Coal workers’ pneumoconiosis (CWP), commonly referred to as black lung, is a chronic lung disease that results from the inhalation and deposition of coal dust in the lungs. While this disease continues to afflict coal miners, its prevalence has steadily declined over three decades since 1970. Based on a voluntary X-ray surveillance program, conducted by the National Institute for Occupational Safety and Health (NIOSH), this downward trend, however, ended in 2000 and has actually begun to rise. The Mine Safety and Health Administration (MSHA) instituted a Comprehensive Initiative to “End Black Lung” to combat the reported upturn in black lung disease. Rulemaking, with the intent of strengthening respirable dust regulations, is a major part of this initiative. This thesis addresses a controversial aspect of the newly proposed rules – single-shift compliance sampling. Establishing new requirements for respirable dust compliance requires an understanding of both the accuracy and variability of measurements. Measurement variability is especially important in underground mining where the workplace is constantly moving and ventilation controls are continually changing. The results of a ventilation study performed in three underground coal mines are presented in this thesis. A total of 600 dust-concentration measurements were obtained in this study using Continuous Personal Dust Monitors (CPDMs). The data was analyzed to determine the variability associated with taking dust measurements in the mining workplace. The Johnson transformation was found to produce the best-fit distribution model for the data. This thesis summarizes the results of this study and presents a statistical procedure for establishing an exposure limit.

The purpose of this thesis is to examine the potential for a more comprehensive method of analysis of coal mine dust. Respirable dust is specifically of interest due to its ability to cause occupational lung disease when miners are overexposed to airborne concentrations. A detailed standard methodology to characterize respirable mine dust is carefully investigated with the use of scanning electron microscopy with energy dispersive x-ray (SEM-EDX). In addition to a thorough description of the developed particle level characterization approach, the method is demonstrated with underground respirable dust samples collected from an underground coal mine in Central Appalachia. Results of this thesis indicate that a comprehensive dust characterization method is possible and can be efficient and effective, when standardized. This analytical approach uses measured compositions, dimensions, and shapes to produce an abundance of data in even a single sample of dust. Verification results show the method is suitable for analysis of respirable particles of common coal mine mineralogy and analysis of many samples in a timely manner. The results obtained from the underground samples in Central Appalachia reveal the quantity of information which can be generated using the developed method. The amount of data which is acquired using the more comprehensive dust characterization method may aid in understanding the health effects of various dust characteristics. / Master of Science

The purpose of this thesis is to examine applications of thermal and laser-based methods to monitor airborne particulates in underground coal mines. Specifically, coal and mixed mineral mine dust, as well as, diesel particulate matter (DPM). These particulates have historically, and continue to have, significant health impacts on underground miners. Chapters 1 and 2 of this thesis concentrate on using a novel method of thermogravimetric analysis (TGA) to characterize respirable coal and mixed mineral mine dust and presents the results of this method being applied to samples collected in Appalachia coal mines. Appalachia has been a geographic "hotspot" for the rise in occupational lung disease amongst underground coal miners, which began in 1990's after decades of steady decline. This has led researchers to propose there could be something unique about the respirable dust composition in Appalachia coal mines, which resulted in the surge of lung disease cases; however, the knowledge base regarding the actual composition of respirable coal mine dust is limited. The results of this thesis show that most of the mass fraction of respirable Appalachia coal mine dust is not coal, but rather carbonates and non-carbonate minerals (i.e. silica and silicates). These findings are significant as many researchers now suspect silica and silicates to be the true culprit in the occupational lung disease of coal miners. DPM presents an additional occupational health hazard to underground coal miners where diesel equipment is used and is difficult to monitor due to its complex nature. In underground metal/non-metal mines, airborne DPM is regulated and monitored using carbon surrogates. However, due to the potential interference from coal-sourced carbon, DPM in coal mines is monitored only by taking samples at the tailpipe of each piece of equipment. This thesis aims to investigate the potential for a laser-based instrument, the FLIR Airtec, to be used in underground coal mines. In particular, what effect the coal dust will have on the instrument, as it measures DPM by way of elemental carbon (EC). The results of this study show that while the Airtec will not over-estimate coal-sourced EC, there could be some sampling artifacts associated with its operation in coal mines, which may inhibit its effectiveness. / Master of Science

The purpose of this thesis is to examine the potential of a novel method for analysis and characterization of coal mine dust. Respirable dust has long been an industry concern due to the association of overexposure leading to the development occupational lung disease. Recent trends of increased incidence of occupational lung disease in miners, such as silicosis and Coal Workers Pneumoconiosis, has shown there is a need for a greater understanding of the respirable fraction of dust in underground coal mines. This study will examine the development of a comprehensive standard methodology for characterization of respirable dust via thermogravimetric analysis (TGA). This method was verified with laboratory-generated respirable dust samples analogous to those commonly observed in underground coal mines. Results of this study demonstrate the ability of the novel TGA method to characterize dust efficiently and effectively. Analysis of the dust includes the determination of mass fractions of coal and non-coal, as well as mass fractions of coal, carbonate, and non-carbonate minerals for larger respirable dust samples. Characterization occurs through the removal of dust particulates from the filter and analysis with TGA, which continuously measures change in mass with specific temperature regions associated with chemical changes for specific types of dust particulates. Results obtained from the verification samples reveal that this method can provide powerful information that may help to increase the current understanding of the health risks linked with exposure to certain types of dust, specifically those found in underground coal mines. / Master of Science

It has long been known that chronic exposures to high concentrations of respirable coal mine dust can lead to the development of lung diseases such as Coal Worker's Pneumoconiosis, commonly referred to as "black lung", and silicosis. Since the mid-1990s, an alarming resurgence of diseases has been documented in central Appalachia, where underground mining often necessitates significant extraction of rock strata along with the thin seams of coal. These circumstances have prompted concern over if or how changing dust composition might be a factor in contemporary disease prevalence. Until now, the total mass concentration and quartz mass fraction of respirable dust have been regulated and monitored in US coal mines. Unfortunately, however, these two metrics alone do not paint a full picture of dust composition. Earlier work in the author's research group established a preliminary thermogravimetric analysis (TGA) method for coal mine dust. The method is intended to allow estimation of three key mass fractions of the dust from separate sources: coal from the coal strata being mined; non-carbonate minerals from the rock strata being mined or drilled; and carbonates that are primarly sourced from application of rock dust products to the mine floor or ribs. However, accuracy of the preliminary method was substantially limited by poor dust recovery from the fibrous filter media used for sample collection. This thesis includes two studies: The first study aims to establish an improved TGA method. It uses smooth polycarbonate (PC) filters for dust sampling and a modified thermal ramping routine. The method is verified using laboratory-generated respirable dust samples. In the second study, the improved TGA method is used to analyze 75 respirable mine dust samples, collected in 15 US mines. Replicate samples are also analyzed by scanning electron microscopy using energy dispersive X-ray (SEM-EDX). TGA and SEM-EDX results are compared to gain insights regarding the analytical methods and general trends in dust composition within and between mines. / Master of Science / It has long been known that chronic exposures to excessive respirable coal mine dust can lead to the development of lung diseases such as Coal Worker’s Pneumoconiosis (“Black Lung”) and silicosis. Disease rates in central Appalachia have shown an alarming and unexpected increase since the mid-1990s, despite declining dust concentrations evident from regulatory compliance monitoring data. Clearly, there is a need to better understand coal mine dust composition, which will require additional analytical methods. Thermogravimetric analysis (TGA) has been proposed as one possible method, because it should allow estimation of three key dust components from separate sources: coal from the coal strata being mined; non-carbonate minerals from the rock strata being mined or drilled; and carbonates from application of rock dust products to the mine floor and ribs. However, preliminary work with TGA showed limited accuracy, mostly due to sampling materials. In this thesis, two studies were performed. The first study aims to establish an improved TGA method using smooth, polycarbonate (PC) filters. The second study demonstrates the method on a large number of mine dust samples, and compares the results to those gained by an alternative method that uses electron microscopy.

No description available.

This thesis describes the development and use of a computer-automated microscopy routine for characterization of respirable dust particles from coal mines. Respirable dust in underground coal mining environments has long been known to pose an occupational health hazard for miners. Typically following years of exposure, coal workers' pneumoconiosis (CWP) and silicosis are the most common disease diagnoses. Although dramatic reductions in CWP and silicosis cases were achieved across the US between about 1970-1999 through a combination of regulatory dust exposure limits, improved ventilation and dust abatement practices, a resurgence in disease incidence has been noted more recently – particularly in parts of Appalachia. To shed light on this alarming trend and allow for better understanding of the role of respirable dust in development of disease, more must be learned about the specific characteristics of dust particles and occupational exposures. This work first sought to develop an automated routine for the characterization of respirable dust using scanning electron microscopy with energy dispersive x-ray (SEM-EDX). SEM-EDX is a powerful tool that allows determination of the size, shape, and chemistry of individual particles, but manual operation of the instrument is very time consuming and has the potential to introduce user bias. The automated method developed here provides for much more efficient analysis – with a data capture rate that is typically 25 times faster than that of the manual method on which it was based – and also eliminates bias between users. Moreover, due to its efficiency and broader coverage of a dust sample, it allows for characterization of a larger and more representative number of particles per sample. The routine was verified using respirable dust samples generated from known materials commonly observed in underground coal mines in the central Appalachian region, as well as field samples collected in this region. This effort demonstrated that particles between about 1-9μm were accurately classified with respect to defined chemical categories, and suggested that analysis of 500 particles across a large area of a sample filter generally provides representative results. The automated SEM-EDX routine was then used to characterize a total of 210 respirable dust samples collected in eight Appalachian coal mines. The mines were located in three distinct regions (i.e., northern, mid-central and south-central Appalachia), which differed in terms of primary mining method, coal seam thickness and mining height, and coal and/or rock mineralogy. Results were analyzed to determine whether number distributions of particle size, aspect ratio, and chemistry classification vary between and within distinct mine regions, and by general sampling location categories (i.e., intake, feeder, production, return). Key findings include: 1) Northern Appalachian mines have relatively higher fractions of coal, carbonate, and heavy mineral particles than the two central Appalachian regions, whereas central Appalachian mines have higher fractions of quartz and alumino-silicate particles. 2) Central Appalachian mines tended to have more mine-to-mine variations in size, shape, and chemistry distributions than northern Appalachian mines. 3) With respect to particle size, samples collected in locations in the production and return categories have the highest percentages of very small particles (i.e., 0.94-2.0μm), followed by the feeder and then the intake locations. 4) With respect to particle shape, samples collected in locations in the production and return categories have higher fractions of particles with moderate (i.e., length is 1.5 to 3x width) to relatively high aspect ratios (i.e., length is greater than 3x width) compared to feeder and intake samples. 5) Samples with relatively high fractions of alumino-silicates have higher fractions of particles with moderate aspect ratios than samples with low alumino-silicate fractions. 6) Samples with relatively high fractions of quartz particles have higher fractions of particles with moderate aspect ratios and higher percentages of very small particles than samples with no identified quartz particles. 7) Samples with high fractions of carbonates have higher percentages of particles with relatively low aspect ratios (i.e., length and width are similar) than samples with no identified carbonate particles. / Master of Science

Yarrowia lipolytica a la particularité de pousser sur de nombreux substrats hydrophobes et de les métaboliser via la β-oxydation ou de les stocker dans des corps lipidiques. Le premier contact entre la cellule et les substrats hydrophobes se déroule sur la surface cellulaire. A cette étape, la composition de la paroi cellulaire en protéines, en β-glucane et en chitine pourrait jouer un rôle important sur l’adhésion et la prise en charge des substrats hydrophobes. Dans le cadre de cette thèse, nous avons étudié l’effet d’une source de carbone hydrophobe (par rapport au glucose) et de la composition du milieu de culture sur la structure pariétale et la production de mélanine chez Y. lipolytica. Les résultats de nos analyses biochimiques et moléculaires ont montré que l’oléate de méthyle (un composé utilisé comme modèle de source de carbone hydrophobe) a induit des modifications au niveau de la paroi cellulaire et que ces modifications ont conduit ou non à l’adhésion de gouttelettes lipidiques à la surface cellulaire ; à une modification du contenu des composés pariétaux ; à la résistance des cellules à des composés toxiques ayant pour cible la paroi cellulaire ; et à la modification des profils d’expression de 27 gènes analysés codant des protéines de paroi cellulaire. L’identité des gènes surexprimés suggère la participation de leurs protéines dans l’intégrité de la paroi cellulaire, dans l’adhésion des gouttelettes lipidiques et dans la réponse au stress par l’oléate de méthyle et/ou le manque d’azote. Nos résultats ont aussi montré que la présence d’oléate de méthyle, huile de ricin et ricinoléate de méthyle a induit la production de DHN-mélanine chez une souche génétiquement modifiée de Y. lipolytica (MTLY40-2p) dont la β-oxydation est affectée / Yarrowia lipolytica can grow on many hydrophobic substrates and metabolize them via the β-oxidation pathway or store them into lipid bodies. The first contact between the cell and the hydrophobic substrates is by the cell wall. In this step, cell-wall proteins (CWP), β-glucane and chitin of the cell wall could play an important role for the adhesion and uptake of hydrophobic substrates. The aim of this work was to study the effect of a hydrophobic carbon source (compared with glucose) and the culture medium composition on the cell wall composition and the melanin production by Y. lipolytica. The results of our biochemical and molecular analysis showed that the presence of methyl oleate and the nutrients composition of the culture media have induced some modifications at the cell wall level. These modifications were linked with the adhesion or not of the lipid droplets to the cell surface; with a modified content of the cell wall components; with a resistance to compounds that are toxic for the cell wall, and with the modification of the expression patterns of 27 CWP genes analyzed in this work. The identity of the over-expressed CWP genes suggests the participation of their proteins in the cell wall integrity, in the adhesion of lipid droplets and in the stress response to methyl oleate and/or the nitrogen starvation. Our results have also shown that the presence of the lipids methyl oleate, castor oil and methyl ricinoleate induced the production of DHN-melanin by a mutant strain in which the β-oxidation is affected / Yarrowia lipolytica tiene la capacidad de crecer sobre numerosos substratos hidrofóbicos y demetabolizarlos a través de la β-oxidación o de almacenarlos en cuerpos lipídicos. El primercontacto entre la célula y los substratos hidrofóbicos se lleva a cabo mediante la pared celular.En esta etapa, la composición en proteínas, β-glucano y quitina de la pared celular podríajugar un papel importante en la adhesión y la toma de substratos hidrofóbicos. En el presentetrabajo se estudió el efecto de una fuente de carbono hidrofóbica (en comparación conglucosa) y la composición del medio de cultivo sobre la composición de la pared celular y laproducción de melanina en Y. lipolytica.Los resultados de nuestros análisis bioquímicos y moleculares mostraron que el oleato demetilo (un compuesto utilizado como fuente de carbono hidrofóbica) indujo modificacionesen la pared celular y estas modificaciones condujeron al aumento o la disminución de laadhesión de gotas lipídicas a la superficie celular; a la modificación del contenido de loscompuestos de la pared celular; a la resistencia de las células a compuestos tóxicos que tienencomo blanco la pared celular; y a la modificación del perfil de expresión de 27 genes quecodifican para proteínas putativas de la pared celular. La expresión de algunos de estos genesse indujo desde las primeras horas de cultivo en presencia de oleato de metilo. La identidad delos genes sobre-expresados sugiere la participación de sus proteínas en mecanismos como laintegridad de la pared celular, la adhesión de las gotas lipídicas a la superficie celular y larespuesta a estrés inducido por oleato de metilo y/o por la falta de nitrógeno en el medio.Nuestros resultados también mostraron que la presencia de oleato de metilo, de aceite dericino y de ricinoleato de metilo en el medio de cultivo, indujo la producción de un pigmentocafé en una cepa mutante de Y. lipolytica (MTLY40-2p) afectada en la β-oxidación. Ademásobservamos que el perfil espectroscópico la pared celular de la mutante MTYL40-2p, semodifica en función del substrato hidrofóbico presente en el medio de cultivo. Nuestrosresultados sugieren que el pigmento café es DHN-melanina y que la ausencia de peptona en elmedio de cultivo afecta la biosíntesis de esta melanina. A partir de un análisis in silicoproponemos los genes putativos para la biosíntesis de DHN-melanina en Y. lipolytica