Sodium Oxybate for Narcolepsy: Explaining Untoward Effects and Recommending New Approaches in Light of Prevailing Receptor Pharmacology.

Objective: Gamma-hydroxybutyrate (GHB) has been an abused and illicit substance for decades, but the antinarcoleptic medication Xyrem (sodium oxybate), the sodium salt of GHB, was approved just in 2002 for increasing wakefulness. We present a case of coma induced by co-ingestion of prescription GHB and ethanol and describe the response to naloxone treatment, by first responders, without evidence of opiate exposure. The purpose of this report is to bridge updated knowledge on GHB and ethanol pharmacology with the clinical sequence of events in a patient co-ingesting these compounds and to theorize on a potentially better pharmacological approach to narcolepsy. Case Summary: The patient was a 25-year-old woman with a history of narcolepsy. She suddenly collapsed at home but became transiently responsive after being administered naloxone in the ambulance. She presented to the emergency department with apnea, poor responsiveness with a Glasgow Coma Score of 7, and urinary incontinence. While undergoing intubation, the patient spontaneously and abruptly awoke. Labs were unremarkable except a blood alcohol concentration of 0.123%. The dosage of, and adherence to, GHB was unknown in this case. Discussion: The case is described in light of the most recent pharmacological advancements on these co-ingestants. A conceptual dose-response curve is shown to facilitate understanding of the complex pharmacology of GHB. Conclusions: Approved and potential alternatives to GHB, for achieving wakefulness, are discussed. Potential new strategies should bear low to no risk of coma with accidental overdose or co-ingestion of ethanol. In addition, promising antidotes for future consideration are discussed.

Organic anion transporter 3 interacts selectively with lipophilic ß-lactam antibiotics.

Transporters are major determinants of the disposition of xenobiotics and endogenous chemicals in the body. Organic anion transporter 3 (Oat3) functions in the kidney and brain to remove metabolic waste, toxins, and drugs, and thus transports diverse chemicals. Some ß-lactam antibiotics interact with Oat3, and penicillin G exhibits a strong dependence on Oat3 for renal elimination. However, over 80 ß-lactams exist, and many have not been assessed for an interaction with Oat3. Moreover, ß-lactams continue to receive U.S. Food and Drug Administration approval. This study identified new ß-lactam-Oat3 interactions, provided a head-to-head comparison with Oat1, and characterized the physicochemical determinants of affinity for Oat3. Cells expressing mouse Oat3 (mOat3) and Oat1 (mOat1), and human OAT3 (hOAT3) were used to test inhibitors, and high-performance liquid chromatography (HPLC) was used to measure transport. Of 26 ß-lactams tested, 12 were clear inhibitors of Oat3, and 14 exhibited poor interactions. Inhibitors exhibited a nearly identical rank-order of potency against mOat3 and hOAT3. Oat1 demonstrated a poor interaction with most ß-lactams. The majority of Oat3 inhibitors were substrates, and there were clear physicochemical differences between inhibitors and noninhibitors. That is, inhibitors had nearly 40% fewer hydrogen bond donors (P < 0.001), a lower total polar surface area (P < 0.05), and greater lipophilicity (LogP of inhibitors, +1.41; noninhibitors, -1.54; P < 0.001). Pharmacophore mapping revealed a prohibitive hydrogen bond donor group in noninhibitors adjacent to a hydrophobic moiety that was important for binding to Oat3. These findings indicate that Oat3 recognizes lipophilic ß-lactams more readily. Moreover, this study has potential implications for designing ß-lactams to avoid renal accumulation or brain efflux via Oat3.

Antitumor and chemopreventive role of major phytochemicals against breast cancer development.

Breast cancer continues to be one of the most commonly diagnosed cancers around the world. Despite the decrease in mortality, there has been a steady increase in its incidence. There is much evidence that naturally occurring phytochemicals could prove to be safer alternatives aimed at prevention and development of breast cancer. In the present review, we discuss important phytochemicals, namely capsaicin, alpha-santalol and diallyl trisulphide that are shown to have chemopreventive and anti-tumour properties against breast cancer development. We examined current knowledge of their bioavailability, safety and modulation of molecular mechanisms including their ability to induce apoptotic cell death, promote cell cycle arrest, and inhibit cellular proliferation in different breast cancer cell lines and in vivo models. This review emphasises the importance of these naturally occurring phytochemicals and their potential of becoming therapeutic options in the arsenal against breast cancer development provided further scientific and clinical validation.

Phenethyl isothiocyanate inhibits oxidative phosphorylation to trigger reactive oxygen species-mediated death of human prostate cancer cells.

Phenethyl isothiocyanate (PEITC), a constituent of edible cruciferous vegetables such as watercress, not only affords significant protection against chemically induced cancer in experimental rodents but also inhibits growth of human cancer cells by causing apoptotic and autophagic cell death. However, the underlying mechanism of PEITC-induced cell death is not fully understood. Using LNCaP and PC-3 human prostate cancer cells as a model, we demonstrate that the PEITC-induced cell death is initiated by production of reactive oxygen species (ROS) resulting from inhibition of oxidative phosphorylation (OXPHOS). Exposure of LNCaP and PC-3 cells to pharmacologic concentrations of PEITC resulted in ROS production, which correlated with inhibition of complex III activity, suppression of OXPHOS, and ATP depletion. These effects were not observed in a representative normal human prostate epithelial cell line (PrEC). The ROS production by PEITC treatment was not influenced by cyclosporin A. The Rho-0 variants of LNCaP and PC-3 cells were more resistant to PEITC-mediated ROS generation, apoptotic DNA fragmentation, and collapse of mitochondrial membrane potential compared with respective wild-type cells. The PEITC treatment resulted in activation of Bax in wild-type LNCaP and PC-3 cells, but not in their respective Rho-0 variants. Furthermore, RNA interference of Bax and Bak conferred significant protection against PEITC-induced apoptosis. The Rho-0 variants of LNCaP and PC-3 cells also resisted PEITC-mediated autophagy. In conclusion, the present study provides novel insight into the molecular circuitry of PEITC-induced cell death involving ROS production due to inhibition of complex III and OXPHOS.

Chemopreventive effects of dietary canola oil on colon cancer development.

Fatty acid composition of dietary fat plays a vital role in colon tumor development in animal models. Fats containing ω-6 fatty acids (e.g., corn oil) enhanced and ω-3 fatty acids (e.g., flaxseed oil) reduced chemically induced colon tumor development in rats. The objective of the present investigation was to study the effects of dietary canola oil, a source of ω-3 fatty acid on azoxymethane-induced colon cancer development in Fischer rats and compare with dietary corn oil. Dietary canola oil significantly (P<0.05) decreased colonic tumor incidence and tumor multiplicity as compared to dietary corn oil in rats. Fatty acid analysis showed that corn oil group had higher levels of ω-6 fatty acid levels, whereas the canola oil groups exhibited higher levels of ω-3 fatty acids from the colon and serum samples of rats. For the mechanistic study, COX-2 expression in the colon samples from the canola oil group was significantly lower (P<0.05) as compared to the corn oil group. Taken together, dietary canola oil may be chemopreventive for colon tumor development in Fischer rats as compared to possibly by increasing ω-3 fatty acid levels and decreasing COX-2 levels.

Autophagy Induction by α-Santalol in Human Prostate Cancer Cells.

BACKGROUND/AIM: Previous studies have shown that the sandalwood oil constituent α-santalol inhibits growth of cultured human prostate cancer cells in vitro and PC-3 prostate cancer xenografts. Along with the studies from our laboratory, it is well established that α-santalol targets the phosphatidylinositol-4,5-bisphosphate 3-kinase-AKT serine/ threonine kinase 1 (AKT) pathway to induce apoptosis but its growth-suppressive effects have not been fully elucidated. The current study was undertaken to investigate the role of autophagy in α-santalol-induced prostate cancer cell death. MATERIALS AND METHODS: Cell lines LNCaP and PC-3 were maintained in an atmosphere of 95% air and 5% CO2 at 37°C. Trypan blue dye exclusion assay was employed to assess the effects of α-santalol with/without 3-methyl adenine on the cell viability of prostate cancer cells. Acidic vesicular organelles induced by α-santalol treatment were detected by staining with acridine orange. Immunofluorescence and immunoblotting were performed to analyze expression of proteins involved in the AKT-mammalian target of rapamycin (mTOR) pathway. RESULTS: LNCaP and PC-3 cells upon treatment with α-santalol resulted in characteristic features analogous to autophagic response, including formation of acidic vesicular organelles, recruitment and cleavage of microtubule-associated protein 1 light chain 3 (LC3) to autophagosomes. Alpha-santalol treatment further suppressed phosphorylation of activated AKT and mTOR, which are critical regulators of autophagic response. In addition, pre-treatment of PC-3 cells with specific inhibitor of autophagy (3-methyladenine) and co-treatment with α-santalol attenuated the expression of LC3-II and phospho-AKT, and significantly reduced the cell viability. CONCLUSION: The present study indicates that α-santalol induces autophagy by targeting the AKT-mTOR pathway in prostate cancer cells, which may serve as a protective mechanism.

Evaluation of long term retention and utilization of the situation, background, assessment, recommendation (SBAR) communication technique.

INTRODUCTION: Effective communication as part of an interprofessional team is a required standard of pharmacy education. The Situation, Background, Assessment, Recommendation (SBAR) communication technique is an evidence-based method shown to improve patient safety, and is embedded in some curricula of pharmacy and other health care professions. The aim of this study is to determine whether students can utilize the SBAR communication technique during an interprofessional skills assessment one year following initial instruction. METHODS: Students are initially trained on the SBAR technique in an interprofessional setting using the Team Strategies and Tools to Enhance Performance and Patient Safety (TeamSTEPPS) method in the fall of the second professional year. One year later, students participated in a simulated interaction with a physician as part of the pulmonary module of the pharmacotherapeutics series. Faculty evaluators noted how many and which components of the SBAR technique students used during the interaction. The simulation was run for two academic years, results of which were compared. RESULTS: There was a significant difference in the number of students who used all four components of SBAR. A significant difference also existed between the use of the "situation" and "background" components. CONCLUSION: The TeamSTEPPS method appears to be an effective method to train students on the SBAR communication technique and results in long term retention. Pharmacy programs should consider the use of the TeamSTEPPS method early in their curricula.

Inhibition of human breast cancer xenograft growth by cruciferous vegetable constituent benzyl isothiocyanate.

Benzyl isothiocyanate (BITC), a constituent of cruciferous vegetables such as garden cress, inhibits growth of human breast cancer cell lines in culture. The present study was undertaken to determine in vivo efficacy of BITC against MDA-MB-231 human breast cancer xenografts. The BITC administration retarded growth of MDA-MB-231 cells subcutaneously implanted in female nude mice without causing weight loss or any other side effects. The BITC-mediated suppression of MDA-MB-231 xenograft growth correlated with reduced cell proliferation as revealed by immunohistochemical analysis for Ki-67 expression. Analysis of the vasculature in the tumors from BITC-treated mice indicated smaller vessel area compared with control tumors based on immunohistochemistry for angiogenesis marker CD31. The BITC-mediated inhibition of angiogenesis in vivo correlated with downregulation of vascular endothelial growth factor (VEGF) receptor 2 protein levels in the tumor. Consistent with these results, BITC treatment suppressed VEGF secretion and VEGF receptor 2 protein levels in cultured MDA-MB-231 cells. Moreover, the BITC-treated MDA-MB-231 cells exhibited reduced capacity for migration compared with vehicle-treated control cells. In contrast to cellular data, BITC administration failed to elicit apoptotic response as judged by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay. In conclusion, the present study demonstrates in vivo anti-cancer efficacy of BITC against MDA-MB-231 xenografts in association with reduced cell proliferation and suppression of neovascularization. These preclinical observations merit clinical investigation to determine efficacy of BITC against human breast cancers.

Akt/survivin pathway inhibition enhances the apoptotic cell death-induced by alpha-santalol in human prostate cancer cells.

We have shown previously that alpha-santalol, a major component of sandalwood oil inhibits growth of cultured prostate cancer cells in vitro by causing apoptosis, but the mechanism of cell death is not fully elucidated. The present study was undertaken to investigate the role of PI3K/Akt/survivin pathway in alpha-santalol-induced apoptosis employing cultured LNCaP and PC-3 human prostate cancer cells. Treatment of prostate cancer cells with alpha-santalol (20, 40 µM) resulted in the down regulation of survivin and p-AKT (s-473) expression and statistically significant reduction in total survivin levels as evidenced by survivin ELISA assay. Furthermore, inhibition of PI3K-Akt pathway by pharmacological inhibitor, LY294002 enhanced the apoptotic cell death induced by alpha-santalol as determined by cell viability, cellular morphology, active caspase-3 activity and expression of cleaved PARP, cleaved caspase-3 levels. In conclusion, the present study provides novel insight into the molecular circuitry of alpha-santalol-induced cell death and reveals that alpha-santalol targets Akt/Survivin pathway to induce cell death and that the cell death is increased in the presence of a known inhibitor of the pathway.

A Copper(II) Macrocycle Complex for Sensing Biologically Relevant Organic Anions in a Competitive Fluorescence Assay: Oxalate Sensor or Urate Sensor?

Fluorescence sensing of oxalate has garnered some attention in the past two decades as a result of this anion's prominence and impact on society. Previous work on oxalate sensors and other divalent anion sensors has led to the conclusion that the sensors are selective for the anion under investigation. However, sensor selectivity is often determined by testing against a relatively small array of "guest" molecules or analytes and studies often exclude potentially interfering compounds. For example, studies on oxalate sensors have excluded compounds such as citrate and urate, which are anions in the biological matrices where oxalate is measured (e.g., urine, blood, and bacterial lysate). In the present study, we reassessed the selectivity of a dinuclear copper(II) macrocycle (Cu2L) in an eosin Y displacement assay using biologically relevant anions. Although previously reported as selective for oxalate, we found greater indicator displacement (fluorescence response) for urate and oxaloacetate and a significant response to citrate. These anions are larger than oxalate and do not appear to fit into the putative binding pocket of Cu2L. Consistent with previous reports, Cu2L did not release eosin Y in the presence of several other dicarboxylates, including adipate, glutarate, malate (except at 10 mM), fumarate, succinate, or malonate (except at 10 mM), and the monocarboxylate acetate. This was demonstrated by the failure of the anions to reverse eosin Y quenching by Cu2L. We also assessed, for the first time, other monocarboxylates, including butyrate, pyruvate, lactate, propionate, and formate. None of these anions were able to displace eosin Y, indicating no interaction with Cu2L that interfered with the eosin Y binding site. Single-crystal X-ray crystallography revealed that nonselective binding of the anions is likely partly caused by readily accessible copper(II) ions on the external surface of Cu2L. In addition, π-π stacking of urate with the aromatic groups of Cu2L cannot be ruled out as a contributor to binding. We conclude that Cu2L is not suitable for oxalate sensing in a biological matrix unless interfering compounds are selectively removed or masked.

Effects of dietary flaxseed on intestinal tumorigenesis in Apc(Min) mouse.

Dietary flaxseed has been shown to prevent azoxymethane (AOM)-induced colorectal cancers in male Fisher rats. The present study was designed to investigate the chemopreventive effects of dietary flaxseed on the development of intestinal tumors in Apc(Min) mice. Apc(Min) mice were divided into five different groups, fed with control (AIN-93M meal), corn meal, flaxseed meal, corn oil, and flaxseed oil supplemented diets. Results showed that dietary flaxseed significantly decreased (P < 0.05) tumor multiplicity and size in the small intestine and colon as compared to control, corn-treated groups. Intestine, colon, and serum samples of corn-treated groups showed higher levels of omega -6 fatty acids, whereas the flaxseed treated groups exhibited higher levels of omega -3 fatty acids. Lignans were detected in the serum, intestine, and colon samples for flaxseed meal group. COX-1 and COX-2 expression in the colon samples from the flaxseed meal group were significantly lower (P < 0.05) as compared to the corn meal group. Dietary flaxseed may be chemopreventive for intestinal tumor development in Apc(Min) mice possibly by increasing omega -3 fatty acid levels, lignans, and decreasing COX-1 and COX-2 levels.

Cellular responses to cancer chemopreventive agent D,L-sulforaphane in human prostate cancer cells are initiated by mitochondrial reactive oxygen species.

PURPOSE: Present study was undertaken to elucidate the mechanism of cellular responses to D,L-sulforaphane (SFN), a highly promising cancer chemopreventive agent. METHODS: Mitochondrial DNA deficient Rho-0 variants of LNCaP and PC-3 cells were generated by culture in the presence of ethidium bromide. Apoptosis was assessed by analysis of cytoplasmic histone-associated DNA fragmentation and activation of caspase-3. Immunoblotting was performed to determine the expression of apoptosis- and cell cycle-regulating proteins. Generation of reactive oxygen species (ROS), mitochondrial membrane potential (MMP), and cell cycle distribution were measured by flow cytometry. RESULTS: The Rho-0 variants of LNCaP and PC-3 cells were significantly more resistant to SFN-induced ROS generation, apoptotic DNA fragmentation, disruption of MMP, cytosolic release of cytochrome c, and G2/M phase cell cycle arrest compared with corresponding wild-type cells. SFN-induced autophagy, which serves to protect against apoptotic cell death in PC-3 and LNCaP cells, was also partially but markedly suppressed in Rho-0 variants compared with wild-type cells. SFN statistically significantly inhibited activities of mitochondrial respiratory chain enzymes in LNCaP and PC-3 cells. CONCLUSION: These results indicate, for the first time, that mitochondria-derived ROS serve to initiate diverse cellular responses to SFN exposure in human prostate cancer cells.

Chemopreventive effects of sarcophine-diol on ultraviolet B-induced skin tumor development in SKH-1 hairless mice.

Sarcophine-diol (SD), one of the structural modifications of sarcophine, has shown chemopreventive effects on 12-dimethylbenz(a)anthracene-initiated and 12-O-tetradecanoylphorbol-13-acetate-promoted skin tumor development in female CD-1 mice. The objective of this study was to determine the chemopreventive effects of SD on UVB-induced skin tumor development in hairless SKH-1 mice, a model more relevant to human skin cancer, and to determine the possible mechanisms of action. Carcinogenesis was initiated and promoted by UVB radiation. Female hairless SKH-1 mice were divided into two groups having 27 mice in each group: control and SD treatment. The control group was topically treated with 100 microL acetone and SD treatment group was topically treated with SD (30 microg/100 microL in acetone) 1 hour before each UVB radiation for 32 weeks. Tumor counts were recorded on a weekly basis for 30 weeks. Effects of SD on the expression of caspases were investigated to elucidate the possible mechanism of action. The proteins from epidermal homogenates of experimental mice were used for SDS-PAGE and Western blotting using specific antibodies against caspase-3, caspase-8 and caspase-9 respectively. TUNEL assay was used for determining DNA fragmented apoptotic cells in situ. Results showed that at the end of experiment, tumor multiplicity in control and SD treatment groups was 25.8 and 16.5 tumors per mouse respectively. Furthermore, Topical treatment of SD induced DNA fragmented apoptotic cells by upgrading the expressions of cleaved caspase-3 and caspase-8. This study clearly suggested that SD could be an effective chemopreventive agent for UVB-induced skin cancer by inducing caspase dependent apoptosis.

Medicinal properties of alpha-santalol, a naturally occurring constituent of sandalwood oil: review.

Alpha-santalol is a naturally occurring sesquiterpene that is derived from sandalwood oil. Its wide range of health benefits have been attributed to the modulation of various signalling pathways involved in the development of a particular disease. For example, the antitumour and cancer preventive properties of alpha-santalol have been shown to involve cell death induction through apoptosis and cell cycle arrest in various cancer models. A marked decrease in inflammatory markers have also been shown with alpha-santalol administration in skin tissue models. The current review is aimed at bringing the most recent advances of alpha-santalol against various disease-specific models and highlighting its associated mechanistic details.

Effects of alpha-santalol on proapoptotic caspases and p53 expression in UVB irradiated mouse skin.

BACKGROUND: Cancer chemoprevention by naturally occurring agents, especially phytochemicals, minerals and vitamins has shown promising results against various malignancies in a number of studies both under in vitro and in vivo conditions. One such phytochemical, alpha-santalol, a major component of sandalwood oil, is effective in preventing skin cancer in both chemically and UVB-induced skin cancer development in CD-1, SENCAR and SKH-1 mice; however, the mechanism of its efficacy is not fully understood. The objective of the present investigation was to study the effects of alpha-santalol on apoptosis proteins and p53 in UVB-induced skin tumor development in SKH-1 mice to elucidate the mechanism of action. MATERIALS AND METHODS: Female SKH-1 mice were divided into two groups: Group 1, which served as control received topical application of acetone (0.1 ml) one hour before UVB treatment; Group 2 received alpha-santalol (0.1 ml, 5% w/v in acetone, topical) one hour prior to UVB treatment. UVB-induced promotion was continued for 30 weeks. RESULTS: Pre-treatment with alpha-santalol one hour prior to UVB exposure significantly (p < 0.05) reduced tumor incidence and multiplicity, and resulted in a significant (p < 0.05) increase in apoptosis proteins, caspase-3 and -8 levels and tumor suppressor protein, p53. CONCLUSION: These results suggest that alpha-santalol prevents skin cancer development by inducing proapoptotic proteins via an extrinsic pathway and increasing p53.

Alpha-Santalol, a Component of Sandalwood Oil Inhibits Migration of Breast Cancer Cells by Targeting the ß-catenin Pathway.

BACKGROUND/AIM: Alpha-santalol, a terpenoid found in sandalwood oil has been shown to inhibit breast cancer cell growth in vitro by inducing apoptosis, but the mechanisms underlying the growth inhibitory effects of alpha-santalol are not fully understood. In this study, we demonstrate that α-santalol treatment targets Wnt/ß-catenin pathway to inhibit migration of cultured breast cancer cells. MATERIALS AND METHODS: Migration assays, immunoblotting and immunofluorescence were used to examine the mechanism of action of a-santalol in breast cancer cells. RESULTS: Exposure of MDA-MB 231 and MCF-7 cells to α-santalol resulted in a significant reduction in their migratory potential and wound healing ability. In addition, α-santalol affected the localization of ß-catenin from cytosol to nucleus in MDA-MB 231 cells. CONCLUSION: Alpha-santalol inhibited migration of breast cancer cells may be mediated, in part, by targeting Wnt//ß-catenin pathway. ß-catenin represents an important target of α-santalol's response for future pre-clinical studies.

Chemoprevention by alpha-santalol on UVB radiation-induced skin tumor development in mice.

Studies have shown the chemopreventive effects of alpha-santalol on chemically and UVB-induced skin cancer in mice. The objective of the present investigation was to find the lowest effective concentration of alpha-santalol for the chemopreventive effects on UVB-induced skin tumor development in mice and to determine antiperoxidant effect of alpha-santalol in order to elucidate its possible mechanism of action. Female SKH-1 mice were divided into different groups receiving either vehicle alone or different concentrations of alpha-santalol. Mice in all the groups were initiated and promoted with UVB radiation for skin tumor development. The promotion phase continued for 30 weeks. Skin tumors were counted once a week for 30 weeks. Lipid peroxidation was assayed in skin and liver microsomes by measuring malonaldehyde formed using thiobarbituric acid method. Topical administration of alpha-santalol reduced UVB-induced skin tumor development in a concentration-dependent manner. Application of alpha-santalol (5%) significantly (p < 0.05) delayed skin tumor development for 25 weeks and reduced tumor multiplicity. alpha-Santalol also inhibited in vitro lipid peroxidation in skin and liver microsomes. alpha-Santalol application prevents UVB-induced skin tumor development possibly by acting as an antiperoxidant.

Chemopreventive effects of sarcotriol on ultraviolet B-induced skin tumor development in SKH-1 hairless mice.

Sarcotriol (ST) has been shown to be chemopreventive on 7,12-dimethyl-benz(a)anthracene (DMBA) initiated and 12-O-tetradecanoylphorbol-13-acetate (TPA)-promoted skin tumor development in CD-1 mice in recent studies from our laboratory. The objective of this study was to determine the chemopreventive effects of ST on ultraviolet B (UVB)-induced skin tumor development in female SKH-1 hairless mice, an experimental model relevant to human skin cancer development, and its possible mechanisms of action. Female SKH-1 mice were divided into two groups: Control and ST treated. Control was topically treated with 100 microliter acetone and ST treated group administered with 30 microgram ST in 100 microliter acetone one hour before UVB exposure. For UVB-induced tumorigenesis, carcinogenesis was initiated and promoted by UVB (180 mJ/cm(2)). Group weights and tumor counts were taken once every week. After 30 weeks, mice were sacrificed and dorsal skin samples were collected. The proteins from the skin sample were further used for SDS-PAGE and Western blotting using specific antibodies against caspase-3, caspase-8, caspase-9 and p53. Tumor multiplicity was found 19.6, 5.2 in the control and ST treated groups respectively. Caspase-3, -8, -9 and p53 were significantly (P < 0.05) upregulated in ST treated group compared to Control group. Together, this study for the first time identifies the chemopreventive effects of ST in UVB-induced carcinogenesis possibly by inducing apoptosis and upregulating p53.

Determination of mammalian lignans in biological samples by modified gas chromatography/mass spectrometry.

Lignans in flaxseed have been part of the human diet for centuries. In 1955, the isolation and structure of the lignan derivative secoisolariciresinol diglucoside (SDG) was reported. The biological role of SDG and mammalian lignan metabolites enterodiol and enterolactone was initially reported 20 years later. Experimental evidences showed the beneficial effects of lignans on breast, colon, and thyroid cancer. A modified gas chromatography/mass spectrometry (GC/MS) assay was developed for lignans in serum and colon samples of rats fed flaxseed meal. The method developed for the analysis of metabolites involves extraction and derivatization of samples and quantitative analysis by selected ion monitoring using GC/MS. The levels of lignan metabolites enterodiol and enterolactone were determined to be 0.013 and 0.23 microM in serum samples and 0.008 and 1.63 microM in colon samples.

Survivin Down-regulation by α-Santalol Is Not Mediated Through PI3K-AKT Pathway in Human Breast Cancer Cells.

BACKGROUND: α-Santalol, a terpenoid found in sandalwood oil, has been shown to inhibit cancer cell growth in vitro by inducing apoptosis. This study was performed to investigate the anticancer properties of α-santalol associated with the induction of apoptosis in cultured MCF-7 [estrogen receptor (ER)-positive, and wild-type p53)] and MDA-MB-231 (ER-negative and mutant p53) breast cancer cells. MATERIALS AND METHODS: Expression of major proteins examined in the study were determined using a standard western blot protocol and analyzed by LICOR-Odyssey infra-red scanner. Total protein levels of survivin were confirmed by survivin enzyme-linked immunosorbent assay (ELISA) kit. Cell viability was assessed by the trypan blue dye exclusion assay, and caspase-3 activity was determined by caspase-3 (active) ELISA kit. RESULTS: Treatment of breast cancer cells for 6 and 9 h with α-santalol (20, and 40 µM) resulted in statistically significant concentration-dependent down-regulation of survivin. Phosphorylated protein kinase B (pAKT) levels were found to be slightly up-regulated despite the down-regulation of survivin. Pharmacological inhibition of the phosphoinositide 3-kinase - protein kinase B (PI3K-AKT) pathway did not result in a synergistic/additive increase in cell death or caspase-3 activity caused by α-santalol. CONCLUSION: The study reveals that survivin down-regulation by α-santalol in breast cancer cells is not mediated through the PI3K-AKT pathway.