Simultaneous determination of irinotecan (CPT-11) and SN-38 in tissue culture media and cancer cells by high performance liquid chromatography: Application to cellular metabolism and accumulation studies

A simple and sensitive HPLC method was developed to simultaneously determine CPT-11 and its major metabolite SN-38 in culture media and cell lysates. Camptothecin (CPT) was used as internal standard (I.S.). Compounds were eluted with acetonitrile–50 mM disodium hydrogen phosphate buffer containing 10 mM sodium 1-heptane-sulfonate, with the pH adjusted to 3.0 using 85% (w/v) orthophosphoric acid (27/73, v/v) by a Hyperclon ODS (C18) column (200 mm × 4.6 mm i.d.), with detection at excitation and emission wavelengths of 380 and 540 nm, respectively. The average extraction efficiencies were 96.9–108.3% for CPT-11 in culture media and 94.3–107.2% for CPT-11 in cell lysates; and 87.7–106.8% for SN-38 in culture media and 90.1–105.6% for SN-38 in cell lysates. Within- and between-day precision and accuracy varied from 0.1 to 10.3%. The limit of quantitation (precision and accuracy <20%) was 5.0 and 2.0 ng/ml for CPT-11 and 1.0 and 0.5 ng/ml for SN-38 in culture media and cell lysates, respectively. This method was successfully applied to quantitate the cellular accumulation and metabolism of CPT-11 and SN-38 in H4-II-E, a rat hepatoma cell line.

Simultaneous determination of the lactone and carboxylate forms of irintecan (CPT-11) and its actie metabolite SN-38 by high-performance liquid chromatography: Application to plasma pharmacokinetic studies in the rat.

Irinotecan (CPT-11) and its main metabolite SN-38 are potent anticancer derivatives of camptothecin (CPT), with active lactone and inactive carboxylate forms coexisting. A simple and sensitive HPLC method using the ion-pairing reagent tetrabutylammonium hydrogen sulfate (TBAHS) was developed to simultaneously determine all four analytes in rat plasma samples. Camptothecin (CPT) was used as internal standard. The mobile phase was 0.1 M potassium dihydrogen phosphate containing 0.01 M TBAHS (pH 6.4)–acetonitrile (75:25, v/v). Separation of the compounds was carried out on a Hypersil C18 column, monitored at 540 nm (excitation wavelength at 380 nm). All four compounds gave linear response as a function of concentration over 0.01–10 μM. The limit of quantitation in rat plasma was 0.01, 0.008, 0.005 and 0.005 μM for CPT-11 lactone, CPT-11 carboxylate, SN-38 lactone and SN-38 carboxylate, respectively. The method was successfully used in the study on the effect of coadministered thalidomide on the plasma pharmacokinetics of CPT-11 and SN-38 in rats. Coadministered thalidomide (100 mg/kg body weight by intraperitoneal injection) significantly increased the AUC_0–10h values of CPT-11 lactone and CPT-11 carboxylate by 32.6% and 30.3 %, respectively, (P < 0.01), but decreased the values by 19.2% and 32.4% for SN-38 lactone and carboxylate, respectively, (P < 0.05). Accordingly, the value of total body clearance (CL) of CPT-11 lactone was significantly lower in combination group compared to the control (1.329 versus 1.837 L/h/kg, P = 0.0002). Plasma t_1/2β values for SN-38 lactone and carboxylate were significantly (P < 0.01) smaller in rats with coadministered thalidomide, as compared to rats receiving CPT-11 alone. Further studies are needed to explore the underlying mechanisms for the observed kinetic interaction between CPT-11 and thalidomide.

St Johns wort modulates the toxicities and parmacokinetics of CPT-11 (Irontecan) in rats

CPT-11 is a DNA topoisomerase I inhibitor for the therapy of colorectal cancer, whereas St. Johnrsquos Wort (Hypericum perforatum, SJW) is a widely used herbal anti-depressant. This study aimed to investigate the effects of co-administered SJW on the toxicities and pharmacokinetics of CPT-11 and the underlying mechanisms. The body weight loss, gastrointestinal and hematological toxicities induced by CPT-11, and the pharmacokinetic parameters of CPT-11 were evaluated in rats pretreated with SJW or vehicle. Rats treated with CPT-11 alone experienced rapid decrease in body weight, whereas co-administration of SJW with CPT-11 resulted in lesser body weight loss. The gastrointestinal and hematological toxicities following CPT-11 injection were both alleviated in the presence of SJW. The rat pharmacokinetics of both CPT-11 and its metabolite SN-38 were significantly altered in presence of SJW. In conclusion, co-administered SJW significantly ameliorated the toxicities induced by CPT-11. The protective effect of SJW may be partially due to pharmacokinetic interaction between CPT-11 and SJW.

Vector Theory of Spontaneous Lorentz Violation

Classical electromagnetism predicts two massless propagating modes, which are known as the two polarizations of the photon. On the other hand, if the Lorentz symmetry of classical electromagnetism is spontaneously broken, the new theory will still have two massless Nambu-Goldstone modes resembling the photon. If the Lorentz symmetry is broken by a bumblebee potential that allows for excitations out of the minimum, then massive modes arise. Furthermore, in curved spacetime, such massive modes will be created through a process other than the usual Higgs mechanism because of the dependence of the bumblebee potential on both the vector field and the metric tensor. Also, it is found that these massive modes do not propagate due to the extra constraints.

Expectancy violation and perceptions of rape victim credibility

Purpose. Rape victims have been found to have a heightened risk of secondary victimization in the legal system through biased perceptions of their credibility. In this study, participants observed a video of a rape victim reporting the crime and evaluated her credibility, to establish the influence of victim emotional expressiveness on evaluations of victim credibility. Methods. The nonverbal (eye-contact, crying) and paralinguistic (tone of voice) behaviour of the rape victim was manipulated such that the emotional presentation viewed by observers was either expressive, or not expressive. One hundred and thirty seven participants were randomly allocated to observe a videotape of either an expressive or a non-expressive victim. Participants' specific expectations about the emotional expressiveness of rape victims in general were also measured. Results. Results revealed no significant main effect of emotional expressiveness on perceptions of credibility. However, among participants with a strong expectation of emotional expressiveness, a rape victim who was emotionally expressive was perceived to be significantly more credible than a victim who was not emotionally expressive. Conclusions. It appears to be expectancy violation rather than emotional expressiveness per se that biases observers' perceptions of rape victim credibility.

Fatty acid-specific alterations in leptin, PPARα, and CPT-1 gene expression in the rainbow trout

It is known that fatty acids (FA) regulate lipid metabolism by modulating the expression of numerous genes. In order to gain a better understanding of the effect of individual FA on lipid metabolism related genes in rainbow trout (Oncorhynchus mykiss), an in vitro time-course study was implemented where twelve individual FA (butyric 4:0; caprylic 8:0; palmitic (PAM) 16:0; stearic (STA) 18:0; palmitoleic16:1n-7; oleic 18:1n-9; 11-cis-eicosenoic 20:1n-9; linoleic (LNA) 18:2n-6; α-linolenic (ALA) 18:3n-3; eicosapentenoic (EPA) 20:5n-3; docosahexaenoic (DHA) 22:6n-3; arachidonic (ARA) 20:4n-6) were incubated in rainbow trout liver slices. The effect of FA administration over time was evaluated on the expression of leptin, PPARα and CPT-1 (lipid oxidative related genes). Leptin mRNA expression was down regulated by saturated fatty acids (SFA) and LNA, and was up regulated by monounsaturated fatty acids (MUFA) and long chain PUFA, whilst STA and ALA had no effect. PPARα and CPT-1mRNA expression were up regulated by SFA, MUFA, ALA, ARA and DHA; and down regulated by LNA and EPA. These results suggest that there are individual and specific FA induced modifications of leptin, PPARα and CPT-1 gene expression in rainbow trout, and it is envisaged that such results may provide highly valuable information for future practical applications in fish nutrition.

Traveling solitons in Lorentz and CPT breaking systems

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Concerning CP violation in 331 models

We consider the implementation of CP violation in the context of 331 models. In particular we treat a model where only three scaler triplets are needed in order to give all fermions a mass while keeping neutrino massless. In this case all CP violation is provided by the scalar sector.

Soft CP violation in K-meson systems

We consider a model with soft CP violation which accommodates the CP violation in the neutral kaons even if we assume that the Cabibbo-Kobayashi-Maskawa mixing matrix is real and the sources of CP violation are three complex vacuum expectation values and a trilinear coupling in the scalar potential. We show that for some reasonable values of the masses and other parameters the model allows us to explain all the observed CP violation processes in the K-0-(K) over bar (0) system.

Spontaneous CP violation in the 3-3-1 model with right-handed neutrinos

We implement the mechanism of spontaneous CP violation in the 3-3-1 model with right-handed neutrinos and recognize their sources of CP violation. Our main result is that the mechanism works already in the minimal version of the model and new sources of CP violation emerges as an effect of new physics at energies higher than the electroweak scale.

Parameter degeneracies in neutrino oscillation measurement of leptonic CP and T violation

The measurement of the mixing angle theta(13), sign of Deltam(13)(2), and the CP or T violating phase delta is fraught with ambiguities in neutrino oscillation. In this paper we give an analytic treatment of the paramater degeneracies associated with measuring the nu(mu)-->nu(e) probability and its CP and/or T conjugates. For CP violation, we give explicit solutions to allow us to obtain the regions where there exist twofold and fourfold degeneracies. We calculate the fractional differences, (Deltatheta/(θ) over bar), between the allowed solutions which may be used to compare with the expected sensitivities of the experiments. For T violation we show that there is always a complete degeneracy between solutions with positive and negative Deltam(13)(2) which arises due to a symmetry and cannot be removed by observing one neutrino oscillation probability and its T conjugate. Thus there is always a fourfold parameter degeneracy apart from exceptional points. Explicit solutions are also given and the fractional differences are computed. The biprobability CP/T trajectory diagrams are extensively used to illuminate the nature of the degeneracies.

Tree-level violation of the equivalence principle

Massive particles of spin 0 and 1 violate the equivalence principle (EP) at the tree level. on the other hand, if these particles are massless, they agree with the EP, which leads us to conjecture that from a semiclassical viewpoint massless particles, no matter what their spin, obey the EP. General relativity predicts a deflection angle of 2.63' for a nonrelativistic spinless massive boson passing close to the Sun, while for a massive vectorial boson of spin 1 the corresponding deflection is 2.62'.