β-Elemene Piperazine Derivatives Induce Apoptosis in Human Leukemia Cells through Downregulation of c-FLIP and Generation of ROS

β-Elemene is an active component of the herb medicine Curcuma Wenyujin with reported antitumor activity. To improve its antitumor ability, five novel piperazine derivatives of β-elemene, 13-(3-methyl-1-piperazinyl)-β-elemene (DX1), 13-(cis-3,5-dimethyl-1-piperazinyl)-β-elemene (DX2), 13-(4-ethyl-1-piperazinyl)-β-elemene (DX3), 13-(4-isopropyl-1-piperazinyl)-β-elemene (DX4) and 13-piperazinyl-β-elemene (DX5), were synthesized. The antiproliferative and apoptotic effects of these derivatives were determined in human leukemia HL-60, NB4, K562 and HP100-1 cells. DX1, DX2 and DX5, which contain a secondary amino moiety, were more active in inhibiting cell growth and in inducing apoptosis than DX3 and DX4. The apoptosis induction ability of DX1 was associated with the generation of hydrogen peroxide (H2O2), a decrease of mitochondrial membrane potential (MMP), and the activation of caspase-8. Pretreatment with the antioxidants N-acetylcysteine and catalase completely blocked DX1-induced H2O2 production, but only partially its activation of caspase-8 and induction of apoptosis. HL-60 cells were more sensitive than its H2O2-resistant subclone HP100-1 cells to DX1-induced apoptosis. The activation of caspase-8 by these compounds was correlated with the decrease in the levels of cellular FLICE-inhibitory protein (c-FLIP). The proteasome inhibitor MG-132 augmented the decrease in c-FLIP levels and apoptosis induced by these derivatives. FADD- and caspase-8-deficient Jurkat subclones have a decreased response to DX1-induced apoptosis. Our data indicate that these novel β-elemene piperazine derivatives induce apoptosis through the decrease in c-FLIP levels and the production of H2O2 which leads to activation of both death receptor- and mitochondrial-mediated apoptotic pathways

Growth of a human mammary tumor cell line is blocked by galangin, a naturally occurring bioflavonoid, and is accompanied by down-regulation of cyclins D3, E, and A

INTRODUCTION: This study was designed to determine if and how a non-toxic, naturally occurring bioflavonoid, galangin, affects proliferation of human mammary tumor cells. Our previous studies demonstrated that, in other cell types, galangin is a potent inhibitor of the aryl hydrocarbon receptor (AhR), an environmental carcinogen-responsive transcription factor implicated in mammary tumor initiation and growth control. Because some current breast cancer therapeutics are ineffective in estrogen receptor (ER) negative tumors and since the AhR may be involved in breast cancer proliferation, the effects of galangin on the proliferation of an ER(-), AhR(high )line, Hs578T, were studied. METHODS: AhR expression and function in the presence or absence of galangin, a second AhR inhibitor, α-naphthoflavone (α-NF), an AhR agonist, indole-3-carbinol, and a transfected AhR repressor-encoding plasmid (FhAhRR) were studied in Hs578T cells by western blotting for nuclear (for instance, constitutively activated) AhR and by transfection of an AhR-driven reporter construct, pGudLuc. The effects of these agents on cell proliferation were studied by (3)H-thymidine incorporation and by flow cytometry. The effects on cyclins implicated in mammary tumorigenesis were evaluated by western blotting. RESULTS: Hs578T cells were shown to express high levels of constitutively active AhR. Constitutive and environmental chemical-induced AhR activity was profoundly suppressed by galangin as was cell proliferation. However, the failure of α-NF or FhAhRR transfection to block proliferation indicated that galangin-mediated AhR inhibition was either insufficient or unrelated to its ability to significantly block cell proliferation at therapeutically relevant doses (IC(50 )= 11 μM). Galangin inhibited transition of cells from the G(0)/G(1 )to the S phases of cell growth, likely through the nearly total elimination of cyclin D3. Expression of cyclins A and E was also suppressed. CONCLUSION: Galangin is a strong inhibitor of Hs578T cell proliferation that likely mediates this effect through a relatively unique mechanism, suppression of cyclin D3, and not through the AhR. The results suggest that this non-toxic bioflavonoid may be useful as a chemotherapeutic, particularly in combination with agents that target other components of the tumor cell cycle and in situations where estrogen receptor-specific therapeutics are ineffective

Cost evaluation of therapeutic drug monitoring of gentamicin at a teaching hospital in Malaysia

Background: Therapeutic drug monitoring (TDM) makes use of serum drug concentrations as an adjunct to decision-making. Preliminary data in our hospital showed that approximately one-fifth of all drugs monitored by TDM service were gentamicin. Objective: In this study, we evaluated the costs associated with providing the service in patients with bronchopneumonia and treated with gentamicin. Methods: We retrospectively collected data from medical records of patients admitted to the Hospital Universiti Sains Malaysia over a 5-year period. These patients were diagnosed with bronchopneumonia and were on gentamicin as part of their treatment. Five hospitalisation costs were calculated; (i) cost of laboratory and clinical investigations, (ii) cost associated with each gentamicin dose, (iii) fixed and operating costs of TDM service, (iv) cost of providing medical care, and (v) cost of hospital stay during gentamicin treatment. Results: There were 1920 patients admitted with bronchopneumonia of which 67 (3.5%) had TDM service for gentamicin. Seventy-three percent (49/67) patients were eligible for final analysis. The duration of gentamicin therapy ranged from 3 to 15 days. The cost of providing one gentamicin assay was MYR25, and the average cost of TDM service for each patient was MYR104. The average total hospitalisation cost during gentamicin treatment for each patient was MYR442 (1EUR approx. MYR4.02). Conclusion: Based on the hospital perspective, in patients with bronchopneumonia and treated with gentamicin, the provision of TDM service contributes to less than 25% of the total cost of hospitalization

CLINICAL PROGNOSTIC FACTORS IN PATIENTS WITH ADVANCED STAGE OF PROSTATE CANCER

Objectives: To determine the prognostic factors that could predict patient outcome in patients with advanced stage prostate cancer. Patients and Methods: In this study we retrospectively evaluated the medical record data of 222 patients with advanced stage prostate cancer treated by hormonal therapy (either castration or total androgen blockade (TAB)). All pre- and post- treatment data records were evaluated with respect to patient age, prostate and tumor size, tumor grade, stage, PSA, alkaline and acid phosphatase and the number of bone lesions. The response to the hormonal treatment was evaluated either early (12 months after treatment) or late (over all follow-up visits until the last visit or death). Descriptive statistics, student T test, multivariate and Kaplan Meier's curve were used for data analysis. Results: Within 12 months of treatment 70% of the cases showed an improvement with a significant decrease of their pre-treatment values after hormonal therapy. Patient age, tumor stage, the number of bone lesions, serum alkaline and acid phosphatase levels in the pre-treatment data were significantly independent predictors of the overall survival outcome (p= 0.0015, 0.002, 0.001, 0.0002 and 0.028, respectively), while the pre-treatment PSA serum level, tumor grade and the type of hormonal treatment used (either castration or TAB) were no predictors of patient outcome (p= 0.18, 0.82 and 0.47, respectively). Importantly, the PSA serum level and the number of bone lesions in the first 12 months of patient follow-up were significant predictors of the overall disease survival status (p=0.001 and 0.028, respectively). The mean follow-up period of alive cases was 39.42 months ranging from 6 – 171 months. Of the 222 cases 110 (51.6%) had overall disease progression during a mean of 59.4 months, while mortality was reported in 118 cases (53.2%) in the course of a mean of 59.9 months.Conclusion: The pre-treatment patient age, tumor stage, serum alkaline and acid phosphatase, as well as the post-treatment PSA level and the number of bone lesions were significant independent predictors of the overall patient outcome inpatients with advanced stage prostate cancer. However, a survival analysis in relation to the treatment type did not reveal a statistically significant difference between the outcomes of castration and TAB. Facteurs Pronostiques Cliniques chez les Patients Atteints de Cancer Prostatique Avancé Objectifs: De déterminer les facteurs pronostiques prédictifs de l'évolution du cancer prostatique chez nos patients atteints de cancer avancé de la prostate. Patients et Méthodes: Dans cette étude nous avons rétrospectivement évalué les données médicales de 222 patients atteints de cancer avancé de la prostate traités par thérapie hormonale (castration ou blocage androgènique total (BAT)). Toutes les données pré et post ont été évaluées en ce qui concerne l'âge des patients, la taille de la prostate et de la tumeur, le score histologique de la tumeur, le stade clinique, le PSA, la phosphatase alkaline et acide et le nombre de lésions osseuses. La réponse au traitement hormonal a été évaluée aussi bien tôt (12 mois après traitement) ou tard (à la dernière visite ou mort). Des statistiques descriptives, les tests T de Student, multivariable et de Kaplan Meier ont été employées pour l'analyse des données. Résultats: Pendant les 12 premiers mois du traitement, 70% des cas ont montré une amélioration avec une régression significative de leurs tumeurs. L'âge des patients, le stade de la tumeur, le nombre de lésions osseuses, les niveaux de phosphatase alcalines et acides sériques préopératoires étaient des facteurs prédictifs de survie indépendants et significatifs (p = 0.0015, 0.002, 0.001, 0.0002 et 0.028, respectivement), tandis que le taux sérique de PSA pré thérapeutique, le grade de la tumeur et le type de traitement hormonal utilisé (castration ou BAT) n'étaient pas significativement prédictifs de l'évolution des patients (p = 0.18, 0.82 et 0.47, respectivement). Essentiellement, le niveau de PSA et le nombre de lésions osseuses pendant les 12 premiers mois de suivi étaient des facteurs prédictifs significatifs du statut global de survie de la maladie (p=0.001 et 0.028, respectivement). La période moyenne de suivi des cas vivants était de 39.42 mois s'étendant de 6 - 171 mois. Parmi les 222 cas 110 cas (51.6%) ont eu une progression de la maladie pendant un intervalle de temps moyen de 59.4 mois, alors que la mortalité était de 118 cas (53.2%) pendant un intervalle moyen de temps de 59.9 mois. Conclusion: L'âge du patient, le stade de la tumeur, le taux sérique de phosphatase alcaline et acide, comme le taux de PSA pré thérapeutique et le nombre de lésions osseuses étaient des facteurs prédictifs indépendants et significatifs de l'évolution du cancer chez les patients présentant un cancer avancé de la prostate. Cependant, une analyse de survie par rapport au type de traitement n'a pas indiqué une différence statistiquement significative entre les résultats de la castration et le BAT. (Af J Urology: 2003 9(2):94-101