Microhomology-mediated end joining is the principal mediator of double-strand break repair during mitochondrial DNA lesions

Mitochondrial DNA (mtDNA) deletions are associated with various mitochondrial disorders. The deletions identified in humans are flanked by short, directly repeated mitochondrial DNA sequences; however, the mechanism of such DNA rearrangements has yet to be elucidated. In contrast to nuclear DNA (nDNA), mtDNA is more exposed to oxidative damage, which may result in double-strand breaks (DSBs). Although DSB repair in nDNA is well studied, repair mechanisms in mitochondria are not characterized. In the present study, we investigate the mechanisms of DSB repair in mitochondria using in vitro and ex vivo assays. Whereas classical NHEJ (C-NHEJ) is undetectable, microhomology-mediated alternative NHEJ efficiently repairs DSBs in mitochondria. Of interest, robust microhomology-mediated end joining (MMEJ) was observed with DNA substrates bearing 5-, 8-, 10-, 13-, 16-, 19-, and 22-nt microhomology. Furthermore, MMEJ efficiency was enhanced with an increase in the length of homology. Western blotting, immunoprecipitation, and protein inhibition assays suggest the involvement of CtIP, FEN1, MRE11, and PARP1 in mitochondrial MMEJ. Knock-down studies, in conjunction with other experiments, demonstrated that DNA ligase III, but not ligase IV or ligase I, is primarily responsible for the final sealing of DSBs during mitochondrial MMEJ. These observations highlight the central role of MMEJ in maintenance of mammalian mitochondrial genome integrity and is likely relevant for deletions observed in many human mitochondrial disorders.

Jesucristo, plenitud de la revelación : el testimonio del Nuevo Testamento

Desde la perspectiva del Nuevo Testamento, el autor se propone un estudio que ayude a comprender y profundizar la enseñanza del Concilio Vaticano II acerca de “Jesucristo, plenitud de la revelación” (DV 2). La exposición se organiza en dos secciones relativas al Antiguo y al Nuevo Testamento: en el primero se presenta cómo Israel fue descubriendo en su historia el proyecto de Dios para sí y para la humanidad; en el segundo se trata sobre Jesús, el “lugar” donde el designio de Dios tuvo y tiene su plena realización. Para todos los discípulos, Jesucristo sigue siendo el camino hacia una vida plena (cf. DA 375).

Tratado de pazes, entre os serenissimos e poderosissimos Principes D. Carlos II, Rey Catholico, e D. Afonso VI, Rey de Portugal, feito, e concluso no Convento de Sancto Eloy da Cidade de Lisboa, aos 13 de fevereiro de 1668 : sendo mediator o serenissimo, e poderosissimo Principe Carlos II, Rey da Gram Bretanha

Analisa esse tratado que põe fim às guerras da Restauração de Portugal, mandando devolver a cada uma das partes as conquistas que à outra tiverem feito. Nas colônias, a paz passaria a vigorar um ano após a sua publicação, dado o tempo necessário para que a notícia chegasse à América e à Ásia.

Escola da doutrina christã, em que se ensina o que he obrigado a saber o christam : ordenada por modo de dialogo entre dous estudantes hum filozofo, por nome Marcelino, e outro theologo, por nome Diodoro : com exemplos accomodados às materias que se tratam : composta Pello Padre M. Joam da Fonseca, religiozo da Companhia de Jesu, dedicada á purissima Virgem, Glorioza Rainha dos Anjos, Maria Santissima Senhora Nossa da Victoria

João da Fonseca nasceu em Viana do Além-Tejo, em 1632, e morreu em Lisboa, em 1701. Foi professor de Filosofia na Universidade de Evora e missionário em várias vilas e cidades e mestre do noviciado de Coimbra. Distinguiu-se em seu tempo na Teologia Ascética, como se comprova nas obras que nos deixou, as quais são ainda apreciadas pela correção e propriedade da linguagem. Em ‘Escola da Doutrina Christã’, o autor ensina o verdadeiro caminho da vida, resumindo nesta leitura tudo o que em muitos volumes ensina a teologia especulativa e moral. Todas as matérias de que trata são confirmadas com muito se notáveis exemplos. Trata-se, aqui, da primeira edição, impressa em 1688. Tem-se noticia de uma segunda edição publicada pela Oficina da Universidade, em 1750.

Io. gottl. heineccii, iureconsulti quondam celeberrimi : recitationes in elementa iuris civilis secundum ordinem institutionum, accedunt io. christ. gottl. heineccii ... commentarius de vita, fatis ac scriptis b. parentis, auctior emendatior christiani thomasii, delineatio historiae juris nec non burc. gottelf. struvii, bibliotheca juris selectissima cum notis hen. contelmanni, ubi praestantiora iurisconsultorum opera ad nostra usque tempora referuntur

Título Uniforme : Corpus Juris Civilis. Institutas.

A simple route to incorporate redox mediator into carbon nanotubes/Nafion composite film and its application to determine NADH at low potential

Through a new and simple ion-exchange route, two-electron redox mediator thionine has been deliberately incorporated into the carbon nanotubes (CNTs)/Nafion composite film due to the fact that there is strong interaction between any of two among the three materials (ion-exchange process between thionine and Nafion, strong adsorption of thionine by CNTs, and wrapping and solubilizing of CNTs with Nation). The good homogenization of electron conductor CNTs in the integrated films provides the possibility of three-dimensional electron conductive network. The resulting integrated films exhibited high and stable electrocatalytic activity toward NADH oxidation with the significant decrease of high overpotential, which responds more sensitively more than those modified by thioine or CNTs alone. Such high electrocatalytic activity facilitated the low potential determination of NADH (as low as -0.1 V), which eliminated the interferences from other easily oxidizable species. In a word, the immobilization approach is very simple, timesaving and effective, which could be extended to the immobilization of other cationic redox mediators into the CNTs/Nafion composite film. And these features may offer potential promise for the design of amperometric biosensors.

AMPEROMETRIC GLUCOSE SENSOR WITH FERROCENE AS AN ELECTRON-TRANSFER MEDIATOR

A glucose oxidase (GOD) electrode with ferrocene (Fc) used as an electron transfer mediator has been described. Using Nafion, Fc was modified on a glassy carbon (GC) electrode surface, and glucose oxidase was then immobilized on the Fc-Nafion film, forming a GOD-Fc-Nafion enzyme electrode. The preparation method was quite simple and rapid. The enzyme electrode showed a reversible reaction of the redox couple (Fc+/Fc), used in a biosensor system, displayed a sensitive catalytic current response (response time was less than 20 s) on variation of the glucose concentration, with a wide linear range up to 16 mM and with good repeatability. The enzyme electrode showed almost no deterioration over the course of three weeks. There was little or no interference from electro-active anions, such as ascorbic acid, to the determination of glucose based on Nafion film and lower oxidizing potentials of the enzyme electrode.

Clergy, crusified within the body of Christ - a challenge to pastoral care

M.A. Thesis / University of Pretoria / Department of Practical Theology / Advised by Prof M Masango

The universal mission of the church of Christ : a discourse / delivered at Hanley, Staffordshire, August 4, 1884, being the Fernley lecture of 1884, by Benjamin Hellier.

http://www.archive.org/details/universalmission00helluoft

Christ versus Krishna : a brief comparison between the chief events, characteristics & mission of the Babe of Bethlehem Judµa and the Babe of Brindabun Mathurapuri : with a concise review of Hindooism, proving its derivation from Christianity / by L.A. Sakes.

http://www.archive.org/details/christversuskris014648mbp