 8> 202 < 1002 for Sep. In the case of control (ethanol UHDs) both R1 and R2 show a negative relationship. CONCLUSION: R1 denotes a relative number of OH groups with strong and weak hydrogen bonds. R2 indicates the relative number of OH groups with broken and weak H-bonds. Therefore, the UHDs of the two drugs and the control are different from each other with respect to hydrogen bond strength of OH groups and the number of free OH groups or non-hydrogen bonded water molecules.

Raman spectroscopy shows difference in drugs at ultrahigh dilution prepared with stepwise mechanical agitation

Abstract Objective: The present study aims at deciphering the nature of the water structure of two ultrahigh diluted (UHD) homeopathic drugs by Laser Raman Spectroscopy. Method: Two homeopathic drugs Sulphur and Natrum mur in three UHD 30cH, 200cH and 1000cH were selected for the study. Raman spectra of the drugs and their medium (90% ethanol) were obtained in the wave number region of 2600-3800 cm-1. The intensity ratio at vibration frequencies between 3200 and 3420 (R1) and that between 3620 and 3420 (R2) was calculated for each UHD as well as the control. Results: Raman spectra shows differences in intensities in different UHDs and their control in the stretching vibrations of CH and OH groups. The three UHDs of each drug show an inverse relationship with respect to the R1 values. However, for R2 the relationship of UHD for each drug is positive. Conclusion: R1 provides information about the relative number of OH groups with strong and weak hydrogen bonds. R2 suggests the relative number of OH groups with broken and weak hydrogen bonds. Judged from R1 values the lower is the rank of UHD, the stronger is the H-bond of the OH groups. In the light of R2 values, the higher is the UHD rank the more abundant is the free OH groups. So, hydrogen bond strength and free OH groups together make an effective UHD rank relating to Sulphur and Natrum mur. (AU)

Raman spectroscopy reveals variation in free OH groups and hydrogen bond strength in ultrahigh dilutions

Objective: To decipher the nature of water structure in two ultrahigh diluted (UHD) homeopathic drugs by Laser Raman Spectroscopy. Method: Two homeopathic drugs Calcarea carbonica (Calc.) and Sepia officinalis (Sep.) in 8cH, 202cH, and 1002cH and their diluent medium 90% ethanol in 8cH and 202cH were used in the present study. Laser Raman spectra of all the samples were obtained in the wave number region of 2400 – 4200 cm-1. The intensity ratio at vibration frequencies between 3200 and 3420 (R1) and that between 3620 and 3420 (R2) were calculated for each UHD of the samples. Results: The spectra show a marked difference in intensities in the stretching vibrations of CH and OH groups of all the samples. R1 values for three UHDs of Calc. and Sep. show negative and positive relationships, respectively. In the case of R2 values, the relationship in three UHDs is 8<202>1002 for Calc., and 8> 202 < 1002 for Sep. In the case of control (ethanol UHDs) both R1 and R2 show a negative relationship. Conclusion: R1 denotes a relative number of OH groups with strong and weak hydrogen bonds. R2 indicates the relative number of OH groups with broken and weak H-bonds. Therefore, the UHDs of the two drugs and the control are different from each other with respect to hydrogen bond strength of OH groups and the number of free OH groups or non-hydrogen bonded water molecules.(AU)

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...

Transfer of the effect of potentized mercuric chloride on á-amylase fromone test tube to another through capillary water

In a series of experiments we showed that treatment of a plant or animal with a dilutedand agitated substance might affect other plants or animals connected to the former by the capillarywater in cotton threads. The aim of the present study was to establish whether drug effect could betransferred in a cell-free medium.Design: Two test tubes, each containing 1 ml of 1% starch solution and 1 ml of á-amylase, wereconnected by means wet cotton threads encased in a polythene tube. One of the tubes alsocontained Mercurius corrosivus (Merc-c) 30 cH and the other ethanol solution (control). After 15min, the enzyme activity was stopped with DNSA, and the breakdown product of starch, maltose,was estimated. A third, separate tube contained all the tested materials except for Merc-c and thecontrol solution. In a second experiment two tubes, one containing 1,200 ppm and the other 200ppm of maltose, were similarly connected over 15 min. Both experiments were repeated 20 times.Results: In the first experiment, the amount of maltose was similar in both connected tubes, but itwas significantly lower in the unconnected tube. In the second experiment, maltose concentrationin both tubes remained unchanged.Conclusion: The information of Merc-c 30 cH was effectively transferred through capillary waterbetween two tubes in cell-free medium. This effect was not due to physical transfer of either solventor solutes. Water seems to the most probable carrier of information in diluted and agitatedsolutions...

Transfer of the effect of potentized mercuric chloride on á-amylase fromone test tube to another through capillary water

Objective: In a series of experiments we showed that treatment of a plant or animal with a dilutedand agitated substance might affect other plants or animals connected to the former by the capillarywater in cotton threads. The aim of the present study was to establish whether drug effect could betransferred in a cell-free medium.Design: Two test tubes, each containing 1 ml of 1% starch solution and 1 ml of á-amylase, wereconnected by means wet cotton threads encased in a polythene tube. One of the tubes alsocontained Mercurius corrosivus (Merc-c) 30 cH and the other ethanol solution (control). After 15min, the enzyme activity was stopped with DNSA, and the breakdown product of starch, maltose,was estimated. A third, separate tube contained all the tested materials except for Merc-c and thecontrol solution. In a second experiment two tubes, one containing 1,200 ppm and the other 200ppm of maltose, were similarly connected over 15 min. Both experiments were repeated 20 times.Results: In the first experiment, the amount of maltose was similar in both connected tubes, but itwas significantly lower in the unconnected tube. In the second experiment, maltose concentrationin both tubes remained unchanged.Conclusion: The information of Merc-c 30 cH was effectively transferred through capillary waterbetween two tubes in cell-free medium. This effect was not due to physical transfer of either solventor solutes. Water seems to the most probable carrier of information in diluted and agitatedsolutions.(AU)

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

Objective: 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. The level of dip at 3630 cm-1 and subsequent rise in intensity in the lower frequency region represent the quantity of free water molecules and strong alcoholic OH bond around 3250 cm-1, respectively. The results of subtraction between two different potencies are not zero but have marked positive or negative values. Conclusion (i) Potencies have stronger intermolecular interactions and a higher number of chemical environments than the control, as revealed by the data on hwhm. (ii) The three potencies of each of the two drugs show distinct variation in the number of free water molecules and strength of hydrogen bonding. (iii) There exists both inter-drug and inter-potency variation as revealed by the difference spectra and results of subtraction between two difference spectra.(AU)

Homeopathic drugs complementary, antidotal and inimical to Nux vomica produce stronger anti-alcoholic effect on toads than Nux

In homeopathy some drugs are known to act as complementary, antidotal or inimical to a particular drug. Practitioners can follow this rule when they apply one drug following another. Potentized Nux vomica can reduce acute hypnotic effect of alcohol on toads. Sulphur and Sepia are reported to be complementary to Nux-vom, while Coffea cruda and Zincum met are antidotal and inimical to Nux, respectively [...] Methods: Five batches of toads, each comprising 20 individuals, were treated by partial immersion in a drug diluted with distilled water 1:500 for 20 min. The control consisted of 90% ethanol diluted with distilled water 1:500. The drugs were Nux vom 200 CH, Sulphur 200 CH, Sepia 200 CH, Coffea 200 CH and Zincum 200 CH. Toads of each batch were separately exposed to 260mM ethanol solution and tested every 10 min to see if they had lost their righting reflex (RR)[...] Results: Toads treated with the five drugs took significantly longer time (P<0.01, one-way ANOVA) to lose RR than those treated with the control. The time taken to lose RR was significantly longer (P<0.01, one-way ANOVA) with Sulphur 200 CH, Sepia 200 CH, Coffea 200 CH and Zincum 200 CH than with Nux vom 200 CH alone. The situation was same when Nux treatment was followed by each of the four drugs. Of the five drugs Coffea showed the strongest anti-hypnotic effect. Conclusion: 1. Drugs complementary, antidotal and inimical to Nux vom showed the same anti-alcoholic effect as Nux in terms of increased tolerance to alcohol anesthesia. 2. The anti-alcoholic effect of Nux vom was markedly superseded by the above four drugs. 3. Of all the drugs tested Coffea showed the strongest anti-alcoholic effect. 4. It appears that the above four drugs produced their individual dominant effect cancelling the individual effect of Nux vom.

Homeopathic drugs complementary, antidotal and inimical to Nux vomica produce stronger anti-alcoholic effect on toads than Nux

In homeopathy some drugs are known to act as complementary, antidotal or inimical to a particular drug. Practitioners can follow this rule when they apply one drug following another. Potentized Nux vomica can reduce acute hypnotic effect of alcohol on toads. Sulphur and Sepia are reported to be complementary to Nux-vom, while Coffea cruda and Zincum met are antidotal and inimical to Nux, respectively [...] Methods: Five batches of toads, each comprising 20 individuals, were treated by partial immersion in a drug diluted with distilled water 1:500 for 20 min. The control consisted of 90% ethanol diluted with distilled water 1:500. The drugs were Nux vom 200 CH, Sulphur 200 CH, Sepia 200 CH, Coffea 200 CH and Zincum 200 CH. Toads of each batch were separately exposed to 260mM ethanol solution and tested every 10 min to see if they had lost their righting reflex (RR)[...] Results: Toads treated with the five drugs took significantly longer time (P<0.01, one-way ANOVA) to lose RR than those treated with the control. The time taken to lose RR was significantly longer (P<0.01, one-way ANOVA) with Sulphur 200 CH, Sepia 200 CH, Coffea 200 CH and Zincum 200 CH than with Nux vom 200 CH alone. The situation was same when Nux treatment was followed by each of the four drugs. Of the five drugs Coffea showed the strongest anti-hypnotic effect. Conclusion: 1. Drugs complementary, antidotal and inimical to Nux vom showed the same anti-alcoholic effect as Nux in terms of increased tolerance to alcohol anesthesia. 2. The anti-alcoholic effect of Nux vom was markedly superseded by the above four drugs. 3. Of all the drugs tested Coffea showed the strongest anti-alcoholic effect. 4. It appears that the above four drugs produced their individual dominant effect cancelling the individual effect of Nux vom. (AU)