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.

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.

Repair Welding Effects on the Bending Fatigue Strength of AISI 4130 Aeronautical Steel used in a Critical to the Flight-Safety Structure

The aim of this study was to analyze the effect of successive TIG (tungsten inert gas) welding repairs on the reverse bending fatigue strength of AISI 4130 steel, which is widely used in components critical to the flight-safety. In order to simulate the abrupt maneuvers, wind bursts, motor vibration and helixes efforts, which generate cyclic bending loadings at the welded joints of a specific aircraft component called motor cradle, experimental reverse bending fatigue tests were carried out on specimens made from hot-rolled steel plate, 1.10 mm (0.043 in) thick, by mean of a SCHENK PWS equipment, with load ratio R = -1, under constant amplitude, at 30 Hz frequency and room temperature. It was observed that the bending fatigue strength decreases after the TIG (Tungsten Inert Gas) welding process application on AISI 4130 steel, with subsequent decrease due to re-welding sequence as well. Microstructural analyses and microhardness measurements on the base material, heat-affected zone (HAZ) and weld metal, as well as the effects of the weld bead geometry on the obtained results, have complemented this study.

Considerations about the welding repair effects on the structural integrity of an airframe critical to the flight-safety

Structures critical to the flight-safety are commonly submitted to several maintenance repairs at the welded joints in order to prolong the in-service life of aircrafts. The aim of this study is to analyze the effects of Tungsten Inert Gas (TIG) welding repair on the structural integrity of the AISI 4130 aeronautical steel by means of experimental fatigue crack growth tests in base-material, heat-affected zone (HAZ) and weld metal. The tests were performed on hot-rolled steel plate specimens, 0.89 mm thick, with load ratio R = 0.1, constant amplitude, at 10 Hz frequency and room temperature. Increase of the fracture resistance was observed in the weld metal but decreasing in the HAZ after repair. The results were associated to microhardness and microstructural changes with the welding sequence. (C) 2010 Published by Elsevier Ltd.

Comparative study between two techniques of incisional hernia repair with polypropylene mesh in rabbits

OBJETIVO: Comparar duas técnicas de tratamento da hérnia incisional em coelhos utilizando a tela de polipropileno apoiando um reforço peritônio - aponeurótico ou suturada nas bordas do anel herniário 'em ponte . MÉTODOS: Foram operados 60 coelhos para a produção de hérnia incisional, em uma incisão mediana de 4 centímetros. Após 30 dias, metade dos animais foram operados com o fechamento primário da parede, com colocação de uma tela de polipropileno apoiando o reforço e a outra metade dos animais com a colocação da tela suturada nas bordas do anel herniárioem ponte . Os animais foram avaliados com 30 (M1), 60 (M2)e 90 (M3) dias de pós-operatório. Os parâmetros analisados foram a evolução clínica, análise da força de ruptura da cicatriz, estudo macroscópico, análise microscópica e morfométrica. RESULTADOS: Não foram observadas diferenças significantes com relação a força de ruptura e estudos histológicos nos dois grupos e vários momentos estudados. Não houve diferença estatística com relação às complicações, embora os animais que receberam a telaem ponte tiveram aderências mais firmes e intensas à parede abdominal. CONCLUSÕES: As duas técnicas utilizadas para correção da hérnia incisional em coelhos não mostraram diferenças significantes quanto a força de ruptura, análise histológica e morfométrica. O número de complicações foi semelhante, porém a aderência de órgãos da cavidade abdominal à área de cicatriz foi muito mais intensa no grupo em que a tela foi colocadaem ponte .

Repair of the anophthalmic cavity of rats with synthetic hydroxyapatite

We placed spheres of synthetic hydroxyapatite (calcium chloride combined with sodium phosphate) in the eviscerated or enucleated orbital cavity of rats in order to evaluate the biocompatibility of this material with the orbital cavity. The study was conducted on 50 albino rats, 25 of which were submitted to enucleation and 25 to evisceration of one eye. The animals were sacrificed 7, 15, 21, 30 and 60 days after surgery and the orbital content was submitted to histopathological examination. A reaction of the young granulation tissue type was observed first. The hydroxyapatite was gradually surrounded by a granulomatous macrophage inflammatory response and covered with dense connective tissue that formed a sort of mesh septating and supporting progressively smaller blocks of the substance. The same type of reaction was observed in the enucleated and eviscerated cavities. We conclude that synthetic hydroxyapatite is an inert nonallergenic material which is appropriate for volume replacement in the anophthalmic cavity

Vegetal polymer in repair of defects of the orbital floor: an experimental study in rabbits

Background: An experimental study was done to assess the ability of the vegetal polymer miniplates and screws to repair defects of the orbital floor.Methods: An artificial standard-sized defect was created in the bony floor of right orbit of 45 albino rabbits. The animals were divided into three experimental groups: control group (G1) involving animals with orbital floor defect and no treatment; titanium group (G2) containing animals with orbital floor defect repaired by titanium miniplates and screws; vegetal polymer group (G3) composed of animals with similar orbital floor defects repaired by vegetal polymer miniplates and screws. Throughout the course of the experiment, the animals were clinically evaluated. At 15, 30 and 60 days after surgery, the animals were killed. They were X-rayed immediately after the floor defect and at the moment of sacrifice. Histological and morphometric evaluation of inflammatory reaction and bone healing was done. Data were statistically evaluated.Results: No implants were extruded. Bone consolidation was similar in G2 and G3 and better than in G1 group animals. Inflammatory reaction was most pronounced in animals of G3 15 days after surgery, and it subsided over time.Conclusion: Vegetal polymer miniplates and screws induces small inflammatory reaction and had the ability to stimulate bone growth with good integration in the orbital floor defect allowing to consider the vegetal polymer adequate option to treat orbital floor defects. Future studies involving long-term follow-up and biomechanical tests to evaluate material resistance to traction are needed.