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.

Accumulation of (5 ` S)-8,5 `-cyclo-2 `-deoxyadenosine in organs of Cockayne syndrome complementation group B gene knockout mice

Cockayne syndrome (CS) is a human genetic disorder characterized by sensitivity to UV radiation, neurodegeneration, premature aging among other phenotypes. CS complementation group B (CS-B) gene (csb) encodes the CSB protein (CSB) that is involved in base excision repair of a number of oxidatively induced lesions in genomic DNA in vivo. We hypothesized that CSB may also play a role in cellular repair of the DNA helix-distorting tandem lesion (5`S)-8,5`-cyclo-2`-deoxyadenosine (S-cdA). Among many DNA lesions. S-cdA is unique in that it represents a concomitant damage to both the sugar and base moieties of the same nucleoside. Because of the presence of the C8-C5` covalent bond, S-cdA is repaired by nucleotide excision repair unlike most of other oxidatively induced lesions in DNA, which are subject to base excision repair. To test our hypothesis, we isolated genomic DNA from brain, kidney and liver of wild type and csb knockout (csb(-/-)) mice. Animals were not exposed to any exogenous oxidative stress before the experiment. DNA samples were analysed by liquid chromatography/mass spectrometry with isotope-dilution. Statistically greater background levels of S-cdA were observed in all three organs of csb(-/-) mice than in those of wild type mice. These results suggest the in vivo accumulation of S-cdA in genomic DNA due to lack of its repair in csb(-/-) mice. Thus, this study provides, for the first time, the evidence that CSB plays a role in the repair of the DNA helix-distorting tandem lesion S-cdA. Accumulation of unrepaired S-cdA in vivo may contribute to the pathology associated with CS. Published by Elsevier B.V.

ESR dating of pleistocene mammal teeth and its implications for the biostratigraphy and geological evolution of the coastal plain, Rio Grande do Sul, southern Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Occupational exposure of physical therapists to electric and magnetic fields and the efficacy of Faraday cages

Objective. Measure physical therapists' exposure to the electric and magnetic fields produced by 17 shortwave diathermy devices in physical therapy clinics in the city of Presidente Prudente, São Paulo State, Brazil. Compare the observed values with the exposure levels recommended by the International Commission on Non-ionizing Radiation Protection (ICNIRP). Observe the efficacy of Faraday cages as a means of protecting physical therapists from exposure to oscillating electric and magnetic fields.Methods. Electric and magnetic field measurements were taken at four points during actual physical therapy sessions: in proximity to the operator's pelvis and head, the devices' electrical cables, and the electrodes. The measuring equipment was a Wandel & Goltermann EMR-200.Results. The values obtained in proximity to the electrodes and cables were 10 to 30 times higher than ICNIRP's recommended occupational reference levels. In the shortwave diathermy treatment rooms with Faraday cages, the fields were even higher than in treatment rooms not so equipped-principally the magnetic field, where the values were more than 100 times higher than the ICNIRP exposure limit.Conclusions. The electric and magnetic field intensities obtained in this study are generally above the exposure levels recommend in ICNIRP standards. It was also observed that the Faraday cage offers physical therapists no protection, and instead, increases their level of exposure.

Um estudo algorítmico para otimização do plano de tratamento da radioterapia conformal

This work performs an algorithmic study of optimization of a conformal radiotherapy plan treatment. Initially we show: an overview about cancer, radiotherapy and the physics of interaction of ionizing radiation with matery. A proposal for optimization of a plan of treatment in radiotherapy is developed in a systematic way. We show the paradigm of multicriteria problem, the concept of Pareto optimum and Pareto dominance. A generic optimization model for radioterapic treatment is proposed. We construct the input of the model, estimate the dose given by the radiation using the dose matrix, and show the objective function for the model. The complexity of optimization models in radiotherapy treatment is typically NP which justifyis the use of heuristic methods. We propose three distinct methods: MOGA, MOSA e MOTS. The project of these three metaheuristic procedures is shown. For each procedures follows: a brief motivation, the algorithm itself and the method for tuning its parameters. The three method are applied to a concrete case and we confront their performances. Finally it is analyzed for each method: the quality of the Pareto sets, some solutions and the respective Pareto curves

Pharmaceutical applications of AC Biosusceptometry

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Measurement of gastric contraction activity in dogs by means of AC biosusceptometry

The mechanical nature of gastric contraction activity (GCA) plays an important role in gastrointestinal motility. The aim of this study was to detect GCA in anaesthetized dogs, using simultaneously the techniques of AC biosusceptometry (ACB) and manometry, analysing the characteristics of frequency and amplitude (motility index) of GCA, modified by drugs such as prostigmine and N-butyl-scopolamine. The ACB method is based on a differential transformer of magnetic flux and the magnetic tracer works as a changeable external nucleus. This magnetic tracer causes a modification in the magnetic flux, which is detected by the coils. The results obtained from the ACB showed a performance comparable to the manometry in measuring the modifications in the frequency and amplitude of the GCA. We concluded that this ACB technique, non-invasive and free of ionizing radiation, is an option for evaluating GCA and can be employed in future clinical studies.

Cytostatic in vitro Effects of DTCM-Glutarimide on Bladder Carcinoma Cells

Bladder cancer is a common malignancy worldwide. Despite the increased use of cisplatin-based combination therapy, the outcomes for patients with advanced disease remain poor. Recently, altered activation of the PI3K/Akt/mTOR pathway has been associated with reduced patient survival and advanced stage of bladder cancer, making its upstream or downstream components attractive targets for therapeutic intervention. In the present study, we showed that treatment with DTCM-glutaramide, a piperidine that targets PDK1, results in reduced proliferation, diminished cell migration and G1 arrest in 5637 and T24 bladder carcinoma cells. Conversely, no apoptosis, necrosis or autophagy were detected after treatment, suggesting that reduced cell numbers in vitro are a result of diminished proliferation rather than cell death. Furthermore previous exposure to 10 mu g/ml DTCM-glutarimide sensitized both cell lines to ionizing radiation. Although more studies are needed to corroborate our findings, our results indicate that PDK1 may be useful as a therapeutic target to prevent progression and abnormal tissue dissemination of urothelial carcinomas.

MEH-PPV hypsochromic shifts in halogenated solvents induced by gamma-rays

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Assessment of Gastrointestinal Motility in Renal Transplant Recipients by Alternate Current Biosusceptometry

Background. Gastrointestinal (GI) complications are common after renal transplantation, mainly owing to immunosuppressive therapy. Assessment of GI transit time can facilitate rational management of these disorders.Objective. We evaluate the GI transit parameters in renal transplant recipients taking tacrolimus, azathioprine, and prednisone with the use of the alternate current biosusceptometry (ACB) technique and compared them with healthy volunteers.Methods. Ten renal transplant recipients and 10 healthy volunteers were enrolled in this study. After an overnight fast, patients and volunteers ingested a standard meal containing magnetic markers. The biomagnetic monitoring was performed at 10-minute intervals for at least 8 hours to obtain gastric emptying as well as the colonic arrival time-intensity curves. Mean gastric emptying time (MGET), mean colon arrival time (MCAT), and mean small intestinal transit time (MSITT) were quantified and compared between control and patient groups with results expressed as mean +/- SD.Results. The MGET measured by the ACB technique was 48 +/- 31 minutes and 197 +/- 50 minutes for patients and healthy subjects, respectively. MSITT and MCAT values calculated for patients versus volunteers were 171 +/- 71 minutes versus 197 +/- 71 minutes and 219 +/- 83 minutes versus 373 +/- 52 minutes, respectively. Renal transplant recipients showed significantly faster; gastric emptying and colon arrival times (P < .001) compared with normal volunteers; however, small intestinal transit time was not significantly different (P = .44).Conclusions. In stable renal transplant recipients, the GI transit parameters were significantly faster than in normal healthy volunteers. ACB sensors are versatile technologies that can be used for clinical research, because they offer an excellent opportunity to evaluate GI transit in a noninvasive manner without the use of ionizing radiation.

Chromosome sensitivity to bleomycin in G2 lymphocytes from Down syndrome patients

Several studies have demonstrated that lymphocytes from patients with Down syndrome (DS) exhibit an increased frequency of chromosome aberrations when they are exposed to ionizing radiation or to chemicals at the G0 or G1 phases of the cell cycle, but not at G2 when compared to normal subjects. To determine the susceptibility of DS lymphocytes at G2 phase, bleomycin, a radiomimetic agent, was used to induce DNA breaks in blood cultures from 24 Down syndrome patients. All the patients with DS showed free trisomy 21 (47,XX + 21 or 47,XY + 21). Individuals that showed an average number of chromatid breaks per cell higher than 0.8 were considered sensitive to the drug. No control child showed susceptibility to bleomycin, and among the 24 patients with DS, only one was sensitive to the drug. No significant difference was observed between the two groups, regarding chromatid break frequencies in treated G2 lymphocytes. The distribution of bleomycin-induced breaks in each group of chromosomes was similar for DS and controls. No significant difference was found in the response to bleomycin between male and female subjects. Probably, the main factor involved in chromosome sensitivity of lymphocytes from patients with DS is the phase of the cell cycle in which the cell is treated.

A novel biomagnetic instrumentation with four magnetoresistive sensors to evaluate gastric motility.

A novel instrumentation using anisotropic magnetoresistive (AMR) sensors associated with magnetic coils excitation was developed to evaluate gastrointestinal tract motility parameters. The susceptometer has four sensors that were used to measure the gastric activity contractions (GAC) in anaesthetized dogs, its performance was evaluated by manometry with good results.

Eficácia do extrato aquoso do Triticum vulgare no tratamento de radiodermatites geradas pelo uso da radioterapia em gatos

The use of ionizing radiation in the treatment of oncological diseases in veterinary medicine has grown in recent years. In general, these animals are very well tolerated by the treatment with radioactive elements; however, the harmful effects that may occur are not always able to be controlled, particularly those relating to skin burns. Then, the purpose of this study was to evaluate the effects of aqueous extract of Triticum vulgare in treat of 10 cats radiodermatitis with squamous cell carcinoma (SCC), which were treated with radiotherapy as single modality treatment. The drug was used topically and daily until complete healing of wounds. Based on the findings, it may concluded beneficial effects of the drug, because it allowed the formation of granulation tissue and epithelization of wounds with epidermal repair not exuberant, good-quality cosmetics and, considering the deleterious effects that can result from cancer treatments.

Image quality evaluation of chest for computed radiography

The daily-to-day of medical practice is marked by a constant search for an accurate diagnosis and therapeutic assessment. For this purpose the doctor serves up a wide variety of imaging techniques, however, the methods using ionizing radiation still the most widely used because it is considered cheaper and above all very efficient when used with control and quality. The optimization of the risk-benefit ratio is considered a major breakthrough in relation to conventional radiology, though this is not the reality of computing and digital radiology, where Brazil has not established standards and protocols for this purpose. This work aims to optimize computational chest radiographs (anterior-posterior projection-AP). To achieve this objective were used a homogeneous phantoms that simulate the characteristics of absorption and scattering of radiation close to the chest of a patient standard. Another factor studied was the subjective evaluation of image quality, carried out by visual grading assessment (VGA) by specialists in radiology, using an anthropomorphic phantom to identify the best image for a particular pathology (fracture or pneumonia). Quantifying the corresponding images indicated by the radiologist was performed from the quantification of physical parameters (Detective Quantum Efficiency - DQE, Modulation Transfer Function - MTF and Noise Power Spectrum - NPS) using the software MatLab®. © 2013 Springer-Verlag.

In vitro targeting of Polo-like kinase 1 in bladder carcinoma: Comparative effects of four potent inhibitors

Despite the improvements in neoadjuvant chemotherapy, the outcome of patients with advanced bladder cancer has changed very little over the past 30 years. In the present study we tested and compared the in vitro antitumor activities of four different inhibitors of Polo-like kinase 1 (PLK1) (BI 2536, BI 6727, GW843682X, and GSK461364), against 3 bladder carcinoma cell lines RT4, 5637 and T24. The impact on radiosensitivity and drug interactions in simultaneous treatments with cisplatin, methotrexate, and doxorubicin were also investigated. Our results showed that PLK1 inhibition prevented cell proliferation and clonogenicity, causing significant inhibition of invasion of tumor cells, though modest differences were observed between drugs. Moreover, all PLK1 inhibitors induced G2/M arrest, with the subsequent induction of death in all 3 cell lines. Drug interactions studies showed auspicious results for all PLK1 inhibitors when combined with the commonly used cisplatin and methotrexate, though combinations with doxorubicin showed mostly antagonistic effects. Comparably, the four PLK1 inhibitors efficiently sensitized cells to ionizing radiation. Our findings demonstrate that irrespective of the inhibitor used, the pharmacological inhibition of PLK1 constrains bladder cancer growth and dissemination, providing new opportunities for future therapeutic intervention. However, further laboratorial and preclinical tests are still needed to corroborate the usefulness of using them in combination with other commonly used chemotherapeutic drugs. © 2013 Landes Bioscience.