The contralateral kidney presents with impaired mitochondrial functions and disrupted redox homeostasis after 14 days of unilateral ureteral obstruction in mice.

In unilateral ureteral obstruction (UUO), both oxidative stress and mitochondrial dysfunction are related to cell death. The aim of this study has been to characterize profiles of enzyme antioxidant activities and mitochondrial functioning of the contralateral (CL) compared to UUO and Sham (false-operated) kidneys of Balb/c mice. Kidneys were resected 14 days after obstruction for immunohistochemical and cortical mitochondrial functioning assays. Antioxidant enzymes activities were investigated in mitochondria and cytosol. Oxygen consumption (QO2) and formation of O2 reactive species (ROS) were assessed with pyruvate plus malate or succinate as the respiratory substrates. QO2 decreased in CL and UUO in all states using substrates for complex II, whereas it was affected only in UUO when substrates for complex I were used. Progressive decrease in mitochondrial ROS formation-in the forward and reverse pathway at complex I-correlates well with the inhibition of QO2 and, therefore, with decreased electron transfer at the level of complexes upstream of cytochrome c oxidase. CL and UUO transmembrane potential responses to ADP were impaired with succinate. Intense Ca2+-induced swelling was elicited in CL and UUO mitochondria. Important and selective differences exist in CL antioxidant enzymes with respect to either Sham or UUO kidneys: CL kidneys had increased mitochondrial glutathione peroxidase and cytosolic catalase activities, indicative of compensatory responses in the face of an early altered ROS homeostasis (as detected by 4-hydroxynonenal), and of a significant tendency to apoptosis. In CL and UUO, upregulation of nuclear (erythroid-derived 2)-like 2 transcription factor (Nrf2), as well as of cytoplasmic and nuclear Kelch-like ECH-associated protein 1 (Keap1) in opposition to decreased heme oxygenase-1 (HO-1), suggest impairment of the Nrf2/Keap1/HO-1 system. It is concluded that chronic obstruction impairs mitochondrial function in CL and UUO, preferentially affecting complex II.

Antitumor Effect of Pomolic Acid in Acute Myeloid Leukemia Cells Involves Cell Death, Decreased Cell Growth and Topoisomerases Inhibition.

BACKGROUND: Acute myeloid leukemia (AML) represents the largest number of annual deaths from hematologic malignancy. In the United States, it was estimated that 21.380 individuals would be diagnosed with AML and 49.5% of patients would die in 2017. Therefore, the search for novel compounds capable of increasing the overall survival rate to the treatment of AML cells is urgent. OBJECTIVES: To investigate the cytotoxicity effect of the natural compound pomolic acid (PA) and to explore the mechanism of action of PA in AML cell lines with different phenotypes. METHODS: Three different AML cell lines, HL60, U937 and Kasumi-1 cells with different mechanisms of resistance were used to analyze the effect of PA on the cell cycle progression, on DNA intercalation and on human DNA topoisomerases (hTopo I and IIα) in vitro studies. Theoretical experiments of the inhibition of hTopo I and IIα were done to explore the binding modes of PA. RESULTS: PA reduced cell viability, induced cell death, increased sub-G0/G1 accumulation and activated caspases pathway in all cell lines, altered the cell cycle distribution and inhibited the catalytic activity of both human DNA topoisomerases. CONCLUSION: Finally, this study showed that PA has powerful antitumor activity against AML cells, suggesting that this natural compound might be a potent antineoplastic agent to improve the treatment scheme of this neoplasm.

Trypanosoma cruzi sensitizes mice to fulminant SEB-induced shock: overrelease of inflammatory cytokines and independence of Chagas' disease or TCR Vbeta-usage.

Trypanosoma cruzi-infected mice display increased susceptibility to shock induced by injection of lipopolysaccharide (LPS), anti-CD3, or resulting from interleukin (IL)-10-defective response to the parasite itself, but the basis of such susceptibility remains unknown. Herein, we tested the susceptibility of mice inoculated with virulent and avirulent T. cruzi to staphylococcal enterotoxins (SE), potent inducers of inflammatory cytokine secretion. Mice infected with T. cruzi CL-strain or inoculated with the avirulent clone CL-14, a clone that does not induce disease or polyclonal lymphocyte activation, succumb suddenly to low doses of staphylococcal enterotoxin B (SEB), but not to staphylococcal enterotoxin A (SEA). High plasma levels of TNF, IFN-gamma, and liver transaminases alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were found in these mice, indicating lethal toxic shock. Sensitization to shock required inoculation of live avirulent trypomastigotes and a time interval before challenge with SEB. We found no prior skewing of T cell receptor (TCR) Vbeta-repertoire in CL-14-inoculated mice that could be responsible for sensitization. Splenocytes from CL-14-inoculated mice proliferated more under anti-Vbeta8 than anti-TCRbeta stimulation when compared with normal mice, but were suppressed to SEB stimulation. Both SEB and anti-Vbeta8 antibodies stimulated splenocytes from T. cruzi-inoculated mice to secrete higher levels of inflammatory cytokines than normal controls. Taken together, our results show that T. cruzi inoculation can sensitize mice to lethal SEB-induced shock even in the absence of tissue damage, polyclonal lymphocyte activation, or previously increased levels of inflammatory cytokines, and they suggest that altered reactivity of Vbeta8 lymphocytes may be involved in the phenomenon.

Effects of oleanolic acid on pulmonary morphofunctional and biochemical variables in experimental acute lung injury.

We analysed the effects of oleanolic acid (OA) on lung mechanics and histology and its possible mechanisms of action in experimental acute lung injury (ALI). BALB/c mice were randomly divided into Control (saline, ip) and ALI (paraquat, 25 mg/kg, ip) groups. At 1 h, both groups were treated with saline (SAL, 50 µl ip), OA (10 mg/kg ip), or dexamethasone (DEXA, 1 mg/kg ip). At 24 h, lung static elastance, viscoelastic pressure, and alveolar collapse reduced more after OA compared to DEXA administration. Tumour necrosis factor-α, macrophage migration inhibitory factor, interleukin-6, interferon-γ, and transforming growth factor-ß mRNA expressions in lung tissue diminished similarly after OA or DEXA. Conversely, only OA avoided reactive oxygen species generation and yielded a significant decrease in nitrite concentration. OA and DEXA restored the reduced glutathione/oxidized glutathione ratio and catalase activity while increasing glutathione peroxidase induced by paraquat. In conclusion, OA improved lung morphofunction by modulating the release of inflammatory mediators and oxidative stress.

Topological polar surface area defines substrate transport by multidrug resistance associated protein 1 (MRP1/ABCC1).

Multidrug resistance-associated protein 1 (MRP1/ABCC1) is a very promiscuous transporter. Herein we used topological polar surface area (TPSA), a descriptor defined as the sum of surfaces of polar atoms in a molecule, to analyze drug transport by MRP1. We suggested that compounds with high TPSA are transported while those with low TPSA are not. The conjugation to GSH increases TPSA values favoring transport. A strong correlation between TPSA and transport properties (K(m)) was also found.

Balanced cytokine-producing pattern in mice immunized with an avirulent Trypanosoma cruzi.

We have previously demonstrated that inoculation of BALB/c mice with trypomastigotes of CL-14, an avirulent Trypanosoma cruzi clone, prevents the development of parasitemia and mortality after challenge with virulent CL strain. In this report, we investigated the cytokine and antibody profiles induced by inoculation with CL-14 clone. Groups of mice were inoculated with trypomastigotes of CL-14 clone and challenged with infective CL strain. Challenged CL-14-inoculated mice had lower levels of IFN-gamma and higher production of IgG1 antibodies as compared to CL strain-infected mice. Previous inoculation with CL-14 clone partially prevented the suppression of IL-2 production caused by CL strain infection. No significant differences were found regarding IL-4 production by splenocytes from CL-14-inoculated or control groups after challenge with CL-strain. Our results show that protection against acute T. cruzi infection induced by CL-14 inoculation correlates with a balanced T1/T2 cytokine production, a profile likely to be beneficial for the host.

Trypanosoma cruzi: requirements for induction and maintenance of protective immunity conferred by immunization.

Immunization with CL-14-trypomastigotes generates efficient humoral and cellular responses against infective challenge. Herein, we investigated the relevance of these mechanisms in vivo. Immunization with live CL-14-trypomastigotes protected only part of beta2m(-/-) mice but efficiently protected perforin-knockout mice. Fixed CL-14-trypomastigotes could successfully immunize BALB/c, though live trypomastigotes lowered the requirements for doses and time intervals. Post-immune depletion of CD4 or CD8 subsets did not affect protection conferred by immunization, but switched the production of anti-Trypanosoma cruzi antibodies to IgG2a. Sublethal irradiation partially broke the resistance of immune mice, leading to development of late parasitemia. Passive serum transfer from immune mice conferred protection to nai;ve mice. Our results indicate that presentation of cytosolic antigens by MHC class I molecules is involved in the generation of immunity and suggest that the humoral response contributes to a great extent to keep CL-14-immunized mice protected against infective challenge.

In vitro activities of iboga alkaloid congeners coronaridine and 18-methoxycoronaridine against Leishmania amazonensis.

In previous studies, we demonstrated the leishmanicide effect of coronaridine, a natural indole alkaloid isolated from stem bark of Peschiera australis (Delorenzi et al., Antimicrob. Agents Chemother. 45:1349-1354, 2001). In this study we show the leishmanicidal effect of the synthetic coronaridine and its racemic 18-methoxylated analog, 18-methoxycoronaridine. Both alkaloids revealed a potent leishmanicide effect against Leishmania amazonensis, a causative agent of cutaneous and diffuse cutaneous leishmaniasis in the New World. Despite their potent leishmanicide effect, both alkaloids were neither toxic to murine macrophages nor did they modulate their oxidative or cytokine production responses.

Balanced cytokine-producing pattern in mice immunized with an avirulent Trypanosoma cruzi

We have previously demonstrated that inoculation of BALB/c mice with trypomastigotes of CL-14, an avirulent Trypanosoma cruzi clone, prevents the development of parasitemia and mortality after challenge with virulent CL strain. In this report, we investigated the cytokine and antibody profiles induced by inoculation with CL-14 clone. Groups of mice were inoculated with trypomastigotes of CL-14 clone and challenged with infective CL strain. Challenged CL-14-inoculated mice had lower levels of IFN-g and higher production of IgG1 antibodies as compared to CL strain-infected mice. Previous inoculation with CL-14 clone partially prevented the suppression of IL-2 production caused by CL strain infection. No significant differences were found regarding IL-4 production by splenocytes from CL-14-inoculated or control groups after challenge with CL-strain. Our results show that protection against acute T. cruzi infection induced by CL-14 inoculation correlates with a balanced T1/T2 cytokine production, a profile likely to be beneficial for the host

Oligodendrogliomas: a genética molecular e o desenvolvimento estratégia terapêutica adjuvante / Oligodendrogliomas: molecular genetics and development of adjuvant therapeutic strategies

A incidência dos oligodendrogliomas tem aumentado provavelmente em razão do progresso na precisão de diagnóstico. Aproximadamente dois terços dos pacientes com a forma mais agressiva, oligodendroglioma anaplásico, mostram resposta substancial à quimioterapia com a associação procarbazina/lomustina/vincristina (PCV). Entretanto, os resultados da quimuiterapia anaplásica dos oligodendrogliomas resulta em grande número de células com ERK/MAPK ativadas. O monoterpeno álcool perílico demonstra atividades quimiopreventiva e quimioterápica em diversos modelos de tumores e sugere-se que estas possam estar associadas com a capacidade de inibir a farmesilação pós-traducional e a sinalização da Ras, assim como a cascata de sinalização por meio da RAF-MEK-ERK. Estudo do nosso grupo observou que pode estar atuando mediante a inibição da fosforilação da extracellular regulated kinase (ERK), uma proteína envolvida na cascata de transdução de sinal através da membrana e proliferação celular induzida pela proteína Ras. Este artigo discute a redução de oligodendroglioma anaplásico recidivado em paciente tratado durante nove meses com álcool perílico por via intranasal.