Gustavo Henrique Bianco de Souza

A non-michaelian behavior of the in vitro metabolism of the pentacyclic triterpene alfa and beta amyrins by employing rat liver microsomes

Pharmacological studies employing alpha and beta amyrin have demonstrated potential application in several biological activities suggesting their application as promising drugs. In the early drug development, metabolism studies may give important parameters regarding the efficacy and safety of the drug candidate. Therefore, the aim of this work was to determine the enzymatic kinetic parameters of these pentacyclic triterpenes. Chromatographic analyzes were performed using a Shimadzu GC-MS system. The resolution of amyrins was achieved with a DB5-MS column of 0.25 μM film thickness, 30.0 cm length and 0.25 mm diameter. At this condition, the retention times of beta- and alpha-amyrin were 21.3 and 20.2 min, respectively. The proposed method showed to be linear over the concentration range of 0.16-42.18 μM for beta amyrin and 0.11-28.12 μM for alpha amyrin. The lowest concentration quantified by the validated method was 0.16 μM for beta and 0.11 μM for alpha amyrin. The stability study showed that amyrins were stable at room temperature for 12h and at 37°C for 1h. The absolute recovery of the amyrin isomers from the rat microsome was 54.3-59.2%. The enzymatic kinetics presented sigmoidal plots. It was observed a Vmax=0.698 ± 0.022 μmol/mg protein/min, S50=4.4 μM and Hill coefficient of 2.7 ± 0.17 for alpha amyrin and a Vmax=0.775 ± 0.034 μmol/mg protein/min, S50=7.0 μM and Hill coefficient of 2.5 ± 0.21 for beta amyrin. The obtained results give the first clues regarding amyrin metabolism and suggests a more detailed study conducted employing isolated CYP isoforms.

Baccharis trimera improves the antioxidant defense system and inhibits iNOS and NADPH oxidase expression in a rat model of inflammation.

Acetaminophen is a common analgesic and antipyretic compound which, when administered in high doses, has been associated with significant morbidity and mortality, secondary to hepatic toxicity. Although this may be due to a direct interaction of reactive acetaminophen metabolites with hepatocyte proteins, recent studies have suggested that reactive species produced by neutrophils also contribute to the pathophysiological process. Researches on the chemical composition of B. trimera show that this plant has bioactive compounds such as flavonoids, related to the organisms protection against free radicals. Therefore, in the present study, using Fischer rats, the effect of B. trimera on the antioxidant defense system, the production of nitric oxide (NO) and on the expression of nitric oxide synthase (iNOS), superoxide dismutase (SOD), catalase (CAT) and of the subunits of the NADPH oxidase in neutrophils was evaluated in a model of phagocytosis induced by zimosan (ZC3b) and in a model of inflammation induced by acetaminophen. The results show that the treatment with B. trimera improves the defense system of antioxidant and restores the balance ROS / NO that is altered in the inflammatory process induced by APAP. In conclusion, B. trimera extracts exert antioxidant properties by scavenging ROS and decrease the expression of genes responsible by reactive species production in neutrophils.

Protective Effect of Baccharis trimera Extract on Acute Hepatic Injury in a Model of Inflammation Induced by Acetaminophen

Background. Acetaminophen (APAP) is a commonly used analgesic and antipyretic. When administered in high doses, APAP is a clinical problem in the US and Europe, often resulting in severe liver injury and potentially acute liver failure. Studies have demonstrated that antioxidants and anti-inflammatory agents effectively protect against the acute hepatotoxicity induced by APAP overdose. Methods. The present study attempted to investigate the protective effect of B. trimera against APAP-induced hepatic damage in rats. The liver-function markers ALT and AST, biomarkers of oxidative stress, antioxidant parameters, and histopathological changes were examined. Results. The pretreatment with B. trimera attenuated serum activities of ALT and AST that were enhanced by administration of APAP. Furthermore, pretreatment with the extract decreases the activity of the enzyme SOD and increases the activity of catalase and the concentration of total glutathione. Histopathological analysis confirmed the alleviation of liver damage and reduced lesions caused by APAP. Conclusions. The hepatoprotective action of B. trimera extract may rely on its effect on reducing the oxidative stress caused by APAP-induced hepatic damage in a rat model. General Significance. These results make the extract of B. trimera a potential candidate drug capable of protecting the liver against damage caused by APAP overdose.

Genotoxic effects of (−)-cubebin in somatic cells of mice

(−)‐Cubebin belongs to the dibenzylbutyrolactone lignan group, which is widely distributed in the plant kingdom. Because this compound shows interesting biological activities, it is extremely important to evaluate its possible genotoxic activity to allow its safe use in humans. Thus, the present study was performed to investigate the genotoxicity potential activity of (−)‐cubebin assessed by two assays: micronucleus in bone marrow cells and comet test in peripheral blood leukocytes of Swiss mice. In the (−)‐cubebin dose range‐finding assays, the maximum tolerated dose was greater than 2000 mg kg−1. The compound was administered by an oral route at single doses of 250, 500 and 2000 mg kg−1 body weight. Cytotoxicity was assessed by scoring 200 consecutive total polychromatic (PCE) and normochromatic (NCE) erythrocytes (PCE/NCE ratio). Under our experimental conditions, micronucleus and comet assays, respectively, showed that (−)‐cubebin caused dose‐related clastogenic and genotoxic effects in the somatic cells investigated. PCE/NCE ratio showed no cytotoxicity for the three doses of the compound. The data suggest caution in the ingestion of (−)‐cubebin by humans, especially at high doses. Copyright

Baccharis trimera inhibits reactive oxygen species production through PKC and down-regulation p47phox phosphorylation of NADPH oxidase in SK Hep-1 cells:

Baccharis trimera, popularly known as “carqueja”, is a native South-American plant possessing a high concentration of polyphenolic compounds and therefore high antioxidant potential. Despite the antioxidant potential described for B. trimera, there are no reports concerning the signaling pathways involved in this process. So, the aim of the present study was to assess the influence of B. trimera on the modulation of PKC signaling pathway and to characterize the effect of the nicotinamide adenine dinucleotide phosphate oxidase enzyme (NOX) on the generation of reactive oxygen species in SK Hep-1 cells. SK-Hep 1 cells were treated with B. trimera, quercetin, or rutin and then stimulated or not with PMA/ionomycin and labeled with carboxy H2DCFDA for detection of reactive oxygen species by flow cytometer. The PKC expression by Western blot and enzyme activity was performed to evaluate the influence of B. trimera and quercetin on PKC signaling pathway. p47 phox and p47 phox phosphorylated expression was performed by Western blot to evaluate the influence of B. trimera on p47 phox phosphorylation. The results showed that cells stimulated with PMA/ionomycin (activators of PKC) showed significantly increased reactive oxygen species production, and this production returned to baseline levels after treatment with DPI (NOX inhibitor). Both B. trimera and quercetin modulated reactive oxygen species production through the inhibition of PKC protein expression and enzymatic activity, also with inhibition of p47 phox phosphorylation. Taken together, these results suggest that B. trimera has a potential mechanism for inhibiting reactive oxygen species production through the PKC signaling pathway and inhibition subunit p47 phox phosphorylation of nicotinamide adenine dinucleotide phosphate oxidase.

Entanglement of self-interacting scalar fields in an expanding spacetime

We evaluate self-interaction effects on the quantum correlations of field modes of opposite momenta for scalar theory in a two-dimensional asymptotically flat Robertson-Walker spacetime. Such correlations are encoded both in the von Neumann entropy defined through the reduced density matrix in one of the modes and in the covariance expressed in terms of the expectation value of the number operators for each mode in the evolved state. The entanglement between field modes carries information about the underlying spacetime evolution.

Alice and Bob in an expanding spacetime

We investigate the teleportation of a qubit between two observers Alice and Bob in an asymptotically flat Robertson-Walker expanding spacetime. We use scalar or fermionic field modes inside Alices and Bobs ideal cavities and show the degradation of the teleportation quality, as measured by the fidelity, through a mechanism governed by spacetime expansion. This reduction is demonstrated to increase with the rapidity of the expansion and to be highly sensitive to the coupling of the field to spacetime curvature, becoming considerably stronger as it reduces from conformal to minimal. We explore a perturbative approach in the cosmological parameters to compute the Bogoliubov coefficients in order to evaluate and compare the fidelity degradation of fermionic and scalar fields.

Antibacterial activity of extract and fractions from branches of Protium spruceanum and cytotoxicity on fibroblasts

Abstract The crude ethanol extract (CEE) and fractions from branches of Protium spruceanum were subjected to antibacterial and cytotoxicity assays. Compounds of the most active fraction were identified by GC-MS and LC-MS. CEE was active against 19 bacteria and the ethyl acetate fraction (EAF) showed the lowest minimum bactericidal concentration (MBC 0.3–80.0 mg/mL). Through time-kill assay was observed that EAF induced rapid bactericidal effect against Staphylococcus saprophyticus. The cytotoxicity tests against L929 fibroblasts showed great potential of EAF on the treatment of infections caused by five bacteria (MBC < IC50). The results provide in vitro scientific support to the possible application of branches of P. spruceanum as antimicrobial agent that may contribute for treatment of infections.

Quantum estimation in neutrino oscillations

In this work, we analyze two-flavor neutrino oscillations within the framework of quantum estimation theory (QET). We compute the quantum Fischer information (QFI) for the mixing angle θ and show that mass measurements are the ones that achieve optimal precision. We also study the Fischer information (FI) associated with flavor measurements and show that they are optimized at specific neutrino times-of-flight. Therefore, although the usual population measurement does not realize the precision limit set by the QFI, it can in principle be implemented with the best possible sensitivity to θ. We investigate how these quantifiers relate to the single-particle, mode entanglement in neutrino oscillations. We demonstrate that this form of entanglement does not enhance either of them. In particular, our results show that in single-particle settings, entanglement is not directly connected with the enhancement of precision in metrological tasks.

Exotic looped trajectories via quantum marking

We provide an analytical and theoretical study of exotic looped trajectories (ELTs) in a double-slit interferometer with quantum marking. We use an excited Rydberg-like atom and which-way detectors such as superconducting cavities, just as in the Scully-Englert-Walther interferometer. We indicate appropriate conditions on the atomic beam or superconducting cavities so that we determine an interference pattern and fringe visibility exclusive from the ELTs. We quantitatively describe our results for Rubidium atoms and propose this framework as an alternative scheme to the double-slit experiment modified to interfere only these exotic trajectories.