Potentiated homeopathic drug Arsenicum Album 30C inhibits intracellular reactive oxygen species generation and up-regulates expression of arsenic resistance gene in arsenite-exposed bacteria Escherichia coli.

OBJECTIVE: To examine if potentiated homeopathic drug Arsenicum Album 30C (Ars Alb 30C) can reduce sodium arsenite-induced toxicity in Escherichia coli. METHODS: E. coli were exposed to low arsenite insult after they grew up to log phase in standard Luria-Bertani medium. E. coli were treated with 1 or 2 mmol/L sodium arsenite alone (control), or Ars Alb 30C was added to the medium of a subset of sodium arsenite-treated bacteria (drug-treated), or homeopathically agitated alcohol was added to the medium containing a subset of sodium arsenite-treated bacteria (placebo-treated). A sub-set of untreated E. coli served as the negative control. Glucose uptake, specific activities of hexokinase, lipid peroxidase (LPO), superoxide dismutase (SOD) and catalase, intra- and extra-cellular sodium arsenite content, cell growth, cell membrane potential, DNA damage, intracellular reactive oxygen species (ROS), adenosine triphosphate (ATP) and free glutathione content and expressions of arsB and ptsG gene in normal control, sodium arsenite-treated, drug-treated and placebo-treated E. coli were analyzed. Treatments were blinded and randomized. RESULTS: In sodium arsenite-treated E. coli, glucose uptake, intracellular ROS, LPO and DNA damage increased along with decrease in the specific activities of hexokinase, SOD and catalase, intracellular ATP and free glutathione contents and cell membrane potential and growth, and there were increases in expression levels of arsB gene and ptsG gene. Ars Alb 30C administration reduced arsenic toxicity in E. coli by inhibiting generation of ROS and increasing tolerance to arsenite toxicity and cell growth. CONCLUSION: Ars Alb 30C ameliorated arsenic toxicity and DNA damage, validating efficacy of ultra-highly diluted remedies used in homeopathy.

Potential of the homeopathic remedy, Arnica Montana 30C, to reduce DNA damage in Escherichia coli exposed to ultraviolet irradiation through up-regulation of nucleotide excision repair genes.

OBJECTIVE: To examine to what degree an ultra-highly diluted homeopathic remedy, Arnica Montana 30C (AM-30C), used in the treatment of shock and injury, can modulate the expression of nucleotide excision repair genes in Escherichia coli exposed to ultraviolet (UV) irradiation. METHODS: E. coli were cultured to their log phase in a standard Luria-Bertani medium and then exposed to sublethal doses of UV irradiation at 25 and 50 J/m(2) for 22.5 and 45 s, respectively. The UV-exposed bacteria were then supplemented with either AM-30C (drug) or placebo (P-30C). The drug-treated and placebo-treated bacteria were subjected to assay for DNA damage and oxidative stress 90 min after UV exposure. Several protocols like comet assay, gel electrophoresis for DNA ladder and intracellular reactive oxygen species (ROS) generation, and biomarker measurement like superoxide dismutase (SOD), catalase (CAT) and reduced glutathione (GSH) were conducted. The mRNA expressions of the excision repair genes like ultraviolet repair uvrA, B and C genes (or also known as excision repair genes) were estimated by reverse transcription-polymerase chain reaction method. RESULTS: The UV-exposed bacteria showed DNA damage and oxidative stress, as revealed by an increase in ROS generation, and a decrease in SOD, CAT and GSH activities. As compared to placebo, the AM-30C-treated bacteria showed less DNA damage and oxidative stress as manifested by a decrease in ROS generation, and an increase in SOD, CAT and GSH activities. AM-30C also up-regulated the expression of repair genes as compared to the control. CONCLUSION: AM-30C helped repair the DNA damage through up-regulation of repair genes and also ameliorated the oxidative stress through the reduction of ROS generation and suitable modulation of anti-oxidative stress enzymes.

Rapid green synthesis of silver nanoparticles from silver nitrate by a homeopathic mother tincture Phytolacca Decandra.

OBJECTIVE: To examine if a homeopathic mother tincture (Phytolacca Decandra) is capable of precipitating silver nanoparticles from silver nitrate (AgNO(3)) and to characterize the biosynthesized nanoparticles for evaluating their biological activities. METHODS: A total of 100 mg of AgNO(3) was added to 20mL of Milli-Q water and stirred vigorously. Then 5mL of the homeopathic mother tincture of Phytolacca Decandra (ethanolic root extract of Phytolacca decandra) was added and stirred continuously. Reduction took place rapidly at 300K and completed in 10 min as shown by stable light greenish-yellow color of the solution which gave colloid of silver nanoparticles. The colloid solution was then centrifuged at 5000×g to separate the nanoparticles for further use. The nanoparticles were characterized by spectroscopic analysis, particle size analysis and zeta potential measurements, and morphology was analyzed by atomic force microscopy. The drug-DNA interaction was determined by circular dichroism spectrophotometry and melting temperature profiles by using calf thymus DNA as the target. The biological activities were determined using a cancer cell line A549 in vitro and using bacteria Escherichia coli and fungus Saccharomyces cerevisiae as test models. RESULTS: Phytolacca Decandra precipitated silver nanoparticles in ambient conditions. The nanoparticles had 91 nm particle size, with polydispersity index of 0.119 and zeta potential of -15.6 mV. The silver nanoparticles showed anticancer and antibacterial properties, but no clear antifungal properties. CONCLUSION: This could be a novel environment-friendly method to biosynthesize silver nanoparticles using a cost-effective, nontoxic manner. The homeopathic mother tincture may utilize this property of nano-precipitation in curing diseases or disease symptoms.

Potentized homeopathic drug Arsenicum Album 30C positively modulates protein biomarkers and gene expressions in Saccharomyces cerevisae exposed to arsenate.

OBJECTIVE: This study examines if homeopathic drug Arsenicum Album 30C (Ars Alb 30C) can elicit ameliorative responses in yeast (Saccharomyces cerevisiae) exposed to arsenate. METHODS: The yeast S. cerevisiae 699 was cultured in a standard yeast extract peptone dextrose broth medium. It was exposed to the final concentration of 0.15 mmol/L arsenate for two intervals, 1 h and 2 h, respectively. The cell viability was determined along with the assessment of several toxicity biomarkers such as catalase (CAT), superoxide dismutase (SOD), total thiol (GSH) and glucose-6-phosphate dehydrogenase (G6PDH), lipid peroxidation, protein carbonylation and DNA damage. Reactive oxygen species (ROS) accumulation, expressions of relevant stress transcription activators like Yap-1 and Msn 2, and mRNA expression of yeast caspase-1 (Yca-1) were also measured. RESULTS: Treatment of arsenate increased lipid peroxidation, protein carbonylation, DNA damage, ROS accumulation and expressions of Yap-1, Msn 2 and Yca-1 and decreased GSH, G6PDH, CAT and SOD. Ars Alb 30C administration decreased lipid peroxidation, protein carbonylation, DNA damage, ROS formation and Msn 2 and Yca-1 expressions and increased cell viability, GSH, G6PDH, CAT and SOD significantly (P<0.05), except for a slight increase in Yap-1 expression. CONCLUSION: Ars Alb 30C triggers ameliorative responses in S. cerevisiae exposed to arsenate.

Analysis of the capability of ultra-highly diluted glucose to increase glucose uptake in arsenite-stressed bacteria Escherichia coli.

OBJECTIVE: Whether ultra-highly diluted homeopathic remedies can affect living systems is questionable. Therefore, this study sees value in the analysis of whether homeopathically diluted glucose 30C has any effect on Escherichia coli exposed to arsenite stress. METHODS: E. coli were cultured to their log phase in standard Luria-Bertani medium and then treated with either 1 mmol/L or 2 mmol/L sodium arsenite, with or without supplementation of either 1% or 3% glucose, an ultra-highly diluted and agitated ethanolic solution (70%) of glucose (diluted 10(60) times), glucose 30C or 70% ethanol (placebo) in the medium. Glucose uptake, specific activities of hexokinase and glucokinase, membrane potential, intracellular adenosine triphosphate (ATP) and expression of glucose permease in E. coli were analyzed at two different time intervals. Arsenic content in E. coli (intracellular) and in the spent medium (extracellular) was also determined. RESULTS: In arsenite-exposed E. coli, the glucose uptake increased along with decreases in the specific activities of hexokinase and glucokinase, intracellular ATP and membrane potential and an increase in the gene expression level of glucose permease. Glucose uptake increased further by addition of 1%, 3% or ultra-highly diluted glucose in the medium, but not by the placebo. CONCLUSION: The results demonstrated the efficacy of the ultra-highly diluted and agitated glucose in mimicking the action of actual glucose supplementation and its ability to modulate expressions of hexokinase and glucokinase enzymes and glucose permease genes, thereby validating the efficacy of ultra-high dilutions used in homeopathy.

An initial report on the efficacy of a millesimal potency Arsenicum Album LM 0/3 in ameliorating arsenic toxicity in humans living in a high-risk arsenic village.

BACKGROUND: Millions of people are at risk of groundwater arsenic contamination, and there is no known remedy that can effectively remove the symptoms of prolonged arsenic poisoning. A potentized homeopathic drug, Arsenicum Album LM 0/3 (Ars Alb LM 0/3), is claimed in homeopathic literature to have the ability to treat symptoms similar to that of arsenic poisoning. OBJECTIVE: This study examines whether Ars Alb LM 0/3 could provide some degree of amelioration for the victims living in an arsenic-affected village where no arsenic-free drinking water is available. DESIGN, SETTING, PARTICIPANTS AND INTERVENTIONS: This study was carried out on volunteers living in an arsenic-affected village where no arsenic-free drinking water is available. Twenty-eight volunteers from the village of Dasdiya, in Haringhata block under Nadia District, West Bengal, India, an arsenic-contaminated village where wells contain 55 to 95 µg/L arsenic, were selected to undertake a double-blind and placebo-controlled trial. The subjects provided samples of blood and urine before and after 2 months of taking either "verum" or "placebo". Another 18 subjects living in an arsenic-free village, served as the negative controls. MAIN OUTCOME MEASURES: Samples of blood and urine from the subjects were assayed for arsenic content, according to various toxicity biomarkers and pathophysiological parameters. RESULTS: Out of the original 28 subjects, only 14 subjects provided samples while the other 14 dropped out. There were elevated levels of arsenic in the blood and urine, alkaline and acid phosphatases, lipid peroxidation, and glutathione activities and increased blood glucose, triacylglycerol, cholesterol, and low-density lipoprotein cholesterol contents, whereas there were decreased levels of aspartate and alanine aminotransferases, gamma glutamyl transferase, glucose-6-phosphate dehydrogenase contents, high-density lipoprotein cholesterol and packed cell volume in the subjects. After 2 months of homeopathic remedy administration, the verum-fed subjects showed positive modulations within these parameters with slight lowering of matrix metalloproteinase activity as compared with the placebo group. CONCLUSION: Ars Alb LM 0/3 shows potential for use in high-risk arsenic villages as an interim treatment for amelioration of arsenic toxicity until more extensive medical treatment and facilities can be provided to the numerous victims of arsenic poisoning.

Biosynthesized silver nanoparticles by ethanolic extracts of Phytolacca decandra, Gelsemium sempervirens, Hydrastis canadensis and Thuja occidentalis induce differential cytotoxicity through G2/M arrest in A375 cells.

The capability of crude ethanolic extracts of certain medicinal plants like Phytolacca decandra, Gelsemium sempervirens, Hydrastis canadensis and Thuja occidentalis used as homeopathic mother tinctures in precipitating silver nanoparticles from aqueous solution of silver nitrate has been explored. Nanoparticles thus precipitated were characterized by spectroscopic, dynamic light scattering, X-ray diffraction, atomic force and transmission electron microscopic analyses. The drug-DNA interactions of silver nanoparticles were analyzed from data of circular dichroism spectroscopy and melting temperature profiles using calf thymus DNA (CT-DNA) as target. Biological activities of silver nanoparticles of different origin were then tested to evaluate their effective anti-proliferative and anti-bacterial properties, if any, by exposing them to A375 skin melanoma cells and to Escherichia coli C, respectively. Silver nanoparticles showed differences in their level of anti-cancer and anti-bacterial potentials. The nanoparticles of different origin interacted differently with CT-DNA, showing differences in their binding capacities. Particle size differences of the nanoparticles could be attributed for causing differences in their cellular entry and biological action. The ethanolic extracts of these plants had not been tested earlier for their possible efficacies in synthesizing nanoparticles from silver nitrate solution that had beneficial biological action, opening up a possibility of having therapeutic values in the management of diseases including cancer.