Alternative Mitochondrial Functions In Cell Physiopathology: Beyond Atp Production.

It is well known that mitochondria are the main site for ATP generation within most tissues. However, mitochondria also participate in a surprising number of alternative activities, including intracellular Ca2+ regulation, thermogenesis and the control of apoptosis. In addition, mitochondria are the main cellular generators of reactive oxygen species, and may trigger necrotic cell death under conditions of oxidative stress. This review concentrates on these alternative mitochondrial functions, and their role in cell physiopathology.33241-5

Inhibition Of Mitochondrial Permeability Transition By Low Ph Is Associated With Less Extensive Membrane Protein Thiol Oxidation.

Ca2+ and inorganic phosphate-induced mitochondrial swelling and membrane protein thiol oxidation, which are associated with mitochondrial permeability transition, are inhibited by progressively decreasing the incubation medium pH between 7.2 and 6.0. Nevertheless, the detection of mitochondrial H2O2 production under these conditions is increased. Permeability transition induced by phenylarsine oxide, which promotes membrane protein thiol cross-linkage in a process independent of Ca2+ or reactive oxygen species, is also strongly inhibited in acidic incubation media. In addition, we observed that the decreased protein thiol reactivity with phenylarsine oxide or phenylarsine oxide-induced swelling at pH 6.0 is reversed by diethyl pyrocarbonate, in a hydroxylamine-sensitive manner. These results provide evidence that the inhibition of mitrochondrial permeability transition observed at lower incubation medium pH is mediated by a decrease in membrane protein thiol reactivity, related to the protonation of protein histidyl residues.19525-3

PARÂMETRO DE INSERÇÃO URBANA: O CASO DA HABITAÇÃO DE INTERESSE SOCIAL EM CAMPINAS

O desenvolvimento urbano e o direito à cidade desempenham um papel fundamental na formação de bairros e na qualidade de vida das pessoas. No entanto, apesar da maior parte da população e suas habitações se concentrarem nas cidades, ações para garantir a Inserção Urbana (IU) de conjuntos habitacionais ainda são inexpressivas. Vários estudos desenvolvidos no Brasil ressaltam problemas da IU de conjuntos habitacionais. Contudo, a avaliação da IU de área de abrangência dos empreendimentos ainda carece de aprofundamentos metodológicos com aplicação de ferramentas para a produção de resultados mensuráveis. Dois objetivos nortearam essa pesquisa. O primeiro visa compreender IU de conjuntos habitacionais e seus parâmetros, a partir da literatura; e o segundo, por meio de um estudo de caso, busca analisar parâmetros na escala metropolitana do empreendimento com o entorno imediato, e os impactos sociais da IU para a sociedade. A ferramenta foi consolidada a partir de dois estudos desenvolvidos pelo Sistema Nacional de Habitação, e LabCidade-FAU/USP. Os resultados da pesquisa podem proporcionar um avanço para a mitigação de impactos sociais na formação de bairros, e direcionar investimentos na produção de habitações, quer seja em ações para espaços urbanos inclusivos, como no desenvolvimento sustentável de longo prazo.Urban development and the right to the city play a vital role in the formation of neighbourhoods and people’s quality of life. However, although most of today's population lives in cities, actions to ensure proper urban insertion (UI) of housing estates are still scarce, especially those considered as social interest developments. Several studies developed in Brazil highlight UI problems for social housing estates. The evaluation of the UI of housing catchment areas still lacks methodological deepening with applications of tools for producing measurable results. Two objectives led to the research: understand the UI of housing estates and parameters based on the literature; deepen the knowledge about UI, through a case study, based on the analysis of parameters on a metropolitan scale, and the spatial relationship of the projects with the immediate environment, considering the social impacts of UI of housing for society. A tool was consolidated based on the parameters of two studies developed by the National Housing System and the LabCidade-FAU/ USP. Results of the research should advance knowledge for the mitigation of social impacts in the formation of neighbourhoods and guide investments in the production of housing, whether as actions for inclusive urban spaces or long-term sustainable development

Cell Therapy Attenuates Cardiac Dysfunction Post Myocardial Infarction: Effect of Timing, Routes of Injection and a Fibrin Scaffold

Background: Cell therapy approaches for biologic cardiac repair hold great promises, although basic fundamental issues remain poorly understood. In the present study we examined the effects of timing and routes of administration of bone marrow cells (BMC) post-myocardial infarction (MI) and the efficacy of an injectable biopolymer scaffold to improve cardiac cell retention and function. Methodology/Principal Findings: (99m)Tc-labeled BMC (6x10(6) cells) were injected by 4 different routes in adult rats: intravenous (IV), left ventricular cavity (LV), left ventricular cavity with temporal aorta occlusion (LV(+)) to mimic coronary injection, and intramyocardial (IM). The injections were performed 1, 2, 3, or 7 days post-MI and cell retention was estimated by gamma-emission counting of the organs excised 24 hs after cell injection. IM injection improved cell retention and attenuated cardiac dysfunction, whereas IV, LV or LV* routes were somewhat inefficient (< 1%). Cardiac BMC retention was not influenced by timing except for the IM injection that showed greater cell retention at 7 (16%) vs. 1, 2 or 3 (average of 7%) days post-MI. Cardiac cell retention was further improved by an injectable fibrin scaffold at day 3 post-MI (17 vs. 7%), even though morphometric and function parameters evaluated 4 weeks later displayed similar improvements. Conclusions/Significance: These results show that cells injected post-MI display comparable tissue distribution profile regardless of the route of injection and that there is no time effect for cardiac cell accumulation for injections performed 1 to 3 days post-MI. As expected the IM injection is the most efficient for cardiac cell retention, it can be further improved by co-injection with a fibrin scaffold and it significantly attenuates cardiac dysfunction evaluated 4 weeks post myocardial infarction. These pharmacokinetic data obtained under similar experimental conditions are essential for further development of these novel approaches

Potent Cardioprotective Effect of the 4-Anilinoquinazoline Derivative PD153035: Involvement of Mitochondrial KATP Channel Activation

Background: The aim of the present study was to evaluate the protective effects of the 4-anilinoquinazoline derivative PD153035 on cardiac ischemia/reperfusion and mitochondrial function. Methodology/Principal Findings: Perfused rat hearts and cardiac HL-1 cells were used to determine cardioprotective effects of PD153035. Isolated rat heart mitochondria were studied to uncover mechanisms of cardioprotection. Nanomolar doses of PD153035 strongly protect against heart and cardiomyocyte damage induced by ischemia/reperfusion and cyanide/aglycemia. PD153035 did not alter oxidative phosphorylation, nor directly prevent Ca(2+) induced mitochondrial membrane permeability transition. The protective effect of PD153035 on HL-1 cells was also independent of AKT phosphorylation state. Interestingly, PD153035 activated K(+) transport in isolated mitochondria, in a manner prevented by ATP and 5-hydroxydecanoate, inhibitors of mitochondrial ATP-sensitive K(+) channels (mitoK(ATP)). 5-Hydroxydecanoate also inhibited the cardioprotective effect of PD153035 in cardiac HL-1 cells, demonstrating that this protection is dependent on mitoK(ATP) activation. Conclusions/Significance: We conclude that PD153035 is a potent cardioprotective compound and acts in a mechanism involving mitoK(ATP) activation

Mitochondrial Uncoupling Inhibits p53 Mitochondrial Translocation in TPA-Challenged Skin Epidermal JB6 Cells

The tumor suppressor p53 is known to be able to trigger apoptosis in response to DNA damage, oncogene activation, and certain chemotherapeutic drugs. In addition to its transcriptional activation, a fraction of p53 translocates to mitochondria at the very early stage of apoptosis, which eventually contributes to the loss of mitochondrial membrane potential, generation of reactive oxygen species (ROS), cytochrome c release, and caspase activation. However, the mitochondrial events that affect p53 translocation are still unclear. Since mitochondrial uncoupling has been suggested to contribute to cancer development, herein, we studied whether p53 mitochondrial translocation and subsequent apoptosis were affected by mitochondrial uncoupling using chemical protonophores, and further verified the results using a siRNA approach in murine skin epidermal JB6 cells. Our results showed that mitochondrial uncoupling blocked p53 mitochondrial translocation induced by 12-O-tetradecanoylphorbol 13-acetate (TPA), a known tumor promoter to induce p53-mediated apoptosis in skin carcinogenesis. This blocking effect, in turn, led to preservation of mitochondrial functions, and eventually suppression of caspase activity and apoptosis. Moreover, uncoupling protein 2 (UCP2), a potential suppressor of ROS in mitochondria, is important for TPA-induced cell transformation in JB6 cells. UCP2 knock down cells showed enhanced p53 mitochondrial translocation, and were less prone to form colonies in soft agar after TPA treatment. Altogether, our data suggest that mitochondrial uncoupling may serve as an important regulator of p53 mitochondrial translocation and p53-mediated apoptosis during early tumor promotion. Therefore, targeting mitochondrial uncoupling may be considered as a novel treatment strategy for cancer

Cardioprotective Effect of Nicorandil, a Mitochondrial ATP-Sensitive Potassium Channel Opener, Prolongs Survival in HSPB5 R120G Transgenic Mice

BACKGROUND: Transgenic (TG) mice with overexpression of an arg120gly (R120G) missense mutation in HSPB5 display desmin-related cardiomyopathy, which is characterized by formation of aggresomes. It is also known that progressive mitochondrial abnormalities and apoptotic cell death occur in the hearts of R120G TG mice. The role of mitochondrial dysfunction and apoptosis in disease progression, however, remains uncertain. METHODS AND RESULTS: Mitochondrial abnormalities and apoptotic cell death induced by overexpression of HSPB5 R120G were analyzed in neonatal rat cardiomyocytes. Overexpression of mutant HSPB5 led to development of aggresomes with a concomitant reduction in cell viability in the myocytes. Overexpression of mutant HSPB5 induced a reduction in the cytochrome c level in the mitochondrial fraction and a corresponding increase in the cytoplasmic fraction in the myocytes. Down-regulation of BCL2 and up-regulation of BAX were detected in the myocytes expressing the mutant HSPB5. Concomitant with mitochondrial abnormality, the activation of caspase-3 and increased apoptotic cell death was observed. Cell viability was dose-dependently recovered in myocytes overexpressing HSPB5 R120G by treatment with nicorandil a mitochondrial ATP-sensitive potassium channel opener. Nicorandil treatment also inhibited the increase in BAX, the decrease in BCL2, activation of caspase-3 and apoptotic cell death by mutant HSPB5. To confirm the results of the in-vitro study, we analyzed the effect of nicorandil in HSPB5 R120G TG mice. Nicorandil treatment appeared to reduce mitochondrial impairment and apoptotic cell death and prolonged survival in HSPB5 R120G TG mice. CONCLUSIONS: Nicorandil may prolong survival in HSPB5 R120G TG mice by protecting against mitochondrial impairments

Hepatoprotektivno djelovanje ekstrakta biljke Calotropis gigantea na oštećenje jetre štakora tetraklormetanom

Ethanolic extract (50 %) of stems of Calotropis gigantea R. Br. (Asclepiadaceae) at doses of 250 and 500 mg kg1 were studied for hepatoprotective activity in male Wistar rats with liver damage induced using carbon tetrachloride, 2 mL kg-1 twice a week. The protective effect of C. gigantea extract was compared with the standard drug silymarin. Various biochemical parameters such as aspartate amino transferase (AST), alanine amino transferase (ALT), glutathione (GSH), lipid peroxide (LPO), superoxidedismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT) were evaluated. The results revealed that the C. gigantea extract significantly decreased AST, ALT (p < 0.001) and lipid peroxide (p < 0.01) levels. The antioxidant parameters GSH, GPx, SOD and catalase levels were increased considerably compared to the levels in groups treated with carbon tetrachloride onlyEtanolni ekstrakt (50 %) stabljika biljke Calotropis gigantea R. Br. (Asclepiadaceae) u dozi 250 i 500 mg kg1 testiran je na hepatoprotektivno djelovanje oštećenje jetre mužjaka Wistar štakora inducirano tetraklormetanom, 2 mL kg1 dva puta tjedno. Zaštitni učinak ekstrakta biljke C. gigantea uspoređivan je sa standarnim lijekom silimarinom. Evaluirani su različiti biokemijski parametri kao što su aspartat amino transferaza (AST), alanin amino transferaza (ALT), glutation (GSH), lipidni peroksidi (LPO), superoksiddismutaza (SOD), glutation peroksidaze (GPx) i katalaza (CAT). Rezultati ukazuju da ekstrakt biljke C. gigantea značajno smanjuje koncentracije AST, ALT (p < 0.001) i lipidnih peroksida (p < 0.01). Koncentracije antioksidativnih parametara GSH, GPx, SOD i katalaze bile su značajno povišene u usporedbi sa skupinom tretiranom samo tetraklormetanom

The Influence of L-Carnitine on Oxidative Modification of LDL In Vitro

Owing to their structure and function, low-density lipoproteins (LDLs) are particularly susceptible to the oxidative modifications. To prevent against oxidative modification of LDL, L-carnitine, with endogenous small water-soluble quaternary amine possessing antioxidative properties, was used. The aim of this paper was to prove the in vitro influence of L-carnitine on the degree of oxidative modification of the lipid part (estimated by conjugated dienes, lipid hydroperoxides, and malondialdehyde levels) and the protein part (estimated by dityrosine and tryptophan levels) of LDL native and oxidized by cooper ions. The level of lipophylic LDL antioxidant—α-tocopherol was also measured