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BACKGROUND AND AIMS: Hypercholesterolemia and oxidative stress are two of the most important risk factors for atherosclerosis. The aim of the present work was to evaluate mandarin (Citrus reticulata) peel oil (MPO) in cholesterol metabolism and lipid synthesis, and its antioxidant capacity. METHODS AND RESULTS: Incubation of hepatic HepG2 cells with MPO (15-60 µL/L) reduced cholesterogenesis and saponifiable lipid synthesis, demonstrated by [14C]acetate radioactivity assays. These effects were associated with a decrease in a post-squalene reaction of the mevalonate pathway. Molecular docking analyses were carried out using three different scoring functions to examine the cholesterol-lowering property of all the components of MPO against lanosterol synthase. Docking simulations proposed that minor components of MPO monoterpenes, like alpha-farnesene and neryl acetate, as well the major component, limonene and its metabolites, could be partly responsible for the inhibitory effects observed in culture assays. MPO also decreased RAW 264.7 foam cell lipid storage and its CD36 expression, and prevented low-density lipoprotein (LDL) lipid peroxidation. CONCLUSION: These results may imply a potential role of MPO in preventing atherosclerosis by a mechanism involving inhibition of lipid synthesis and storage and the decrease of LDL lipid peroxidation.
Assuntos
Antioxidantes/farmacologia , Aterosclerose/prevenção & controle , Colesterol/metabolismo , Citrus , Dislipidemias/tratamento farmacológico , Células Espumosas/efeitos dos fármacos , Frutas , Hepatócitos/efeitos dos fármacos , Hipolipemiantes/farmacologia , Lipoproteínas LDL/metabolismo , Óleos de Plantas/farmacologia , Animais , Antioxidantes/isolamento & purificação , Aterosclerose/etiologia , Aterosclerose/metabolismo , Antígenos CD36/metabolismo , Citrus/química , Dislipidemias/complicações , Dislipidemias/metabolismo , Células Espumosas/metabolismo , Frutas/química , Células Hep G2 , Hepatócitos/metabolismo , Humanos , Hipolipemiantes/isolamento & purificação , Transferases Intramoleculares/antagonistas & inibidores , Transferases Intramoleculares/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Camundongos , Simulação de Acoplamento Molecular , Óleos de Plantas/isolamento & purificação , Células RAW 264.7RESUMO
Las lesiones dentales no cariosas comprenden un conjunto de procesos que se caracterizan por la pérdida o el desgaste patológico de los tejidos duros del diente, como ser el esmalte y la dentina. En la etapa inicial es difícil el diagnóstico, cuando la lesión va en aumento y si no es tratada a tiempo ni se modifican los factores de riesgo pueden llegar afectar de manera progresiva la pulpa dental. Estas lesiones han aumentado en los últimos años, debido al mayor tiempo de dientes en boca, acompañado de hábitos parafuncionales y dietas altamente acidas, a esto se le agrega el estrés que también provoca perdida de estructura dental. Por lo que no toda perdida de tejido mineralizado de los diente es provocada por una carga bacteriana. Estas lesionesno cariosas difieren en su etiología, se debe ser muy minucioso en su evaluación clínica y descripción, ya que muchas características clínicas pueden generar un diagnóstico erróneo y por ende un tratamiento no predecible, se debe tomar en consideración la existencia de combinaciones entre lesiones. Objetivo: Conocer la etiología y características clínicas propias de cada lesión, para poder realizar un diagnóstico oportuno y seguro. La revisión se realizó por medio de una búsqueda en internet en los buscadores de Hinari, Science Direct, Google académico, revisando artículos originales y revisiones bibliográficas entre los años de 2014-2019. Conclusiones: Las lesiones dentales no cariosas tienen etiologías multi factoriales, características clínicas diferenciales y puede haber una combinación de varias patologías...(AU)
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Humanos , Abrasão Dentária/diagnóstico , Esmalte Dentário/lesões , Erosão Dentária , Atrito Dentário/complicaçõesRESUMO
Geraniol (GOH), like other plant-derived natural bioactive compounds, has been found to possess antiproliferative properties that are essential to cope with malignant tumors. However, the mechanisms of molecular action of GOH are not fully elucidated. The aim of this study was to evaluate the effect of GOH on some oxidative parameters in human tumor cell lines (HepG2 and A549). Cytotoxicity evaluated in cell lines by the MTT assay, genotoxicity by the comet assay, and lipid peroxidation by the TBARS. The activities of antioxidant the enzymes, superoxide dismutase (SOD), catalase (CAT) and glutathione-S-transferase (GST), were also analyzed. Additionally, intracellular reactive oxygen species (ROS), nitric oxide, and lactate production were determined in HepG2 cells. Both tumor cell lines showed a clear concentration-dependent response to GOH in several of the parameters evaluated. Lipids turned out to be more sensitive than DNA to oxidative damage induced by GOH. TBARS levels increased with respect to control (p < 0.05) by 33% and 122% in HepG2 and A549 cells, respectively treated with 200 µM GOH. However, GOH caused a statistically significant decrease in SOD and CAT activities in HepG2 cells only. GST was not affected in any cell lines. GOH induced the production of ROS but not nitric oxide in HepG2, which shows that ROS were mainly responsible for oxidative damage. Lactate release increased statistically significantly compared to control (p < 0.001), by 41% and 86% at 200 and 800 µM GOH respectively, showing that this monoterpene also affected the glycolytic pathway in HepG2 cells. These results suggest that oxidative stress could mediate the anti-proliferative effects of GOH in HepG2 and A549 cells.
Assuntos
Monoterpenos Acíclicos/farmacologia , Proliferação de Células , Estresse Oxidativo/efeitos dos fármacos , Células A549 , Monoterpenos Acíclicos/administração & dosagem , Monoterpenos Acíclicos/química , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Células Hep G2 , Humanos , Estrutura MolecularRESUMO
Cytochrome c is a prototypical multifunctional protein that is implicated in a variety of processes that are essential both for sustaining and for terminating cellular life. Typically, alternative functions other than canonical electron transport in the respiratory chain are associated to alternative conformations. In this work we apply a combined experimental and computational study of Cyt c variants to assess whether the parameters that regulate the canonical electron transport function of Cyt c are correlated with those that determine the transition to alternative conformations, using the alkaline transition as a model conformational change. The results show that pKa values of the alkaline transition correlate with the activation energies of the frictionally-controlled electron transfer reaction, and that both parameters are mainly modulated by the flexibility of the Ω-loop 70-85. Reduction potentials and non-adiabatic ET reorganization energies, on the other hand, are both modulated by the flexibilities of the Ω-loops 40-57 and 70-85. Finally, all the measured thermodynamic and kinetic parameters that characterize both types of processes exhibit systematic variations with the dynamics of the hydrogen bond between the axial ligand Met80 and the second sphere ligand Tyr67, thus highlighting the critical role of Tyr67 in controlling canonical and alternative functions of Cyt c.
Assuntos
Citocromos c/química , Animais , Transporte de Elétrons , Cavalos , Ligação de Hidrogênio , Cinética , Oxirredução , Conformação Proteica , TermodinâmicaRESUMO
BACKGROUND: In Mexico, wheat and corn flour fortification with folic acid (FA) was implemented in 2001 and mandated in 2008, but without direct enforcement. Current Mexican nutrient-content tables do not account for FA contained in bakery bread and corn masa-based foods, which are dietary staples in Mexico. OBJECTIVE: The objective of this study was to examine the impact of FA fortification of dietary staples on the proportion of the population consuming below the Estimated Average Requirement (EAR) for folate or above the Tolerable Upper Intake Level (UL) for FA. METHODS: We measured FA and folate content in dietary staples (bakery bread and tortillas) using microbial assays and MS, and we recalculated FA intake from 24-h recall dietary intake data collected in the 2012 Mexican National Health and Nutrition Survey (Encuesta Nacional de Salud y Nutrición) utilizing estimates from our food measurements, using nutrient concentrations from tortillas to approximate nutrient content of other corn masa-derived foods. The revised FA intake estimates were used to examine population-level intake of FA and dietary folate equivalent (DFE) accounting for geographic differences in FA content with statistical models. RESULTS: FA content in dietary staples was variable, whereas use of FA-fortified flour in corn masa tortillas increased with population size in place of residence. Accounting for dietary staples' FA fortification increased population estimates for FA and DFE intake, resulting in a lower proportion with intake below the EAR and a higher proportion with intake above the UL. Despite accounting for FA-fortified staple foods, 9-33% of women of childbearing age still have intake below the EAR, whereas up to 12% of younger children have intake above the UL. CONCLUSIONS: Unregulated FA fortification of dietary staples leads to unpredictable total folate intake without adequately impacting the intended target. Our findings suggest that monitoring, evaluation, and enforcement of mandatory fortification policies are needed. Without these, alternate strategies may be needed in order to reach women of childbearing age while avoiding overexposing children.
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Pão/análise , Ácido Fólico/metabolismo , Inquéritos Nutricionais , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Feminino , Farinha/análise , Alimentos Fortificados/análise , Humanos , Lactente , México , Pessoa de Meia-Idade , Inquéritos Nutricionais/estatística & dados numéricos , Necessidades Nutricionais , Triticum/química , Triticum/metabolismo , Adulto Jovem , Zea mays/química , Zea mays/metabolismoRESUMO
Here we investigated the effect of electrostatic interactions and of protein tyrosine nitration of mammalian cytochrome c on the dynamics of the so-called alkaline transition, a pH- and redox-triggered conformational change that implies replacement of the axial ligand Met80 by a Lys residue. Using a combination of electrochemical, time-resolved SERR spectroelectrochemical experiments and molecular dynamics simulations we showed that in all cases the reaction can be described in terms of a two steps minimal reaction mechanism consisting of deprotonation of a triggering group followed by ligand exchange. The pKaalk values of the transition are strongly modulated by these perturbations, with a drastic downshift upon nitration and an important upshift upon establishing electrostatic interactions with a negatively charged model surface. The value of pKaalk is determined by the interplay between the acidity of a triggering group and the kinetic constants for the forward and backward ligand exchange processes. Nitration of Tyr74 results in a change of the triggering group from Lys73 in WT Cyt to Tyr74 in the nitrated protein, which dominates the pKaalk downshift towards physiological values. Electrostatic interactions, on the other hand, result in strong acceleration of the backward ligand exchange reaction, which dominates the pKaalk upshift. The different physicochemical conditions found here to influence pKaalk are expected to vary depending on cellular conditions and subcellular localization of the protein, thus determining the existence of alternative conformations of Cyt in vivo.
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Álcalis/química , Citocromos c/metabolismo , Nitratos/metabolismo , Eletricidade Estática , Tirosina/metabolismo , Animais , Cavalos , Concentração de Íons de Hidrogênio , Ligantes , Simulação de Dinâmica Molecular , OxirreduçãoRESUMO
Cytochrome c (cyt c) is a small soluble heme protein characterized by a relatively flexible structure, particularly in the ferric form, such that it is able to sample a broad conformational space. Depending on the specific conditions, interactions, and cellular localization, different conformations may be stabilized, which differ in structure, redox properties, binding affinities, and enzymatic activity. The primary function is electron shuttling in oxidative phosphorylation, and is exerted by the so-called native cyt c in the intermembrane mitochondrial space of healthy cells. Under pro-apoptotic conditions, however, cyt c gains cardiolipin peroxidase activity, translocates into the cytosol to engage in the intrinsic apoptotic pathway, and enters the nucleus where it impedes nucleosome assembly. Other reported functions include cytosolic redox sensing and involvement in the mitochondrial oxidative folding machinery. Moreover, post-translational modifications such as nitration, phosphorylation, and sulfoxidation of specific amino acids induce alternative conformations with differential properties, at least in vitro. Similar structural and functional alterations are elicited by biologically significant electric fields and by naturally occurring mutations of human cyt c that, along with mutations at the level of the maturation system, are associated with specific diseases. Here, we summarize current knowledge and recent advances in understanding the different structural, dynamic, and thermodynamic factors that regulate the primary electron transfer function, as well as alternative functions and conformations of cyt c. Finally, we present recent technological applications of this moonlighting protein.
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Citocromos c/metabolismo , Animais , Técnicas Biossensoriais , Transporte de Elétrons , Humanos , Cinética , Mitocôndrias/enzimologia , Oxirredução , TermodinâmicaRESUMO
ABSTRACT The leaves extracts of some species of Bauhinia L. s.l. are consumed to treat diabetes, inflammation, pains and several disorders in traditional medicine in austral South America. Despite its wide use and commercialization, sale is not controlled, and botanical quality of samples is not always adequate because of plant misidentification and adulteration. Here, we characterized leaf vein pattern in nineteen taxa to contribute to the recognition and commercial quality control of plant material commercially available. The vein characters intercostal tertiary and quinternary vein fabric, areole development and shape, free ending veinlet branching and marginal ultimate venation allowed to distinguish the main medicinal species in the region.
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We report a spectroscopic, electrochemical and spectroelectrochemical characterization of the soluble cytochrome c domain (Cyt-D) from the Rhodothermus marinus caa3 terminal oxygen reductase and its putative electron donor, a high potential [4Fe-4S] protein (HiPIP). Cyt-D exhibits superior stability, particularly at the level of the heme pocket, compared to archetypical cytochromes in terms of thermal and chemical denaturation, alkaline transition and oxidative bleaching of the heme, which is further increased upon adsorption on biomimetic electrodes. Therefore, this protein is proposed as a suitable building block for electrochemical biosensing. As a proof of concept, we show that the immobilized Cyt-D exhibits good electrocatalytic activity towards H2O2 reduction. Relevant thermodynamic and kinetic electron transfer parameters for Cyt-D and HiPIP are also reported, including reorganization energies of 0.33 eV and 0.42 eV, respectively.
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Citocromos/metabolismo , Catálise , Citocromos/química , Técnicas Eletroquímicas , Estabilidade Enzimática , Cinética , Oxirredução , TermodinâmicaRESUMO
Here we report a spectroscopic, electrochemical and computational study of cytochrome c showing that nitration of Tyr74 induces Tyr deprotonation, which is coupled to Met/Lys axial ligand exchange, and results in concomitant gain of peroxidatic activity at physiological pH.
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Citocromos c/química , Nitratos/química , Prótons , Tirosina/química , Simulação por Computador , Concentração de Íons de Hidrogênio , Ligantes , Modelos Moleculares , Oxirredução , TermodinâmicaRESUMO
We report on the synthesis of two series of 1,4-naphthohydroquinone derivatives conjugated with amino acids (Gly, Ala, Phe, and Glu) and with substituted purines linked by an aliphatic chain. The compounds were obtained through Diels-Alder cycloaddition between myrcene and 1,4-benzoquinone and evaluated in vitro for their cytotoxicity (GI50 ) against cultured human cancer cells of A-549 lung carcinoma, HT-29 colon adenocarcinoma, and MCF-7 breast carcinoma. The GI50 values found for some hydroquinone-amino acid and hydroquinone-purine hybrids against MCF-7 are in an activity range comparable to that of the reference drug doxorubicin.
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Aminoácidos/síntese química , Aminoácidos/farmacologia , Antineoplásicos/síntese química , Antineoplásicos/farmacologia , Hidroquinonas/síntese química , Hidroquinonas/farmacologia , Purinas/síntese química , Purinas/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Doxorrubicina/farmacologia , Células HT29 , Humanos , Concentração Inibidora 50 , Células MCF-7 , Estrutura MolecularRESUMO
We have employed a combination of protein film voltammetry, time-resolved vibrational spectroelectrochemistry and molecular dynamics simulations to evaluate the electron-transfer reorganization free energy (λ) of cytochrome c (Cyt) in electrostatic complexes that mimic some basic features of protein-protein and protein-lipid interactions. The results reveal the existence of two native-like conformations of Cyt that present significantly different λ values. Conversion from the high to the low λ forms is triggered by electrostatic interactions, and involves the rupture of a weak H-bond between first- (M80) and second-sphere (Y67) ligands of the heme iron, as a distinctive feature of the conformational switch. The two flexible Ω loops operate as transducers of the electrostatic signal. This fine-tuning effect is abolished in the Y67F Cyt mutant, which presents a λ value similar to the WT protein in electrostatic complexes. We propose that interactions of Cyt with the natural redox partner proteins activate a similar mechanism to minimize the reorganization energy of interprotein electron transfer.
Assuntos
Citocromos c/química , Animais , Citocromos c/genética , Técnicas Eletroquímicas , Transporte de Elétrons , Cavalos , Ligação de Hidrogênio , Simulação de Dinâmica Molecular , Mutação Puntual , Análise Espectral Raman , Eletricidade Estática , Tirosina/química , Tirosina/genéticaRESUMO
Nitrogen (N) remobilization in wheat (Triticum aestivum) plants is crucial because it determines the grain protein concentration and the baking quality of flour. In order to evaluate the influence of cytokinins on N remobilization during N starvation, we analyzed various N remobilization parameters in wheat plants that were watered with 6-benzylaminopurine (BAP) either with or without KNO(3). Besides, the effects of BAP on protein synthesis were evaluated, and the size and ultrastructure of chloroplasts of BAP-treated plants were studied. BAP supply inhibited N remobilization of plants independently of N supply as shown by the increase in protein, Rubisco, chlorophyll, sugar and starch concentrations in the older leaves, the decrease in amino acid and sugar export to the phloem, and the decrease in protein, Rubisco and chlorophyll concentrations in the younger leaves. Besides, BAP supply increased nitrate reductase activity and decreased nitrate concentration, thus suggesting an increased assimilatory capacity. The increase in protein concentration could be explained mainly by a significant decrease in protein degradation and, to a lesser extent, by an increase in protein synthesis. Finally, an increase both in the size of the chloroplast and in the plastoglobuli and starch contents in BAP-supplied plants was observed. We propose that cytokinins retain the sink activity of the older leaves by inhibiting amino acid and sugar export to the phloem and stimulating assimilate accumulation in the chloroplasts of the older leaves. Besides, BAP may increase protein concentration of the older leaves both by decreasing protein degradation and maintaining protein synthesis even under stress conditions.
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Cloroplastos/ultraestrutura , Citocininas/fisiologia , Nitrogênio/metabolismo , Triticum/metabolismo , Sequência de Bases , Primers do DNA , Microscopia Eletrônica , Reação em Cadeia da Polimerase , Triticum/crescimento & desenvolvimento , Triticum/ultraestruturaRESUMO
INTRODUCTION: The aim of this report was to experimen-tally demonstrate the biological actions of acupuncture in an animal model of immune-mediated inflammation associated with a deposition of collagen. MATERIALS AND METHODS: Male Wistar rats were sensitized by a subcu-taneous implant of heat-solidified hen egg-white and divided into 4 groups: acupuncture, sham acupuncture, immobilized, and control. Acupuncture was initiated the day after sensitization and repeated twice a week for 3 weeks. The dorsal acupoints chosen were GV-14 and BL-13, the ventral acupuncture points were LU-1, CV-17, ST-36 and SP-6. The dorsal points were stimulated manually and the ventral ones by electroacupuncture. On day 14, animals were challenged through the tail vein with Sepharose(R)beads coupled with ovalbumin. One week later, animals were bled, plasma corticoster-one concentrations were measured and the lungs were removed for histological evaluation. RESULTS: Measurement of the areas of pulmonary lesion on hematoxylin-eosin stained slides showed a significant decrease (p < 0.05) in the inflammatory infiltrate in the acupuncture group, compared to the other 3 groups. Utilization of Litt and Picrosirius staining methods, in order to better visualize the infiltrate of eosinophils and the deposition of collagen, respectively, showed that both were much less intense in the acupuncture group. Corticosterone plasma levels were similar in all groups. CONCLUSION: Point-specific acupuncture treatment effectively reduced the inflammatory process and the deposition of collagen around ovalbumin-Sepharose beads intravenously embolized to the lungs of rats previously sensitized with the same protein that was administered subcutaneously.