Thesis (M.S.)--Virginia Polytechnic Institute and State University, 1991. / Vita. Abstract. Includes bibliographical references (leaves 30-37). Also available via the Internet.

The purpose of this study was to determine the factors affecting the acid degradation of chrysotile asbestos (Mg_3Si_2O_5(OH_4)) . Millions of tons of asbestos have found use in this country as insulative or ablative material. More than 95 percent of the asbestos in use is of the chrysotile variety. The remaining 5 percent is composed of various types of fibrous amphiboles. The inhalation of asbestos can lead to several diseases in humans. Asbestosis, lung cancer and mesothelioma are the most common afflictions associated with asbestos inhalation, and they may occur up to 40 years after the initial exposure. It has previously been reported that if more than 50 percent of the magnesium is removed from a chrysotile sample its carcinogenicity is reduced to nil. Several inorganic acids were studied to determine their ability to leach magnesium from chrysotile. It was found that the ability to leach magnesium was dependent upon the acidic anion in addition to the concentration of the acid. The ordering of the efficiency of the acids in their ability to remove magnesium from chrysotile was found to be HCl > H_2SO_4 > H_3PO_4 > HNO_3. Predictive equations were developed to allow the calculation of the amount of magnesium removed under various acid concentrations as a function of time and acid species. The effects of temperature and dissolved spectator cations upon the degradation process were also examined. There was no major effect on the amount of magnesium removed as a function of spectator cation concentration. An infrared method was also developed to allow the determination of the percent degradation of a chrysotile sample directly. The shifts in the positions of three silicate stretching peaks (1068 cm^-1, 948 cm^-1 and 715 cm^-1) and one magnesium oxygen stretching peak (415 cm"1) as a function of the percent magnesium removed were correlated to allow this determination.

Thesis (Ph. D.)--Virginia Polytechnic Institute and State University, 1985. / Vita. Abstract. Includes bibliographical references (leaves 195-205). Also available via the Internet

A aneuploidia é uma característica dos tumores sólidos. Ela pode ser resultado de diferentes erros durante a mitose, como a amplificação centrossômica, mitoses multipolares, e anormalidades durante a citocinese. Hoje se sabe que a aneuploidia pode estar relacionada à supressão ou progressão tumoral dependendo do grau da aneuploidia e do contexto genético das células, e assim esforços vem sendo feitos a fim de elucidar quais erros durante a mitose estão relacionados à formação de células aneuploides viáveis e inviáveis. Estudos prévios do nosso grupo mostraram que tratamentos de células em cultura com fibras de crisotila e com vincristina levam a formação de células aneuploides. Agora direcionados nossos esforços para elucidar os mecanismos envolvidos na formação dessas células, investigando alterações nos centrossomos, número de cromossomos, e origens e destinos de mitoses multipolares após o tratamento com crisotila e com vincristina. As fibras de crisotila, em linhagens de células tumorais e normais, levaram a padrões de localização alterados de proteínas relacionadas à abscisão durante a citocinese, e ocorreu a regressão deste processo e consequente formação de apenas uma célula-filha com o dobro do conteúdo de cromossomos e de centrossomos. Nas duas linhagens estudadas essas células tetraploides progrediram no ciclo celular, gerando mitoses multipolares e consequente formação de células aneuploides. O tratamento com vincristina levou a respostas similares e também distintas em células normais e tumorais. Durante a retenção em metáfase ocorreu a fragmentação da matriz pericentriolar, e as células foram encaminhadas à morte celular ou à saída da mitose sem a ocorrência de divisão celular. Entretanto, células de origem normal tetraploides não progrediram no ciclo celular e não formaram mitoses multipolares, enquanto na linhagem tumoral as células apresentaram aumento da expressão de Aurora A e células com conteúdo cromossômico aumentado e aneuploide em mitoses multipolares. As mitoses multipolares formaram uma, duas ou três células e apresentaram diversas anormalidades no processo de divisão. As alterações observadas no número e composição dos centrossomos após o tratamento com as duas linhagens indicaram que processos de amplificação centrossômica ocorreram após o tratamento. Os dados foram compatíveis com a reduplicação dos centrossomos e com a formação de centríolos a partir do aumento da matriz pericentriolar. Os dados reunidos mostram que apenas células tumorais foram capazes de proliferar mesmo após diferentes erros mitóticos, enquanto células normais puderam apenas superar os erros ocasionados pelas fibras de crisotila / Aneuploidy is a feature of solid tumors. Aneuploid cells result from errors during mitosis, such as centrosome amplification, multipolar mitosis and cytokinesis abnormalities. The capability of aneuploidy to promote and to suppress tumorigenesis has driven the efforts to characterize mitotic errors that form viable and not viable aneuploid cells. We have previously shown that chrysotile, an asbestos fiber, and vincristine, a chemotherapeutic agent, are able to induce aneuploidy. Now we directed our focus to discover possible mechanisms involved in aneuploid cell formation. Herein we evaluated centrosome morphology, chromosome number, and origins and fates of multipolar mitosis after chrysotile and vincristine treatment. Chrysotile fibers, in normal and cancer cells, led to mislocalization of proteins involved in abscission, which resulted in cytokinesis regression and tetraploid cells. These cells were able to enter cell cycle, giving rise to multipolar mitosis and aneuploid cells. Vincristine treatment led to specific and common responses in normal and cancer cells. During metaphase arrest, pericentrosomal matrix was fragmented, and the cells could be conducted to mitotic slippage in both lineages. However, normal tetraploid cells could not progress through cell cycle and neither to form multipolar mitosis, while cancer tetraploid cells showed Aurora A overexpression, structural and numerical centrosome abnormalities, multipolar mitosis and high levels of aneuploidy. The results showed that cancer cells could proliferate even after several mitotic errors, while normal cells could only overcome errors induced by chrysotile treatment

Asbesto é um nome geral dado a seis tipos de fibras minerais encontradas naturalmente na crosta terrestre. Estas fibras vêm sendo exploradas industrialmente desde 1970, porém diversos trabalhadores expostos às fibras apresentaram patologias no trato respiratório, como fibroses e carcinomas. Alguns tipos de fibra foram banidos do mercado, porém o tipo de asbesto crisotila ainda pode ser comercializado na maioria dos países. Estudos in vivo e in vitro tentam elucidar as alterações causadas pela exposição à asbesto nos tecidos e nas células que possam estar relacionadas ao aparecimento de doenças, e foi verificado que a exposição às fibras leva a quebras na dupla fita de DNA, estresse oxidativo, formação de células micronucleadas e células aneuploides. O presente estudo teve como objetivo verificar a presença de alterações causadas em células em cultura expostas à crisotila por 48 h e recuperadas em meio livre de fibras por 48 h, 4 dias e 8 dias, além de observar por microscopia em tempo real divisões aberrantes após a exposição as fibras por 24 e 48h. Foram verificadas alterações que permaneceram na cultura mesmo após 8 dias de recuperação, quando não foram mais observadas fibras na cultura, como formação de células aneuploides, diminuição de frequência de células em G0/G1, aumento de células em G2/M e aumento relativo de células em metáfase quanto à porcentagem de células em fases mais tardias da fase M do ciclo. Já aumento da frequência de células micronucleadas ocorreu apenas nos períodos quando foram observadas fibras na cultura. Para análise da formação de mitoses multipolares e destinos destas células foram construídos vetores para expressão de tubulinas fusionadas a proteínas fluorescentes RFP e GFP, padronizadas as condições de transfecção e de aquisição de imagens para que as células tratadas com crisotila fossem observadas por time-lapse. Alguns destinos de mitoses multipolares causadas pelo tratamento com crisotila foram observados, como morte em metáfase, divisão gerando duas ou três células filhas, fusão de células durante a telófase e retenção em metáfase. Os dados sugerem também a indução da amplificação centrossômica, que parece ocorrer inicialmente em células interfásicas, e também devido à fusão de células. / Asbestos is a general name given to six different fibrous silicate minerals found naturally in the earth\'s crust. These fibers are being exploited industrially since 1970, but several workers exposed to the fibers developed diseases in the respiratory tract, such as fibrosis and carcinomas. Some types of fiber were banished from the market, but the type of asbestos chrysotile can still be marketed in most countries. Studies in vivo and in vitro are trying to elucidate the asbestos effects in tissues and cells that could be related to the development of diseases, and these studies verified that asbestos exposure lead to DNA double strand breaks, oxidative stress, multinucleated and aneuploid cell formation. The present work aimed to verify the alterations in culture cells exposed to chrysotile for 48 h and recovered in fiber-free medium for 48 h, 4 days and 8 days, and also observe aberrant mitosis using time-lapse microscopy after 24 h and 48 h of chrysotile exposure. Some alterations were observed and remained in cell culture even after 8 days of recovery when chrysotile fibers were no longer observed - such as aneuploid cell formation, increased frequencies of G2/M cell, decreased frequencies of G1 cells, and increased frequencies of cells in early M phases as metaphase. The induction of micronuclei occurred only during the periods that fibers were observed in cell culture. For the analysis of multipolar mitosis formation and destinies of these cells after chrysotile treatment, DNA vectors for the expression of tubulins fused to fluorescent proteins (GFP and RFP) were constructed, and the conditions for cells transfection and image acquisition for time-lapse microscopy were established. The fate of some multipolar metaphases was observed: cell retention on metaphase, cell cycle progression generating two or three daughter cells, cell fusion during cytokinesis or during telophase after a multipolar anaphase, and cell death. The centrosome amplification was not observed during the M phase of cell cycle, and may occur in interphase, and also despite cell fusion.

Asbesto é um nome geral dado a seis tipos de fibras minerais encontradas naturalmente na crosta terrestre. Estas fibras vêm sendo exploradas industrialmente desde 1970, porém diversos trabalhadores expostos às fibras apresentaram patologias no trato respiratório, como fibroses e carcinomas. Alguns tipos de fibra foram banidos do mercado, porém o tipo de asbesto crisotila ainda pode ser comercializado na maioria dos países. Estudos in vivo e in vitro tentam elucidar as alterações causadas pela exposição à asbesto nos tecidos e nas células que possam estar relacionadas ao aparecimento de doenças, e foi verificado que a exposição às fibras leva a quebras na dupla fita de DNA, estresse oxidativo, formação de células micronucleadas e células aneuploides. O presente estudo teve como objetivo verificar a presença de alterações causadas em células em cultura expostas à crisotila por 48 h e recuperadas em meio livre de fibras por 48 h, 4 dias e 8 dias, além de observar por microscopia em tempo real divisões aberrantes após a exposição as fibras por 24 e 48h. Foram verificadas alterações que permaneceram na cultura mesmo após 8 dias de recuperação, quando não foram mais observadas fibras na cultura, como formação de células aneuploides, diminuição de frequência de células em G0/G1, aumento de células em G2/M e aumento relativo de células em metáfase quanto à porcentagem de células em fases mais tardias da fase M do ciclo. Já aumento da frequência de células micronucleadas ocorreu apenas nos períodos quando foram observadas fibras na cultura. Para análise da formação de mitoses multipolares e destinos destas células foram construídos vetores para expressão de tubulinas fusionadas a proteínas fluorescentes RFP e GFP, padronizadas as condições de transfecção e de aquisição de imagens para que as células tratadas com crisotila fossem observadas por time-lapse. Alguns destinos de mitoses multipolares causadas pelo tratamento com crisotila foram observados, como morte em metáfase, divisão gerando duas ou três células filhas, fusão de células durante a telófase e retenção em metáfase. Os dados sugerem também a indução da amplificação centrossômica, que parece ocorrer inicialmente em células interfásicas, e também devido à fusão de células. / Asbestos is a general name given to six different fibrous silicate minerals found naturally in the earth\'s crust. These fibers are being exploited industrially since 1970, but several workers exposed to the fibers developed diseases in the respiratory tract, such as fibrosis and carcinomas. Some types of fiber were banished from the market, but the type of asbestos chrysotile can still be marketed in most countries. Studies in vivo and in vitro are trying to elucidate the asbestos effects in tissues and cells that could be related to the development of diseases, and these studies verified that asbestos exposure lead to DNA double strand breaks, oxidative stress, multinucleated and aneuploid cell formation. The present work aimed to verify the alterations in culture cells exposed to chrysotile for 48 h and recovered in fiber-free medium for 48 h, 4 days and 8 days, and also observe aberrant mitosis using time-lapse microscopy after 24 h and 48 h of chrysotile exposure. Some alterations were observed and remained in cell culture even after 8 days of recovery when chrysotile fibers were no longer observed - such as aneuploid cell formation, increased frequencies of G2/M cell, decreased frequencies of G1 cells, and increased frequencies of cells in early M phases as metaphase. The induction of micronuclei occurred only during the periods that fibers were observed in cell culture. For the analysis of multipolar mitosis formation and destinies of these cells after chrysotile treatment, DNA vectors for the expression of tubulins fused to fluorescent proteins (GFP and RFP) were constructed, and the conditions for cells transfection and image acquisition for time-lapse microscopy were established. The fate of some multipolar metaphases was observed: cell retention on metaphase, cell cycle progression generating two or three daughter cells, cell fusion during cytokinesis or during telophase after a multipolar anaphase, and cell death. The centrosome amplification was not observed during the M phase of cell cycle, and may occur in interphase, and also despite cell fusion.

No description available.

A aneuploidia é uma característica dos tumores sólidos. Ela pode ser resultado de diferentes erros durante a mitose, como a amplificação centrossômica, mitoses multipolares, e anormalidades durante a citocinese. Hoje se sabe que a aneuploidia pode estar relacionada à supressão ou progressão tumoral dependendo do grau da aneuploidia e do contexto genético das células, e assim esforços vem sendo feitos a fim de elucidar quais erros durante a mitose estão relacionados à formação de células aneuploides viáveis e inviáveis. Estudos prévios do nosso grupo mostraram que tratamentos de células em cultura com fibras de crisotila e com vincristina levam a formação de células aneuploides. Agora direcionados nossos esforços para elucidar os mecanismos envolvidos na formação dessas células, investigando alterações nos centrossomos, número de cromossomos, e origens e destinos de mitoses multipolares após o tratamento com crisotila e com vincristina. As fibras de crisotila, em linhagens de células tumorais e normais, levaram a padrões de localização alterados de proteínas relacionadas à abscisão durante a citocinese, e ocorreu a regressão deste processo e consequente formação de apenas uma célula-filha com o dobro do conteúdo de cromossomos e de centrossomos. Nas duas linhagens estudadas essas células tetraploides progrediram no ciclo celular, gerando mitoses multipolares e consequente formação de células aneuploides. O tratamento com vincristina levou a respostas similares e também distintas em células normais e tumorais. Durante a retenção em metáfase ocorreu a fragmentação da matriz pericentriolar, e as células foram encaminhadas à morte celular ou à saída da mitose sem a ocorrência de divisão celular. Entretanto, células de origem normal tetraploides não progrediram no ciclo celular e não formaram mitoses multipolares, enquanto na linhagem tumoral as células apresentaram aumento da expressão de Aurora A e células com conteúdo cromossômico aumentado e aneuploide em mitoses multipolares. As mitoses multipolares formaram uma, duas ou três células e apresentaram diversas anormalidades no processo de divisão. As alterações observadas no número e composição dos centrossomos após o tratamento com as duas linhagens indicaram que processos de amplificação centrossômica ocorreram após o tratamento. Os dados foram compatíveis com a reduplicação dos centrossomos e com a formação de centríolos a partir do aumento da matriz pericentriolar. Os dados reunidos mostram que apenas células tumorais foram capazes de proliferar mesmo após diferentes erros mitóticos, enquanto células normais puderam apenas superar os erros ocasionados pelas fibras de crisotila / Aneuploidy is a feature of solid tumors. Aneuploid cells result from errors during mitosis, such as centrosome amplification, multipolar mitosis and cytokinesis abnormalities. The capability of aneuploidy to promote and to suppress tumorigenesis has driven the efforts to characterize mitotic errors that form viable and not viable aneuploid cells. We have previously shown that chrysotile, an asbestos fiber, and vincristine, a chemotherapeutic agent, are able to induce aneuploidy. Now we directed our focus to discover possible mechanisms involved in aneuploid cell formation. Herein we evaluated centrosome morphology, chromosome number, and origins and fates of multipolar mitosis after chrysotile and vincristine treatment. Chrysotile fibers, in normal and cancer cells, led to mislocalization of proteins involved in abscission, which resulted in cytokinesis regression and tetraploid cells. These cells were able to enter cell cycle, giving rise to multipolar mitosis and aneuploid cells. Vincristine treatment led to specific and common responses in normal and cancer cells. During metaphase arrest, pericentrosomal matrix was fragmented, and the cells could be conducted to mitotic slippage in both lineages. However, normal tetraploid cells could not progress through cell cycle and neither to form multipolar mitosis, while cancer tetraploid cells showed Aurora A overexpression, structural and numerical centrosome abnormalities, multipolar mitosis and high levels of aneuploidy. The results showed that cancer cells could proliferate even after several mitotic errors, while normal cells could only overcome errors induced by chrysotile treatment

Chrysotile can be linked to three diseases: lung cancer, asbestosis, and mesothelioma. The duration and intensity of exposure along with fiber size appear to play an important role in the development of the diseases. Chrysotile is part of the serpentine group which has the general composition of Mg₃Si₂O₅(OH)₄. The fluids in lung tissue contain very low concentrations of magnesium and silicon. As a result they are quite undersaturated with respect to chrysotile and chrysotile will dissolve. Its persistence in lung tissue is simply a result of its dissolution kinetics. The purpose of this study was to estimate the lifetime of a respirable size fiber of chrysotile in lung tissue. The dissolution reaction for chrysotile for pH's less than nine is: Mg₃Si₂O₅(OH)₄ + 6H⁺ = 3Mg²⁺+ 2H₄4SiO₄ + H₂0 This reaction proceeds in two steps. First, the magnesium hydroxide layer of the serpentine dissolves leaving behind the silica structure of the fiber. Then the silica dissolves. Therefore, the fiber lifetime depends upon the rate of silica release. Over the range of undersaturation expected for lung tissue, the rate of silica release was found to be independent of pH with the average rate being 5.94 (±3.05) x 10⁻¹⁰ moles m⁻² sec⁻¹. A shrinking fiber model was used to determine the relationship between dissolution time and fiber diameter. It was found that the most hazardous sized fiber of chrysotile (1 μm) would completely dissolve in about 9 months, consideration of one standard deviation above and below the mean of the rate constant gives estimates of the lifetime of a fiber ranging from 6 to 19 months. / Master of Science

Examination of Chrysotile Asbestos from the Eastern Townships of Quebec using optical and electron microscopic methods has shown that Chrysotile fibres are intimately associated with minute particles of magnetite. / Measurements were made using a fluxgate magnetometer of the remnant magnetic fields of bulk commercial fibres and of airborne total dust and of the respirable fraction of airborne dust collected in Quebec asbestos mines and mills using a Hexhlet horizontal eleutriator. The validity of the Hexhlet horizontal eleutriator for respirable fibre collection in the mill environment was determined. It was observed that the remnant magnetic field intensities of airborne dust in laboratory generated dust clouds, and in the work places were related in a meaningful way to the magnetite content of the dust and to the total mass and respirable mass fractions of airborne dust, and also, to the concentrations of fibre greater than 5 micrometre length (aspect ratio > 3:1) measured using phase contrast microscopy. Calibration curves were developed for each of 9 Quebec mills. Repeat calibrations showed the lines to be reproducible within (+OR-) 10 percent. The accuracy of the method for mass dust determination was dependent on sample size with errors as low as (+OR-) 7.5 percent for samples larger than 3 milligrams, and as large as (+OR-) 40 percent for samples of mass smaller than 0.5 milligrams. The relationships between fibre concentrations and remnant magnetic fields were less reproducible and the lower detection limit was limited by the sensitivity of the fluxgate magnetometer. Although the measured remnant magnetic field varied as the magnetite content varied between different mills and commercial fibre products, magnetite to chrysotile ratio remained constant for a given mill and commercial product. Measurements of the remnant magnetic fields of airborne dusts collected in mill environments can thus provide the possibility of the rapid determination of airborne chrysotile fibre concentrations both in terms of mass and number of fibres per litre of air sampled as well as their iron content. / Measurement of the magnetic content of specimens of autopsied lung tissue from asbestos exposed workers indicated that the residual estimated fibre content measured by electron microscopy was less than estimated using magnetic field measurements. . . . (Author's abstract exceeds stipulated maximum length. Discontinued here with permission of author.) UMI