Effects of oxidative stress during human and animal reproductions: A review

Given its high ability to damage important cellular components (lipids, proteins and deoxyribonucleic acid), oxidative stress is now recognized as one of the most common mechanisms associated with development of a variety of diseases and natural events such as pregnancy. During reproduction period, there is a change in the pro-oxidant and antioxidant balance due to the body and circulation modifications that are inherent to the pregnancy process. The present paper discusses the role of oxidative stress on the reproduction process. More effective defense strategies are needed to decrease the deleterious effects of oxidative-stress-induced gestation. This approach could be achieved by antioxidant status alteration. Further clinical and experimental studies are needed for better understanding of oxidative stress mechanism and the impact of antioxidant supplementation on reproduction

DNA methylation patterns in bladder cancer and washing cell sediments: a perspective for tumor recurrence detection

<p>Abstract</p> <p>Background</p> <p>Epigenetic alterations are a hallmark of human cancer. In this study, we aimed to investigate whether aberrant DNA methylation of cancer-associated genes is related to urinary bladder cancer recurrence.</p> <p>Methods</p> <p>A set of 4 genes, including <it>CDH1 </it>(E-cadherin), <it>SFN </it>(stratifin), <it>RARB </it>(retinoic acid receptor, beta) and <it>RASSF1A </it>(Ras association (RalGDS/AF-6) domain family 1), had their methylation patterns evaluated by MSP (Methylation-Specific Polymerase Chain Reaction) analysis in 49 fresh urinary bladder carcinoma tissues (including 14 cases paired with adjacent normal bladder epithelium, 3 squamous cell carcinomas and 2 adenocarcinomas) and 24 cell sediment samples from bladder washings of patients classified as cancer-free by cytological analysis (control group). A third set of samples included 39 archived tumor fragments and 23 matched washouts from 20 urinary bladder cancer patients in post-surgical monitoring. After genomic DNA isolation and sodium bisulfite modification, methylation patterns were determined and correlated with standard clinic-histopathological parameters.</p> <p>Results</p> <p><it>CDH1 </it>and <it>SFN </it>genes were methylated at high frequencies in bladder cancer as well as in paired normal adjacent tissue and exfoliated cells from cancer-free patients. Although no statistically significant differences were found between <it>RARB </it>and <it>RASSF1A </it>methylation and the clinical and histopathological parameters in bladder cancer, a sensitivity of 95% and a specificity of 71% were observed for <it>RARB </it>methylation (Fisher's Exact test (p < 0.0001; OR = 48.89) and, 58% and 17% (p < 0.05; OR = 0.29) for <it>RASSF1A </it>gene, respectively, in relation to the control group.</p> <p>Conclusion</p> <p>Indistinct DNA hypermethylation of <it>CDH1 </it>and <it>SFN </it>genes between tumoral and normal urinary bladder samples suggests that these epigenetic features are not suitable biomarkers for urinary bladder cancer. However, <it>RARB </it>and <it>RASSF1A </it>gene methylation appears to be an initial event in urinary bladder carcinogenesis and should be considered as defining a panel of differentially methylated genes in this neoplasia in order to maximize the diagnostic coverage of epigenetic markers, especially in studies aiming at early recurrence detection.</p

Gemcitabine/cisplatin treatment induces concomitant sertad1, cdkn2b and gadd45a modulation and cellular changes in bladder cancer cells regardless of the site of TP53 mutation.

Simultaneous use of cisplatin (CIS) and gemcitabine (GEN) for treating bladder cancer has increased because of their complementary effects. However, the molecular mechanisms underlying the activities of these two antineoplastic drugs are not fully known. Here, molecular biology techniques and microscopy were used to investigate transcriptomic and morphological changes in low and highgrade urinary bladder transitional carcinoma cell lines [RT4 - wild type TP53; 5637 - two TP53 mutations, one in codon 72 (Arg-Pro) and other in codon 280 (Arg-Thr) and T24 - inframe deletion of tyrosine 126 in the TP53 allele] simultaneously treated with CIS/GEN. Gene expression profile was evaluated by PCR arrays; cell morphology by scanning and transmission electron microscopy, and apoptosis was analyzed using fluorescent dye. Results showed concomitantly upregulation of CDKN2B (G1/S transition), GADD45A (DNA repair and apoptosis) and SERTAD1 (regulation of transcription) gene, increased number of nuclear chamfers and apoptotic cells, and reduced number of microfilaments, organelles and in the size of the nucleus in 5637 and T24 cells after simultaneous treatment with CIS/GEN. In conclusion, independently of the TP53 mutation status and tumor grade, CIS/ GEN induced gene modulation accompanied by changes in cell morphologies, which confirm the antiproliferative activity of the treatment protocol. These findings help to understand the pathways modulated by these antineoplastic agents and may provide insights for anti-cancer chemotherapy

Cell adhesion and growth on surfaces modified by plasma and ion implantation

In this study, we show and discuss the results of the interaction of living CHO (Chinese Hamster Ovary) cells, in terms of adhesion and growth on glass, SU-8 (epoxi photoresist), PDMS (polydimethylsiloxane), and DLC (hydrogen free diamond-like carbon) surfaces. Glass, SU-8, and DLC but not PDMS showed to be good surfaces for cell growth. DLC surfaces were treated by oxygen plasma (DLC-O) and sulfur hexafluoride plasma (DLC-F). After 24 h of cell culture, the number of cells on DLC-O was higher than on DLC-F surface. SU-8 with silver implanted, creating nanoparticles 12 nm below the surface, increased significantly the number of cells per unit area. (C) 2014 AIP Publishing LLC.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Portal de Contratação Utilities

Num mundo cada vez mais competitivo e não alheio a crises económicas, a margem de erro é mínima para as entidades empresariais. Deste modo, têm estas instituições que estar dotadas de mecanismos de resposta focadas nas linhas da inovação e criatividade, não podendo os seus líderes descurar a prossecução da estabilidade económico-financeira. A tais circunstancialismos, alia-se a ideia da globalização o que tudo somado, leva à obrigatoriedade das empresas em adotarem os valores de eficácia, eficiência e competitividade nas áreas de angariação de novos clientes e da sua manutenção, devendo os seus métodos organizacionais ser sinónimo destes ideais chaves. No que respeita aos serviços de utilidade pública, a competitividade na aquisição e expansão da clientela, não deixa de ser uma realidade inegável, pelo que as empresas têm o dever de investir na melhoria e desenvolvimento dos instrumentos tecnológicos de gestão, captação e cativação de novos consumidores, para que tais organizações possam conhecer um desenvolvimento sustentável e harmonioso.In an increasingly competitve world and not unrelated to economic crises, the margin for error is minimal for business entities. Therefore, these institutions have to be equipped with response mechanisms focused on innovation and creativity, and their leaders must not neglect the pursuit of economic and financial stability. In fact, the impositions of the market, the demands of clients, the pressure from investors, the competition from other peers, generate ferocity in the fight for competitiveness and control of public utility markets. In addition to these circumstances, the idea of globalisation, which all adds up, leads companies to adopt the values of effectiveness, efficiency and competitiveness in the areas of attracking new clients and maintaining them, and their organisational methods should be synonymous with these key ideals. As far as public utility services are concerned, competitiveness in acquiring and expanding customers is an undeniable reality, and companies have a duty to invest in the improvement and development of technological instruments for managing, attracting and captivating new consumers, so that such organizations can experience a sustainable and harmonious development

MRE11A and SKP2 genes are associated with the increased cytotoxicity induced by the synergistic effects of cisplatin and gemcitabine in bladder cancer cells.

The combination of gemcitabine and cisplatin has been shown previously to elicit a synergistic therapeutic effect on bladder cancer cell lines and result in reduced cell survival. However, the precise mechanism by which cells die has not been elucidated. Cell cycle-related genes are the predominant targets of chemotherapeutic protocols. Therefore, molecular biomarkers that are predictive of therapeutic outcomes associated with tumor sensitivity might be important for optimal treatment protocol selection. The aim of this study was to investigate the changes in gene expression in cell cycle-related genes that were induced by cisplatin, gemcitabine or a combined treatment using both agents in a low-grade urinary bladder transitional carcinoma cell line (RT4). The following three treatment protocols were used: 1.0 lM cisplatin, 1.56 lM gemcitabine and a combination of 1.0 lM cisplatin and 1.56 lM gemcitabine. Cytometry and morphology analysis (by phase-contrast photomicrography) were performed in addition to pathway-specific gene expression analysis using quantitative RT-PCR gene arrays. The following results were observed after 1.0 lM cisplatin treatment: (1) a decrease in cell number, (2) an increased percentage of scattered cells and (3) downregulated expression of genes related to cell cycle arrest, G1/S-to-mitotic cell cycle transition, DNA repair, apoptosis, transcription and mitosis. Treatment with 1.56 lM gemcitabine, or with both drugs simultaneously, induced the following effects: (1) a decrease in cell number, (2) an increased percentage of scattered and elongated cells, (3) the modulation of genes that are predominantly involved in DNA repair and (4) a significant upregulation of genes related to cell cycle arrest. Reduced cell density was observed after the combined treatment compared to the two other single-agent protocols. The downregulation of MRE11A and SKP2 was observed only in cells subjected to the combined treatment. In conclusion, cisplatin, gemcitabine and the combination of both drugs elicited distinct toxicogenomic effects in the RT4 bladder transitional carcinoma cell line, although disruptions in the expression of cell cycle control-related genes and other pathways responsible for cell survival were observed for all of the protocols. MRE11A and SKP2 downregulation appeared to be responsible for the synergistic therapeutic effects elicited by cisplatin and gemcitabine

Cell growth on 3D microstructured surfaces.

Chinese Hamster Ovary (CHO) cell cultures were grown on surfaces lithographed with periodic 3D hexagonal microcavity array morphology. The range of microcavity size (inscribed circle diameter) was from 12 ?m to 560 ?m. CHO cells were grown also on flat surfaces. The characterization was performed with respect to cell growth density (number of nuclei per unit area) by fluorescence opticalmicroscopy and evaluated by correlation function analysis. We found that optimum microcavity radius was 80 ?m, concerning to the maximum cell growth density, being even greater than the growth density on a flat (unstructured) substrate of the same material. This finding can be important for optimization of biotechnological processes and devices

Effects of lycopene, synbiotic and their association on early biomarkers of rat colon carcinogenesis

This study evaluated whether a synergy exists for the combined treatment with lycopene and synbiotic on early biomarkers; of colon carcinogenesis. Male Wistar rats received a diet containing 300 mg/kg of lycopene and/or synbiotic (Bifidobacterium lactis plus oligofructose/inulin) or their combination 2 weeks before and during carcinogen treatment with 1.2-dimethylhydrazine (DMH). Twenty-four hours after the last DMH application, the colons were processed for immunohistochemical analysis of proliferating cell nuclear antigen (PCNA), p53 protein, hematoxylin-eosin staining for apoptosis analysis and genotoxicity of fecal water by comet assay. Eight weeks after the last DMH application, the colons were analyzed for development of classical aberrant crypt foci (ACF) and mucin-negative ACF. Treatment with lycopene, synbiotic or their combination significantly increased apoptosis, reduced the PCNA and p53 labeling indexes and the development of classical ACF and mucin-negative ACF. Furthermore, a lower genotoxicity of fecal water was also detected in the groups treated with the chemopreventive agents. An additive/synergistic effect of the combined treatment with lycopene/synbiotic was observed only for the fecal water genotoxicity and mucin-negative ACF parameters. These results indicate that an additive/synergistic of the combination of chemopreventive agents on the initiation phase of colon carcinogenesis can be detected using selective early biomarkers. (C) 2009 Elsevier Ltd. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Cigarette smoke affects apoptosis in rat tongue mucosa: Role of bcl-2 gene family

While it has been clearly demonstrated that smoking is the most significant exogenous factor involved in oral carcinogenesis, little is known about the global molecular and cellular changes that occur prior to the appearance of clinically detectable symptoms. Thus, the aim of this study was to investigate the expressivity of bcl-2, bax and PCNA in the rat tongue mucosa exposed to cigarette smoke by means of immunohistochemistry. A total of twelve male Wistar rats were distributed into 2 groups: negative control and experimental group exposed to cigarette smoke during 75 days. After experimental period, no histopathological changes in the tongue mucosa were detected in the negative control and the experimental group. on the other hand, an overexpression of bcl-2 was detected (p 0.05). Also, the labeling index for bcl-2 and bax showed an increase 75 days after cigarette exposure (p < 0.01). PCNA-labeling index did not show remarkable changes between groups. Taken together, our results show that bcl-2 is overexpressed in the rat tongue keratinocytes after cigarette smoke exposure