Transcription coupled repair of 8-oxoguanine in murine cells: The Ogg1 protein is required for repair in nontranscribed sequences but not in transcribed sequences

To assess the role of the Ogg1 DNA glycosylase in the transcription-coupled repair (TCR) of the mutagenic lesion, 7,8-dihydro-8oxoguanine (8-OxoG), we have investigated the removal of this lesion in wild-type and ogg1−/− null mouse embryo fibroblast (MEF) cell lines. We used nonreplicating plasmids containing a single 8-OxoG·C base pair in a different assay that allowed us to study the removal of 8-OxoG located in a transcribed sequence (TS) or in a nontranscribed sequence (NTS). The results show that the removal of 8-OxoG in a wild-type MEF cell line is faster in the TS than in the NTS, indicating TCR of 8-OxoG in murine cells. In the homozygous ogg1−/− MEF cell line, 8-OxoG was not removed from the NTS whereas there was still efficient 8-OxoG repair in the TS. Expression of the mouse Ogg1 protein in the homozygous ogg1−/− cell line restored the ability to remove 8-OxoG in the NTS. Therefore, we have demonstrated that Ogg1 is essential for the repair of 8-OxoG in the NTS but is not required in the TS. These results indicate the existence of an Ogg1-independent pathway for the TCR of 8-OxoG in vivo.

Stimulation of human 8-oxoguanine-DNA glycosylase by AP-endonuclease: potential coordination of the initial steps in base excision repair

8-Oxoguanine-DNA glycosylase 1 (OGG1), with intrinsic AP lyase activity, is the major enzyme for repairing 7,8-dihydro-8-oxoguanine (8-oxoG), a critical mutagenic DNA lesion induced by reactive oxygen species. Human OGG1 excised the damaged base from an 8-oxoG·C-containing duplex oligo with a very low apparent kcat of 0.1 min–1 at 37°C and cleaved abasic (AP) sites at half the rate, thus leaving abasic sites as the major product. Excision of 8-oxoG by OGG1 alone did not follow Michaelis–Menten kinetics. However, in the presence of a comparable amount of human AP endonuclease (APE1) the specific activity of OGG1 was increased ∼5-fold and Michaelis–Menten kinetics were observed. Inactive APE1, at a higher molar ratio, and a bacterial APE (Nfo) similarly enhanced OGG1 activity. The affinity of OGG1 for its product AP·C pair (Kd ∼ 2.8 nM) was substantially higher than for its substrate 8-oxoG·C pair (Kd ∼ 23.4 nM) and the affinity for its final β-elimination product was much lower (Kd ∼ 233 nM). These data, as well as single burst kinetics studies, indicate that the enzyme remains tightly bound to its AP product following base excision and that APE1 prevents its reassociation with its product, thus enhancing OGG1 turnover. These results suggest coordinated functions of OGG1 and APE1, and possibly other enzymes, in the DNA base excision repair pathway.

Excision of 8-oxoguanine within clustered damage by the yeast OGG1 protein

Clustered damages are formed in DNA by ionising radiation and radiomimetic anticancer agents and are thought to be biologically severe. 7,8-dihydro-8-oxoguanine (8-oxoG), a major DNA damage resulting from oxidative attack, is highly mutagenic leading to a high level of G·C→T·A transversions if not previously excised by OGG1 DNA glycosylase/AP lyase proteins in eukaryotes. However, 8-oxoG within clustered DNA damage may present a challenge to the repair machinery of the cell. The ability of yeast OGG1 to excise 8-oxoG was determined when another type of damage [dihydrothymine, uracil, 8-oxoG, abasic (AP) site or various types of single-strand breaks (SSBs)] is present on the complementary strand 1, 3 or 5 bases 5′ or 3′ opposite to 8-oxoG. Base damages have little or no influence on the excision of 8-oxoG by yeast OGG1 (yOGG1) whereas an AP site has a strong inhibitory effect. Various types of SSBs, obtained using either oligonucleotides with 3′- and 5′-phosphate termini around a gap or through conversion of an AP site with either endonuclease III or human AP endonuclease 1, strongly inhibit excision of 8-oxoG by yOGG1. Therefore, this large inhibitory effect of an AP site or a SSB may minimise the probability of formation of a double-strand break in the processing of 8-oxoG within clustered damages.

Measurement of 8-hydroxy-2′-deoxyguanosine in DNA by high-performance liquid chromatography-mass spectrometry: comparison with measurement by gas chromatography-mass spectrometry

Measurement of 8-hydroxy-2′-deoxyguanosine (8-OH-dGuo) in DNA by high-performance liquid chromatography/mass spectrometry (LC/MS) was studied. A methodology was developed for separation by LC of 8-OH-dGuo from intact and modified nucleosides in DNA hydrolyzed by a combination of four enzymes: DNase I, phosphodiesterases I and II and alkaline phosphatase. The atmospheric pressure ionization-electrospray process was used for mass spectral measurements. A stable isotope-labeled analog of 8-OH-dGuo was used as an internal standard for quantification by isotope-dilution MS (IDMS). Results showed that LC/IDMS with selected ion-monitoring (SIM) is well suited for identification and quantification of 8-OH-dGuo in DNA at background levels and in damaged DNA. The sensitivity level of LC/IDMS-SIM was found to be comparable to that reported previously using LC-tandem MS (LC/MS/MS). It was found that approximately five lesions per 106 DNA bases can be detected using amounts of DNA as low as 2 µg. The results also suggest that this lesion may be quantified in DNA at levels of one lesion per 106 DNA bases, or even lower, when more DNA is used. Up to 50 µg of DNA per injection were used without adversely affecting the measurements. Gas chromatography/isotope-dilution MS with selected-ion monitoring (GC/IDMS-SIM) was also used to measure this compound in DNA following its removal from DNA by acidic hydrolysis or by hydrolysis with Escherichia coli Fpg protein. The background levels obtained by LC/IDMS-SIM and GC/IDMS-SIM were almost identical. Calf thymus DNA and DNA isolated from cultured HeLa cells were used for this purpose. This indicates that these two techniques can provide similar results in terms of the measurement of 8-OH-dGuo in DNA. In addition, DNA in buffered aqueous solution was damaged by ionizing radiation at different radiation doses and analyzed by LC/IDMS-SIM and GC/IDMS-SIM. Again, similar results were obtained by the two techniques. The sensitivity of GC/MS-SIM for 7,8-dihydro-8-oxoguanine was also examined and found to be much greater than that of LC/MS-SIM and the reported sensitivity of LC/MS/MS for 8-OH-dGuo. Taken together, the results unequivocally show that LC/IDMS-SIM is well suited for sensitive and accurate measurement of 8-OH-dGuo in DNA and that both LC/IDMS-SIM and GC/IDMS-SIM can provide similar results.

Escherichia coli Nth and human hNTH1 DNA glycosylases are involved in removal of 8-oxoguanine from 8-oxoguanine/guanine mispairs in DNA

The spectrum of DNA damage caused by reactive oxygen species includes a wide variety of modifications of purine and pyrimidine bases. Among these modified bases, 7,8-dihydro-8-oxoguanine (8-oxoG) is an important mutagenic lesion. Base excision repair is a critical mechanism for preventing mutations by removing the oxidative lesion from the DNA. That the spontaneous mutation frequency of the Escherichia coli mutT mutant is much higher than that of the mutM or mutY mutant indicates a significant potential for mutation due to 8-oxoG incorporation opposite A and G during DNA replication. In fact, the removal of A and G in such a situation by MutY protein would fix rather than prevent mutation. This suggests the need for differential removal of 8-oxoG when incorporated into DNA, versus being generated in situ. In this study we demonstrate that E.coli Nth protein (endonuclease III) has an 8-oxoG DNA glycosylase/AP lyase activity which removes 8-oxoG preferentially from 8-oxoG/G mispairs. The MutM and Nei proteins are also capable of removing 8-oxoG from mispairs. The frequency of spontaneous G:C→C:G transversions was significantly increased in E.coli CC103mutMnthnei mutants compared with wild-type, mutM, nth, nei, mutMnei, mutMnth and nthnei strains. From these results it is concluded that Nth protein, together with the MutM and Nei proteins, is involved in the repair of 8-oxoG when it is incorporated opposite G. Furthermore, we found that human hNTH1 protein, a homolog of E.coli Nth protein, has similar DNA glycosylase/AP lyase activity that removes 8-oxoG from 8-oxoG/G mispairs.

Atividade antioxidante in vitro de extrato de folhas de oliveira (Olea europaea L.) e seu efeito protetor sobre danos oxidativos em eritrócitos humanos

O objetivo do presente estudo foi avaliar a atividade antioxidante de extrato de folhas de oliveira (EFO) (Olea europaea L.) por diferentes metodologias analíticas in vitro e in situ, para verificação de efeito em sistemas biológicos. O extrato foi obtido a partir de folhas secas de oliveira, previamente micronizadas, em metanol/água (80/20%) na proporção 1:20 (m/v), após remoção de compostos solúveis em n-hexano. Após liofilização, no EFO foi avaliado o poder redutor por Folin-Ciocalteau, conteúdo de flavonoides totais, teor de oleuropeina, poder de redução do íon férrico (FRAP) e atividade antioxidante sobre DPPHo, ABTSo+, ânion superóxido (O2o-), ácido hipocloroso (HOCl) e óxido nítrico (NOo). O extrato foi também avaliado quanto ao efeito protetor sobre danos oxidativos em eritrócitos humanos. O ácido ascórbico foi utilizado como referência. O experimento foi repetido seis vezes (n = 6) e os ensaios realizados em duplicata. O poder redutor do extrato e o conteúdo de flavonoides totais e oleuropeína foram 131,7 ± 9,4 mg equivalente de ácido gálico/g extrato seco (ms), 19,4 ± 1,3 mg equivalente de quercetina/g ms e 25,5 ± 5,2 mg oleuropeína/g ms, respectivamente. O ensaio de FRAP apresentou 281,8 ± 22,8 mg equivalente de trolox/g ms. O EFO foi efetivo na inibição dos radicais DPPHo e ABTSo+, dependente da concentração de extrato, com valores de IC50 de 13,8 ± 0,8 e 16,1 ± 1,2 µg/mL, respectivamente. Com relação à atividade antioxidante sobre espécies reativas de importância biológica, o EFO apresentou forte capacidade de inibição de O2o- (IC50 = 52,6 ± 2,1 µg/mL) e NOo (IC50 = 48,4 ± 6,8 µg/mL), quando comparado ao ácido ascórbico. Porém, a inibição de HOCl não foi tão eficiente (IC50 = 714,1 ± 31,4 µg/mL). O EFO inibiu a hemólise induzida em eritrócitos de maneira dependente da concentração (IC50 = 7,8 ± 1,1 µg/mL), assim como a peroxidação lipídica e a formação de meta-hemoglobina, com valores de IC50 de 38,0 ± 11,7 e 186,3 ± 29,7 µg/mL, respectivamente. Os resultados obtidos neste estudo sugerem que extrato de folhas de oliveira possui efetiva atividade antioxidante em sistemas biológicos, pelo efeito sequestrador de determinadas espécies reativas que participam dos processos bioquímicos, e pela prevenção de danos oxidativos em eritrócitos humanos. Portanto, sua ingestão pode estar relacionada com a prevenção de estresse oxidativo in vivo, com consequentes benefícios à saúde.

Tunisia. Supervising Tunisian Elections by civil society: How to improve it? Arab Citizenship Review No. 7. January 2015

Introduction. On October 26, 2014, Tunisia held its second democratic legislative elections. Participation among more than 5 million registered voters was at about 60%, a relatively good turnout for the country, compared to the 52% voters in 2011. Preliminary results for the 33 constituencies (27 within the country and 6 for expatriates) reveal that secular frontrunner Nidaa Tounes (Call of Tunisia) won around 37% percent of votes while moderate Islamist party Ennahdha, winner of the 2011 elections and leader of Tunisia’s post-revolution government, received 27% of votes. Other parties with notable percentages are the Free Patriotic Union (French: UPL) with 4.4% and the leftist party, Popular Front, with 3.7%. Legislative were immediately followed by two round presidential elections the first one held on November 23, the second one after one month. Conversely to what was expected, people were more attracted by presidential elections even though president has notably less prerogatives than the parliament: representing the state, he is mainly responsible for determining the general state policies in the domains of defense, foreign relations and national security (article 76.) This paradox is ascribed to national imaginary more confident in a “Zaïm” (leader) rather than a collective institution such as a parliament. The turnout was at about 64% within the national 27 constituencies. Out of 70 candidates (including 5 female), 27 (with only one female) met the legal requirements to run for the presidency. The result confirms the legislative trend and Beji Caid Essebsi, leader of Nidaa, was proclaimed the third President of Tunisia. He gained 39.46% of the votes at the first round elections. Essebsi was followed by Moncef Marzouki who received an unexpected score (33.43%) at the first round, thanks to the support of Ennahdha audience and to an active and insistent campaign focused on the idea that revolution is threatened by old regime guard “come-back.” Rewarded for his long militant live, the extreme leftist Hamma Hammami in a new look gained 7,8% of the votes while the new comer Slim Riahi received 5,5% despite rumors circulating on his personal reputation. Notably, Kalthoum Kennou gained 0,55% (18.287 votes) but listed eleventh out of 27.

Amendment to Interstate Commerce Act (trip leasing). Hearing before the Committee on Interstate and Foreign Commerce, United States Senate, 83d Congress, 1st session on H.R.3203. July 8-June 25, 1954.

pt.1. July 8 and 9, 1953. 191 p pt.2. May 10, 11, June 7, 8, and 25, 1954. pp. 193-511.

En lagom lätt text, det är så lagom lätt! : En kvalitativ undersökning av tio elevers textrörlighet inför texter från 8 SIDOR

Myndigheten för tillgängliga medier ger ut tidningen 8 SIDOR med nyheter på lättläst svenska,  som riktar sig till alla med lässvårigheter. Uppsatsens syfte är att ta reda på om läsare i  målgruppen och skribenter till texter i tidningen har samma syn på vad som är en lättläst och  begriplig text, och vad de tycker förenklar och försvårar en text. Tio elever i årskurs 7-8 i grundsärskolan och grundskolan har fått samtala i tre fokusgrupper om  tre texter från tidningens webbsida. Samtalen har analyserats utifrån begreppet textrörlighet,  som är ett verktyg som analyserar läsarens relation till texten. Textrörlighet delas upp i  textbaserad rörlighet, rörlighet utåt och interaktiv rörlighet. De två journalister som har skrivit  texterna har intervjuats om begreppet lättläst, mottagaranpassning och texterna. I många fall resonerar elever och journalister lika, men eleverna önskar tydligare rubriker och i  viss mån ämnen som är mer anpassade till dem själva. Det som fungerar bäst för elevernas  textbaserade rörlighet är när texten är logiskt uppbyggd och har lagom svåra ord samt en  välvald bild. För textrörligheten utåt fungerar det bäst när eleverna har intresse och  förkunskaper i ämnet. Den interaktiva rörligheten fungerar inte särskilt bra inför någon av  texterna, vilket kan bero på ovana vid att läsa tidningsartiklar och/eller ovana vid att reflektera  över texter. Slutsatsen är att en tydlig mottagaranpassning är viktig och ger resultat, men är  svår att genomföra när tidningen har ett stort urval av ämnen och många olika målgrupper. 

Fragm. lat. I 84 - Psalterium (Ps. 17,7 - 17; 115,16 - 117,11)

Trägerband: 'Nicolaus de Lyra, Praeceptorium'

On Binary Self-Dual Codes of Length 62 with an Automorphism of Order 7

Николай Янков - Класифицирани са с точност до еквивалетност всички оптимални двоични самодуални [62, 31, 12] кодове, които притежават автоморфизъм от ред 7 с 8 независими цикъла при разлагане на независими цикли. Използвайки метода за конструиране на самодуални кодове, притежаващи автоморфизъм от нечетен прост ред е доказано, че съществуват точно 8 нееквивалентни такива кода. Три от получените кодове имат тегловна функция, каквато досега не бе известно да съществува.

Tudás és menedzsment – Tudásmenedzsment. 7. európai tudásmenedzsment-konferencia, Budapest

A Budapesti Corvinus Egyetem adott otthont 2006 szeptemberében az évente megrendezett Európai Tudásmenedzsment Konferenciának, melynek megrendezésében az egyetem részéről az Információrendszerek tanszék dolgozói vettek részt. A tudásmenedzsment mint téma az 1990-es évektől kezdődően jelenik meg egyre markánsabban a menedzsment és informatikai tudományterületeken. A tudás mint az erőforrások kezelésére irányuló törekvés ugyanakkor évezredek-évszázadok óta jelen van a gazdasági és politikai gondolkodásban.

The Beacon, September 8, 2010

Vol. 24, Issue 7, 8 pages

A REPRESENTATIVE OUTSIDER AND THE INCLUSION OF THE OUTSIDER IN ACTS 8:26-40

In Luke’s two volumes, Luke is not interested only in Gentiles and those with high social status but also in the marginalized and those who are outsiders. This dissertation seeks to read Luke’s concern for outsiders and the theme of the inclusion of outsiders in the new kingdom of God in Luke’s narrative of the Ethiopian eunuch’s conversion in Acts 8:26-40. This paper examines the Ethiopian eunuch’s complex identity from the perspectives of the Greco-Roman world, Old Testament (LXX) allusions to the Elijah-Elisha narratives, and Luke’s interpretation of the Isaianic quotation of the Suffering Servant in Acts 8:32-33 (cf. Isaiah 53:7-8). This study pays close attention to the correlations between the theme of outsiders and three key characters in Acts 8:26-40: the Ethiopian eunuch, Philip, and the Suffering Servant. First, Luke depicts the Ethiopian eunuch as the consummate outsider—geographically, morally, socially, ethnically, and in terms of gender—and indicates that the eunuch represents other marginalized outsiders. The eunuch shows no one can prevent outsiders like him from inclusion in the kingdom of God. Second, Luke portrays Philip as a prophet, specifically a prophet like Elijah and Elisha. Philip emulates Elijah and Elisha by reaching out to the outsider (in this instance, the Ethiopian eunuch). Third, Luke presents the Isaianic Suffering Servant as a religious and social outsider and identifies the character with Jesus and the Ethiopian eunuch. The indescribable descendants of the Suffering Servant signify a universally inclusive messianic community and fulfill the outsiders’ inclusion within the people of God as Isaiah prophesied (Isaiah 56:3-8). This thesis finally suggests ways to read the story of the Ethiopian eunuch today and concludes that it is imperative to include those outsiders among us within the community of Jesus’s followers.

¿Fue la crisis política del Occidente musulmán del siglo XIII debida a un cambio climático? Una aproximación histórica al fin del Periodo Cálido Medieval

Durante el siglo XIII se produjo una sucesión de revueltas que supuso la desaparición del Imperio almohade y su sustitución por poderes regionales en al-Andalus, el Magreb y el Magreb al-Aqsà. La historiografía ha presentado el surgimiento y pugna entre estos poderes como un fenómeno social, político e, incluso, cultural y religioso, con el que se ha podido explicar su aniquilación o marginalización. Este trabajo pretende contextualizar los hechos desde una perspectiva medioambiental, de forma que la desintegración del califato almohade, el surgimiento de aquellos poderes y la progresión de los reinos cristianos en la península ibérica puedan entenderse desde una visión global de cambio climático y una posible crisis agrícola.