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Background: Use of ultra-high diluted remedies in homeopathy and their claimed efficacy in curing diseases has been challenged time and again by non-believers despite many evidence-based positive results published in favor of their efficacy in curing/ameliorating disease symptoms. Aims: To test the ability of ultra-high diluted homeopathic remedies beyond Avogadro’s limit, if any, in manifesting gene modulating effects in controlled in vitro experimental model. Methods: Since cancer cells manifest aberrant epigenetic gene expressions, we conducted global microarray gene expression profiling of HeLa cells (an established epigenetic model of HPV18 positive cell line) treated with two different potentized homeopathic remedies, namely, Condurango 30c and Hydrastis canadensis 30C (used in the treatment of cancer), as compared to that of placebo (succussed alcohol 30c). Results: Data revealed distinctly different expression patterns of over 100 genes as a consequence of treatment with both homeopathc remedies compared to placebo. Conclusion: Results indicate that action of the potentized drugs was “more than placebo” and these ultra-highly diluted drugs acted primarily through modulation of gene expression.

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.

Sodium salts of dimethyldithiocarbamate, diethyldithiocarbamate and pyrrolidinedithiocarbamate react with the triply bonded dirhenium (II,II) complex Re2(μ-Ph2PCH2PPh2)2Cl4 (1) in refluxing ethanol to afford the products of the type Re2(μ-Ph2PCH2PPh2)(LR)4 (2(LR)) where LR represents the dithiocarbamato ligands [LR = S2CNMe2, 2(LMe); S2CNEt2, 2(LEt) and S2CN(CH2)4, 2(LPyr)]. These are the first examples of dithiocarbamato chelated dirhenium complexes with a Re24+ core containing Ph2PCH2PPh2 (dppm) ligand. The spectral (IR, UV-vis, NMR, ESI-MS) and electrochemical properties of the complexes are reported. The identity of 2(LEt) has been established by single-crystal X-ray structure determination (Re–Re distance 2.3196(8) A). The electronic structure and the absorption spectra of the complexes are scrutinized by density functional theory (DFT) and time-dependent DFT (TD-DFT) analyses. DFT analysis shows that the highest occupied molecular orbitals are mainly metal δ* based orbitals. The complex 2(LEt) was also screened in vitro for its antiproliferative properties against human non-small cell lung carcinoma cells (A549), the cervical cancer cell line (HeLa) and peripheral blood mononuclear cells (PBMCs) by MTT assay which indicates its preferential cytotoxicity against the cancer cells (A549, HeLa).

Background: Groundwater arsenic affects millions of people in about 20 countries. In West Bengal (India) and Bangladesh alone over 100 million people are exposed. The arsenic concentration in contaminated groundwater in Bangladesh was above the maximum permissible level of 0.05 mg/l as recommended by WHO for developing countries [1]. Drinking water is not the only source of poisoning. In arsenic contaminated areas, crops, vegetables, cereals, poultry, cattle, etc, also contain traces of arsenic. Chronic arsenic intoxication has been associated with several diseases such as melanosis, leuco-melanosis, hyperkeratosis, oedema, skin cancer… Cazin et al [2], have demonstrated the effect of high dilutions of arsenic compounds. They noted increased arsenic elimination from blood through urine and faeces in intoxicated rats. According to these research, the aim of Khuda Buksh studies [3-4-5] was to investigate whether high dilution Arsenicum album have any effect on arsenic accumulation in different tissues and to understand also how this high dilution could produce a protective effect on all the different organs. Methodology: Firstly, the effect of Arsenicum album 30 cH on the amount of arsenic accumulation was determined by spectrophotometric analysis in four tissues namely liver, kidney and testis in mice intoxicated by arsenic. The protective effect in chronic and acute arsenic intoxicated mice of Arsenicum Album 6cH, 30cH and 200cH has been evaluated using not only the activities of enzymatic and biomarker toxicity (aspartate amino transferase (AST), alanine amino transferase (ALT), acid phosphatase (AcP), alkaline phosphatase (AlkP), lipid peroxidation (LPO) and reduced glutathione (GSH)) but also the cytogenetical parameters (chromosome aberrations (CA), mitotic index (MI), sperm head anomaly (SHA) etc., ). Because, it is well demonstrated that these enzymes biomarkers reflect the degree of hepatotoxicity and oxidative stress caused by arsenic intoxication. Results: Compared to controls, Arsenicum album 30cH induced a significant decrease in accumulation of arsenic in 4 tissues namely liver, spleen, kidney and testis in intoxicated mice. In addition, both Arsenicum album 6cH, 30cH and 200cH reduced chromosome aberrations, sperm head abnormality frequencies and activities of acid and alkaline phosphatases, aspartate and alanine aminotransferases and lipid peroxidation, while mitotic index and activities of glutathione, catalase and succinate dehydrogenase were increased compared to controls. Conclusion: Altogether, theses results provide evidence of protective potentials of the Arsenicum album dilution against acute and chronic arsenic intoxication in mice. They also offer a new hypothesis that the mechanism of the homeopathic dilution could act through regulation of expression of certain genes. This explanation seems to be plausible because all biomarker tests are regulated by specific genetic regulatory mechanisms [6]. keywords: Arsenicum album, arsenic intoxication, enzymatic and biomarker toxicity. References: [1] WHO. WHO Guidelines for Drinking Water Quality, Vol. 2, 2nd edition. Geneva: WHO, 1996, 940–9. [2] Cazin JC, Cazin M, Gaborit JL, Chaoui A, Boiron J, Belon P, et al. A study of the effect of decimal and centesimal dilutions of arsenic on the retention and mobilization of arsenic in the rat. Hum Toxicol 1987;6:315–20. [3] Mitra K, Kundu SN, Khuda-Bukhsh AR. Efficacy of a potentized homoeopathic drug (Arsenicum Album-30) in reducing toxic effects produced by of arsenic trioxide in mice. I. On rate accumulation of arsenic in certain vital organs. Comp Ther Med 1998;6:178–84. [4] Pathikrit Banerjee. Evidences of Protective Potentials of Microdoses of Ultra-high Diluted Arsenic Trioxide in Mice Receiving Repeated Injections of Arsenic Trioxide. eCAM 2009; 1-10. [5] Pathikrit Banerjee, Comparative Efficacy of Two Microdoses of a Potentized Homeopathic Drug, Arsenicum Album, to Ameliorate Toxicity Induced by Repeated Sublethal Injections of Arsenic Trioxide in Mice. Pathobiology 2008;75:156–170. [6]/ Khuda-Bukhsh AR. Potentized homeopathic drugs act through regulation of gene expression: a hypothesis to explain their mechanism and pathways of action in vivo. Comp Ther Med 1997;5:43–6

Stem cells are those cells of the body (somatic cells) that can divide and become differentiated into different cell types as the living organism grows. They may be used at appropriate time to replace cells and tissues that have been damaged or lost due to disease and to perform specialized functions of the particular cells they replenished. Aging refers to the biological process involved in becoming older, and the term is applicable to virtually all living organisms from bacteria through man and some perennial plants. There are several ways through which stem cells play a role in the process of aging. Spontaneous or induced damage and/or errors take place and accumulation of these products in the genetic material has been implicated in the process of aging. The replacement mechanism of the damaged tissues assisted by different stem cells is reported to be very efficient in young people than that of the older ones. Thus, aging is not always associated with the accumulation of damage but may also reflect inability of the stem cells to efficiently maintain the replenishment mechanism of the differentiation process needed for controlling the repair of specific damaged tissue or organ. Maintaining a subtle and dynamic balance of the stem cell pool is a prerequisite for optimum homeostasis of proliferation and propagation of stem cell niche, which in turn favors the elemental factors for their differentiation into different progenies or lineages, thereby implicating their possible role in stalling the overall process of aging.

The human body is constantly exposed to a variety of oxidizing agents from naturally occurring processes, environmental stimuli/pollutants, and through lifestyle-related stress/activities. The key factor in a biological system is to maintain a balance between oxidants and antioxidants to sustain optimal physiological conditions. Imbalance of the process leads to oxidative stress which causes DNA damage and genomic instability resulting in an increased risk of cancer. Immune cells are one of the major components of the tumor microenvironment and can be affected by both oxidative stress and subsequent DNA damage response (DDR) in the tumor cells. This review summarizes the current knowledge of DDR and immune microenvironment in therapeutic cancer research and role of some food plants and their phytochemicals as antioxidants in targeted cancer therapy.

Objective: Psorinum therapy is claimed to combat/ameliorate a variety of human cancers, but for obvious reasons these studies are devoid of any untreated/placebo-treated controls. Therefore, if Psorinum 6x administration to cancer cells of different origin can show visible anti-cancer effects in a controlled in vitro study has been examined. Materials & Methods: Psorinum 6x was provided by Hahnemann Publishing Company (HAPCO), 165 BB Ganguly Street, Kolkata for our research. It was prepared by HAPCO by following standard homoeopathic guidelines from authentic pus cells obtained from an eczema patient being treated at National Institute of Homeopathy, Salt Lake, Kolkata. MTT assay was initially done on several cancer cell lines like A549 (lung cancer), HeLa (cervix cancer), HepG2 (liver cancer) and MCF7 (breast cancer). Psorinum 6x showed strongest anticancer effect against A549 though it also had lesser effect against other cell lines tested. Therefore, A549 was chosen as the model cell line for further study using several relevant protocols. Effects of Psorinum 6x were compared with that of "Placebo 6x" control made of the same stock of "vehicle" used for preparation of Psorinum 6x. Protocols like analysis of cell cycle progression, generation of reactive oxygen species (ROS), change in mitochondrial membrane potential (MMP) and actual cell death (apoptosis), if any, were analysed flow-cytometrically. Whether Psorinum 6x could damage DNA and induce morphological changes were also determined microscopically. Expression of different signal proteins related to cell death (apoptosis) and survival were critically studied by western blot analysis and confocal microscopy. Further, to determine if Psorinum 6x could interact directly with DNA to induce any conformational changes was also determined by circular dichroism (CD) spectroscopy. Results: Psorinum 6x treatment reduced cell viability and inhibited cell proliferation at 24h after treatment, arresting cell cycle at sub-G stage. Upon Psorinum treatment, there were increase of ROS and MMP depolarization, morphological changes and DNA damage typical of apoptosis in A549 cells, along with externalization of phosphatidyl serine. Further, an increase in p53 level, Bax translocation into mitochondria, cytochrome c release into cytosol along with reduction of Bcl2 level and caspase 3 activation were noted which eventually drove A549 cells towards apoptosis; the apoptotic signalling was found to be through mitochondria-mediated caspase 3 dependent pathway. Evidence of direct interaction of Psorinum with cellular DNA was revealed from CD-spectroscopy. Conclusion: Thus, Psorinum 6 x had positive anti-cancer effects against a number of cancer cells, of which it appeared to have strongest effect against the lung cancer cell, A549.

ZusammenfassungIn den letzten Jahren wurden mithilfe modernster Elektronenmikroskopie durch mehrere Autoren Nanopartikel von den Ausgangssubstanzen von Arzneien nachgewiesen. Während man annimmt, dass diese Nanopartikel von Arzneien in Hochpotenzen von der homöopathischen Potenzierung herrühren, haben wir mit einer wissenschaftlich anerkannten Methode Nanokapseln aus den Extrakten homöopathischer Urtinkturen und ihren biologisch aktiven Inhaltsstoffen produziert, wofür wir das nicht-toxische, biologisch abbaubare Polymer Poly(lactid-co-glycolid) nutzten sowie Mechanismus und Wirkungspfade ihrer biologischen Aktivität untersuchten.

Homoeopathy has been confronted with certain major issues such as (i) how is medicinal property of homoeopathic drug transferred to and retained by the vehicle; (ii) how can it act in biological system in the absence of any original drug substance in ultra high diluted remedies? and (iii) plausible mechanism and pathways of biological action including mysteries of ‘like cured by likes’. We demonstrated through controlled studies the ability of ultra-highly diluted homoeopathic drugs in rendering protection/repair of cytogenetic damages inflicted by whole-body X-irradiation in mammalian model mice in vivo. We examined cytogenetic damages in arsenic, cadmium, mercury and stannum intoxicated mice and their remediation by homoeopathic drugs derived from agitated ultra-high dilutions of their respective salt, deploying many scientific protocols. We made a human trial on efficacy of Arsenicum album against groundwater arsenic-contaminated victims. We tested efficacy of some homoeopathic remedies in cancer of mice and thalassemia in human. We produced nano-capsules of homoeopathic mother tinctures and their bioactive components and induced nano-precipitation of silver from silver nitrate by homoeopathic mother tinctures and characterised them for their physicochemical properties and biological action. We studied tissue distribution of nanoparticles, precise mechanism and pathways of their action that involved certain signal proteins and their pathways, both in vivo and in vitro. This approach made a significant contribution towards elucidating the role of drug nanoparticles in inducing ‘memory of water’ and mechanism and pathways of action of homoeopathic remedies through epigenetic modifications that supported ‘gene regulatory hypothesis’.

Probiotic Pediococcus pentosaceus GS4 produces conjugated linoleic acid (CLA) through biohydrogenation, which may be vital for colon cancer mitigation. Here, we have appraised the anti-cancer potential of probiotic GS4 against colon cancer. CLA produced by probiotic GS4 reduced the proliferation of HCT-116 in vitro rendering towards apoptosis and subsequently abrogated NF-κB and p-Akt. Probiotic GS4 efficiently mitigated colon cancer in azoxymethane (AOM)-induced colon cancer mice model, which was evident from reduced disease severity, alleviated oxidative stress and neoplastic characters. Further delineation of underlying consequences at the level of host–microbe interface, the probiotic GS4 arbitrated the gut microflora-associated biohydrogenation, which subsequently triggered apoptosis in colonocytes, apparent from PARP cleavage, caspase 3 activity, DNA fragmentation and inhibition of HDAC activity. Conclusively, modulation of host–microbe interface with selective colonizing strategies combined with unique biohydrogenation ability of probiotic GS4 promises to be a therapeutic/functional food supplement responsible for mitigation of colon cancer.

Objectives This study examined the relative efficacies of a derivative of betulinic acid (dBA) and its poly (lactide- co-glycolide) (PLGA) nano-encapsulated form in A549 lung cancer cells in vivo and in co-mutagen [sodium arsenite (SA) + benzo]undefined[a]pyrene (BaP)]-induced lung cancer in mice in vivo. Methods dBA was loaded with PLGA nanoparticles by using the standard solvent displacement method. The sizes and morphologies of nano-dBA (NdBA) were determined by using transmission electron microscopy (TEM), and their intracellular localization was verified by using confocal microscopy. The binding and interaction of NdBA with calf thymus deoxyribonucleic acid (CT-DNA) as a target were analyzed by using conventional circular dichroism (CD) and melting temperature (Tm) profile data. Apoptotic signalling cascades in vitro and in vivo were studied by using an enzyme-linked immunosorbent assay (ELISA); the ability of NdBA to cross the blood-brain barrier (BBB) was also examined. The stage of cell cycle arrest was confirmed by using a fluorescence-activated cell-sorting (FACS) data analysis. Results The average size of the nanoparticles was ~ 110 nm. Confocal microscopy images confirmed the presence of NdBA in the cellular cytoplasm. The bio-physical properties of dBA and NdBA ascertained from the CD and the Tm profiles revealed that NdBA had greater interaction with the target DNA than dBA did. Both dBA and NdBA arrested cell proliferation at G0/G1, NdBA showing the greater effect. NdBA also induced a greater degree of cytotoxicity in A549 cells, but it had an insignificant cytotoxic effect in normal L6 cells. The results of flow cytometric, cytogenetial and histopathological studies in mice revealed that NdBA caused less nuclear condensation and DNA damage than dBA did. TEM images showed the presence of NdBA in brain samples of NdBA fed mice, indicating its ability to cross the BBB. Conclusion Thus, compared to dBA, NdBA appears to have greater chemoprotective potential against lung cancer.

To provide in vitro evidence of Psorinum treatment against cancer cells in a controlled study.Effects of homeopathic Psorinum 6× on cell viability were initially determined in several cancer cell lines, including A549, HepG2 and MCF-7, using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, and an ethanol 6× control. The cell line that exhibited highest inhibition was selected and used in the following experiments. A range of Psorinum 6× doses was used to explore treatment effects on cell cycle arrest, cell death (apoptosis), generation of reactive oxygen species (ROS) and change in mitochondrial membrane potential (MMP) using flow cytometry and fluorescence microscopy, respectively. Expression of several signal proteins related to apoptosis and cell survival were quantified with Western blotting and confocal microscopy. Further, circular dichroism (CD) spectroscopy was used to determine possible drug-DNA interactions, as well as the induction of conformational changes.Treatment of cancer cell lines with Psorinum showed greater anticancer effects in A549 cells than in others. In A549 cells Psorinum treatment inhibited cell proliferation at 24 h after treatment, and arrested cell cycle at sub-G1 stage. It also induced ROS generation, MMP depolarization, morphological changes and DNA damage, as well as externalization of phosphatidyl serine. Further, increases in p53 expression, Bax expression, cytochrome c release, along with reduction of Bcl-2 level and caspase-3 activation were observed after Psorinum 6× treatment, which eventually drove A549 cells towards the mitochondria-mediated caspase-3-dependent pathway. CD spectroscopy revealed direct interaction of Psorinum with DNA, using calf thymus-DNA as target.Psorinum 6× triggered apoptosis in A549 cells via both up- and down-regulations of relevant signal proteins, including p53, caspase-3, Bax and Bcl-2.

Removal of bio-accumulated pesticides in edible fish is a global problem. In this study, we tested protective capability of a phytochemical pelargonidin-loaded non-toxic, biodegradable poly-lactide-co-glycolide nano-particles (NPG) against toxicity induced by a pesticide cypermethrin (CM) in a fish model (Oreochromis mossambica) in vivo and also in L6 muscle cell line, in vitro. First we assessed potential sustainable release of nanoparticles following oral administration of NPG to fish, their ability to cross sub-cellular membranes in several tissues and efficacy to cross blood-brain-barrier. Next, protective ability of NPG, if any, against CM in fish was evaluated deploying parameters like % cell viability, DNA damage in muscle cells and modulation of anti-oxidative-enzymes like superoxide dismutase, catalase and lipid peroxidase. Modulation of reactive oxygen species generation, nuclear condensation and alteration in stress related protein signalling cascade were assessed in L6 cells. Results revealed that NPG had nano-size range (~10–12 nm) and negative zeta potential (−17 mV). Bioavailability and distribution of NPG could be followed by spectrophotometric absorbance of pelargonidin at 293 nm from 6 h onward till 24 h in all important tissues including the brain. Thus, 0.5 mg/g b.w. NPG could demonstrate protective ability in CM-intoxicated fish muscle cells in respect of % cell viability, DNA damage and stress related enzymes. Similar alterations could also be found in signalling protein cascade in L6 cells in response to treatment of 5 μg/ml NPG against CM-induced toxicity and depletion of overall ROS generation and nuclear condensation. Therefore, NPG could be used as a potential drug in management of pesticide toxicity in cultured edible fish.

Garlic (

Proper fusion of fluorescence mechanisms in a single fluorophore unit is highly desirable to obtain better sensitivity, as well as selectivity towards a particular metal ion. In this regard, a Zn2+ chemosensor exhibiting strong fluorescence through synergistic CHEF-PET-ESIPT fluorescence mechanism has not received substantial deployment. This type of chemosensor provides an ample avenue to detect metal ions with a large stokes shift and are useful in living cell imaging study. Herein, we report an unparalleled Zn2+ chemosensor (L) based on these three types of fluorescence mechanisms by condensing 2,6-diformyl-4-methylphenol and 2-hydrazinyl-4,6-dimethylpyrimidine. All the three types of mechanism were experimentally verified by UV-Vis, fluorescence and time resolved photoluminescence (TRPL) spectroscopic evaluation. The sensor L and Zn2+ formed a 1 : 1 ratio complex demonstrating a high binding constant value (Ka = 4.812 × 105 M−1) and L showed clear discrimination from the Cd2+ ion. The limit of detection (LOD) and limit of quantification (LOQ) were calculated to be 9.727 × 10−7 (M) and 3.24 × 10−6 (M), respectively. We have also shown that cell imaging can be performed using L without many cytotoxicity concerns.

The dipicolylamine (DPA) motif which is known as a binder of Zn(II) ions, has been utilized in devising rhodamine labelled compounds 1 and 2. Compound 1 acts as a FRET sensor and shows excellent selectivity for Cu(II) ions over a series of other cations in CH3CN/H2O by exhibiting a colour change (colourless to pink) of the solution. The spectral and colour changes are recovered in the presence of CN− ions and thus, the ensemble 1·Cu2+ in CH3CN/H2O is established as the medium for selective detection of CN− ions. In contrast, the modified compound 2 with the dipicolylamine motif as the principal binding site has been established as the colorimetric sensor of Cu(II) ions and the fluorometric sensor of Hg(II), Zn(II) and Cd(II) ions. Both the compounds 1 and 2 are cell permeable and are successfully employed for the detection of intercellular metal ions through bright field and fluorescence imaging.

A new fluorene–pyridine linked imine conjugate, HAFPA, has been synthesized and characterized. HAFPA has been successfully found to recognize Zn2+ and Cd2+ selectively among a wide range of biologically relevant metal ions. The chemosensing behaviour of the ligand has been demonstrated through 1H NMR, ESI-MS, emission, absorption, colour change, luminescence spectra. The probe HAFPA displays a prominent fluorescence enhancement along with a large red shift (∼60 nm and 65 nm), which may be because of the CHEF (chelation-enhanced fluorescence) processes after its interaction with Zn2+ and Cd2+ respectively. The binding of the ligand with Zn2+ and Cd2+ was investigated through ESI-MS, Job's plot and reversibility studies. The fluorescence property of the ligand after complexation with Zn2+ and Cd2+ can be restored in the presence of EDTA and sulphide ions. This reversibility phenomenon is utilized in the construction of a logic gate. The DFT, TDDFT and triplet excited state calculations were performed in order to demonstrate the structure of the ligand and its complexes and their electronic properties. Importantly, our chemosensor could be used to detect intracellular Zn2+ and Cd2+ with no cytotoxicity in A549 cells, and a colorimetric test strip of the ligand for the detection of both the ions could be useful for all practical purposes.

A novel fluorescent chemosensor L has been designed and synthesized based on the rhodamine-B moiety. The dye L exhibits selective fluorescence enhancement towards the Al3+ ion over other biologically relevant metal ions in aqueous ethanolic media (EtOH–H2O, 2 : 3, v/v). The structure of the probe L has been established by 1H and 13C-NMR spectroscopy, single crystal X-ray diffraction, ESI-mass spectrometry and elemental analysis. The cleavage of the spirolactam bond of the rhodamine moiety induced by the Al3+ ion generates the delocalized xanthene fluorophore that is responsible for the emission enhancement of the probe L. The recognition behavior of the receptor L has been investigated experimentally with supports from theoretical DFT studies. Furthermore, the efficacy of L in cell imaging studies is also probed by confocal microscopy.

Bismuth sulphide nanoparticles are prepared by sol–gel method using gelatine as capping agent. Synthesized nanoparticles are well dispersed in water and the medium is nearly neutral, both essential for biological application. Triethanolamine is used as a complexing agent for solubilization of Bi3+ in water and sodium sulphide as a sulphur source. Nanoparticles are well characterized by standard methods and show marked cytotoxicity and alter membrane potentials of both mitochondria and HeLa cells. Schematic route to synthesize water-soluble Bi2S3 nanoparticles

OBJECTIVES: Condurango (Gonolobus condurango) extract is used by complementary and alternative medicine (CAM) practitioners as a traditional medicine, including homeopathy, mainly for the treatment of syphilis. Condurango bark extract is also known to reduce tumor volume, but the underlying molecular mechanisms still remain unclear. METHODS: Using a cervical cancer cell line (HeLa) as our model, the molecular events behind condurango extract's (CE's) anticancer effect were investigated by using flow cytometry, immunoblotting and reverse transcriptase-polymerase chain reaction (RT-PCR). Other included cell types were prostate cancer cells (PC3), transformed liver cells (WRL-68), and peripheral blood mononuclear cells (PBMCs). RESULTS: Condurango extract (CE) was found to be cytotoxic against target cells, and this was significantly deactivated in the presence of N-acetyl cysteine (NAC), a scavenger of reactive oxygen species (ROS), suggesting that its action could be mediated through ROS generation. CE caused an increase in the HeLa cell population containing deoxyribonucleic acid (DNA) damage at the G zero/Growth 1 (G0/G1) stage. Further, CE increased the tumor necrosis factor alpha (TNF-I±) and the fas receptor (FasR) levels both at the ribonucleic acid (RNA) and the protein levels, indicating that CE might have a cytotoxic mechanism of action. CE also triggered a sharp decrease in the expression of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-IoB ) both at the RNA and the protein levels, a possible route to attenuation of B-cell lymphoma 2 (Bcl-2), and caused an opening of the mitochondrial membrane's permeability transition (MPT) pores, thus enhancing caspase activities. CONCLUSION: Overall, our results suggest possible pathways for CE mediated cytotoxicity in model cancer cells.

A pyridoxal-based fluorescent probe HL was synthesized for the detection of Cu(2+) in methanol with moderate selectivity. Upon addition of Cu(2+), to the solution of the probe in methanol exhibited a remarkable change in emission at 500 nm. With the limit of detection of 10 μM, the probe could well meet the recommended (less than 32 μM in drinking water) of the World Health Organization (WHO). The intracellular Cu(2+) imaging behaviour of HL was carried out on HeLa cells.

An 8-aminoquinoline based fluorescent Zn2+ sensor, 2-methoxy-6-((quinolin-8-ylimino)methyl)phenol (HL), was synthesized. It has been found to display quick responses through fluorogenic properties on selective 2:1 binding to Zn2+, as delineated by absorption and fluorescence titrations as well as by Job’s method and X-ray crystallography studies. This probe features visible light excitation (461 nm) and emission (594 nm) profiles, excellent selectivity responses for Zn2+ over other competing biological metal ions with high binding constant and about 17-fold enhancement in fluorescence quantum yield (Φ) upon half equivalent of Zn2+ binding under physiological pH window. The low detection limit in micromolar range (1.3 × 10−7 M) for Zn2+, makes HL a suitable candidate for the development of a potential probe. It also exhibits cell permeability and intracellular Zn2+ sensing in HeLa cells from human cervical cancer cell and rat hippocampal slices.

Controlled release of medications remains the most convenient way to deliver drugs. In this study, we precipitated gold nanoparticles with quercetin. We loaded gold-quercetin into poly(DL-lactide-co-glycolide) nanoparticles (NQ) and tested the biological activity of NQ on HepG2 hepatocarcinoma cells to acquire the sustained release property. We determined by circular dichroism spectroscopy that NQ effectively caused conformational changes in DNA and modulated different proteins related to epigenetic modifications and cell cycle control. The mitochondrial membrane potential (MMP), reactive oxygen species (ROS), cell cycle, apoptosis, DNA damage, and caspase 3 activity were analyzed by flow cytometry, and the expression profiles of different anti- and pro-apoptotic as well as epigenetic signals were studied by immunoblotting. A cytotoxicity assay indicated that NQ preferentially killed cancer cells, compared to normal cells. NQ interacted with HepG2 cell DNA and reduced histone deacetylases to control cell proliferation and arrest the cell cycle at the sub-G stage. Activities of cell cycle-related proteins, such as p21(WAF), cdk1, and pAkt, were modulated. NQ induced apoptosis in HepG2 cells by activating p53-ROS crosstalk and induces epigenetic modifications leading to inhibited proliferation and cell cycle arrest.

Objectives: Quercetin, a flavonoid compound, has been reported to induce apoptosis in cancer cells, but its anti-inflammatory effects, which are also closely linked with apoptosis, if any, on non-small-cell lung cancer (NSCLC) have not so far been critically examined. In this study, we tried to determine if quercetin had any demonstrable anti-inflammatory potential, which also could significantly contribute to inducing apoptosis in a NSCLC cell line, A549. Methods: In this context, several assays, including cytotoxicity, flow cytometry and fluorimetry, were done. Gene expression was analyzed by using a western blot analysis. Results: Results revealed that quercetin could induce apoptosis in A549 cells through mitochondrial depolarization by causing an imbalance in B-cell lymphoma 2/ Bcl2 Antagonist X (Bcl2/Bax) ratio and by down-regulating the interleukine-6/signal transducer and activator of transcription 3 (IL-6/STAT3) signaling pathway. An analysis of the data revealed that quercetin could block nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) activity at early hours, which might cause a down-regulation of the IL-6 titer, and the IL-6 expression, in turn, could inhibit p-STAT3 expression. Down-regulation of both the STAT3 and the NF-κB expressions might, therefore, cause down-regulation of Bcl2 activity because both are major upstream effec tors of Bcl2. Alteration in Bcl2 responses might result in an imbalance in the Bcl2/Bax ratio, which could ultimately bring about mitochondria mediated apoptosis in A549 cells. Conclusion: Overall, the finding of this study indicates that a quercetin induced anti-inflammatory pathway in A549 cells appeared to make a significant contri bution towards induction of apoptosis in NSCLC and, thus, may have a therapeutic use such as a strong apoptosis inducer in cancer cells.

Garlic (Allium sativum L.), a popular spice, has been used for decades in treating several medical conditions. Although Allicin, an active ingredient of garlic has been extensively studied on carci...

Background: DNA hypermethylation induces cancer progression involving CpG island of DNA and causes inactivation of tumour suppressor genes. In this study, DNA hypermethylation status of lung cancer and ability of ultra-highly diluted Condurango 30C to modulate DNA methylation were ascertained by analysis of lung cancer-specific tumour suppressor genes in respect to placebo. Materials and Methods: DNA methylation status, if any, was determined by PCR-SSCP analyses in lung cancer-specific tumour suppressor genes (p15, p16 and p53) using H460-NSCLC cell and BaP-induced lung cancer of rats. The ability of Condurango 30C to modulate DNA methylation, if any, was verified against placebo control in blinded manner. Results: Condurango 30C-treated DNA showed significant decrease in band intensity of p15 and p53 genes especially in methylated condition in vitro, at IC50 dose (2.43µl/100µl). SSCP analysis of p15 and p53 genes in Condurango 30C-treated DNA also suggests that Condurango 30C can decrease methylation, in vitro. Inhibition of p15 hypermethylation was observed in post-cancer treatment of rats with Condurango 30C. SSCP results gave a better indication of differences in band position of p15 and p53 in Condurango 30C-treated lung samples. Conclusion: Condurango 30C could trigger epigenetic modification in lung cancer via modulation of DNA hypermethylation.

Pyridinium-based chemosensor 1 built on tris(aminomethyl)amine (tren) has been designed, synthesized and established as a chemosensor for ATP over ADP, AMP and a series of other anions in aqueous CH3CN at pH 6.5. Compound 1 exhibits a significant change in emission upon complexation of ATP. In CH3CN, the sensor selectively binds H2PO4− and forms an excimer with significant intensity. Furthermore, the intracellular ATP detection using 1 was possible through fluorescent confocal imaging.

Objectives: To examine if HIV nosode in 30c dilution (HIV 30c) has therapeutic potential against lung cancer cells (A549) as compared to WRL-68 normal cells and to elucidate its possible molecular mechanism of action on DNA replication and apoptosis. Methods: Effects of HIV 30c were thoroughly tested for its possible anticancer potential on A549 cells (lung cancer); WRL-68 normal liver cells served as control. Three doses, one at LD50 and two below LD-50, were used. Proliferation, migration and senescence assays were made and generation of reactive oxygen species (ROS) studied by routine techniques. The ability of HIV 30c to induce apoptosis in A549 cells and its possible signalling pathway were determined using immunoblots of relevant signal proteins and confocal microscopy, including studies on telomerase reverse transcriptase (TERT) and topoisomerase II (Top II) activities, intimately associated with cell division and DNA replication. Results: HIV 30c prevented cancer cell proliferation and migration, induced pre-mature senescence, enhanced pro-apoptotic signal proteins like p53, bax, cytochrome c, caspase-3 and inhibited anti-apoptotic signal proteins Bcl2, TERT and Top II, changed mitochondrial membrane potential and caused externalization of phosphatidyl serine. Thus, it induced apoptosis as also evidenced from increase in cells with distorted membrane morphology, nuclear condensation, DNA fragmentation, and ROS, typical of apoptosis in progress. Conclusion: HIV 30c nosode has therapeutic potential for inducing cytotoxic effects on A549 cells as manifested by changes in nuclear condensation, DNA fragmentation, ROS generation and MMP, and for its inhibitory action on cell proliferation, cell migration, expression of telomerase reverse transcriptase and Top II genes, and increasing expression of pro-apoptotic genes. • HIV nosode in 30th centesimal dilution induces apoptosis in cancer cells. • It acts by inhibiting cell proliferation and migration in A549 cells. • It down-regulates expression of TERT and topoisomerase II enzymes. • It induces conformational changes in DNA with associated senescence.

Increased evidence of chemo-resistance, toxicity and carcinogenicity necessitates search for alternative approaches for determining next generation cancer therapeutics and targets. We therefore tested the efficacy of plant alkaloid berberine on human papilloma virus (HPV) -18 positive cervical cancer cell HeLa systematically-involving certain cellular, viral and epigenetic factors. We observed disruptions of microtubule network and changes in membrane topology due to berberine influx through confocal and atomic force microscopies (AFM). We examined nuclear uptake, internucleosomal DNA damages, mitochondrial membrane potential (MMP) alterations and cell migration assays to validate possible mode of cell death events. Analytical data on interactions of berberine with pBR322 through fourier transform infrared (FTIR) and gel migration assay strengthen berberine׳s biologically significant DNA binding abilities. We measured cellular uptake, DNA ploidy and DNA strand-breaks through fluorescence activated cell sorting (FACS). To elucidate epigenetic modifications, in support of DNA binding associated processes, if any, we conducted methylation-specific restriction enzyme (RE) assay, methylation specific-PCR (MSP) and expression studies of histone proteins. We also analyzed differential interactions and localization of cellular tumor suppressor p53 and viral oncoproteins HPV-18 E6-E7 through siRNA approach. We further made in-silico approaches to determine possible binding sites of berberine on histone proteins. Overall results indicated cellular uptake of berberine through cell membrane depolarization causing disruption of microtubule networks and its biological DNA binding abilities that probably contributed to epigenetic modifications. Results of modulation in p53 and viral oncoproteins HPV-18 E6-E7 by berberine further proved its potential as a promising chemotherapeutic agent in cervical cancer.

The well known rhodamine framework offers an ideal model for the development of fluorescence enhanced chemosensors. Herein, a novel and simple molecule chemosensor, (E)-2-((benzo[d][1,3]dioxol-4-ylmethylene)amino)-3′,6′-bis(diethylamino)spiro[isoindoline-1,9′-xanthen]-3-one ( L ), has been designed by combining a rhodamine B hydrazide and a benzo[d][1,3]dioxole-4-carbaldehyde in a single molecule to prove the selectivity and sensitivity for Al 3+ and Cr 3+ in a dual-channel mode (fluorescence emission and UV–Vis). The signal transduction occurred by the increase of conjugation in the ring-open form than in the ring-closed form. Furthermore, the chemosensor L could also be used as an imaging probe without cytotoxicity for uptake and detection of Al 3+ ion in HeLa cells.

The water-soluble heteronuclear metallomacrocyclic sandwich clusters [NaCu6(hpnbpda)3(OH)3(OH2)3](NO3)·4H2O (1), [NaCu6(hpnbpda)3(OH)3(OH2)3](ClO4) (2), and [NaCu6(hpnbpda)3(OH)3(OH2)3](PF6) (3) [H3hpnbpda = N,N′-bis(2-pyridylmethyl)-2-hydroxy-1,3-propanediamine-N,N′-diacetic acid] have been synthesized in methanol at room temperature and fully characterized using several analytical techniques including single-crystal X-ray diffraction. The molecular architecture of complex 1 is built from the template assembly of three dinuclear [Cu2(hpnbpda)]+ fragments through their weak oxophillic interactions with a central sodium(I) cation. A close analysis of the single-crystal X-ray structure reveals that the metallic core of cluster 1 consists of six highly distorted octahedral CuII ions arranged at the corners of a trigonal prism that encapsulates the sodium(I) ion. Complex 1 displays a rare μ3:η2:η1:η1 bridging mode of six carboxylate groups of three hpnbpda3– ligands with each bridging between two copper(II) atoms and the sodium(I) center. Variable-temperature magnetic-susceptibility measurements (2–300 K) on a powdered microcrystalline sample reveal that the paramagnetic copper centers in complex 1 are antiferromagnetically coupled to one another. Systematic biological investigation such as cytotoxicity assessment, DNA content analysis in terms of cell-cycle distribution by means of the fluorescence-activated cell sorting (FACS) method, DNA binding, DNA cleavage, and the underlying mechanism of possible apoptotic cell-death events in human cervical cancer cells (HeLa) were carried out in detail using complex 1. The in vitro assays using complex 1 in HeLa cells provided new findings that indicate its possible future therapeutic application. Theoretical calculations were carried out to find the Fukui functions at the metal sites in complex 1 to predict the possible metal centers involved in the DNA binding.

Objective Use of cisplatin, a conventional anticancer drug, is restricted because it generates strong hepatotoxicity by accumulating in liver. Therefore its anticancer potential can only be fully exploited if its own toxicity is considerably reduced. Towards this goal, ethanolic extract of the plant, Boldo ( Peumus boldus ), known for its antihepatotoxic effects, was used simultaneously with cisplatin, to test its ability to reduce cisplatin's cytotoxicity without affecting its anticancer potential. Methods The cytotoxicity of Boldo extract (BE) and cisplatin, administered alone and in combination, was determined in three cancer cell lines (A549, HeLa, and HepG2) and in normal liver cells (WRL-68). Drug-DNA interaction, DNA damage, cell cycle, apoptosis, reactive oxygen species (ROS) and mitochondrial membrane potential (MMP, ΔΨ) were also studied. Hepatotoxicity and antioxidant activity levels were determined by alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase and glutathione assays in mice. The cytotoxicity of related proteins was tested by Western blotting. Results Co-administration of BE and cisplatin increased viability of normal cells, but had no effect on the viability of cancer cells. Boldo protected liver from damage and normalized different antioxidant enzyme levels in vivo and also reduced ROS and re-polarized MMP in vitro. Bax and cytochrome c translocation was reduced with caspase 3 down-regulation. Further, a drug-DNA interaction study revealed that BE reduced cisplatin's DNA-binding capacity, resulting in a reduction in DNA damage. Conclusion Results indicated that a low dose of BE could be used beneficially in combination with cisplatin to reduce its toxicity without hampering cisplatin's anticancer effect. These findings signify a potential future use of BE in cancer therapy.

Chronic intake of alcohol is known to be the third most harmful risk factor for some serious diseases including cancer. Though chronic abuse of alcohol in high dose is reported to cause progressive damage and organ failure mainly affecting liver, whether the same in a low dose can produce any hepatotoxic or genotoxic effect is not precisely known. Therefore, in this study, effects of chronic feeding of low dose alcohol were critically assessed at both genotoxic and hepatotoxic levels in a mammalian model—mice. Mice were chronically fed 0.6 ml of 0.85 M ethanol daily for 7 days onward through 120 days and sacrificed at six fixation intervals, namely at 7, 15, 30, 60, 90 and 120 days. Cytogenetical endpoints like chromosome aberrations, micronuclei, mitotic index from bone marrow cells and sperm head abnormality from epididymis of males were studied and for assessment of genotoxicity and several toxicity markers like acid and alkaline phosphatases, alanine and aspartate amino transferases, lipid peroxidation, reduced glutathione, glutathione reductase, succinate dehydrogenase and catalase were analyzed for assessment of hepatotoxicity. Additionally, liver histology, lymphocyte viability and hepatic cell viability were also studied. In this study, although chronic consumption of the low dose of alcohol did not produce significant changes in parameters like histology, percentages of hepatic cell and lymphocyte viability but certain clastogenic changes were observed at longer intervals (60 day onward). Thus, overall results suggested possible generation of hepatotoxicity and oxidative stress in chronically fed low alcohol consumption group, which could also increase cancer risk.

Toxic effects of cadmium (Cd) were examined on a cadmium-resistant strain of Aspergillus foetidus isolated from wastewater. The Cd removal potential was analyzed. The results indicated that the strain could tolerate up to 25 mM and 63 mM Cd in liquid and solid Czapek-Dox media, respectively. It efficiently removed Cd from liquid growth media and industrial wastewater by mycelial biosorption. The strain produced oxalic acid for the purpose of Cd bioleaching as confirmed by the presence of cadmium oxalate crystals on the mycelial surface. Intracellular proline contents and the antioxidative enzyme activities increased up to a certain level to detoxify the overproduced free radicals. These data indicate that the strain has inherent mechanisms to grow in Cd contaminated environment, tolerate high Cd doses and high Cd uptake potential which are pre-requisite for acting as a suitable candidate for Cd bioremediation.

In homeopathy, ultra-low doses of drugs at ultra-high dilutions are often used with great benefits to patients although at such dilutions physical existence of even a single molecule of the original drug substance is highly improbable. Despite serious challenges thrown by scientists and rationalists from time to time, homeopathy has managed to survive over 200 years now, and is no more considered a myth. Research activities on homeopathy in recent years, at clinical, physical, chemical, biological and medical levels with acceptable scientific norms and approach have paved the way for more rigorous research, particularly at the molecular level to understand the physico-chemical nature and mechanism of action of ultra-high dilutions. Although major breakthrough has been made in understanding many physical aspects and interactions between the “drug” and “aquatic ethanol” used as vehicle/solvent/diluents, certain aspects in regard to structure of water/aquatic ethanol and the latter’s changing structural organization still remain unclear. In recent years, the quest for understanding the mechanism of biological action of the ultra-high dilutions has made homeopathy a hot bed of research. Much progress has been made in understanding the molecular mechanism in the light of the “gene regulatory hypothesis” that can explain the action of the homeopathic high dilutions in all living organisms, both in higher and lower animals as well as in plants. The present review focuses mainly on research in support of the gene regulatory hypothesis, and mention has been made of some relevant physical and biological aspects at cellular and molecular levels.