Deep vein thrombosis cured by homeopathy: A case report.

Venous thrombosis (VT) of deep vein is a life-threatening condition which may lead to sudden death as an immediate complication due to formation of thrombo-embolism. VT is associated with various risk factors such as prolonged immobilization, inflammation, and/or coagulation disorders including muscular or venous injury. Deep venous thrombosis (DVT) frequently occurs in the lower limb. Successful treatment of DVT exclusively with homeopathic remedies has rarely been recorded in peer-reviewed journals. The present case report intends to record yet another case of DVT in an old patient totally cured exclusively by the non-invasive method of treatment with micro doses of potentized homeopathic drugs selected on the basis of the totality of symptoms and individualization of the case. Since this report is based on a single case of recovery, results of more such cases are warranted to strengthen the outcome of the present study.

Flavonol isolated from ethanolic leaf extract of Thuja occidentalis arrests the cell cycle at G2-M and induces ROS-independent apoptosis in A549 cells, targeting nuclear DNA.

OBJECTIVES: The K-ras gene mutation commonly found in lung adenocarcinomas contributes to their non-invasive expansion. Our main objective here was to develop a chemopreventive agent against K-ras-mutated lung adenocarcinoma cell line like-A549. MATERIALS AND METHODS: We isolated flavonol from ethanolic leaf extract of Thuja occidentalis, and evaluated its apoptotic potentials on A549 cells. They were treated with 1-10 µg/ml of flavonol and viability was tested retaining normal lung cells L-132 as control. We performed assays such as TUNEL, annexin V, cell-cycle and mitochondrial membrane potentials, by FACS analysis. ROS-mediated oxidative stress and drug-DNA interactions were analysed along with gene expression studies for p53, Bax-Bcl2, cytochrome c, the caspase cascade genes and PARP. RESULTS: Flavonol reduced A549 cell viability in a dose- and time-dependent manner (IC50 value = 7.6 ± 0.05 µg/ml following 48 h incubation) sparing normal L-132 cells. It effected G2-M phase cell cycle arrest and apoptosis, as indicated by progressive increase in the sub-G1, annexin V and TUNEL-positive cell populations. Apoptotic effects appeared to be mitochondria-dependent, caspase-3-mediated, but ROS-independent. Analysis of circular dichroism data revealed that flavonol intercalated with nuclear DNA. In vivo studies on non small cell lung carcinoma (NSCLC)-induced mice confirmed anti-cancer potential of flavonol. CONCLUSION: Flavonol-induced apoptosis apparently resulted from intercalation of cells' nuclear DNA. Flavonol inhibited growth of induced lung tumours in the mice, indicating its potential as an effective agent against NSCLC.

Quercetin induces cytochrome-c release and ROS accumulation to promote apoptosis and arrest the cell cycle in G2/M, in cervical carcinoma: signal cascade and drug-DNA interaction.

OBJECTIVES: Small aromatic compounds like flavonoids can intercalate with DNA molecules bringing about conformational changes leading to reduced replication and transcription. Here, we have examined one dietary flavonoid, quercetin (found in many fruit and vegetables), for possible anti-cancer effects, on HeLa cells originally derived from a case of human cervical cancer. MATERIAL AND METHODS: By circular dichroism spectroscopy we tested whether quercetin effectively interacted with DNA to bring about conformational changes that would strongly inhibit proliferation and migration of the HeLa cells. Cytotoxic effects of quercetin on cancer/normal cells, if any, were determined by MTT assay and such depolarization of mitochondrial membrane potential, as a consequence of quercetin treatment, and accumulation of reactive oxygen species (ROS) also were studied, by FACS analysis and expression profiles of different anti- and pro-apoptotic genes and their products were determined. RESULTS: Quercetin intercalated with calf thymus cell DNA and HeLa cell DNA and inhibition of anti-apoptotic AKT and Bcl-2 expression were observed. Levels of mitochondrial cytochrome-c were elevated and depolarization of mitochondrial membrane potential occurred with increase of ROS; upregulation of expression of p53 and caspase-3 activity were also noted. These alterations in signalling proteins and externalization of phosphotidyl serine residues were involved with initiation of apoptosis. Reduced AKT expression suggested reduction in cell proliferation and metastasis potential, with arrest of the cell cycle at G2/M. CONCLUSION: Quercetin would have potential for use in cervical cancer chemotherapy.

Efficacy of a plant extract (Chelidonium majus L.) in combating induced hepatocarcinogenesis in mice.

Ethanolic whole plant extract of Chelidonium majus, extensively used in traditional systems of medicine against various liver ailments, has been tested for its possible anti-tumor, hepato-protective and anti-genotoxic effects in p-dimethylaminoazobenzene (p-DAB) induced hepatocarcinogenesis in mice through multiple assays: cytogenetical, biochemical, histological and electron microscopical. Different sets of mice, 5 (for 7, 15 and 30 days' treatment) or 10 (for 60, 90 and 120 days) each, were chronically fed a diet suitably mixed with p-DAB and phenobarbital to develop liver tumors. One sub-group of carcinogen fed mice was also fed C. majus extract; 0.1 ml daily (drug-treated) while the other equal amount of dilute ethyl alcohol ("vehicle" of plant extract) (positive control). A separate group of mice was maintained with normal diet without any carcinogen treatment (negative control). Data of several cytogenetical endpoints and biochemical assay of some toxicity marker enzymes at all fixation intervals and histology of liver sections through ordinary, scanning and transmission electron microscopy at 60 and 120 days and that of spleen and kidney at 90 days were critically analyzed in the treated lots vis-a-vis controls. The results suggest anti-tumor, anti-genotoxic and hepato-protective effects of the plant extract, showing potentials for use in cancer therapy.

A potentized homeopathic drug, Arsenicum Album 200, can ameliorate genotoxicity induced by repeated injections of arsenic trioxide in mice.

Groundwater arsenic contamination has become a menacing global problem. No drug is available until now to combat chronic arsenic poisoning. To examine if a potentized homeopathic remedy, Arsenicum Album-200, can effectively combat chronic arsenic toxicity induced by repeated injections of Arsenic trioxide in mice, the following experimental design was adopted. Mice (Mus musculus) were injected subcutaneously with 0.016% arsenic trioxide at the rate of 1 ml/100 g body weight, at an interval of 7 days until they were killed at day 30, 60, 90 or 120 and were divided into three groups: (i) one receiving a daily dose of Arsenicum Album-200 through oral administration, (ii) one receiving the same dose of diluted succussed alcohol (Alcohol-200) and (iii) another receiving neither drug, nor succussed alcohol. The remedy or the placebo, as the case may be, was fed from the next day onwards after injection until the day before the next injection, and the cycle was repeated until the mice were killed. Two other control groups were also maintained: one receiving only normal diet, and the other receiving normal diet and succussed alcohol. Several toxicity assays, such as cytogenetical (chromosome aberrations, micronuclei, mitotic index, sperm head anomaly) and biochemical (acid and alkaline phosphatases, lipid peroxidation), were periodically made. Compared with controls, the drug fed mice showed reduced toxicity at statistically significant levels in respect of all the parameters studied, thereby indicating protective potentials of the homeopathic drug against chronic arsenic poisoning.

Preponderance of GC-rich sites in silver-stained nucleolus organizing regions of Rita rita (Hamilton) and Mystus gulio (Hamilton) (Bagridae, Pisces), as revealed by chromomycin A3-staining technique and scanning electron microscopic studies.

The karyotypes of two species of catfish, Rita rita (Hamilton) (2n = 54; 14m + 34sm + 6st; NF = 102) and Mystus gulio (Hamilton) (2n = 58; 30m + 12sm + 2st + 14t, NF = 100) were studied through Giemsa-, silver- and chromomycin A(3)-staining techniques. The silver-stained karyotypes in both sexes of R. rita and M. gulio revealed that the nucleolus organizing regions were located terminally at the shorter arms (Tp) of one pair of submetacentric chromosomes, placed at positions Nos. 2 and 1, respectively, which was confirmed by scanning electron microscopy. Staining with a GC-specific fluorochrome, chromomycin A(3), produced bright fluorescence in the Ag-positive nucleolus organizer regions, suggesting thereby that nucleolus organizing regions actually included GC-rich sites of active r-RNA genes in metaphase chromosomes of these two bagrids. Further such studies are needed due to the extreme paucity of data on fish.

Response to high LET radiation 12C (LET, 295 keV/microm) in M5 cells, a radio resistant cell strain derived from Chinese hamster V79 cells.

PURPOSE: To study the effects of 12C-beam of 295 keV/microm (57.24 MeV) on M5 and Chinese hamster V79 cells by using cytogenetic assays like micronuclei (MN) induction, chromosomal aberrations (CA) and apoptosis. Additionally, the relative survival of these two cell lines was tested by the colony forming ability of the cells, with a view to understanding the mechanism of cellular damages that lead to difference in cell survival. MATERIALS AND METHODS: Confluent cells were irradiated with 12C-beam at various doses using 15UD Pelletron accelerator. Cell survival was studied by the colony forming ability of cells. MN assay was done by fluorescent staining. Different types of chromosomal aberrations in metaphase cells were scored at 12 h after irradiation. Apoptosis was measured at different post irradiation times as detected by nuclear fragmentation and DNA ladder was prepared after 48 h of incubation. RESULTS: Dose-dependent decrease in surviving fractions was found in both the cell lines. However, the surviving fractions were higher in M5 cells in comparison to V79 cells when exposed to the same radiation doses. On the other hand, induced MN frequencies, CA frequencies and apoptosis percentages were less in M5 cells than V79 cells. Very good correlations between surviving fractions and induced MN frequencies or induced total CA or induced apoptosis percentages were obtained in this study. CONCLUSIONS: The cell strain M5 showed relatively more radio-resistance to 12C-beam compared to Chinese hamster V79 cells in this study. As the MN formation, CA and apoptosis induction were less in M5 cells as compared to parental V79 cells, the higher cell survival in the former could possibly be attributed to their better repairing ability leading to higher cell survival.

Preponderance of GC-rich sites in silver-stained nucleolus organizing regions of Rita rita (Hamilton) and Mystus gulio (Hamilton) (Bagridae, Pisces), as revealed by chromomycin A3-staining technique and scanning electron microscopic studies

The karyotypes of two species of catfish, Rita rita (Hamilton) (2n = 54; 14m + 34sm + 6st; NF = 102) and Mystus gulio (Hamilton) (2n = 58; 30m + 12sm + 2st + 14t, NF = 100) were studied through Giemsa-, silver- and chromomycin A(3)-staining techniques. The silver-stained karyotypes in both sexes of R. rita and M. gulio revealed that the nucleolus organizing regions were located terminally at the shorter arms (Tp) of one pair of submetacentric chromosomes, placed at positions Nos. 2 and 1, respectively, which was confirmed by scanning electron microscopy. Staining with a GC-specific fluorochrome, chromomycin A(3), produced bright fluorescence in the Ag-positive nucleolus organizer regions, suggesting thereby that nucleolus organizing regions actually included GC-rich sites of active r-RNA genes in metaphase chromosomes of these two bagrids. Further such studies are needed due to the extreme paucity of data on fish.

Genotoxic effects of cadmium chloride and azadirachtin treated singly and in combination in fish.

The genotoxic effects of cadmium chloride (CdCl(2)) and azadirachtin (Aza) were assessed singly and conjointly in a fish, Oreochromis mossambicus, with endpoints such as chromosome aberrations, abnormal red cell nuclei, abnormal sperm morphology, and protein content (both qualitative and quantitative) of selected tissues, namely, muscle, heart, eye, brain, gill, liver, spleen, and kidney. The primary objectives were, first, to examine if CdCl(2), a common pollutant, and Aza, a natural product of the neem plant used extensively as an 'ecofriendly' agent for many purposes, had any genotoxic effect of their own on nontarget aquatic organisms of economic importance; and second, if Aza could have any ameliorating effect on CdCl(2)-induced genotoxicity in O. mossambicus tissues. As compared with distilled water-treated controls, both CdCl(2) and Aza induced genotoxicity in O. mossambicus, the former in greater quantity than that produced by Aza. However, Cd-induced toxicity in O. mossambicus appeared to be ameliorated to some extent by Aza.

Ameliorating effect of microdoses of a potentized homeopathic drug, Arsenicum Album, on arsenic-induced toxicity in mice.

BACKGROUND: Arsenic in groundwater and its accumulation in plants and animals have assumed a menacing proportion in a large part of West Bengal, India and adjoining areas of Bangladesh. Because of the tremendous magnitude of the problem, there seems to be no way to tackle the problem overnight. Efforts to provide arsenic free water to the millions of people living in these dreaded zones are being made, but are awfully inadequate. In our quest for finding out an easy, safe and affordable means to combat this problem, a homeopathic drug, Arsenicum Album-30, appears to yield promising results in mice. The relative efficacies of two micro doses of this drug, namely, Arsenicum Album-30 and Arsenicum Album-200, in combating arsenic toxicity have been determined in the present study on the basis of some accepted biochemical protocols. METHODS: Mice were divided into different sets of control (both positive and negative) and treated series (As-intoxicated, As-intoxicated plus drug-fed). Alanine amino transferase (ALT) and aspartate amino transferase (AST) activities and reduced glutathione (GSH) level in liver and blood were analyzed in the different series of mice at six different fixation intervals. RESULTS: Both Arsenicum Album-30 and Arsenicum Album-200 ameliorated arsenic-induced toxicity to a considerable extent as compared to various controls. CONCLUSIONS: The results lend further support to our earlier views that microdoses of potentized Arsenicum Album are capable of combating arsenic intoxication in mice, and thus are strong candidates for possible use in human subjects in arsenic contaminated areas under medical supervision.

Karyotype, Ag-NOR, CMA3 and SEM studies in a fish (Mystus tengara, Bagridae) with indication of female heterogamety.

Somatic karyotypes in M. tengara contained 54 chromosomes, comprising 26 homomorphic pairs in both sexes and one pair of heteromorphic nature in female (one big submetacentric and one small subtelocentric chromosomes), while in males this pair was homomorphic (with two big sub-metacentric chromosomes). The Nucleolus Organizer Regions (NORs) were located at one arm of the suspected sex elements in both sexes, while another pair of metacentric chromosomes (No.7) also showed Ag-positive arm. The CMA3 technique revealed relatively bright fluorescing zones in the regions of chromosomes that showed Ag-positive staining, revealing thereby the preponderance of GC-rich active sites of rRNA genes in NORs. SEM studies revealed clear heteromorphism to exist in the elements suspected as sex chromosomes in females.

Efficacy of vitamin-C (L-ascorbic acid) in reducing genotoxicity in fish (Oreochromis mossambicus) induced by ethyl methane sulphonate.

The genotoxic effects of ethyl methane sulphonate (EMS) have been assessed in a fish, Oreochromis mossambicus with endpoints including chromosome aberrations, abnormal red blood cell nuclei, abnormal sperm morphology, and protein content (both qualitative and quantitative) of selected tissues, namely, muscle, heart, eye, brain, gill, liver, spleen and kidney. EMS caused chromosomal aberrations, nuclear anomalies in red blood cells, abnormal sperm morphology, and alteration of protein synthesis in various tissues. Some of the EMS toxicity appeared to be modulated and ameliorated in this fish by vitamin-C treatment.

Assessment of EMS-induced genotoxicity in the Indian climbing perch, Anabas testudineus: cytogenetical vis-à-vis protein endpoints.

Genotoxic effects of EMS have been assessed in fish, A. testudineus, using widely accepted cytogenetic protocols like chromosome aberrations, nuclear anomalies in red blood cells and abnormal sperm head morphology. In addition, gel electrophoretic protein profiles and total protein contents in nine selected tissues were analysed for evaluating their utility as potential indicators of genotoxicity. EMS not only caused chromosomal aberrations in somatic cells, nuclear anomalies in red blood cells, and increased incidence of sperm with abnormal head morphology, but also altered significantly both protein profiles and total protein contents in all tissues tested vis-à-vis suitable controls, indicating relevance of protein data in genotoxicity assessment.

Comparative efficacy of two microdoses of a potentized homoeopathic drug, Cadmium Sulphoricum, in reducing genotoxic effects produced by cadmium chloride in mice: a time course study.

BACKGROUND: Cadmium poisoning in the environment has assumed an alarming problem in recent years. Effective antimutagenic agents which can reverse or combat cadmium induced genotoxicity in mice have not yet been reported. Therefore, in the present study, following the homeopathic principle of "like cures like", we tested the efficacy of two potencies of a homeopathic drug, Cadmium Sulphoricum (Cad Sulph), in reducing the genotoxic effects of Cadmium chloride in mice. Another objective was to determine the relative efficacy of three administrative modes, i.e. pre-, post- and combined pre and post-feeding of the homeopathic drugs. For this, healthy mice, Mus musculus, were intraperitoneally injected with 0.008% solution of CdCl2 @ 1 ml/100 gm of body wt (i.e. 0.8 mcg/gm of bw), and assessed for the genotoxic effects through such studies as chromosome aberrations (CA), micronucleated erythrocytes (MNE), mitotic index (MI) and sperm head anomaly (SHA), keeping suitable succussed alcohol fed (positive) and CdCl2 untreated normal (negative) controls. The CdCl2 treated mice were divided into 3 subgroups, which were orally administered with the drug prior to, after and both prior to and after injection of CdCl2 at specific fixation intervals and their genotoxic effects were analyzed. RESULTS: While the CA, MNE and SHA were reduced in the drug fed series as compared to their respective controls, the MI showed an apparent increase. The combined pre- and post-feeding of Cad Sulph showed maximum reduction of the genotoxic effects. CONCLUSIONS: Both Cad Sulph-30 and 200 were able to combat cadmium induced genotoxic effects in mice and that combined pre- and post-feeding mode of administration was found to be most effective in reducing the genotoxic effect of CdCl2 followed by the post-feeding mode.

Cytogenetical effects of sonication in mice and their modulations by actinomycin D and a homeopathic drug, Arnica 30.

Experiments were designed to examine if Actinomycin D, an antibiotic, and Amica 30, a homeopathic drug used against shock and injury, can ameliorate cytogenetic damage induced by single or multiple exposures to ultrasonication. Separate sets of healthy mice were directly exposed to sonication for two minutes either once or they received multiple exposures at an interval of 20 days. The mice were then assessed at different intervals, against suitable controls, using parameters like chromosome aberrations (CA), mitotic index (MI), sperm head anomaly (SHA) and micronucleated erythrocytes (MNE). Separate groups of sonicated mice were either orally administered with Arnica 30 (alcohol 30 in control) or injected intramuscularly with Actinomycin-D (AMD). Elevated frequencies of CA, MI, MNE and SHA were noted in sonicated series. AMD had genotoxic effects of its own and also had additive effects on sonication induced genotoxicity. Sonicated mice fed with Arnica 30 showed appreciably reduced genotoxicity as against alcohol 30 and distilled water fed controls, thereby showing ameliorating effect which may have human application.

Efficacy of a potentized homeopathic drug (Arsenicum-Aalbum-30) in reducing cytotoxic effects produced by arsenic trioxide in mice: IV. Pathological changes, protein profiles, and content of DNA and RNA.

OBJECTIVE: To examine if the potentized homeopathic drug Arsenicum Album-30 can help restore the damage produced in protein profiles, DNA and RNA contents in liver and testis as a result of arsenic treatment in mice. DESIGN: Sets of mice were injected with arsenic trioxide, one set was fed with Ars. Alb-30, another with Alcohol-30 and the final set was fed neither. The gel electrophoretic protein profiles and DNA and RNA contents in these three sets were studied. METHODS: Protein profiles were studied by SDS-PAGE method; the DNA and RNA contents were assayed by the standard methods through diphenylamine and orcinol reactions respectively. RESULTS: arsenic trioxide injection produced some pathological conditions, drastic changes (mainly reduction of protein bands) in protein sub-fractions, reduced DNA and RNA contents in both liver and testis; Ars. Alb-30-fed arsenic-intoxicated mice showed revival and restoration in both liver and testis as revealed by gel patterns and quantitative assay of DNA and RNA. CONCLUSION: Efficacy of the homeopathic drug Ars. Alb-30 in reducing arsenic-induced damage to protein and nucleic acids is substantiated and the mechanism of action of the homeopathic drug through expression of regulatory genes inferred.

Differential C-heterochromatin distribution in two species of freshwater fish, Anabas testudineus (Bloch.) and Puntius sarana (Hamilton.).

Anabas testudineus (2n = 46) had the more conserved pattern of its C-heterochromatin distributed mainly in the centromeric region, whereas Puntius sarana (2n = 50) exhibited a rather unorthodox pattern, many chromosomes showing interstitial, some telomeric and a few chromosomes showing centromeric C-band localization. Further, lateral asymmetry in distribution of heterochromatin was also noted in two pairs of chromosomes in P. sarana. The possible implications of the differential distribution noted in these two species has been discussed.

Chromosome banding studies in an Indian mullet: evidence of structural rearrangements from NOR locations.

C-, G- and NOR bands have been studied in the female sex of Rhinomugil corsula. (Mugilidae, Pisces) by deploying the conventional methodologies with suitable modifications of minor nature. The diploid metaphase complements contained 48 acrocentric chromosomes. The localization of C-band heterochromatin was found to be mostly at or near the centromeric regions of the acrocentric chromosomes. The G-type bands were not so well defined, but some of the G-banded chromosomes also contained C-bands. Interestingly, silver-positive NORs were found at the telomeric ends of five acrocentric chromosomes, including one homologous pair having NORs in both chromatids, while one chromosome showed NORs in both of its chromatids and the other two had only one NOR localized at one of its chromatids. This would suggest that one homologue of the second pair of NOR-bearing chromosomes possibly underwent a chromatid exchange with a non-NOR bearing chromosome. This is quite a unique situation not reported earlier in any species of fish., though some other form of NOR-polymorphism/heteromorphism has rarely been reported. Therefore, further exploration in natural populations of this species to examine the other sex and to verify if there also exists other chromosomally polymorphic races (in respect of NOR-polymorphism) of this species, would be rewarding.