Orientador: Alexandre Nunes Ponezi / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Civil, Arquitetura e Urbanismo / Made available in DSpace on 2018-08-21T07:25:38Z (GMT). No. of bitstreams: 1 Torres_DaniladeLeoneFrancaeFreitas_M.pdf: 2036503 bytes, checksum: 068ab775d9c675592e34d0bfdd2ca1e9 (MD5) Previous issue date: 2012 / Resumo: O uso de efluente tratado na agricultura irrigada constitui uma alternativa adequada para a preservação de mananciais, além de ser uma importante fonte de nutrientes. No entanto, os riscos de toxicidade devem ser avaliados. Testes ecotoxicológicos apresentam-se como uma ferramenta apropriada, devido à sensibilidade dos organismos e sua ampla aceitação por órgãos governamentais e científicos. O objetivo do presente estudo foi avaliar a toxicidade aguda na fotobactéria marinha Vibrio fischeri e no microcrustáceo Daphnia similis (CE50) e do potencial citotóxico, genotóxico e mutagênico em Allium cepa (cebola) de um efluente sanitário tratado usado para irrigação. Os ensaios foram realizados no efluente bruto e nos tratados utilizados para irrigação: anaeróbio e pos-tratamento anaeróbio (nitrificado). Os resultados evidenciaram que o efluente bruto, apresentou uma maior variação dos valores de CE50 para ambos os organismos. O efluente anaeróbio apresentou um efeito tóxico agudo maior para V. fischeri que para D. similis. Já o nitrificado (pos-tratamento anaeróbio) apresentou uma toxicidade aguda menor para ambos os organismos testados. Na avaliação das aberrações cromossômicas em A. cepa, os efluentes tratados apresentaram uma redução do potencial citotóxico (p<0,05) em relação ao efluente bruto. Pode-se concluir que os testes escolhidos foram adequados para o monitoramento do efluente. Recomenda-se que o uso de efluente tratado na agricultura seja realizado de forma regrada, atentando para os padrões de Boas Práticas Agrícolas (GAP) e de toxicidade. Além disso, em escala real é necessário um estudo de monitoramento periódico que avalie impactos a longo prazo, incluindo testes de toxicidade crônica / Abstract: The use of the treated effluent in agriculture has proven to be an important alternative, since its preservation of water and its potential as source of nutrients, with low potential for negative impact on the environment. However, toxicity remains an important issue. Ecotoxicological tests are appropriate tools to evaluate the toxicity of effluents. The aim of this study was to evaluate acute toxicity on marine bacteria Vibrio fischeri, on micro crustacean Daphnia similis (EC50) and on Allium cepa (onion) (assessment of cytotoxic, mutagenic and genotoxic potencial) in treated sanitary effluent used for irrigation. We tested the raw effluent, and in its treatments, used for irrigation: anaerobic and anaerobic posttreatment (nitrified). Our results showed that the raw effluent had a greater variation of EC50 for both organisms, due to an intrinsic variability of its composition. The anaerobic effluent had a greater acute toxic effect on V. fischeri than for D. similis. Yet, the nitrified (anaerobic post-treatment stage) showed a lower toxicity for both tested organisms. We found a reduction in the citotoxical potential in A. cepa as compared to the raw efluente. We concluded that the tests chosen were adequate for monitoring the effluent since the organisms showed adequate sensitivity. It is therefore suggested that the use of treated wastewater in agriculture should be standardized, noting Good Agricultural Practices (GAP) and toxicity stardards. Furthermore for larger scale assumption it is necessary a periodic monitoring study to assess long term impacts, including chronic toxicity tests / Mestrado / Saneamento e Ambiente / Mestra em Engenharia Civil

L’objectif de ce travail est l’étude de la plante face aux molécules issues de la phototransformation de la sulcotrione. La recherche d’anomalies a été menée sur des extrémités racinaires d’Allium cepa L. var. aggregatum par un dénombrement au microscope des anomalies chromosomiques causées par l’ajout des différents xénobiotiques. L’étude a été réalisée en deux temps. Tout d’abord l’impact génotoxique d’un mélange constitué de la sulcotrione et de ses photoproduits a été évalué. Puis les caractéristiques de chaque produit de dégradation ont été analysées indépendamment. Il s’est avéré que la sulcotrione et ses produits de transformation CMBA et XDD ont des caractéristiques de toxicité qui sont spécifiques de chacun d’entre eux, avec une toxicité plus importante pour XDD. Il a été observé que XDD peut s’hydrolyser. Les études sur les produits de seconde génération ont permis d’identifier HMBA qui présente de nombreuses similitudes avec l’acide salicylique et provoque notamment une croissance racinaire accrue chez les espèces monocotylédones testées. / Cytotoxicity and genotoxicity of non irradiated and irradiated sulcotrione were investigated in Allium cepa test. Then, the cytotoxic effects of 2-chloro-4-mesylbenzoic acid (CMBA) and xanthene-1,9-dione-3,4-dihydro-6-methylsulfonyl (XDD), the two main photoproducts of sulcotrione were investigated at different concentrations. We analyze plant response treated with sulcotrione phototransformation by-products. Chromosomal aberrations were observed in Allium cepa root meristems. First, we investigated the genotoxic impact of a sulcotrione mixture and its photoproducts. Then each by-product was independently tested. Sulcotrione CMBA and XDD have specific toxicity, and XDD has a greater toxicity. In our latest analysis we observed that XDD can be hydrolyzed. HMBA was identified as a salicylic acid derivative and can increase root growth in monocotyledonous species.

Doctorat en Sciences / info:eu-repo/semantics/nonPublished

A form of familial sex-linked mental retardation has been associated with the expression of a fragile site near the terminal end of the long arm of the X chromosome. Previous reports on the fragile X chromosome showed expression of the fragile site to be limited to chromosome preparations from peripheral blood lymphocytes of mentally retarded males and their female relatives in families in which the disorder was segregating. Fragile site expression has also been shown to be a function of the medium employed in cell culture. The fragile X chromosome could only be demonstrated in lymphocytes cultured in medium 199 or media deprived of folic acid. This study was undertaken to develop a method for demonstrating the fragile X chromosome in cultured skin fibroblasts. Fibroblast cell lines from five patients (two mentally retarded males, two obligate carrier females, and a potential carrier female) from a family in which familial sex-linked mental retardation was known to be segregating were established and routinely maintained in a complete culture medium. Forty-three hours prior to chromosome harvest, cells from each patient were transferred to media deficient in folic acid. Under conditions of folic acid deprivation, it was possible to elicit expression of the fragile X chromosome in skin fibroblasts from all five patients studied. No fragile X chromosomes were detected in fibroblasts from three normal control subjects. In a preliminary assessment of the reliability of the fibroblast method, three patients (two mentally retarded males and a potential carrier female) from a second unrelated family in which the disorder is known to be segregating were studied with this method. The fragile X chromosome could be demonstrated in fibroblasts from both of the retarded male patients but could not be. demonstrated in fibroblast chromosome preparations from the potential carrier female. Lymphocytes for all patients studied were grown under similar folate deprived conditions for the purpose of comparing the effectiveness of fibroblast culture with lymphocyte culture in demonstrating the expression of the fragile X chromosome. Neither tissue was shown to consistently provide a higher frequency of expression of the fragile X chromosome. In addition to folate deprivation, it was shown that two other features of the fibroblast method influenced the frequency of expression of the fragile X chromosome. The fragile site was expressed at a significantly higher frequency in chromosome preparations in which the chromosomes were not severely contracted. The frequency of expression in fibroblasts was also shown to be significantly higher with a hypotonic treatment at chromosome harvest using 1% NaCitrate rather than 0.075M KC1. Because fragile site expression was shown to be a function of the degree of chromosome condensation, two agents, 5-BrdU and actinomycin-D, were studied to examine their decondensation effects on the frequency of expression. Neither BrdU nor actinomycin D proved effective in accentuating the frequency of expression. Since fibroblasts behave much like amniocytes in terms of cell culture and chromosome harvest, the development of a method for demonstrating the fragile X chromosome in cultured skin fibroblasts is a step toward the prospect of reliable antenatal diagnosis of familial sex-linked mental retardation associated with a fragile X chromosome. / Medicine, Faculty of / Medical Genetics, Department of / Graduate

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第21995号 / 医博第4509号 / 新制||医||1037(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 戸井 雅和, 教授 万代 昌紀, 教授 武藤 学 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Bei der Untersuchung von Dequalinium Chloride wurden alle relevanten genetischen Endpunkte berücksichtigt. Einer absolut negativen Datenlage steht lediglich ein spezifischer mutagener In vitro-Effekt im Ames-Test entgegen: In einem Bakterienstamm wurden eine bestimmte Art von Genmutationen, Frameshift-Mutationen, ausgelöst. Die Relevanz dieses Ergebnisses wurde in vivo im Spot Test überprüft. Der Spot Test war deshalb ausgewählt worden, da er die Erfassung von Mutagenen erlaubt, welche spezifisch Frameshift-Mutationen auslösen (Fahrig, 1995). Dies war eine notwendige Voraussetzung für den Einsatz des Tests, da im Ames-Test gerade diese Art von Mutationen ausgelöst worden war und deshalb dieser Endpunkt auch verfolgt werden musste. Die Verfolgung dieses genetischen Endpunkts in vivo im Spot Test mit Mäusen brachte trotz Verabreichung toxischer Dosen (5 und 7 mg/kg) ein negatives Ergebnis. Da zuvor schon der HPRT-Test ein negatives Ergebnis gezeigt hatte, stehen der spezifischen Frameshift-induzierenden Wirkung von Dequalinium Chloride in Bakterien eindeutig negative Ergebnisse im Säuger in vitro und in vivo gegenüber. Für die Relevanz der Ergebnisse im Spot Test und damit für die Wertung ist entscheidend, dass Dequalinium Chloride die Zielzellen in genügenden Konzentrationen erreicht, um eine Wirkung ausüben zu können. Bedenkt man, dass Dequalinium Chloride toxische Wirkung auf Muttertiere und Neugeborene ausübte und daß die Häufigkeit von gelben Missdifferenzierungssflecken und weißen, auf der Abtötung von Pigmentzellen beruhenden Bauchflecken gegenüber der Kontrolle erhöht war, so kann davon ausgegangen werden, dass Dequalinium Chloride die Zielzellen erreicht, aber weder mutagene noch rekombinogene Wirkung ausgeübt hat. Isoliert dastehenden positiven In-vitro-Befunden sollte von vornherein keine große Bedeutung beigemessen werden. Im vorliegenden Falle konnte gezeigt werden, dass eine mutagene Wirkung nur bei Bakterien und weder in vivo noch in vitro in Säugerzellen auftrat. Aus diesem Grunde kann dem Befund mit Bakterien keine Bedeutung für den Menschen beigemessen werden.:I. Datenlage 1. Genmutationen in vitro 1.1 Salmonella-Mikrosomen (Ames)-Test 1.2. HPRT-Test mit V79 Zellen des Chinesischen Hamsters 2. Chromosomenaberrationen in vitro 2.1 Chromosomenaberrationen in menschlichen Lymphozyten 3. Genmutationen in vivo 3.1 Spot Test mit Mäusen II. Wertung

Carcinogen-induced DNA base modification and subsequent DNA lesions are the critical events for the expression of premalignant phenotype of the cell. We have therefore investigated the chemopreventive efficacy of a vanadium salt against diethylnitrosamine (DEN)-induced early DNA and chromosomal damages in rat liver. Hepatocarcinogenesis was induced in male Sprague-Dawley rats with a single, necrogenic, intraperitoneal injection of DEN (200 mg/kg body weight). 8-Hydroxy-2′-deoxyguanosines (8-OHdGs), strand-breaks and DNA-protein crosslinks (DPCs) were measured by HPLC, comet assay and spectrofluorimetry, respectively. There was a significant and steady elevation of modified bases 8-OHdGs along with substantial increments of the extent of single-strand-breaks (SSBs), DPCs and chromosomal aberrations (CAs) following DEN exposure. Supplementation of vanadium as ammonium metavanadate (NH4VO3, +V oxidation state) at a dose of 0.5 ppm in terms of the salt weight throughout the experiment abated the formations of 8-OHdGs (P < 0.0001; 79.54%), tailed DNA (P < 0.05; 31.55%) and length:width of DNA mass (P < 0.02; 61.25%) in preneoplastic rat liver. Vanadium treatment also inhibited DPCs (P < 0.0001; 58.47%) and CAs (P < 0.001; 45.17%) studied at various time points. The results indicate that the anticlastogenic potential of vanadium in vivo might be due to the observed reductions in liver-specific 8-OHdGs, SSBs and/or DPCs by this trace metal. We conclude that, vanadium plays a significant role in limiting DEN-induced genotoxicity and clastogenicity during the early stages of hepatocarcinogenesis in rats.

Somatic Copy Number Alterations (CNAs) are common events in human cancers. Identifying CNAs and Significant Copy number Aberrations (SCAs) in cancer genomes is a critical task in searching for cancer-associated genes. Advanced genome profiling technologies, such as SNP array technology, facilitate copy number study at a genome-wide scale with high resolution. However, due to normal tissue contamination, the observed intensity signals are actually the mixture of copy number signals contributed from both tumor and normal cells. This genetic confounding factor would significantly affect the subsequent copy number analyses. In order to accurately identify significant aberrations in contaminated cancer genome, we develop a Java AISAIC package (Accurate Identification of Significant Aberrations in Cancer) that incorporates recent novel algorithms in the literature, BACOM (Bayesian Analysis of Copy number Mixtures) and SAIC (Significant Aberrations in Cancer). Specifically, BACOM is used to estimate the normal tissue contamination fraction and recover the "true" copy number profiles. And SAIC is used to detect SCAs using large recovered tumor samples. Considering the popularity of modern multi-core computers and clusters, we adopt concurrent computing using Java Fork/Join API to speed up the analysis. We evaluate the performance of the AISAIC package in both empirical family-wise type I error rate and detection power on a large number of simulation data, and get promising results. Finally, we use AISAIC to analyze real cancer data from TCGA portal and detect many SCAs that not only cover majority of reported cancer-associated genes, but also some novel genome regions that may worth further study. / Master of Science

Nepřesnosti při výrobě a sestavení rotačně souměrné čočky a deflektrou a jejich přesného zařazení do elektronově optického systému se projeví jako dodatečné pole příslušné symetrie, které deformuje ideální zobrazení. Tato dodatečná pole dokážeme spočítat pomocí metody konečných prvků v programu EOD. Toleranční analýza spočívá ve stanovení požadavků na rozměry a sestavení jednotlivých prvků a jejich částí. Korekce vad seřízení pak spočívá v určení typu a polohy korekčních vychylovacích cívek a multipólů tak, aby se tyto dodatečné vady odstranily, nebo aby se minimalizoval jejich vliv. Cílem disertační práce je analýza projevů vad seřízení a chování neseřízených systémů prozařovacích elektronových mikroskopů.

This thesis consists of two parts: a report on the use of a binocular Shack-Hartmann (SH) sensor to study the dynamic correlation of ocular aberrations; and the application of an adaptive optics (AO) system to investigate the effect of the manipulation of aberrations on the accommodation control. The binocular SH sensor consists of one laser source and one camera to reduce system cost and complexity. Six participants took part in this study. Coherence function analysis showed that coherence values were dependent on the subject, aberration and frequency component. Inter-ocular correlations of the aberration dynamics were fairly weak for all participants. Binocular and monocular viewing conditions produced similar wavefront error dynamics. The AO system has a dual wavefront sensing channel. The extra sensing channel permits direct measurement of the eye¿s aberrations independent of the deformable mirror. Dynamic correction of aberrations during steady-state fixation did not affect the accommodation microfluctuations, possibly due to the prior correction of the static aberration level and/or the limited correction bandwidth. The inversion of certain aberrations during dynamic accommodation affected the gain and latency of accommodation response (AR), suggesting that the eye used the aberrations to guide its initial path of accommodative step response. Corrections of aberrations at various temporal locations of AR cycle produced subject- and aberration-dependent results. The gain and phase lag of the AR to a sinusoidally moving target were unaffected by aberration correction. The predictable nature of the target had been suggested as the reason for its failure to produce any significant effect on the AR gain and phase lag.