The Recombination Protein RAD52 Cooperates with the Excision Repair Protein OGG1 for the Repair of Oxidative Lesions in Mammalian Cells

Oxidized bases are common types of DNA modifications. Their accumulation in the genome is linked to aging and degenerative diseases. These modifications are commonly repaired by the base excision repair (BER) pathway. Oxoguanine DNA glycosylase (OGG1) initiates BER of oxidized purine bases. A small number of protein interactions have been identified for OGG1, while very few appear to have functional consequences. We report here that OGG1 interacts with the recombination protein RAD52 in vitro and in vivo. This interaction has reciprocal functional consequences as OGG1 inhibits RAD52 catalytic activities and RAD52 stimulates OGG1 incision activity, likely increasing its turnover rate. RAD52 colocalizes with OGG1 after oxidative stress to cultured cells, but not after the direct induction of double-strand breaks by ionizing radiation. Human cells depleted of RAD52 via small interfering RNA knockdown, and mouse cells lacking the protein via gene knockout showed increased sensitivity to oxidative stress. Moreover, cells depleted of RAD52 show higher accumulation of oxidized bases in their genome than cells with normal levels of RAD52. Our results indicate that RAD52 cooperates with OGG1 to repair oxidative DNA damage and enhances the cellular resistance to oxidative stress. Our observations suggest a coordinated action between these proteins that may be relevant when oxidative lesions positioned close to strand breaks impose a hindrance to RAD52 catalytic activities.

Base Excision Repair Activities Differ in Human Lung Cancer Cells and Corresponding Normal Controls

Oxidative damage to DNA is thought to play a role in carcinogenesis by causing Mutations, and indeed accumulation of oxidized DNA bases has been observed in samples obtained from tumors but not from surrounding tissue within the same patient. Base excision repair (BER) is the main pathway for the repair of oxidized modifications both in nuclear and mitochondrial, DNA. In order to ascertain whether diminished BER capacity might account for increased levels of oxidative DNA damage in cancer cells, the activities of BER enzymes in three different lung cancer cell lines and their non-cancerous counterparts were measured using oligonucleotide substrates with single DNA lesions to assess specific BER enzymes. The activities of four BER enzymes, OGG1, NTH1, UDG and APE1, were compared in mitochondrial and nuclear extracts. For each specific lesion, the repair activities were similar among the three cell lines used. However, the specific activities and cancer versus control comparison differed significantly between the nuclear and mitochondrial compartments. OGG1 activity, as measured by 8-oxodA incision, was upregulated in cancer cell mitochondria but down-regulated in the nucleus when compared to control cells. Similarly, NTH1 activity was also up-regulated in mitochondrial extracts from cancer cells but did not change significantly in the nucleus. Together, these results support the idea that alterations in BER capacity are associated with carcinogenesis.

The mitochondrial transcription factor A functions in mitochondrial base excision repair

Mitochondrial transcription factor A (TFAM) is an essential component of mitochondrial nucleoids TFAM plays an important role in mitochondrial transcription and replication TFAM has been previously reported to inhibit nucleotide excision repair (NER) in vitro but NER has not yet been detected in mitochondria, whereas base excision repair (BER) has been comprehensively characterized in these organelles The BER proteins are associated with the inner membrane in mitochondria and thus with the mitochondrial nucleoid, where TFAM is also situated However, a function for TFAM in BER has not yet been investigated This study examines the role of TFAM in BER In vitro studies with purified recombinant TFAM indicate that it preferentially binds to DNA containing 8-oxoguanines, but not to abasic sites, uracils, or a gap in the sequence TFAM inhibited the in vitro incision activity of 8-oxoguanine DNA glycosylase (OGG1), uracil-DNA glycosylase (UDG), apurinic endonuclease 1 (APE1), and nucleotide incorporation by DNA polymerase gamma (pol gamma) On the other hand, a DNA binding-defective TFAM mutant, L58A, showed less inhibition of BER in vitro Characterization of TFAM knockdown (KD) cells revealed that these lysates had higher 8oxoG incision activity without changes in alpha OGG1 protein levels TFAM KD cells had mild resistance to menadione and increased damage accumulation in the mtDNA when compared to the control cells In addition, we found that the tumor suppressor p53, which has been shown to interact with and alter the DNA binding activity of TFAM, alleviates TFAM-Induced inhibition of BER proteins Together, the results suggest that TFAM modulates BER in mitochondria by virtue of its DNA binding activity and protein interactions Published by Elsevier B V

The Xanthomonas citri effector protein PthA interacts with citrus proteins involved in nuclear transport, protein folding and ubiquitination associated with DNA repair

P>Xanthomonas axonopodis pv. citri utilizes the type III effector protein PthA to modulate host transcription to promote citrus canker. PthA proteins belong to the AvrBs3/PthA family and carry a domain comprising tandem repeats of 34 amino acids that mediates protein-protein and protein-DNA interactions. We show here that variants of PthAs from a single bacterial strain localize to the nucleus of plant cells and form homo- and heterodimers through the association of their repeat regions. We hypothesize that the PthA variants might also interact with distinct host targets. Here, in addition to the interaction with alpha-importin, known to mediate the nuclear import of AvrBs3, we describe new interactions of PthAs with citrus proteins involved in protein folding and K63-linked ubiquitination. PthAs 2 and 3 preferentially interact with a citrus cyclophilin (Cyp) and with TDX, a tetratricopeptide domain-containing thioredoxin. In addition, PthAs 2 and 3, but not 1 and 4, interact with the ubiquitin-conjugating enzyme complex formed by Ubc13 and ubiquitin-conjugating enzyme variant (Uev), required for K63-linked ubiquitination and DNA repair. We show that Cyp, TDX and Uev interact with each other, and that Cyp and Uev localize to the nucleus of plant cells. Furthermore, the citrus Ubc13 and Uev proteins complement the DNA repair phenotype of the yeast Delta ubc13 and Delta mms2/uev1a mutants, strongly indicating that they are also involved in K63-linked ubiquitination and DNA repair. Notably, PthA 2 affects the growth of yeast cells in the presence of a DNA damage agent, suggesting that it inhibits K63-linked ubiquitination required for DNA repair.

Hesitations and repair in German

The occurrence of pauses and hesitations in spontaneous speech has been shown to occur systematically, for example, "between sentences, after discourse markers and conjunctions and before accented content words." (Hansson [15]) This is certainly plausible in English, where pauses and hesitations can and often do occur before content words such as nominals, for example, "uh, there's a … man." (Chafe [8]) However, if hesitations are, in fact, evidence of "deciding what to talk about next," (Chafe [8]) then the complex grammatical system of German should render this pausing position precarious, since pre-modifiers must account for the gender of the nominals they modify.In this paper, I present data to test the hypothesis that pre-nominal hesitation patterns in German are dissimilar to those in English. Hesitations in German will be shown, in fact, to occur within noun phrase units. Nevertheless, native speakers most often succeed in supplying a nominal which conforms to the gender indicated by the determiner or pre-modifier. Corrections, or repairs, of infelicitous pre-modifiers indicate that the speaker was unable to supply a nominal of the same gender which the choice of pre-modifier had committed him/her to. The frequency of such repairs is shown to vary according to task, with fewest repairs occurring in elicited speech which allows for linguistic freedom and therefore is most like spontaneous speech. The data sets indicate that among German native speakers, hesitations occurring before noun phrase units (pre-NPU hesitations) indicate deliberation of what to say, while hesitations within or before the head of the noun phrase (pre-NPH hesitations) indicate deliberation of how to say what has already been decided (cf. Chafe [8]).

Improved Road Design for Future Maintenance - Analysis of Road Barrier Repair Costs

The cost of a road construction over its service life is a function of the design, quality of construction, maintenance strategies and maintenance operations. Unfortunately, designers often neglect a very important aspect which is the possibility to perform future maintenance activities. The focus is mainly on other aspects such as investment costs, traffic safety, aesthetic appearance, regional development and environmental effects. This licentiate thesis is a part of a Ph.D. project entitled “Road Design for lower maintenance costs” that aims to examine how the life-cycle costs can be optimized by selection of appropriate geometrical designs for the roads and their components. The result is expected to give a basis for a new method used in the road planning and design process using life-cycle cost analysis with particular emphasis on road maintenance. The project started with a review of literature with the intention to study conditions causing increased needs for road maintenance, the efforts made by the road authorities to satisfy those needs and the improvement potential by consideration of maintenance aspects during planning and design. An investigation was carried out to identify the problems which obstruct due consideration of maintenance aspects during the road planning and design process. This investigation focused mainly on the road planning and design process at the Swedish Road Administration. However, the road planning and design process in Denmark, Finland and Norway were also roughly evaluated to gain a broader knowledge about the research subject. The investigation was carried out in two phases: data collection and data analysis. Data was collected by semi-structured interviews with expert actors involved in planning, design and maintenance and by a review of design-related documents. Data analyses were carried out using a method called “Change Analysis”. This investigation revealed a complex combination of problems which result in inadequate consideration of maintenance aspects. Several urgent needs for changes to eliminate these problems were identified. Another study was carried out to develop a model for calculation of the repair costs for damages of different road barrier types and to analyse how factors such as road type, speed limits, barrier types, barrier placement, type of road section, alignment and seasonal effects affect the barrier damages and the associated repair costs. This study was carried out using a method called the “Case Study Research Method”. Data was collected from 1087 barrier repairs in two regional offices of the Swedish Road Administration, the Central Region and the Western Region. A table was established for both regions containing the repair cost per vehicle kilometre for different combinations of barrier types, road types and speed limits. This table can be used by the designers in the calculation of the life-cycle costs for different road barrier types.

Product verification and defect repair - status and considerations

Product verifications have become a cost-intensive and time-consuming aspect of modern electronics production, but with the onset of an ever-increasing miniaturisation, these aspects will become even more cumbersome. One may also go as far as to point out that certain precision assembly, such as within the biomedical sector, is legally bound to have 0 defects within production. Since miniaturisation and precision assembly will soon become a part of almost any product, the verifications phases of assembly need to be optimised in both functionality and cost. Another aspect relates to the stability and robustness of processes, a pre-requisite for flexibility. Furthermore, as the re-engineering cycle becomes ever more important, all information gathered within the ongoing process becomes vital. In view of these points, product, or process verification may be assumed to be an important and integral part of precision assembly. In this paper, product verification is defined as the process of determining whether or not the products, at a given phase in the life-cycle, fulfil the established specifications. Since the product is given its final form and function in the assembly, the product verification normally takes place somewhere in the assembly line which is the focus for this paper.

Barriers to entry in monopoly markets: automobile distribution in Brazil

O Objetivo deste Trabalho é Analisar os Efeitos da Entrada de uma Segunda Concessionária de Automóveis em Mercados Previamente Monopolizados. para Tanto, Construiu-Se um Banco de Dados com a Localização de Concessionárias de Automóveis em Microrregiões e Características Demográficas e Econômicas Destas Microrregiões. a Partir Desse Banco de Dados e de Modelos de Escolha Binária, Foram Identificadas Variáveis que Condicionam a Existência e o Número de Concessionárias em Microrregiões. Utilizando-Se de um Modelo Adaptado de Bresnahan e Reiss (1990), Foram Estimados os Custos Fixos de Entrada de Concessionárias em Mercados Monopolizados. os Resultados Obtidos Sugerem que as Barreiras À Entrada não são Significativas, o que Aumenta a Probabilidade de que a Cláusula de Exclusividade nos Contratos de Concessão não Cause Danos À Concorrência no Mercado Brasileiro de Distribuição de Automóveis

Um modelo exploratório para o processo de compra de MRO baseadas na internet Michelin Brasil

A presente dissertação tem por objetivo criar um modelo de compras de rMRO (manutenção, reparos e operações) baseado na Internet, para a Michelin no Brasil. Atualmente, a empresa Michelin está participando de uma comunidade virtual, sob a forma de um "vertal", criada pelas maiores empresas da indústria de pneu e borracha (Michelin, Goodyear, Pirelli, Continental, Cooper, Sumitomo, entre outras), e cuja intenção é desenvolver uma praça de mercado global para realização das suas compras e aprovisionamentos. A Rubbernetwork.com (RNC) foi fundada como uma empresa independente, reunindo o conjunto das compras da indústria de pneu e borracha, representando um valor de mais de cinqüenta bilhões de dólares por ano gastos em matérias-primas, equipamentos, maquinário, bens e serviços. Esta central de compras permitirá a seus membros aumentar a eficácia de suas operações e realizar economias substanciais nos custos de transação. Cada transação será protegida por apropriados sistemas advindos da implementação da tecnologia da informação, a fim de garantir a confidencialidade de todas as atividades comerciais de cada empresa individualmente. Os membros participantes estão convencidos de que a iniciativa possibilitará economias significativas e melhorará a eficiência de suas cadeias de valor. A comunidade virtual será aberta a todos os fabricantes e fornecedores relacionados à indústria de pneu e borracha, sendo também bem vindas oportunidades 2 de interação com outras comunidades virtuais, especialmente aquelas associadas à indústria automotiva. É dentro do contexto de implementação do projeto Rubbernetwork.com (RNC) que a dissertação se desenvolverá, tecendo estratégias e modelos de negócios concernentes aos processos de compras relacionados a MRO (manutenção, reparos e operações). O desenvolvimento de um "vertal" na Internet não impede que portais horizontais complementares, e não excludentes, sejam também acrescidos como alternativa de negócios. Há de se dar um tratamento específico às compras diárias que envolvem um grande número de diferentes fornecedores, várias áreas internas na empresa e consomem tempo desnecessário em trabalhos burocráticos, sem fins estratégicos. Cabe ressaltar que tal categoria de compras, denominada MRO, apesar de seu baixo valor por produto ou serviço adquirido, gera um elevado volume de transações e um alto impacto operacional. Pelo fato de tratar-se de uma experiência inovadora em setor de características próprias - mercado de pneu e borracha -, e de estar-se criando uma expectativa de cooperação entre empresas concorrentes, em busca da racionalização do processo de compras e divisão de custos operacionais, a metodologia utilizada, quanto aos seus fins, será exploratória e aplicada I. Quanto aos meios de investigação, a pesquisa realizada será: de campo, estudo de caso e bibliográfica. O universo da pesquisa restringir-se-á à empresa Michelin, onde o processo de compras de MRO baseado na Internet será contemplado, através de levantamento de dados nos E.UA e França, a fim de propor-se um modelo para a Michelin Brasil.

Os impactos da internet no varejo automobilístico brasileiro

Ainda considerada uma tecnologia emergente, a Internet tem provocado importantes transformações na comercialização, logística de suprimentos e distribuição, modelagem organizacional, produção e nos recursos humanos no setor do varejo automobilístico nacional. Outrossim, a Internet tem mudado a forma de atuação no mercado por parte das empresas de veículos automotores brasileiros, as quais devem passar a dispor de novos métodos de gestão, práticas comerciais e recursos da tecnologia on-line, visto que a tendência do tipo tradicional de vendas face a face é tomar-se obsoleto em relação à venda" virtual". o presente estudo foi desenvolvido com a finalidade de analisar quais os impactos e as mudanças causadas pela Internet nas empresas de varejo de veículos automotores nacionais, bem como apontar possíveis soluções para adaptação das concessionárias de automóveis brasileiras frente a esse novo panorama - a era da tecnologia. Para tanto, além da revisão de literatura, foram realizadas duas pesquisas - uma de abrangência nacional e outra regional, que teve como base a cidade do Rio de Janeiro. Através da coleta de dados, foram obtidas informações sobre o que influi no uso e não-uso da Internet no segmento de varejo de automotores.

Oral and maxillofacial reconstruction in a cat using wire and acrylic

A 15-month-old DSH cat was presented with facial deformity secondary to multiple oral and maxillofacial fractures after being struck by an automobile. Multiple wires and dental acrylics were used for fracture repair.

Repair of lamellar scleral lesions in dogs with preserved equine renal capsule: short report

OBJETIVO: Avaliar o uso da cápsula renal de eqüino preservada em glicerina 98% no reparo de lesões lamelares esclerais em cães. MÉTODOS: Foram utilizados 12 cães, machos e fêmeas, com peso médio de 12kg. Foram realizadas avaliações clínica e morfológica aos 1, 3, 7, 15, 30 e 60 dias de pós-operatório. Após anestesia geral e procedimentos padrões de preparo do campo operatório, foi realizada cantotomia temporal, seguida de incisão conjutival e escleral com área de 0,5x0,5 cm na posição de 1hora, próxima ao limbo. em seguida, um fragmento de mesma dimensão de cápsula renal de eqüino preservada em glicerina, previamente hidratado em solução salina, foi aplicado ao defeito escleral criado sendo fixado com pontos simples isolados com vicryl 7-0®. RESULTADOS: A avaliação clínica revelou blefaroespasmo/fotofobia até o sétimo dia de pós-operatório. Foi observado edema conjuntival até o quinto dia, acompanhado de secreção ocular mucóide, que persistiu até o décimo dia de pós-operatório. Não foram observados sinais clínicos de rejeição do enxerto em todos os animais, em todos os períodos avaliados. Os segmentos anterior e posterior do bulbo ocular não apresentaram sinais de inflamação. A análise morfológica revelou exsudação inflamatória aguda nos períodos precoces e intermediários da avaliação e inflamação crônica nos períodos tardios da observação. Houve incorporação do enxerto ao leito receptor. CONCLUSÃO: Os resultados sugerem que a cápsula renal de eqüino preservada pode ser mais uma alternativa de membrana biológica para o reparo de lesões esclerais lamelares em cães e no homem.

Mechanism of anticlastogenicity of Agaricus blazei Murill mushroom organic extracts in wild type CHO (K-1) and repair deficient (xrs5) cells by chromosome aberration and sister chromatid exchange assays

Agaricus blazei Murill is a medicinal mushroom native to Brazil. The present work assessed the clastogenic and anticlastogenic potential of organic extracts (ethanol and chloroform/methanol) from the lineage AB97/11 in chinese hamster CHO-K-1 (wild type) and CHO-xrs5 (repair deficient) cells using the chromosome aberration (CA) and sister chromatid exchange (SCE) assays. In these experimental conditions were observed: (a) anticlastogenic effect at concentrations of 0.06 and 0.09% of the EtOH extract and at the 0.03 and 0.06% concentrations of the C/MetOH extract in CHO-K-1; (b) absence of protector effect on CHO-xrs5 cells; and (c) absence of protector effect in the SCE assay. These results indicate that organic extracts of A. blazei lineage AB97/11 present bio-antimutagenic type protective activity. (C) 2003 Elsevier B.V. B.V. All rights reserved.

Considerations on corrosion and weld repair effects on the fatigue strength of a steel structure critical to the flight-safety

The aim of this study is to analyze the effects of corrosion and successive tungsten inert gas (TIC) welding repairs on the reverse bending fatigue strength of AISI 4130 steel used in components critical to the flight-safety. The tests were performed on hot-rolled steel plate specimens, 1.10 mm and 1.60 mm thick, by means of a SCHENK PWS equipment, with load ratio R = -1, constant amplitude, 30 Hz frequency and room temperature. It was observed that the reverse bending fatigue strength of AISI 4130 steel decreases due to the corrosion and the TIC welding and re-welding processes. (C) 2010 Elsevier Ltd. All rights reserved.