Allergens of the urushiol family promote mitochondrial dysfunction by inhibiting the electron transport at the level of cytochromes b and chemically modify cytochrome c1

BACKGROUND: Urushiols are pro-electrophilic haptens that cause severe contact dermatitis mediated by CD8+ effector T-cells and downregulated by CD4+ T-cells. However, the molecular mechanism by which urushiols stimulate innate immunity in the initial stages of this allergic reaction is poorly understood. Here we explore the sub-cellular mechanisms by which urushiols initiate the allergic response. RESULTS: Electron microscopy observations of mouse ears exposed to litreol (3-n-pentadecyl-10-enyl-catechol]) showed keratinocytes containing swollen mitochondria with round electron-dense inclusion bodies in the matrix. Biochemical analyses of sub-mitochondrial fractions revealed an inhibitory effect of urushiols on electron flow through the mitochondrial respiratory chain, which requires both the aliphatic and catecholic moieties of these allergens. Moreover, urushiols extracted from poison ivy/oak (mixtures of 3-n-pentadecyl-8,11,13 enyl/3-n-heptadecyl-8,11 enyl catechol) exerted a higher inhibitory effect on mitochondrial respiration than did pentadecyl catechol or litreol, indicating that the higher number of unsaturations in the aliphatic chain, stronger the allergenicity of urushiols. Furthermore, the analysis of radioactive proteins isolated from mitochondria incubated with 3H-litreol, indicated that this urushiol was bound to cytochrome c1. According to the proximity of cytochromes c1 and b, functional evidence indicated the site of electron flow inhibition was within complex III, in between cytochromes bL (cyt b566) and bH (cyt b562). CONCLUSION: Our data provide functional and molecular evidence indicating that the interruption of the mitochondrial electron transport chain constitutes an important mechanism by which urushiols initiates the allergic response. Thus, mitochondria may constitute a source of cellular targets for generating neoantigens involved in the T-cell mediated allergy induced by urushiols.

Advances in enzyme bioelectrochemistry

ABSTRACT Bioelectrochemistry can be defined as a branch of Chemical Science concerned with electron-proton transfer and transport involving biomolecules, as well as electrode reactions of redox enzymes. The bioelectrochemical reactions and system have direct impact in biotechnological development, in medical devices designing, in the behavior of DNA-protein complexes, in green-energy and bioenergy concepts, and make it possible an understanding of metabolism of all living organisms (e.g. humans) where biomolecules are integral to health and proper functioning. In the last years, many researchers have dedicated itself to study different redox enzymes by using electrochemistry, aiming to understand their mechanisms and to develop promising bioanodes and biocathodes for biofuel cells as well as to develop biosensors and implantable bioelectronics devices. Inside this scope, this review try to introduce and contemplate some relevant topics for enzyme bioelectrochemistry, such as the immobilization of the enzymes at electrode surfaces, the electron transfer, the bioelectrocatalysis, and new techniques conjugated with electrochemistry vising understand the kinetics and thermodynamics of redox proteins. Furthermore, examples of recent approaches in designing biosensors and biofuel developed are presented.

Effects of different frequencies of physical training on electron transport chain and oxidative damage in healthy mice

The present study investigated the effect of different frequencies (three and five times a week) on electron transport chain and oxidative stress after 8 weeks of run training. Methods: Eighteen male mice (CF1, 30-35g) were distributed into the following groups (n=6): untrained (UT); trained three-time per week (T3) and trained five- time per week (T5). All training sessions were at the same intensity and duration (45min/day) in a treadmill for small animals. Forty-eight hours after the last training session, the animals were killed by decapitation and quadriceps (red portion) was removed and stored at -70ºC. Succinate dehydrogenase (SDH), complexes I, II, II-III, IV and hydroperoxides were measured. Results: Training sessions for five times per week were more effective in increasing the mitochondrial respiratory chain enzyme activities (SDH, complexes I, II, II-III, IV) as well as in decreasing the formation hydroperoxides than sessions performed for three times training per week (p<0.05). Conclusion: Our findings clearly showed that a higher the frequency of training session promotes a greater activity of the electron transport chain and consequently reduces the oxidative stress in healthy animals.(AU)

Lactobacillus plantarum: Effect of a protective biofilm on the surface of olives during storage

Abstract The aim of the present study was to investigate the effect of Lactobacillus plantarum adhesion to the surface of olives during storage through studying the interaction between the surfaces of the olives and L. plantarum. The results showed that the total number of adherent L. plantarum increased exponentially from 1.2 × 106 to 1.3 × 108 cfu/g. Images obtained using environmental scanning electron microscopy (ESEM) after 4 days of storage revealed that the olive surface was covered with a uniform and compact biofilm constituted of L. plantarum and yeast. Physicochemical analysis of surface of L. plantarum revealed that it was hydrophilic (Giwi > 0 mJ/m2). The surface of the olives also appeared to be hydrophilic (Giwi = 3.28 mJ/m2). The electron-donor characteristics of the surfaces of L. plantarum and olive were γ− = 53.1 mJ/m2 and γ− = 28.1 mJ/m2, respectively. The formation of a protective biofilm of L. plantarum increased the hydrophilicity (from 3.28 to 46.14 mJ/m2) and the electron-donor capacity (from 28.1 to 67.2 mJ/m2) of the olive surface by 1 day of storage. Analysis of the impact of the biofilm that formed on the surface of the olives during storage showed a reduction in the content of undesirable planktonic microorganisms, such as fungi, which could have occurred due to competition for nutrients and oxygen or modifications in the physicochemical properties of the olives. Thus, coating the surface of olives with a natural material, such as L. plantarum, may be a first step in developing strategies to prevent their microbial colonization.

Coenzyme Q10 defects may be associated with a deficiency of Q10-independent mitochondrial respiratory chain complexes

BACKGROUND: Coenzyme Q10 (CoQ10 or ubiquinone) deficiency can be due either to mutations in genes involved in CoQ10 biosynthesis pathway, or to mutations in genes unrelated to CoQ10 biosynthesis. CoQ10 defect is the only oxidative phosphorylation disorder that can be clinically improved after oral CoQ10 supplementation. Thus, early diagnosis, first evoked by mitochondrial respiratory chain (MRC) spectrophotometric analysis, then confirmed by direct measurement of CoQ10 levels, is of critical importance to prevent irreversible damage in organs such as the kidney and the central nervous system. It is widely reported that CoQ10 deficient patients present decreased quinone-dependent activities (segments I + III or G3P + III and II + III) while MRC activities of complexes I, II, III, IV and V are normal. We previously suggested that CoQ10 defect may be associated with a deficiency of CoQ10-independent MRC complexes. The aim of this study was to verify this hypothesis in order to improve the diagnosis of this disease. RESULTS: To determine whether CoQ10 defect could be associated with MRC deficiency, we quantified CoQ10 by LC-MSMS in a cohort of 18 patients presenting CoQ10-dependent deficiency associated with MRC defect. We found decreased levels of CoQ10 in eight patients out of 18 (45 %), thus confirming CoQ10 disease. CONCLUSIONS: Our study shows that CoQ10 defect can be associated with MRC deficiency. This could be of major importance in clinical practice for the diagnosis of a disease that can be improved by CoQ10 supplementation.

Desempenho fotossintético de folhas jovens e maduras de Vochysia cinnamomea (Vochysiaceae) em áreas de cerrado rupestre intactas e pós-queimada / Photosynthetic performance of young and mature leaves of Vochysia cinnamomea (Vochysiaceae) at intact and after fire areas of cerrado rupestre

O fogo exerce um papel importante como modelador dos ecossistemas de diversas formações vegetais, especialmente o Cerrado. Sua ação induz a brotação em diversas espécies, muitas vezes formando folhas mais vigorosas e fisiologicamente mais ativas. Nesse contexto, o presente trabalho investigou o desempenho fotossintético de folhas jovens e maduras de Vochysia cinnamomea Pohl (Vochysiaceae) em resposta à ação do fogo natural sobre uma área de cerrado rupestre da Serra da Canastra, Minas Gerais, comparando-se; (1) folhas jovens de área queimada e não queimada e (2) folhas jovens e maduras de um mesmo indivíduo. Foram analisadas folhas maduras de indivíduos de área não queimada cuja brotação ocorreu anterior à queimada, e folhas jovens de indivíduos de áreas queimadas e não queimadas cuja brotação ocorreu após a passagem do fogo. Foram coletados e analisados dados relativos às taxas fotossintéticas, rendimento quântico potencial e efetivo, taxa relativa de transporte de elétrons, assimilação líquida de CO2, condutância estomática, transpiração e teores de clorofila. Não houve diferenças significativas entre as folhas jovens da área queimada e da área não queimada em relação aos processos fisiológicos testados. Desta forma não há nenhum tipo de resposta rápida em relação à passagem do fogo. Entretanto, foram encontradas diferenças significativas quando comparadas folhas jovens e maduras. É fato que folhas jovens de V. cinnamomea são estruturalmente distintas de folhas maduras, são mais suculentas e pilosas enquanto as folhas maduras são mais coriáceas, glabras e com cutícula evidente. Estas diferenças estruturais e os estágios fisiológicos de maturação distintos se refletem nas características fisiológicas estudadas nestas folhas que, quando maduras apresentam-se com menor grau de fotoinibição, maior teor de clorofilas e maior assimilação líquida de CO2.

Post-fire effect plays an important role as a modulator of plant ecosystems, especially of the Cerrado. It induces leaf sprouting in several species, often forming vigorous and more physiologically active leaves. In the present study it was investigated the photosynthetic performance of young and mature leaves of Vochysia cinnamomea Pohl (Vochysiaceae) in response to the fire action in a "cerrado rupestre" area of Serra da Canasta, Minas Gerais, comparing; (1) young leaves from fired and intact area, and (2) young and mature leaves from the same plant. Mature leaves from intact area plants which leaf sprouting occurred before fire action in the adjacent area, and young leaves from intact and fired area which leaf sprouting occurred after fire action were analyzed. Data were collected and analyzed on photosynthetic rates, potential and effective quantum yield, electron transport rate, CO2 assimilation, stomatal conductance, transpiration and chlorophyll contents. There were no significant differences between the young leaves of the burned and unburned area in relation to physiological parameters tested. Thus there is no kind of quick response in relation to fire effects. However, significant differences between young and mature leaves were found. Young leaves are structurally distinct of the mature leaves, been more succulent and pilous while mature leaves are more coriaceous, glabrous and with thicked cuticle. The structural differences between young and mature leaves are reflected in distinct physiological performance. Mature leaves present lower degree of photoinhibition, higher pigment content and CO2 assimilation.

Novo papel da proteína XPC na regulação dos complexos da cadeia de transporte de elétrons e desequilíbrio redox / New role of XPC protein in regulating the electron transport chain complexes and redox unbalance

Espécies reativas de oxigênio (EROs) são normalmente e continuamente geradas em mitocôndrias, majoritariamente na cadeia de transporte de elétrons (CTE). Harman (1956, 1972 e 1992) teorizou que os radicais livres gerados nas mitocôndrias seriam a principal causa do envelhecimento. De fato, durante o envelhecimento é observado um desequilíbrio entre formação e remoção de EROs, que resulta em estresse redox. Essa condição favorece a formação de lesões oxidadas no DNA, acarretando em mutagênese ou morte celular. Diversos mecanismos moleculares cooperam para o reparo de DNA. Duas vias de reparo de DNA lidam com a maioria das lesões: o reparo por excisão de base (BER) e o reparo por excisão de nucleotídeos (NER). A via BER corrige pequenas modificações de bases que surgem de reações de desaminação, alquilação e oxidação. A via NER é mais versátil, reconhecendo lesões que distorcem a dupla hélice de DNA, como danos induzidos por luz UV e adutos volumos. Pacientes xeroderma pigmentoso (XP-A a XP-G) herdam mutações em um de sete genes que codificam proteínas envolvidas na via NER, ou em um gene que codifica uma polimerase translesão (XP-V). A doença é caracterizada por fotosensibilidade e incidência elevada de neoplasias cutâneas. A proteína XPC atua na etapa de reconhecimento da lesão de DNA na subvia de reparo global do genoma (GG-NER), e sua mutação dá origem aos sintomas clássicos de XP. Novas funções de XPC foram recentemente descritas: i) atuando como cofator na via BER auxiliando as DNA glicosilases OGG1, TDG e SMUG; ii) atuando como cofator transcricional de elementos responsivos a Oct4/Sox2, RXR e PPARα; e iii) na adaptação metabólica na transformação de queratinócitos. Então, propusemo-nos a investigar as relações entre XPC e a manutenção da integridade do DNA mitocondrial, a sensibilidade celular a estresse redox mitocondrial e possíveis alterações bioenergéticas e redox. Para tal, padronizamos um ensaio in vitro de cinética de incisão em DNA plasmidial a fim de investigarmos o possível papel de XPC no reparo de lesões oxidadas em mtDNA. Porém, nossos dados revelaram que XPC não se encontra em mitocôndrias. Apesar disso, células XP-C são mais sensíveis ao tratamento com azul de metileno (AM), antimicina A (AA) e rotenona (ROT), que geram estresse redox mitocondrial. A sensibilidade à AA foi completamente revertida em células corrigidas. Células XP-C apresentaram alterações quanto ao uso dos complexos mitocondriais, com diminuição da taxa de consumo de oxigênio (OCR) via complexo I e um aumento da OCR via complexo II, dependente da presença de XPC. Ademais, a linhagem XP-C apresentou um desequilíbrio redox mitocondrial com maior produção de EROs e menor atividade de GPx. O DNA mitocondrial de células XP-C apresentou níveis elevados de lesão e deleção, que no entanto não retornaram aos níveis encontrados em células selvagens na linhagem XP-C corrigida. Observamos uma acentuada diminuição da expressão de PPARGC1A, um importante regulador de biogênese mitocondrial. Contudo, não foi possível determinar o mecanismo de supressão da expressão de PPARGC1A. Por fim, identificamos que o tipo de mutação em XPC pode estar associado a expressão de PPARGC1A. Esse estudo abre novas possibilidade na investigação do papel de proteína XPC, à parte da instabilidade genômica, na adaptação metabólica e desequilíbrio redox em direção da progressão tumoral

Mitochondria continuously produce reactive oxygen species (ROS), mainly at the electron transport chain. Harman (1956, 1972 e 1992) proposed that normal aging is driven by increased mitochondrially generated free radicals. Indeed, during the course of aging there is an increased imbalance between formation and removal of ROS, leading to redox stress. This condition favours the formation of oxidized DNA lesions, given rise to mutations and cell death. Several molecular mechanisms cooperates to repair the DNA. Two DNA repair pathways deal with the majority of lesions: base excision repair (BER) and nucleotide excision repair (NER). The BER pathway corrects small base modifications that arise from deamination, alkylation and oxidation reactions. The NER pathway is more versitile, recognizing helix-distorting lesions, such as UV-induced damage and bulky adducts. Xeroderma pigmentosum (XP-A to XP-G) patients inherit mutations in one of seven protein-coding genes involved in NER pathway, or in a gene coding a translesion DNA polymerase (XP-V). Photosensitivity and a thousand-fold increased in the risk of developing cutaneous neoplasms are the main clinical features of XP. XPC protein functions in the recognition step of global genome NER (GG-NER) sub-pathway, and mutations in this gene lead to classical XP symptoms. Recently, it has been described that XPC acts: i) as a cofactor in BER pathway through functional interaction with DNA glycosylases OGG1, TDG and SMUG1; ii) as coactivator in transcription at Oct4/Sox2, RXR and PPARα responsive elements; iii) in metabolic shift during keratinocytes transformation. Thus, we sought to investigate a possible role for XPC in the maintenance of mtDNA integrity, cellular sensitivity to mitochondrial redox stress and eventual bioenergetic and redox changes. For this purpose, we established an in vitro plasmid incision assay to investigate the possible role of XPC in the repair of oxidized lesions in mitochondrial DNA. However, our data revealed that XPC did not localized in mitochondria. Nonetheless, XP-C cells are more sensitive to methylene blue, antimycin A (AA) and rotenone treatment, which induce mitochondrial redox stress. The XP-C sensitivity to AA was completely reverted in XPC-corrected cells. XP-C cells presented altered usage of mitochondrial complexes, with decreased oxygen consumption rate (OCR) via complex I and increased OCR through complex II, an XPC-dependent phenomenon. Furthermore, the XP-C cell line showed mitochondrial redox imbalance with increased ROS production and decrease GPx activity. MtDNA from XP-C cells accumulate lesions and deletions, which, however, were found at similar levels in the corrected cell line. We identified a sharp decrease in the expression of PPARGC1A, a master regulator of mitochondrial biogenesis. Nevertheless, it was not possible to determine the mechanism of suppression of PPARGC1A expression. Finally, our results suggest a possible link between the type of XPC mutation and PPARGC1A expression. This study unfolds new possible roles for XPC, aside from its established roles in genomic instability, in metabolic adaptation and redox imbalance towards tumour progression

Root Canal Penetration of a Sodium Hypochlorite Mixture Using Sonic or Ultrasonic Activation

The purpose of this ex vivo study was to determine, in "open" and "closed" systems, whether the design has an influence on the penetration length of sodium hypochlorite mixed with a radiopaque contrast medium, measured in millimeters, when delivered using positive pressure (PP) and using sonic (SI) or passive ultrasonic (PUI) activation. Sixty single-rooted teeth were divided into two groups: open and closed systems (n=30). Root canal shaping was performed to a working length of 17 mm. The samples were divided into three sub-groups (n=10) according to irrigant delivery and activation: PP, and SI or PUI activation. By using radiographs, penetration length was measured, and vapor lock was assessed. For the closed group, the penetration distance means were: PP 15.715 (±0.898) mm, SI 16.299 (±0.738) mm and PUI 16.813 (±0.465) mm, with vapor lock occurring in 53.3% of the specimens. In the open group, penetration to 17 mm occurred in 97.6% of the samples, and no vapor lock occurred. Irrigant penetration and distribution evaluation using open and closed systems provide significantly different results. For closed systems, PUI is the most effective in delivering the irrigant to working length, followed by SI.

O objetivo deste estudo in vivo foi determinar, para os sistemas "abertos" e "fechados", se o design tem influência na penetração, em milímetros, do hipoclorito de sódio misturado com um meio radiopaco quando empregado na ativação com pressão positiva (PP) e ativação sônica (SI) ou ultrassônica passiva (PUI). Sessenta dentes unirradiculares foram divididos em dois grupos: sistema aberto e sistema fechado (n=30). Os canais radiculares foram trabalhados até um comprimento de trabalho de 17 mm. Os grupos foram subdivididos em três subgrupos (n=10) de acordo com a solução irrigadora e a ativação: PP, e ativação SI ou PUI. Usando radiografias, a distância de penetração foi medida e avaliado o vapor contido. Para o grupo fechado, as distâncias médias de penetração foram PP 15,715 (±0,898) mm, SI 16,299 (±0,738) mm e PUI 16,813 (±0,465) mm e houve vapor contido em 53,3% das amostras. No grupo aberto, houve penetração de 17 mm em 97.6% das amostras, sem contenção de vapor. A penetração do irrigante e avaliação da distribuição usando sistemas aberto e fechado produziram resultados significativamente diferentes. Para os sistemas fechados, PUI é mais eficaz para levar o irrigante até preencher o comprimento de trabalho, seguido por SI.

Fluvoxamine alters the activity of energy metabolism enzymes in the brain

Objective: Several studies support the hypothesis that metabolism impairment is involved in the pathophysiology of depression and that some antidepressants act by modulating brain energy metabolism. Thus, we evaluated the activity of Krebs cycle enzymes, the mitochondrial respiratory chain, and creatine kinase in the brain of rats subjected to prolonged administration of fluvoxamine. Methods: Wistar rats received daily administration of fluvoxamine in saline (10, 30, and 60 mg/kg) for 14 days. Twelve hours after the last administration, rats were killed by decapitation and the prefrontal cortex, cerebral cortex, hippocampus, striatum, and cerebellum were rapidly isolated. Results: The activities of citrate synthase, malate dehydrogenase, and complexes I, II-III, and IV were decreased after prolonged administration of fluvoxamine in rats. However, the activities of complex II, succinate dehydrogenase, and creatine kinase were increased. Conclusions: Alterations in activity of energy metabolism enzymes were observed in most brain areas analyzed. Thus, we suggest that the decrease in citrate synthase, malate dehydrogenase, and complexes I, II-III, and IV can be related to adverse effects of pharmacotherapy, but long-term molecular adaptations cannot be ruled out. In addition, we demonstrated that these changes varied according to brain structure or biochemical analysis and were not dose-dependent. .

Fiebre chikungunya en México: caso confirmado y apuntes para la respuesta epidemiológica / Chikungunya fever in Mexico: confirmed case and notes on the epidemiologic response

La fiebre chikungunya (CHIK) es una enfermedad viral transmitida al ser humano por el mismo vector del dengue, el mosquito Aedes. Además de fiebre y fuertes dolores articulares, produce otros síntomas como mialgias, cefalea, náuseas, cansancio y exantema. No tiene tratamiento específico; el manejo terapéutico de los pacientes se enfoca en el alivio de los síntomas. Históricamente se han reportado brotes de grandes proporciones; incluso desde 2010 se llegó a considerar como una potencial epidemia emergente. En 2013 se introdujo a las islas del Caribe y recientemente se ha reportado en el continente americano. En este trabajo se describe el primer caso confirmado de chikungunya en México, en el municipio de Tlajomulco de Zúñiga, Jalisco, en mayo de 2014, importado de la isla Antigua y Barbuda, en el Caribe, por una mujer de 39 años de edad.

Chikungunya fever (CHIK) is a viral disease transmitted to human beings by the same vector as dengue -the Aedes mosquito. Besides fever and severe pain in the joints, it produces other symptoms such as myalgias, headache, nausea, fatigue and exanthema. There is no specific treatment for it; the therapeutic management of patients focuses on symptom relief. Historically, outbreaks of large proportions have been reported; even since 2010 it was considered to be a potential emerging epidemic. In 2013 it was introduced into the islands of the Caribbean, and it has recently been reported in the American continent. This paper describes the first confirmed case of chikungunya in Mexico -in the municipality of Tlajomulco de Zúñiga, Jalisco, in May, 2014-, which was imported from the Caribbean island of Antigua and Barbuda by a 39 year-old woman.

Effects of 5-aminolevulinic acid (ALA)-containing supernatants from selected Rhodopseudomonas palustris strains on rice growth under NaCl stress, with mediating effects on chlorophyll, photosynthetic electron transport and antioxidative enzymes

Background: Rice is globally one of the most important food crops, and NaCl stress is a key factor reducing rice yield. Amelioration of NaCl stress was assessed by determining the growth of rice seedlings treated with culture supernatants containing 5-aminolevulinic acid (ALA) secreted by strains of Rhodopseudomonas palustris (TN114 and PP803) and compared to the effects of synthetic ALA (positive control) and no ALA content (negative control). Results: The relative root growth of rice seedlings was determined under NaCl stress (50 mM NaCl), after 21 d of pretreatment. Pretreatments with 1 μM commercial ALA and 10X diluted culture supernatant of strain TN114 (2.57 μM ALA) gave significantly better growth than 10X diluted PP803 supernatant (2.11 μM ALA). Rice growth measured by dry weight under NaCl stress ordered the pretreatments as: commercial ALA N TN114 N PP803 N negative control. NaCl stress strongly decreased total chlorophyll of the plants that correlated with non-photochemical quenching of fluorescence (NPQ). The salt stress also strongly increased hydrogen peroxide (H2O2) concentration in NaCl-stressed plants. The pretreatments were ordered by reduction in H2O2 content under NaCl stress as: commercial ALA N TN114 N PP803 N negative control. The ALA pretreatments incurred remarkable increases of total chlorophyll and antioxidative activities of catalase (CAT), ascorbate peroxide (APx), glutathione reductase (GR) and superoxide dismutase (SOD); under NaCl stress commercial ALA and TN114 had generally stronger effects than PP803. Conclusions: The strain TN114 has potential as a plant growth stimulating bacterium that might enhance rice growth in saline paddy fields at a lower cost than commercial ALA.

Estudo das alterações bioenergéticas induzidas por compostos sintéticos em Leishmania spp / Study of Bioenergetic Changes Induced by Synthetic Compounds in Leishmania spp

A leishmaniose é uma doença negligenciada, com especial incidência nos países em desenvolvimento; hoje em dia, 350 milhões de pessoas estão em risco de contrair esta doença, com 2 milhões de novos casos notificados anualmente. O tratamento é reduzido à quimioterapia, com uma escassez alarmante de medicamentos em uso clínico atual e diversos efeitos colaterais, além de relatos de resistência associados a muitos desses medicamentos. Todo esse contexto leva a necessidade da busca urgente de novos fármacos. Neste trabalho avaliou-se o potencial leishmanicida de 63 benzofenonas sintéticas, juntamente com o fármaco buparvaquona, uma naftoquinona usada atualmente para o tratamento da theileriose, sendo uma alternativa no estudo de reposicionamento de fármacos, sendo essa uma estratégia próspera e de baixo custo para a busca de novas drogas leishmanicidas...

Comparação do treinamento físico de quatro e oito semanas sobre atividade da cadeia transportadora de elétrons e marcadores de estresse oxidativo em fígado de camundongos / Comparison between four- and eight- week physical trainings on the mitochondrial respiratory chain enzyme activities and oxidative stress markers in liver of mice

O presente estudo investigou o efeito de quatro e oito semanas de treinamento físico sobre a atividade dos complexos da cadeia transportadora de elétrons (CTE) e os marcadores de estresse oxidativo em fígado de camundongos. Vinte e um camundongos (CF1, 30-35g) foram distribuídos nos seguintes grupos: não treinado (NT); treinado quatro semanas (T4); treinado oito semanas (T8). Quarenta e oito horas após a última sessão de treinamento os animais foram mortos por decapitação e o fígado foi retirado e estocado em -70ºC para posterior análise. Atividade da succinato desidrogenase (SDH), dos complexos I,II,III e IV da CTE, carbonilação de proteína, conteúdo total de tióis e a atividade da superóxido dismutase foram mensurados. Os resultados demonstram que apenas oito semanas de treinamento aumentam a atividade da SDH, dos quatro complexos da CTE, da superóxido dismutase, e o conteúdo total de tióis em relação ao grupo não treinado. Houve ainda diminuição na carbonilação de proteína no respectivo grupo em relação ao NT. Em conclusão, são necessárias oito semanas de treinamento para que ocorram aumento no funcionamento mitocondrial e melhora nos marcadores de estresse oxidativo em fígado de camundongos.

The present study investigated mitochondrial adaptations and oxidative stress markers after four and eight weeks of running training in liver of mice. Twenty-one male mice (CF1, 30-35g) were distributed into the following groups (n=7): untrained (UT); trained - four weeks (T4); trained - eight weeks (T8). Forty-eight hours after the last training session the animals were killed by decapitation and livers were removed and stored at -70ºC. Succinate dehydrogenase (SDH), complexes I, II, II-III and IV, protein carbonyls (PC), total thiol content and superoxide dismutase activity were measured. The results show that endurance training (8-wk) increases the SDH activity and complexes (I, II, III, IV), superoxide dismutase and total thiol content in liver when compared to untrained animals. Decrease in protein carbonylation in the respective group in relation to UT was also observed. It could be concluded that eight weeks of running training are necessary for mitochondrial respiratory chain enzyme activities increase and improvement in oxidative stress markers in liver of mice.

Respiration, oxidative phosphorylation, and uncoupling protein in Candida albicans

The respiration, membrane potential (Dy), and oxidative phosphorylation of mitochondria in situ were determined in spheroplasts obtained from Candida albicans control strain ATCC 90028 by lyticase treatment. Mitochondria in situ were able to phosphorylate externally added ADP (200 µM) in the presence of 0.05 percent BSA. Mitochondria in situ generated and sustained stable mitochondrial Dy respiring on 5 mM NAD-linked substrates, 5 mM succinate, or 100 µM N,N,N',N'-tetramethyl-p-phenylenediamine dihydrochloride plus 1 mM ascorbate. Rotenone (4 µM) inhibited respiration by 30 percent and 2 µM antimycin A or myxothiazole and 1 mM cyanide inhibited it by 85 percent. Cyanide-insensitive respiration was partially blocked by 2 mM benzohydroxamic acid, suggesting the presence of an alternative oxidase. Candida albicans mitochondria in situ presented a carboxyatractyloside-insensitive increase of Dy induced by 5 mM ATP and 0.5 percent BSA, and Dy decrease induced by 10 µM linoleic acid, both suggesting the existence of an uncoupling protein. The presence of this protein was subsequently confirmed by immunodetection and respiration experiments with isolated mitochondria. In conclusion, Candida albicans ATCC 90028 possesses an alternative electron transfer chain and alternative oxidase, both absent in animal cells. These pathways can be exceptional targets for the design of new chemotherapeutic agents. Blockage of these respiratory pathways together with inhibition of the uncoupling protein (another potential target for drug design) could lead to increased production of reactive oxygen species, dysfunction of Candida mitochondria, and possibly to oxidative cell death.

Tratamiento de la enfermedad de Chagas / Treatment of Chagas disease

En la actualidad se acepta que la enfermedad de Chagas humana debe ser tratada en cualquier período de su evolución con la única excepción del período crónico terminal. En el período agudo clínico, infección de menos de 2 meses así como en el biológico: pesquisa de parásitos al fresco, frotis, gota gruesa y con serología convencional positiva e IgM(+). El ideal de tratamiento es con nifurtimox (NF) 8-10 mg/kg día en adultos y 15 mg/kg día en niños por 60-90 días. La dosis se reparte en tres tomas. La curación clínica y serológica es de un 60 por ciento. En Brasil donde se utiliza este fármaco se trata con benznidazol (BNZ) 5 mg/kg día (adultos) y 5-10 mg/kg día en niños por 60 días. En las infecciones congénitas la terapia debe ser precoz en cuanto se realice el diagnóstico por clínica y pesquisa del parásito al fresco, frotis, microstraut etc. Muchas veces el diagnóstico se efectúa por persistenica de la serología por más de 6 meses y el recién nacido ya esta en etapa crónica de la infección. Es necesario efectuar seguimiento clínico serológico y parasitológico de los casos. Las infecciones accidentales deben ser tratadas por 10 días. En los trasplantes de órganos en que el receptor o dador es chagásico se debe indicar terapia con NF o BNZ a igual dosis y tiempo que la señalada anteriormente. Las reactivaciones en los casos crónicos ejem: que adquieren un SIDA o que presentan depresiones del sistema inmunidad celular como leucemias, Hodgkin, etc se deben tratar como cuadros agudos con NF ó NBZ por períodos prolongados de 5 ó más meses. En estos casos obviamente la prevención es lo ideal: hacer serología para Chagas a los pacientes con SIDA, etc

Energy production in Entamoeba histolytica: new perspectives in rational drugs design

The amebicidal action of metronidazol is activated when the enzyme pyruvate: ferredoxin oxido reductase transfers reducing equivalents to the nitro group of the drug. The enzyme is present in Entanoeba histolytica and other anaerobic parasites like Giardia and Trichomonas that lack mitochondria. The selectivity of the drug can be ascribed to the absence of the reductasa in the human host. E. histolytica possesses other enzymes involved in glucose catabolism that are interesting for the rational desing of new drugs. It has glycolytic enzymes that are important for the production of energy like phosphofructokinase, pyruvate phosphate dikinase, phosphoenolpyruvate carboxytransphosphorylase and thiokinase, which use pyrophosphate as a phosphate donor and have no human counterparts. The first part of this article describes the reactions by which E. histolytica obtains energy from glucose degradation, and includes recent advances in thecloning of genes for the various participating enzymes. The second part shows an alternative view for the study of target enzymes that are unique to the parasite, and indicates their importance in therapautic research

Thyroid calorigenesis and oxidative stress: modification of the respiratory burst activity in polymorphonuclear leukocytes

1. Recent research on experimental animals indicates that thyroid hormone administration is associated with greater electron flow through the microsomal, mitochondrial, and peroxisomal electron transport systems of the liver, as indicated by the enhanced production of superoxide radicals (O2.-) and/or hydrogen proxide (H2O2). 2. Hyperthyroidism induces an increased respiratory burst activity in rat and human polymorphonuclear leukocytes, an effect that seems to be related to changes in the myeloperoxidase-H2O2 system of the cell, rather than to the direct actions of the hormone or changes in the opsonic capacity of plasma. 3. These findings indicate that thyroid calorigenesis involves an elevated prooxidant activity in target cells, which may be relevant in determining toxic effects within the tissue or in neighboring cells

Transporte de eletrolitos no epitelio respiratorio dos portadores de fibrose cistica / Eletrolyte transport by airway epithelia in patients with cystic fibrosis

Nas vias aereas de conducao, o transporte de ions tem papel importante na composicao das secrecoes. O canal de 'Cl POT -' e o canal de 'Na POT +' existentes na membrana apical controlam a secrecao e absorcao das celulas epiteliais. Fibrose cistica, o defeito fundamental e a regulacao do canal de'Cl POT -'. Estudos complementares ao nivel de regulacao molecular deste canal poderao levar ao melhor entendimento da doenca e as novas abordagens terapeuticas.

Ultrastructural and metabolic alteration in dyskinetoplastic Trypanosoma cruzi

El cultivo del Trypanosoma cruzi (agente causal de la enfermedad de Chagas; Kinetoplstidae; cepa Tulahuen) en presencia de etidio, una droga de efecto selectivos sobre el DNA mitocondrial, produjo la inhibición del crecimiento del parásito (50% con 1 µg/ml de droga) y alteraciones típicas en el DNA cinetoplástico, verificadas por microscopía electrónica. Los kinetoplastos se vuelven irregulares y erráticos en la ubicación, pierden la forma de arco y las diferencias de las membranas (densidad y crestas), la matriz se vuelve floculenta y filamentosa y el k DNA se compacta y pierde la configuración fibrilar. La espectrofotometría diferencial de los epimastigotes discinetoplásticos demostró la disminución de los citocromos aa3, b y o, mientras que el citocromo c no fue afectado. El efecto del etidio fue maximo sobre aa3; mínimo sobre o y en todos los casos proporcional a la concentración de la droga. Con 1 µg/ml de bromuro de etidio la disminución del citocromo aa3 fue de 93% (promedio de 4 determinaciones ñ E.S.). La respuesta de los epimastigotes discinetoplásticos a los inhibidores de la cadena respiratoria mitocondrial, a saber, cianuro (concentraciones hasta 200 µM), azida y antimicina fue negativa, en contraste con los epismatigotes normales, cuya respiración fue inhibida 75-85% por los mismos inhibidores. El cianuro 1-3 mM inhibió, sin embargo, la respiración endógena de los epimastigotes discineto-plasticos y también la respiración antimicina o azida-insensible de los epimastigotes normales. Resultados similares se obtuvieron con la fracción mitocondrial de T. cruzi, que contiene el citocromo o. Teniendo en cuenta esta circunstancia se postula la operación de una xidasa terminal accesoria, inhibida por el cianuro en concentraciones relativamente altas. El ácido salicihidromáxico, un inhibidor de la respiración dependiente de la oxidasa Cu-dependiente de otros tripanosomatideos, no afectó la respiración del T. cruzi normal o discinetoplástico