Sample records for digitoxin

  1. Digitoxin medication and cancer; case control and internal dose-response studies

    International Nuclear Information System (INIS)

    Haux, Johan; Klepp, Olbjørn; Spigset, Olav; Tretli, Steinar


    Digitoxin induces apoptosis in different human malignant cell lines in vitro. In this paper we investigated if patients taking digitoxin for cardiac disease have a different cancer incidence compared to the general population. Computer stored data on digitoxin concentrations in plasma from 9271 patients with cardiac disease were used to define a user population. Age and sex matched controls from the Norwegian Cancer Registry were used to calculate the number of expected cancer cases. The population on digitoxin showed a higher incidence of cancer compared to the control population. However, an additional analysis showed that the population on digitoxin had a general increased risk of cancer already, before the start on digitoxin. Leukemia/lymphoma were the cancer types which stood out with the highest risk in the digitoxin population before starting on digitoxin. This indicates that yet unknown risk factors exist for cardiovascular disease and lymphoproliferative cancer. An internal dose-response analysis revealed a relationship between high plasma concentration of digitoxin and a lower risk for leukemia/lymphoma and for cancer of the kidney/urinary tract. Morbidity and mortality are high in the population on digitoxin, due to high age and cardiac disease.These factors disturb efforts to isolate an eventual anticancer effect of digitoxin in this setting. Still, the results may indicate an anticancer effect of digitoxin for leukemia/lymphoma and kidney/urinary tract cancers. Prospective clinical cancer trials have to be done to find out if digitoxin and other cardiac glycosides are useful as anticancer agents

  2. Digitoxin medication and cancer; case control and internal dose-response studies

    Directory of Open Access Journals (Sweden)

    Spigset Olav


    Full Text Available Abstract Background Digitoxin induces apoptosis in different human malignant cell lines in vitro. In this paper we investigated if patients taking digitoxin for cardiac disease have a different cancer incidence compared to the general population. Methods Computer stored data on digitoxin concentrations in plasma from 9271 patients with cardiac disease were used to define a user population. Age and sex matched controls from the Norwegian Cancer Registry were used to calculate the number of expected cancer cases. Results The population on digitoxin showed a higher incidence of cancer compared to the control population. However, an additional analysis showed that the population on digitoxin had a general increased risk of cancer already, before the start on digitoxin. Leukemia/lymphoma were the cancer types which stood out with the highest risk in the digitoxin population before starting on digitoxin. This indicates that yet unknown risk factors exist for cardiovascular disease and lymphoproliferative cancer. An internal dose-response analysis revealed a relationship between high plasma concentration of digitoxin and a lower risk for leukemia/lymphoma and for cancer of the kidney/urinary tract. Conclusion Morbidity and mortality are high in the population on digitoxin, due to high age and cardiac disease.These factors disturb efforts to isolate an eventual anticancer effect of digitoxin in this setting. Still, the results may indicate an anticancer effect of digitoxin for leukemia/lymphoma and kidney/urinary tract cancers. Prospective clinical cancer trials have to be done to find out if digitoxin and other cardiac glycosides are useful as anticancer agents.

  3. Binding of digitoxin and digoxin to normal human β-lipoproteins

    International Nuclear Information System (INIS)

    Brock, A.


    The binding of digitoxin and digoxin to purified β-lipoprotein, obtained from pooled normal human serum, was studied under equilibrium conditions. Even with as high concentrations of unbound digitoxin or digoxin as 4 μmol/l, the preparations of β-lipoproteins, containing cholesterol 1.98-3.95 mmol/l, showed no signs of saturation. The binding affinity of digitoxin was about ten times as high as that of digoxin. Gel filtration chromatography, performed on native serum after addition of 3 H-digitoxin or 3 H-digoxin, showed a minor fraction of the cardiac glycosides to be associated with te protein fraction of highest molecular weight. This phenomenon disappeared after precipitation of the β-lipoproteins. In clinical relations the contribution of protein-bound digitoxin caused by the lipoprotein interaction is immaterial compared to that caused by the albumin interaction. (author)

  4. Stofmonografie digoxine en digitoxine

    NARCIS (Netherlands)

    Cremers PMA; Sangster B


    Deze monografie besteedt ruim aandacht aan de kinetiek en effecten van digitalispreparaten en waarnemingen gedaan bij mensen en dieren na gebruik van deze preparaten. Verder worden de klinische symptomatologie en de therapeutische mogelijkheden beschreven van een digitalis-intoxicatie.

  5. Fluorescence spectrometric studies on the binding of puerarin to human serum albumin using warfarin, ibuprofen and digitoxin as site markers with the aid of chemometrics

    International Nuclear Information System (INIS)

    Zhang Guowen; Zhao Nan; Wang Lin


    The interaction of puerarin with human serum albumin (HSA) in pH 7.4 Tris-HCl buffer has been investigated by fluorescence, Fourier transform infrared (FT-IR) and circular dichroism (CD) spectroscopy. The results revealed the presence of static type of quenching mechanism in the binding of puerarin to HSA. The association constants (K a ) between puerarin and HSA were obtained according to Modified Stern-Volmer equation. The calculated thermodynamic parameters indicated that the binding of puerarin to HSA was driven mainly by hydrophobic interaction. The competitive experiments of site markers suggested that the binding site of puerarin to HSA was located in the region of subdomain IIA (sudlow site I). Further, a chemometrics approach, parallel factor analysis (PARAFAC), was applied to resolve the measured three-way synchronous fluorescence spectra data of the competitive interaction between puerarin and warfarin with HSA. The concentration information for the three reaction components, warfarin, puerarin and puerarin-HSA, in the system at equilibrium was obtained simultaneously. The PARAFAC analysis indicated that puerarin in the puerarin-HSA complex was displaced by warfarin, which confirmed the binding site of puerarin to HSA was located in site I. Moreover, the results of CD and FT-IR spectra demonstrated that the secondary structure of HSA was changed in the presence of puerarin. - Highlights: → Puerarin can quench the fluorescence of human serum albumin (HSA). → The HSA fluorescence is quenched by puerarin through a static quenching mechanism. → The binding of puerarin to HSA is driven mainly by hydrophobic interaction. → The parallel factor analysis confirms that puerarin is located in site I of HSA. → The binding of puerarin to HSA induces changes in the secondary structure of HSA.

  6. Influence of polar and non-polar digoxin and digitoxin metabolites on the /sup 86/Rb-uptake of human erythrocytes and the contractility of guinea pig papillary muscles

    Energy Technology Data Exchange (ETDEWEB)

    Belz, G G; Heinz, N [Bundeswehr-Zentralkrankenhaus, Koblenz (Germany, F.R.). Medizinische Abt.; Beiersdorf A G, Hamburg Pharma-Forschung [Germany, F.R.


    The potency of various digoxigenin and digitoxigenin derivatives with different polarity was tested in two biological systems: First, in an /sup 86/Rb-erythrocyte assay which allows to determine the influence on active cation transport (measured as the glycoside concentration exerting half maximal inhibition of /sup 86/Rb-uptake of human erythrocytes = IC/sub 50/). Second, with isolated guinea pig papillary muscle, which allows to determine glycoside effects on contractile force (measured as the glycoside concentration exerting a 100% increase of contractile force = C+/sub 100/B). The IC/sub 50/ of the substances covered a range from 3.2 to 4800 x 10/sup -9/M, the C+/sub 100/B from 0.7 to 978 x 10/sup -6/ M. In both assay systems the glucuronides of glycosides and genins were between 1.4 and 11 times less potent than the original substances. A highly significant correlation (p < 0.0001) was found between IC/sub 50/ and C+/sub 100/B (r = 0.9996) and between log IC/sub 50/ and log C+/sub 100/B (r = 0.9819), the slope for the latter correlation being nearly unity (= 0.9912). The results support the hypothesis that inhibition of active cation transport is an important step in glycoside induced positive-inotropic effect. (orig.) 891 VJ 892 AP.

  7. A novel inexpensive murine model of oral chronic digitalization. (United States)

    Helber, Izo; Kanashiro, Rosemeire M; Alarcon, Ernesto A; Antonio, Ednei L; Tucci, Paulo J F


    A novel inexpensive murine model of oral administration of digitoxin (100 micro g/kg per day) added to routine chow is described. Serum digitoxin levels achieved after oral (n = 5; 116 +/- 14 ng/mL) and subcutaneous (n = 5; 124 +/- 11 ng/mL) administration were similar. A significant increase in the maximal left ventricular pressure rise of treated (n = 9) compared with control (n = 6) rats (dP/dt: 8956 +/- 233 vs 7980 +/- 234 mmHg/s, respectively; P = 0.01) characterized the positive inotropic action of digitoxin. In addition, no differences were observed in treated compared with control rats with regard to the electrocardiogram and systolic and diastolic left ventricular pressures.

  8. Radioimmuoassay study of antidigitoxin antibodies in liquid phase and after coupling on a solid phase

    International Nuclear Information System (INIS)

    Collignon, A.; German, A.; Scherrmann, J.M.; Bourdon, R.


    Antidigitoxin antibodies prepared by immunizing rabbits with a digitoxin-bovine serum albumin conjugate have been studied by radioimmunoassay in the native serum (homogeneous phase antibodies) and after coupling on glass beads (heterogeneous phase antibodies). Homogeneous phase antibodies present a satisfactory titer and affinity constant and react very specifically with digitoxin. Fixation of antibodies on a solid phase induce a loss of their immunoreactivity as it is showed by modification of the inhibition curves, by a greater sensitivity to the chemical structure of the tracer and by a decrease of the affinity constant. Reactionnal kinetic and sensitivity to the incubation temperature are not modified. Heterogeneous phase antibodies present a greater stability. Both antibodies types can be used for a digitoxin radioimmunoassay [fr

  9. Synthesis and evaluation of cardiac glycoside mimics as potential anticancer drugs

    DEFF Research Database (Denmark)

    Jensen, Marie; Schmidt, Steffen; Fedosova, Natalya


    recent years cardiac glycosides have furthermore been suggested to possess valuable anticancer activity. To mimic the labile trisaccharide of digitoxin with a stabile carbohydrate surrogate, we have used sulfur linked ethylene glycol moieties of varying length (mono-, di-, tri- or tetra-ethylene glycol...... the shortest mimics were found to have highest efficacy, with the best ligand having a monoethylene glycol unit (IC(50) 0.24 μM), which was slightly better than digitoxigenin (IC(50) 0.64 μM), while none of the novel cardiac glycoside mimics display an in vitro effect as high as digitoxin (IC(50) 0.02 μM)....

  10. Effect of prophylactic digitalization on the development of myocardial hypertrophy. (United States)

    Cutilletta, A F; Rudnik, M; Arcilla, R A; Straube, R


    The effect of prophylactic digitalization on the development of left ventricular hypertrophy was studied in adult rats. Digitoxin, 0.1 mg/100 g body wt or solvent was given daily for 1 wk prior to either aortic constriction or sham operation and was continued until the animals were killed, either 1 or 4 wk after surgery. A hemodynamic study was done in those animals killed 1 wk after surgery; hearts of all animals were examined for evidence of myocardial hypertrophy. Constriction of the ascending aorta had no significant effect on cardiac output but did reduce peak flow velocity and flow acceleration. An increase in left ventricular mass, RNA, and hydroxyproline was found in the animals with aortic constriction. Digitoxin treatment did not alter peak flow velocity or flow acceleration, but did significantly increase isovolumic (dP/dt)P-1. Digitoxin had no effect on body weight, heart weight, RNA, or hydroxyproline in either the sham-operated animals or in the animals with aortic constriction. Therefore, despite plasma digitoxin levels sufficient to affect myocardial contractility, left ventricular hypertrophy still developed after aortic constriction.

  11. Variability in the occurence of formylated diditalose glycosides in Digitalis purpurea L.

    NARCIS (Netherlands)

    Lugt, Charles Bruno


    The mutual synergistic activity of digitoxin and verdoxin has in recent years led to an interest in Digitalis pururea L. races with a high content of the verdoxin complex. In order to select such races, answers were required to the following quistions: Is the total verdoxin complex (glucoverodoxin +

  12. Reagents for the assay of cardenolide glycosides and aglycones

    International Nuclear Information System (INIS)

    Wilkinson, S.


    Some novel reagents are described for use in the radioimmunoassay of the 3-glycone derivatives of cardenolides (cardiac glycosides) and more especially digoxin, digitoxin, gitoxin, periplocin and lanatosides. Using these reagents these cardenolides and their derivatives may be assayed both in aqueous solution and in urine. A method is also described for performing such assays, including a suitable kit. (U.K.)

  13. The rate of uptake of cardiac glycosides into human cultured cells and the effects of chloroquine on it. (United States)

    Algharably, N; Owler, D; Lamb, J F


    HeLa cells grown on Petri dishes were either pulse labelled with various cardiac glycosides or grown in low concentrations of them for up to 2 days; either in the presence of chloroquine or not. The cells were then homogenised and the cell free homogenate layered on a continuous sucrose gradient; and the glycoside content and that of various markers measured. In another series of experiments HeLa cells were grown on plastic beads under the above conditions and then the content of glycosides and of some marker enzymes measured. The rate of internalisation of ouabain, digoxin and digitoxin from the plasma membrane preparation produced by the bead method is at 9% hr-1, similar to the rate of loss of digoxin and digitoxin from whole cells but much faster than that of ouabain. In the sucrose gradient experiments it was found that [3H]ouabain, digoxin and digitoxin all initially co-distribute with the plasma membrane marker, 5'-nucleotidase, and then leave this fraction of the homogenate at a fast rate when kept at 37 degrees, to co-distribute with the lysosomal marker, beta-hexosaminidase. At 2 degrees the ouabain remains co-distributed with the plasma membrane marker. The rate of transfer is estimated to be some 90% hr-1, much faster than previously thought. Chloroquine causes an increased retention of digoxin and digitoxin in the lysosomal fraction of the homogenate. These results are best explained by supposing that the sodium pump-glycoside complex rapidly enters a region of the peripheral cytoplasm, and that this region then controls the subsequent exit of digoxin and digitoxin from the cell. The main barrier for ouabain occurs at a stage later than this. The consequences of this model on other aspects of pump activity is discussed.

  14. In vitro propagation and production of cardiotonic glycosides in shoot cultures of Digitalis purpurea L. by elicitation and precursor feeding. (United States)

    Patil, Jitendra Gopichand; Ahire, Mahendra Laxman; Nitnaware, Kirti Manik; Panda, Sayantan; Bhatt, Vijay P; Kishor, Polavarapu B Kavi; Nikam, Tukaram Dayaram


    Digitalis purpurea L. (Scrophulariaceae; Foxglove) is a source of cardiotonic glycosides such as digitoxin and digoxin which are commercially applied in the treatment to strengthen cardiac diffusion and to regulate heart rhythm. This investigation deals with in vitro propagation and elicited production of cardiotonic glycosides digitoxin and digoxin in shoot cultures of D. purpurea L. In vitro germinated seedlings were used as a primary source of explants. Multiple shoot formation was achieved for three explant types (nodal, internodal, and leaf) cultured on Murashige and Skoog (MS) medium with several treatments of cytokinins (6-benzyladenine-BA; kinetin-Kin; and thidiazuron-TDZ) and auxins (indole-3-acetic acid-IAA; α-naphthaleneacetic acid-NAA; and 2,4-dichlorophenoxy acetic acid-2,4-D). Maximum multiple shoots (12.7 ± 0.6) were produced from nodal explants on MS + 7.5 μM BA. Shoots were rooted in vitro on MS containing 15 μM IAA. Rooted plantlets were successfully acclimatized. To further maintain the multiple shoot induction, mother tissue was cut into four equal parts and repeatedly sub-cultured on fresh shoot induction liquid medium after each harvest. On adaptation of this strategy, an average of 18 shoots per explant could be produced. This strategy was applied for the production of biomass and glycosides digitoxin and digoxin in shoot cultures on MS medium supplemented with 7.5 μM BA and several treatments with plant growth regulators, incubation period, abiotic (salicylic acid, mannitol, sorbitol, PEG-6000, NaCl, and KCl), biotic (Aspergillus niger, Helminthosporium sp., Alternaria sp., chitin, and yeast extract) elicitors, and precursors (progesterone, cholesterol, and squalene). The treatment of KCl, mycelial mass of Helminthosporium sp., and progesterone were highly effective for the production of cardenolides. In the presence of progesterone (200 to 300 mg/l), digitoxin and digoxin accumulation was enhanced by 9.1- and 11.9-folds

  15. Chronic digitalis therapy in patients before heart transplantation is an independent risk factor for increased posttransplant mortality

    Directory of Open Access Journals (Sweden)

    Rivinius R


    Full Text Available Rasmus Rivinius,1 Matthias Helmschrott,1 Arjang Ruhparwar,2 Ann-Kathrin Rahm,1,3 Fabrice F Darche,1 Dierk Thomas,1 Tom Bruckner,4 Philipp Ehlermann,1 Hugo A Katus,1 Andreas O Doesch1,5 1Department of Cardiology, Angiology and Pneumology, Heidelberg University Hospital, Heidelberg, 2Department of Cardiac Surgery, Heidelberg University Hospital, Heidelberg, 3Faculty of Medicine, University of Heidelberg, Heidelberg, 4Institute for Medical Biometry and Informatics, University of Heidelberg, Heidelberg, 5Asklepios Klinik Bad Salzungen GmbH, Department of Pneumology and Oncology, Bad Salzungen, Germany Objectives: Digitalis therapy (digoxin or digitoxin in patients with heart failure is subject to an ongoing debate. Recent data suggest an increased mortality in patients receiving digitalis. This study investigated the effects of chronic digitalis therapy prior to heart transplantation (HTX on posttransplant outcomes.Patients and methods: This was a retrospective, observational, single-center study. It comprised 530 adult patients who were heart-transplanted at Heidelberg University Hospital between 1989 and 2012. Patients with digitalis prior to HTX (≥3 months were compared to those without (no or <3 months of digitalis. Patients with digitalis were further subdivided into patients receiving digoxin or digitoxin. Primary outcomes were early posttransplant atrial fibrillation and mortality.Results: A total of 347 patients (65.5% had digitalis before HTX. Of these, 180 received digoxin (51.9% and 167 received digitoxin (48.1%. Patients with digitalis before HTX had a significantly lower 30-day (P=0.0148 and 2-year (P=0.0473 survival. There was no significant difference between digoxin and digitoxin in 30-day (P=0.9466 or 2-year (P=0.0723 survival. Multivariate analysis for posttransplant 30-day mortality showed pretransplant digitalis therapy as an independent risk factor (hazard ratio =2.097, CI: 1.036–4.248, P=0.0397. Regarding atrial

  16. Potency of Gamma ray, Electric Current and Elicitor Application, as a Novel Practical Technique, to Improve Biomass Production and Glycoside Quality for Digitalis purpurea L. Grown in Sandy Soil Irrigated with Brackish Water

    International Nuclear Information System (INIS)

    Bosila, H.A.; Afifi, L.M.A.; Ahmed, T.E.S.


    Digitalis purpurea L seeds were treated before sowing with gamma ray (G:0, 2.5, 5, 7.5 KR, and electric current (E:O, 100, 150, 200 mA) then grown in sandy soil irrigated with brackish water (900 ppm) , in splite-splite plot design for 3 replicat (R) at two subsequent seasons , through surface drip irrigation system. Plants at 4- month old and monthly until before flowering were foliar sprayed with MnSO 4 as abiotic elicitor (M :O, 3 ppm). Biomass/ Feddan, percentage of total glycosides and percentage of bioactive glycosides, digitoxin and gitoxin were quantitated. Statistical analysis for the obtained data revealed that G, E and M achieved significant in biomass yield and its quality traits. Moreover, interactions ; GE, GM, EM and GEM achieved synergistic and significant increment for this traits. At such G dose the trait was increased by increasing E dose and M concentration. Hence, G 2.5, 5,7.5 KR E200 mA M3 ppm achieved significant increment, as percent over that of control, in biomass production / Feddan by 22, 29, 32%, total glycoside by 27, 40, 30%, digitoxin 27, 40, 30% for both first and second seasons, respectively. Whereas, increment for gitoxin were 27, 41, 30% at first season and 26, 38, 30% at second season, respectively. Overall, these finding strongly confirm the reliability of GEM as a novel practical technique for overproduction biomass/Fed. and quality improvement bioactive cardiac glycosides, digitoxin and gitoxin in Digitalis purpurea L.

  17. Kinetic properties of /sup 3/H-16. cap alpha. -gitoxin in the isolated guinea-pig atrium

    Energy Technology Data Exchange (ETDEWEB)

    Haustein, K O; Glusa, E [Medizinische Akademie, Erfurt (German Democratic Republic)


    The accumulation and the loss of (/sup 3/H)-16..cap alpha..-gitoxin were estimated in correlation with its positive inotropic effect using the isolated spontaneously beating guinea-pig atrium. The properties of 16..cap alpha..-gitoxin were compared with those of (/sup 3/H)-ouabain and (/sup 3/H)-digitoxin. The experiments were performed with positive inotropic equieffective glycoside concentrations. During a 2-hr incubation period, 16..cap alpha..-gitoxin was accumulated to a low extent (2.05 nmoles per g w.w., tissue/medium (T/M) ratio =0.66). Similar results were obtained with ouabain (47.5 pmoles per g w.w.; T/M ratio = 0.51), while digitoxin was accumulated to a high extent (266 pmoles per g w.w.; T/M ratio = 2.98). Wash-out experiments demonstrated that the inotropic effect disappears more rapidly than the loss of tissue radioactivity occurred, the effects of 16..cap alpha..-gitoxin were diminished more easily after wash-out than those of ouabain and digitoxin. In the presence of 1.0 instead of 0.1 vol-% dimethylformamide, both the accumulation of radioactivity (5.0 nmoles per g w.w.; T/M ratio = 1.58) and the positive inotropic effect are enhanced. From the experiments it was concluded: (1) 16..cap alpha..-gitoxin has similar kinetic properties as ouabain. (2) The concentration of dimethylformamide used influences the kinetic behaviour on the tissue level. (3) No correlation exists neither between the extent of the positive inotropic effect and theamount of accumulated glycoside, nor between the loss of tissue radioactivity and the disappearance of positive inotropic effect.

  18. Studies on digoxin derivatives in different biological media by radioimmunoassay

    International Nuclear Information System (INIS)

    Barmasch, Martha; Perez, L.N.; Altschuler, Noe.


    The fundamental aim of this study was to develop a methodology to be applied to pharmacological studies, directed to demonstrate the passage of digitalic compounds through the blood-brain barrier. This study was a comparative one between β methyl digoxin (βMD) and digitoxin (Dt). A methodology of radioimmunoassay was developed for different biological media: serum (S), plasma (P) and cerebro spinal fluid (CSF). In addition, the immunochemical behaviour (affinity, displacement, etc.) of βMD was studied when reacted with digoxin (D) and digoxin-antisera, supplied by the commercial kits utilized in these studies. (author) [es

  19. Diagnostic reagent system

    International Nuclear Information System (INIS)


    A solid phase reagent for use in radioimmunoassay of antigens and antibodies is described. The reagent is prepared by mixing specific antibody or radiolabeled antigen with polyethylene glycol (4000-6000) and gamma globulin in a buffer at pH 4-10 and lyophilizing, the antigens being thyroxine, triiodothyronine, digoxin and digitoxin (1-1000 μCi 125 I/μg). The buffer consists of a 0.08 molar sodium barbital solution containing 0.1% of ox-serum albumin and 0.35% of 8-anilino-1-naftalene sulfonic acid

  20. Radioimmunoassay method for the determination of cardiotonic glycosides

    International Nuclear Information System (INIS)


    A kit method for the in vitro determination of digoxin and digitoxin is described. The blood serum is mixed with the reagent which consists of an aqueous buffer solution containing a radiolabeled hapten for the glycoside. Antiserum with specific antibodies is added and the mixture is incubated. Thereafter, a thin strip of membrane mainly consisting of an ionexchanger is brought into contact with the mixture to separate the antibody bound hapten from the unbound hapten. The ratios of both are determined by counting the radioactive hapten

  1. Studies concerning the effect of X-rays on electrolytic shifts and on the metabolism of the myocardium. Pt. 7

    International Nuclear Information System (INIS)

    Prignitz, R.; Froehlich, D.; Hoffmeister, G.


    The behavior of a Mg 2+ -dependent microsomal ATPase, which may be activated by Na + and K + , was investigated in the myocardium of Guinea pigs after local irradiation. The part which can be activated by Na + and K + (transport-ATPase) is already slightly inhibited thirty minutes after a dose of 250 R; the inhibition rate with 1,500 R amounts to about 60%, the maximal inhibition effect with a dose of 3,000 R was about 70%. Four hours after a local irradiation with 1,500 R, a restitution of the inhibition effect occurs with overshoot reaction; 15 and 24 hours following irradiation, a significant inhibition of the transport-ATPase activity can again be verified. The k-strophanthidine and digitoxin are inhibiting the concentration ranges analyzed (2 x 10 -5 , 2.5 x 10 -7 M) the activity of the transport-ATPase. The above-mentioned radiation-induced inhribition effect upon the ATPase activated by Na + and K + is additively increased by adding k-strophanthidine (2 x 10 -5 M) and digitoxin (2.5 x 10 -7 M). (orig.) [de

  2. Bilary and urinary excretion of five cardiac glycosides and its correlation with their physical and chemical properties. (United States)

    Marzo, A; Ghirardi, P


    Biliary and urinary excretion of five tritium-labelled cardiac glycosides, i.e. Ouabain, K-strophanthoside, Digoxin, Digitoxin and Deslanatoside C, were investigated in anaesthetized guinea-pigs 5 h after i.v. or enteral administration. Urinary excretion is the main route of elimination in the case of Ouabain and Deslanatoside C. Conversely, biliary excretion is predominant in the case of Digoxin and Digitoxin. K-strophanthoside is excreted both via bile and urine. In conscious guinea-pigs treated i.v. with the same cardiac glycosides the highest levels were observed in urine, bile, kidneys and liver. The relative values of those levels were in agreement with the excretion pattern observed in anaesthetized animals. An inverse linear relation (P less than 0.05) was encountered between biliary excretion rate and polarity of glycoside molecula. This correlation has been previously observed by other authors in other species, but not in the rabbit. This suggests that the correlation may not be considered generally applicable at present.

  3. Problems of rapid digitalization in severe congestive heart failure. (United States)

    Haustein, K O; Assmann, I; Fiehring, H


    The pharmacodynamic effects (changes of systolic time intervals, STI, reaction of pulmonary arterial pressure) of digitoxin were studied in 7 patients with severe congestive heart failure in comparison with the corresponding plasma level. STI indicated glycoside-dependent changes, i.e. shortening of LVETc and QS2c and normalization of prolonged PEPc, while ICT shortening was less observed. In 2 patients with cor pulmonale a pulmonary oedema occurred accompanied with prolonged LVETc. During the early period of glycoside-dependent recompensation no significant correlation between STI shortening and glycoside plasma level was observed. Because of the retarded normalization of the haemodynamics of the pulmonary circulation and because of possible side-effects, rapid digitalization has to be reconsidered.

  4. Trial Watch: Immunogenic cell death inducers for anticancer chemotherapy. (United States)

    Pol, Jonathan; Vacchelli, Erika; Aranda, Fernando; Castoldi, Francesca; Eggermont, Alexander; Cremer, Isabelle; Sautès-Fridman, Catherine; Fucikova, Jitka; Galon, Jérôme; Spisek, Radek; Tartour, Eric; Zitvogel, Laurence; Kroemer, Guido; Galluzzi, Lorenzo


    The term "immunogenic cell death" (ICD) is now employed to indicate a functionally peculiar form of apoptosis that is sufficient for immunocompetent hosts to mount an adaptive immune response against dead cell-associated antigens. Several drugs have been ascribed with the ability to provoke ICD when employed as standalone therapeutic interventions. These include various chemotherapeutics routinely employed in the clinic (e.g., doxorubicin, epirubicin, idarubicin, mitoxantrone, bleomycin, bortezomib, cyclophosphamide and oxaliplatin) as well as some anticancer agents that are still under preclinical or clinical development (e.g., some microtubular inhibitors of the epothilone family). In addition, a few drugs are able to convert otherwise non-immunogenic instances of cell death into bona fide ICD, and may therefore be employed as chemotherapeutic adjuvants within combinatorial regimens. This is the case of cardiac glycosides, like digoxin and digitoxin, and zoledronic acid. Here, we discuss recent developments on anticancer chemotherapy based on ICD inducers.

  5. Comfrey herbal remedy causing second-degree heart block: do not be outfoxed by digitalis. (United States)

    Vithayathil, Mathew Kurian; Edwards, Matthew


    A previously well woman aged 63 years presents to the emergency department with vomiting, palpitations and 3 presyncopal episodes. She had no previous medical or cardiac history, with the patient stating that she tried a herbal remedy of boiled comfrey leaves for insomnia 18 hours before arrival to the department. Her ECG showed multiple abnormalities, including bradycardia, second-degree atrioventricular node block, Mobitz Type 2, a shortened QT interval, downsloping ST depression and presence of U waves. After viewing the images of comfrey and foxglove, it highlighted the possibility of mistaken ingestion of Digitalis, containing the organic forms of cardiac glycosides, such as digoxin and digitoxin. Raised serum digoxin levels confirmed this. The patient was haemodynamically stable, and given digoxin-binding antibodies. After 5 days of cardiac monitoring, her ECG returned to normal rhythm, and she was discharged home. 2016 BMJ Publishing Group Ltd.

  6. A critical appraisal of a further three new commercial digoxin radioimmunoassay kits with reference to cross-reacting substances

    International Nuclear Information System (INIS)

    Wood, W.G.; Wachter, C.


    A further 3 digoxin radioimmunoassay (RIA) kits have been evaluated for performance and cross-reaction with digitoxin, spironolactone, canrenone and furosemide (Lasix-Hoechst). Effects of serum protein concentrations have also been tested. The kits tested were from the following manufacturers: A) Diagnostic Products Corporation Digoxin RIA Kit. B) Byk-Mallinckrodt SPAC Digoxin Kit. C) Boehringer-Mannheim Digoxin RIA Kit. All kits used a 125 I-labelled tracer. Kit A used a conventional liquid phase system using double-antibody separation for bound and free drug. Kits B and C used a solid-phase antibody coated tube method. All kits showed a lower cross-reaction to digitoxin than quoted by the manufacturer. Cross-reaction to spironolactone (Aldactone - Boehringer-Mannheim) was less than 1.50 nmol/l at a serum concentration of 125 mg/l Aldactone in all 3 kits. The cross-reaction to canrenone was somewhat higher, 5.2 nmol/l 'digoxin' being measured in one kit at a serum canrenone concentration of 125 mg/l. There was no cross-reaction with furosemide in any kit, even at a serum concentration of 5 g/l. The coated-tube assays were affected by serum albumin and globulin concentration changes, one kit showing a difference of over 50% binding in the range 1-20% albumin. The double-antibody kit did not show dependence on the concentration of these proteins. All kits measured digoxin with good reproducibility in the range 0.40-10.0 nmol/l. (orig.) [de

  7. Tissue distribution of tritiated digoxin and quabain in mice

    International Nuclear Information System (INIS)

    Klopper, J.F.; Atkins, H.L.


    Tissue concentration of tritiated digoxin in human subjects appears to be selective in that after a single dose of the radioactive tracer a myocardial concentration of digoxin on an average 24 to 30 times higher than the serum concentration is reached. In patients studied 3.5 to 84 hours after administration of tritiated digoxin, the dose in the heart was found to vary between 1.95 to 4.83 percent. Should it be possible to label digoxin or another cardiac glycoside with a suitable gamma-emitting tracer, its use as a cardiac imaging agent should thus be feasible. This study was undertaken to determine the tissue distribution in mice of tritiated digoxin at various time intervals post injection; and to determine if these values showed any correlation to the previously determined human data. A preliminary study in mice using 3 H-digitoxin revealed no selective uptake in the heart with high uptakes in the liver and gut. Since ouabain has a more rapid clinical onset of action than digoxin, its tissue distribution was included in the study and compared to that of digoxin

  8. Induced responses to herbivory and jasmonate in three milkweed species. (United States)

    Rasmann, Sergio; Johnson, M Daisy; Agrawal, Anurag A


    We studied constitutive and induced defensive traits (latex exudation, cardenolides, proteases, and C/N ratio) and resistance to monarch caterpillars (Danaus plexippus) in three closely related milkweed species (Asclepias angustifolia, A. barjoniifolia and A. fascicularis). All traits showed significant induction in at least one of the species. Jasmonate application only partially mimicked the effect of monarch feeding. We found some correspondence between latex and cardenolide content and reduced larval growth. Larvae fed cut leaves of A. angustifolia grew better than larvae fed intact plants. Addition of the cardenolide digitoxin to cut leaves reduced larval growth but ouabain (at the same concentration) had no effect. We, thus, confirm that latex and cardenolides are major defenses in milkweeds, effective against a specialist herbivore. Other traits such as proteases and C/N ratio additionally may be integrated in the defense scheme of those plants. Induction seems to play an important role in plants that have an intermediate level of defense, and we advocate incorporating induction as an additional axis of the plant defense syndrome hypothesis.

  9. Evaluation of the binding interaction between bovine serum albumin and dimethyl fumarate, an anti-inflammatory drug by multispectroscopic methods (United States)

    Jattinagoudar, Laxmi; Meti, Manjunath; Nandibewoor, Sharanappa; Chimatadar, Shivamurti


    The information of the quenching reaction of bovine serum albumin with dimethyl fumarate is obtained by multi-spectroscopic methods. The number of binding sites, n and binding constants, KA were determined at different temperatures. The effect of increasing temperature on Stern-Volmer quenching constants (KD) indicates that a dynamic quenching mechanism is involved in the interaction. The analysis of thermodynamic quantities namely, ∆H° and ∆S° suggested hydrophobic forces playing a major role in the interaction between dimethyl fumarate and bovine serum albumin. The binding site of dimethyl fumarate on bovine serum albumin was determined by displacement studies, using the site probes viz., warfarin, ibuprofen and digitoxin. The determination of magnitude of the distance of approach for molecular interactions between dimethyl fumarate and bovine serum albumin is calculated according to the theory of Förster energy transfer. The CD, 3D fluorescence spectra, synchronous fluorescence measurements and FT-IR spectral results were indicative of the change in secondary structure of the protein. The influence of some of the metal ions on the binding interaction was also studied.

  10. Characterisation of the sarcolemmal calcium pump in guinea pig hearts and its influence through g-strophanthin. Charakterisierung der sarkolemmalen Calciumpumpe des Meerschweinchenherzens und ihre Beeinflussung durch g-Strophanthin

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    Lell, R.


    In this work the sarcolemmal calcium transport of the guinea pig heart is studied with the help of the micropore filtration method on the sarcolemmal vesicles. The calcium ATPase could be better characterised by its ability to accumulate calcium in the sarcolemmal vesicles. The calcium pump is magnesium-dependent and positively activated by sodium and potassium. The pH-optimum of the activity lies around 6.8-7.0 depending on intracellular conditions. Up until a concentration of 30 micrograms protein per ml of reaction mixture the calcium accumulation increased proportionally. With higher concentrations it came to a relative decrease. The addition of digitoxin and g-strophanthin to the reaction mixture did not change the activity of the calcium pump. With a perfusion of the heart with a toxic concentration of 10/sup 06/ M g-strophanthin a glycoside intoxication on the beating heart was caused and a time-dependent inhibition of the calcium pump of sarcolemmal vesicles was detectable.

  11. Plant-derived cardiac glycosides: Role in heart ailments and cancer management. (United States)

    Patel, Seema


    Cardiac glycosides, the cardiotonic steroids such as digitalis have been in use as heart ailment remedy since ages. They manipulate the renin-angiotensin axis to improve cardiac output. However; their safety and efficacy have come under scrutiny in recent times, as poisoning and accidental mortalities have been observed. In order to better understand and exploit them as cardiac ionotropes, studies are being pursued using different cardiac glycosides such as digitoxin, digoxin, ouabain, oleandrin etc. Several cardiac glycosides as peruvoside have shown promise in cancer control, especially ovary cancer and leukemia. Functional variability of these glycosides has revealed that not all cardiac glycosides are alike. Apart from their specific affinity to sodium-potassium ATPase, their therapeutic dosage and behavior in poly-morbidity conditions needs to be considered. This review presents a concise account of the key findings in recent years with adequate elaboration of the mechanisms. This compilation is expected to contribute towards management of cardiac, cancer, even viral ailments. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  12. Posterior encephalopathy with vasospasm: MRI and angiography

    International Nuclear Information System (INIS)

    Weidauer, S.; Gaa, J.; Lanfermann, H.; Zanella, F.E.; Sitzer, M.; Hefner, R.


    Posterior encephalopathy is characterised by headache, impairment of consciousness, seizures and progressive visual loss. MRI shows bilateral, predominantly posterior, cortical and subcortical lesions with a distribution. Our aim was to analyse the MRI lesion pattern and angiographic findings because the pathophysiology of posterior encephalopathy is incompletely understood. We report three patients with clinical and imaging findings consistent with posterior encephalopathy who underwent serial MRI including diffusion-weighted imaging (DWI) and construction of apparent diffusion coefficient (ADC) maps, and four-vessel digital subtraction angiography (DSA). DWI revealed symmetrical subcortical and cortical parieto-occipital high signal. High and also low ADCs indicated probable vasogenic and cytotoxic oedema. On follow-up there was focal cortical laminar necrosis, while the white-matter lesions resolved almost completely, except in the arterial border zones. DSA revealed diffuse arterial narrowing, slightly more marked in the posterior circulation. These findings suggest that posterior encephalopathy may in some cases be due to diffuse, severe vasospasm affecting especially in the parieto-occipital grey matter, with its higher vulnerability to ischemia. Cerebral vasospasm due to digitoxin intoxication, resulting in posterior encephalopathy, has not yet been described previously. (orig.)

  13. Performance of radioimmunoassays for digoxin as evaluated by a group experiment

    International Nuclear Information System (INIS)

    Dwenger, A.; Friedel, R.; Trautschold, I.


    In order to gather informations on the performance of routinely employed test system for the radioimmunological determination of digoxin in serum a group experiment was set up in which 36 laboratories in the Federal Republic of Germany took part. They were asked to determine the digoxin content in 25 specimens including a recovery curve, sera with pathologic composition and known concentrations of digoxin, sera from a pharmacokinetic study and commercial control sera. This identity of the specimens was withheld from the participants. As far as accuracy and precision are concerned the results reported for a total of 54 assays were better than those obtained in group experiments on radioimmunological determination of hormones. Recovery ranged from 90% to 110% in 57%, from 80% to 120% in 85% of the assays. Reproducibility in the series expressed as coefficient of variation was better than 5% in 54%, better than 10% in 85% of the assays. Considerable differences were found for the cross-reactivities of antibodies with digitoxin and metabolites of spironolactone. Dysproteinemia seems to be an unsolved problem whereas moderate hemolysis and hyperlipemia did not lead to severe errors. Improvements, especially with respect to inter-laboratory variances can be expected mainly from a further standardization of standard preparations. (orig.) [de

  14. Human breast tumor cells are more resistant to cardiac glycoside toxicity than non-tumorigenic breast cells.

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    Rebecca J Clifford

    Full Text Available Cardiotonic steroids (CTS, specific inhibitors of Na,K-ATPase activity, have been widely used for treating cardiac insufficiency. Recent studies suggest that low levels of endogenous CTS do not inhibit Na,K-ATPase activity but play a role in regulating blood pressure, inducing cellular kinase activity, and promoting cell viability. Higher CTS concentrations inhibit Na,K-ATPase activity and can induce reactive oxygen species, growth arrest, and cell death. CTS are being considered as potential novel therapies in cancer treatment, as they have been shown to limit tumor cell growth. However, there is a lack of information on the relative toxicity of tumor cells and comparable non-tumor cells. We have investigated the effects of CTS compounds, ouabain, digitoxin, and bufalin, on cell growth and survival in cell lines exhibiting the full spectrum of non-cancerous to malignant phenotypes. We show that CTS inhibit membrane Na,K-ATPase activity equally well in all cell lines tested regardless of metastatic potential. In contrast, the cellular responses to the drugs are different in non-tumor and tumor cells. Ouabain causes greater inhibition of proliferation and more extensive apoptosis in non-tumor breast cells compared to malignant or oncogene-transfected cells. In tumor cells, the effects of ouabain are accompanied by activation of anti-apoptotic ERK1/2. However, ERK1/2 or Src inhibition does not sensitize tumor cells to CTS cytotoxicity, suggesting that other mechanisms provide protection to the tumor cells. Reduced CTS-sensitivity in breast tumor cells compared to non-tumor cells indicates that CTS are not good candidates as cancer therapies.

  15. [No effect of digitalis on sex and adrenal hormones in healthy subjects and in patients with congestive heart failure]. (United States)

    Kley, H K; Abendroth, H; Hehrmann, R; Müller, A; Keck, E; Schneitler, H; Elsässer, H; Krüskemper, H L


    Digoxin was studied to see whether it impairs adrenal function and feminizes male subjects by changing plasma sexual hormones; both have been reported on previously. In eight healthy male subjects neither estrone (38.7 +/- 7.7 vs 35.4 +/- 3.2 pg/ml) nor estradiol (35.8 +/- 6.4 vs 32.2 +/- 3.9 pg/ml) nor testosterone (6.32 +/- 0.74 vs 6.45 +/- 0.73 ng/ml) were found to be altered by digoxin administration (plasma levels 1.55 +/0- 0.27 ng/ml) lasting 35 days. The same was true of free testosterone (147 +/- 24 vs 142 +/- 19 pg/ml) and free estradiol (657 +/- 77 vs 615 +/- 78 fg/ml). Even maximal stimulation of the adrenal and gonadal glands by adrenocorticotropic hormone (ACTH) and human chorionic gonadotropin (hCG) did not exhibit any digoxin-induced alterations in the synthesizing capacity of steroid hormones, as shown by plasma cortisol (increase from 128 +/- 18 to 389 +/- 18 ng/ml) and testosterone (from 5.96 +/- 0.90 to 10.33 +/- 1.19 ng/ml). Furthermore, seven subjects on digoxin were observed over a period of 150-210 days; they did not show any increase of estrogens. This was also found in three subjects when estrogen levels were elevated initially due to extreme obesity. Also, 35 patients who took beta-methyldigoxin (n = 8), beta-acetyldigoxin (n = 20) and digitoxin (n = 7) from 1 to 9 (mean: 1.9) years demonstrated normal plasma concentrations of gonadal and adrenal steroids, irrespective of duration of application or the digitalis compound.(ABSTRACT TRUNCATED AT 250 WORDS)

  16. Studies on the comparability of the results from different methods for the radioimmunological determination of digoxin

    International Nuclear Information System (INIS)

    Dwenger, A.; Trautschold, I.


    Three iodine-125-digoxin radioimmunoassay kits (A Amersham Buchler; B Boehringer Mannheim; C Schwarz Mann/Becton Dickinson) were evaluated with respect to assay quality and comparability of the results. Intra- and interassay variances were calculated for the following types of samples: Three media (a pool serum; b artificial human serum; c buffer solution with albumin and globulin) containing pure digoxin, sera from a pharmacokinetic study, sera with different concentrations of proteins, a hemolytic serum and sera with digitoxin and metabolites of spironolactone. The intra-assay precision depended on the medium of the sample and was higher for samples with identical digoxin concentrations in an identical medium (e.g. CV for 2 μg/l in medium a for kit A: 4.3% for kit B: 7.0%; for kit C: 2.2%) than for samples with identical antigen concentrations in different media (CV for 2 μg/l in media a, b and c for kit A: 6.4%; for kit B: 9.1%; for kit C: 4.3%). The mean recovery in the range 0.5-4 μg/l depended on the kind of medium (a, b or c) and varied for kit A from 84.4% to 100.8%, for kit B from 112.0% to 119.6%, and for kit C from 98.0% to 104.5%. Decreasing serum protein concentrations to less than one half of the physiological concentration gave false negative results for kit A and fals positive results for kit C; for kit B this dependency was not be observed, but there was a decrease of reproducibility. (orig./AJ) [de

  17. Cardiac glycoside activities link Na(+)/K(+) ATPase ion-transport to breast cancer cell migration via correlative SAR. (United States)

    Magpusao, Anniefer N; Omolloh, George; Johnson, Joshua; Gascón, José; Peczuh, Mark W; Fenteany, Gabriel


    The cardiac glycosides ouabain and digitoxin, established Na(+)/K(+) ATPase inhibitors, were found to inhibit MDA-MB-231 breast cancer cell migration through an unbiased chemical genetics screen for cell motility. The Na(+)/K(+) ATPase acts both as an ion-transporter and as a receptor for cardiac glycosides. To delineate which function is related to breast cancer cell migration, structure-activity relationship (SAR) profiles of cardiac glycosides were established at the cellular (cell migration inhibition), molecular (Na(+)/K(+) ATPase inhibition), and atomic (computational docking) levels. The SAR of cardiac glycosides and their analogs revealed a similar profile, a decrease in potency when the parent cardiac glycoside structure was modified, for each activity investigated. Since assays were done at the cellular, molecular, and atomic levels, correlation of SAR profiles across these multiple assays established links between cellular activity and specific protein-small molecule interactions. The observed antimigratory effects in breast cancer cells are directly related to the inhibition of Na(+)/K(+) transport. Specifically, the orientation of cardiac glycosides at the putative cation permeation path formed by transmembrane helices αM1-M6 correlates with the Na(+) pump activity and cell migration. Other Na(+)/K(+) ATPase inhibitors that are structurally distinct from cardiac glycosides also exhibit antimigratory activity, corroborating the conclusion that the antiport function of Na(+)/K(+) ATPase and not the receptor function is important for supporting the motility of MDA-MB-231 breast cancer cells. Correlative SAR can establish new relationships between specific biochemical functions and higher-level cellular processes, particularly for proteins with multiple functions and small molecules with unknown or various modes of action.

  18. A structural view on the functional importance of the sugar moiety and steroid hydroxyls of cardiotonic steroids in binding to Na,K-ATPase. (United States)

    Cornelius, Flemming; Kanai, Ryuta; Toyoshima, Chikashi


    The Na,K-ATPase is specifically inhibited by cardiotonic steroids (CTSs) like digoxin and is of significant therapeutic value in the treatment of congestive heart failure and arrhythmia. Recently, new interest has arisen in developing Na,K-ATPase inhibitors as anticancer agents. In the present study, we compare the potency and rate of inhibition as well as the reactivation of enzyme activity following inhibition by various cardiac glycosides and their aglycones at different pH values using shark Na,K-ATPase stabilized in the E2MgPi or in the E2BeFx conformations. The effects of the number and nature of various sugar residues as well as changes in the positions of hydroxyl groups on the β-side of the steroid core of cardiotonic steroids were investigated by comparing various cardiac glycoside compounds like ouabain, digoxin, digitoxin, and gitoxin with their aglycones. The results confirm our previous hypothesis that CTS binds primarily to the E2-P ground state through an extracellular access channel and that binding of extracellular Na(+) ions to K(+) binding sites relieved the CTS inhibition. This reactivation depended on the presence or absence of the sugar moiety on the CTS, and a single sugar is enough to impede reactivation. Finally, increasing the number of hydroxyl groups of the steroid was sterically unfavorable and was found to decrease the inhibitory potency and to confer high pH sensitivity, depending on their position on the steroid β-face. The results are discussed with reference to the recent crystal structures of Na,K-ATPase in the unbound and ouabain-bound states.

  19. Ouabain, a cardiac glycoside, inhibits the Fanconi anemia/BRCA pathway activated by DNA interstrand cross-linking agents.

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    Dong Wha Jun

    Full Text Available Modulation of the DNA repair pathway is an emerging target for the development of anticancer drugs. DNA interstrand cross-links (ICLs, one of the most severe forms of DNA damage caused by anticancer drugs such as cisplatin and mitomycin C (MMC, activates the Fanconi anemia (FA/BRCA DNA repair pathway. Inhibition of the FA/BRCA pathway can enhance the cytotoxic effects of ICL-inducing anticancer drugs and can reduce anticancer drug resistance. To find FA/BRCA pathway inhibitory small molecules, we established a cell-based high-content screening method for quantitating the activation of the FA/BRCA pathway by measuring FANCD2 foci on DNA lesions and then applied our method to chemical screening. Using commercial LOPAC1280 chemical library screening, ouabain was identified as a competent FA/BRCA pathway inhibitory compound. Ouabain, a member of the cardiac glycoside family, binds to and inhibits Na(+/K(+-ATPase and has been used to treat heart disease for many years. We observed that ouabain, as well as other cardiac glycoside family members--digitoxin and digoxin--down-regulated FANCD2 and FANCI mRNA levels, reduced monoubiquitination of FANCD2, inhibited FANCD2 foci formation on DNA lesions, and abrogated cell cycle arrest induced by MMC treatment. These inhibitory activities of ouabain required p38 MAPK and were independent of cellular Ca(2+ ion increase or the drug uptake-inhibition effect of ouabain. Furthermore, we found that ouabain potentiated the cytotoxic effects of MMC in tumor cells. Taken together, we identified an additional effect of ouabain as a FA/BRCA pathway-inhibiting chemosensitization compound. The results of this study suggest that ouabain may serve as a chemosensitizer to ICL-inducing anticancer drugs.

  20. Identification and quantification of cardiac glycosides in blood and urine samples by HPLC/MS/MS. (United States)

    Guan, F; Ishii, A; Seno, H; Watanabe-Suzuki, K; Kumazawa, T; Suzuki, O


    Cardiac glycosides (CG) are of forensic importance because of their toxicity and the fact that very limited methods are available for identification of CG in biological samples. In this study, we have developed an identification and quantification method for digoxin, digitoxin, deslanoside, digoxigenin, and digitoxigenin by high-performance liquid chromatography tandem mass spectrometry (HPLC/MS/MS). CG formed abundant [M + NH4]+ ions and much less abundant [M + H]+ ions as observed with electrospray ionization (ESI) source and ammonium formate buffer. Under mild conditions for collision-induced dissociation (CID), each [M + NH4]+ ion fragmented to produce a dominant daughter ion, which was essential to the sensitive method of selected reaction monitoring (SRM) quantification of CG achieved in this study. SRM was compared with selected ion monitoring (SIM) regarding the effects of sample matrixes on the methodology. SRM produced lower detection limits with biological samples than SIM, while both methods produced equal detection limits with CG standards. On the basis of the HPLC/MS/MS results for CG, we have proposed some generalized points for conducting sensitive SRM measurements, in view of the property of analytes as well as instrumental conditions such as the type of HPLC/MS interface and CID parameters. Analytes of which the molecular ion can produce one abundant daughter ion with high yield under CID conditions may be sensitively measured by SRM. ESI is the most soft ionization source developed so far and can afford formation of the fragile molecular ions that are necessary for sensitive SRM detection. Mild CID conditions such as low collision energy and low pressure of collision gas favor production of an abundant daughter ion that is essential to sensitive SRM detection. This knowledge may provide some guidelines for conducting sensitive SRM measurements of very low concentrations of drugs or toxicants in biological samples.

  1. Effects of hypoxia on human cancer cell line chemosensitivity (United States)


    Background Environment inside even a small tumor is characterized by total (anoxia) or partial oxygen deprivation, (hypoxia). It has been shown that radiotherapy and some conventional chemotherapies may be less effective in hypoxia, and therefore it is important to investigate how different drugs act in different microenvironments. In this study we perform a large screening of the effects of 19 clinically used or experimental chemotherapeutic drugs on five different cell lines in conditions of normoxia, hypoxia and anoxia. Methods A panel of 19 commercially available drugs: 5-fluorouracil, acriflavine, bortezomib, cisplatin, digitoxin, digoxin, docetaxel, doxorubicin, etoposide, gemcitabine, irinotecan, melphalan, mitomycin c, rapamycin, sorafenib, thalidomide, tirapazamine, topotecan and vincristine were tested for cytotoxic activity on the cancer cell lines A2780 (ovarian), ACHN (renal), MCF-7 (breast), H69 (SCLC) and U-937 (lymphoma). Parallel aliquots of the cells were grown at different oxygen pressures and after 72 hours of drug exposure viability was measured with the fluorometric microculture cytotoxicity assay (FMCA). Results Sorafenib, irinotecan and docetaxel were in general more effective in an oxygenated environment, while cisplatin, mitomycin c and tirapazamine were more effective in a low oxygen environment. Surprisingly, hypoxia in H69 and MCF-7 cells mostly rendered higher drug sensitivity. In contrast ACHN appeared more sensitive to hypoxia, giving slower proliferating cells, and consequently, was more resistant to most drugs. Conclusions A panel of standard cytotoxic agents was tested against five different human cancer cell lines cultivated at normoxic, hypoxic and anoxic conditions. Results show that impaired chemosensitivity is not universal, in contrast different cell lines behave different and some drugs appear even less effective in normoxia than hypoxia. PMID:23829203

  2. Screening approach by ultra-high performance liquid chromatography-tandem mass spectrometry for the blood quantification of thirty-four toxic principles of plant origin. Application to forensic toxicology. (United States)

    Carlier, Jérémy; Guitton, Jérôme; Romeuf, Ludovic; Bévalot, Fabien; Boyer, Baptiste; Fanton, Laurent; Gaillard, Yvan


    Plant poisonings have left their mark on history and still cause many deaths, whether intentional or accidental. The means to show toxicological evidence of such poisonings should be implemented with great care. This article presents a technique for measuring thirty-nine toxic principles of plant origin in the blood, covering a large amount of toxins from local or exotic plants: α-lobeline, α-solanine, aconitine, ajmaline, atropine, brucine, cephalomannine, colchicine, convallatoxin, cymarine, cytisine, digitoxin, digoxin, emetine, gelsemine, ibogaine, jervine, kavain, lanatoside C, lupanine, mitragynine, neriifolin, oleandrin, ouabain, paclitaxel, physostigmine, pilocarpine, podophyllotoxin, proscillaridin A, reserpine, retrorsine, ricinine, scopolamine, senecionine, sparteine, strophanthidin, strychnine, veratridine and yohimbine. Analysis was carried out using an original ultra-high performance liquid chromatography separation coupled with tandem mass spectrometry detection. Extraction was a standard solid phase extraction performed on Oasis(®) HLB cartridge. Thirty-four of the thirty-nine compounds were put through a validation procedure. The assay was linear in the calibration curve range from 0.5 or 5 μg/L to 1000 μg/L according to the compounds. The method is sensitive (LOD from 0.1 to 1.6 μg/L). The within-day precision of the assay was less than 22.5% at the LLOQ, and the between-day precision was less than 21.5% for 10 μg/L for all the compounds included. The assay accuracy was in the range of 87.4 to 119.8% for the LLOQ. The extraction recovery and matrix effect ranged from 30 to 106% and from -30 to 14%, respectively. It has proven useful and effective in several difficult forensic cases. Copyright © 2014 Elsevier B.V. All rights reserved.

  3. Pharmaceutical wastewater being composite mixture of environmental pollutants may be associated with mutagenicity and genotoxicity. (United States)

    Sharif, Ali; Ashraf, Muhammad; Anjum, Aftab Ahmed; Javeed, Aqeel; Altaf, Imran; Akhtar, Muhammad Furqan; Abbas, Mateen; Akhtar, Bushra; Saleem, Ammara


    Pharmaceutical industries are amongst the foremost contributor to industrial waste. Ecological well-being is endangered owing to its facile discharge. In the present study, heavy metals and organic contaminants in waste water were characterized using atomic absorption spectrophotometer and GC-MS, respectively. Mutagenicity and genotoxic potential of pharmaceutical waste water were investigated through bacterial reverse mutation assay and in vitro comet assay, respectively. Ames test and comet assay of first sample were carried out at concentrations of 100, 50, 25, 12.5, 6.25 % v/v effluent with distilled water. Chromium (Cr), lead (Pb), arsenic (As), and cadmium (Cd) were found in high concentrations as compared to WHO- and EPA-recommended maximum limits. Arsenic was found to be the most abundant metal and its maximum concentration was 0.8 mg.L(-1). GC-MS revealed the presence of lignocaine, digitoxin, trimethoprim, caffeine, and vitamin E in waste water. Dose-dependent decrease in mutagenic index was observed in both strains. Substantial increase in mutagenicity was observed for TA-100, when assay was done by incorporating an enzyme activation system, whereas a slight increase was detected for TA-102. In vitro comet assay of waste water exhibited decrease in damage index and percentage fragmentation with the increase in dilution of waste water. Tail length also decreased with an increase in the dilution factor of waste water. These findings suggest that pharmaceutical waste water being a mix of different heavy metals and organic contaminants may have a potent mutagenic and genotoxic effect on exposed living organisms.

  4. Determination of Radiotracer Stability of Tritium-Labelled Compounds in Biological Studies; Determination de la Stabilite des Composes Trities Employes comme Indicateurs en Biologie; 041e 043f 0440 0435 0434 0414 043b ; Determinacion de la Estabilidad de los Compuestos Tritiados en los Estudios Biologicos Mediante Indicadores

    Energy Technology Data Exchange (ETDEWEB)

    Okita, G. T.; Spratt, J. L. [Argonne Cancer Research Hospital and Department of Pharmacology, University of Chicago, Chicago, IL (United States)


    The recent extensive use of tritium-labelled compounds in biological studies makes it imperative that investigators verify the radiotracer stability of tritiated compounds. Even purification of labelled compounds to constant specific activity does not preclude the possibility of the tritium atom exchanging with hydrogen within a biological system. Radiotracer stability can be demonstrated by various methods once, meticulous radiochemical purification of the labelled material has been effected. In this report three different methods for establishing radiotracer stability of tritiated compounds are described. One approach is to compare the biological half-life of a H{sup 3}-compound to that of a similar compound labelled with C{sup 14}. This method is especially applicable to endogenous substances which undergo isotopic dilution when administered to animals. Stability of exogenous compounds can be verified by a second method. Here it is only necessary to demonstrate no diminution in specific activity when the labelled material is re-isolated from biological samples. A third method, less time-consuming than the first, and applicable to both endogenous and exogenous material is the determination of H{sup 3} to C{sup 14} isotope ratio in a mixture of the same compound labelled with both isotopes. Identical isotope ratios before administration of the double-labelled material and after re-isolation from organs or excreta demonstrate radiotracer stability. This method is particularly applicable where isolation of minute amounts of material necessitates the use of non-radioactive carrier. Data demonstrating the use of these methods for the verification of radiotracer stability will be presented with special reference to labelled cholesterol, morphine and digitoxin as examples for the three respective methods. (author) [French] L'emploi, repandu depuis peu, de composes trities dans les etudes biologiques, impose aux chercheurs la necessite de verifier la stabilite de ces