Search Documents

By M. J. Reasor, M. DiMatteo, J. M. Antonini, K. Van Dyke, Z. Ye

"The inhalation of silica has been shown to produce a dramatic inflammatory and toxic response within the lungs of humans and laboratory animals. A variety of cellular and biochemical parameters are used to assess the silica-induced lung injury. The purpose of this paper is to introduce the use of luminok:Jependent chemiluminescence as a new method to study inflammation in both phagocytic cells and lung tissue recovered from silicaexposed animals. Chemiluminescence, or the emission of light, accompanies the release of reactive forms of oxygen when phagocytic cells are challenged. In this study, male Fischer 344 rats were intratracheally instilled with either silica (10 mg/100 g bw) or saline vehicle. One day after the instillations. a marked increase in the chemiluminescence was observed in the lung tissue and bronchoalveolar lavage cells recovered from the silica- treated animals when compared with the saline controls. The light reaction was markedly decreased by either superoxide dismutase or N-nitro-Larginine methyl ester hydrochloride. Superoxide dismutase is involved in the enzymatic breakdown of superoxide anion. while N-nitro-L-arginine methyl ester hydrocholoride, a nitric oxide synthase inhibitor, prevents the formation of nitric oxide. When superoxide anion and nitric oxide react. they form the highly oxidizing substance peroxynitrite. This study then implicates peroxynitrite as an agent that may be responsible for some of the oxidant lung injury that is associated with silica exposure. The use of luminol-dependent chemiluminescence may prove valuable as a method to measure the earliest events in the inflammatory process. and may be an adjunct in studying the mechanisms that produce inflammation. - Environ Health Perspect 102(Suppl 10):37-42 (1994) IntroductionResearch on inflammation has not produced a method that links the initial events of the process to the controlling events. Rather, a variety of parameters are measured that are the result of and not the basis of the inflammatory reaction. Parameters such as tissue weight, edema, blood flow, enzyme and protein release, or production of metabolites (prostaglandins, leukotrienes) have been used extensively. A method is needed that could be applied to cells of a particular type, groups of cells of different types, and even tissue, in an attempt to study the kinetics of the inflammation as well as being sensitive enough to monitor the early events of the process . Since inflammation is so complicated, the relationship between the control of the initial events and the oxidation process has never been explained nor studied in any derail."

Dec 1, 1995

By A. W. Khair, L. D. Gehl

"This paper describes the study of bit wear characteristics on three types of rock namely Tennessee sandstone, Berea sandstone and Indiana limestone. High content of quartz with quartz cement in Tennessee sandstone causes removal of material from the bit body and dam2ge to the tip of the bit resulting into excessive weight and height loss as compared to other two types of rock. The drum speed and water spray pressure do not contribute significant effect on bit wear. The polycrystalline compact diamond (PCD) tipped bits are more wear resistant than the ordinary tungsten carbide (WC) tipped bits, but, due to wider body of PCD bits, the wear rate increases particularly at greater depth of cut. INTRODUCTIONThe impact of bit-rock interaction during the cutting process in underground coal mines has always attracted the-interests of mining operators, researchers and the manufacturers. Coal cutting bears directly on airborne coal dust generation which causes ""black lung"" in coal workers. On the other hand, rock cutting generates frictional sparks causing face ignitions. The bit wear not only produces airborne rcspirable coal dust , but also affects the productivity, safety and economy. Hun and Evans (J) concluded that the point attack bits have greater ignition potential than the conventional wedge shaped bits. The results of a study (2) suggested that after 7% of weight loss, the bit should be replaced. In a survey study (3), it was found that the bit consumption of bit had an effect on the productivity and utilization of the continuous miners. The mining industry is yet to receive a solution which could provide them with optimum operating parameters for coal cutting machines in underground C9al mines.The study was, therefore, taken up to determine wear characteristics as influenced by the rock propenies and varying operating parameters. For the purpose of this study, three types of rock, Tennessee sandstone, Berea sandstone and Indiana limestone were used. The machine operating parameters included (i) drum speed, (ii) water spray pressure and (iii) bit type. Besides studying the wear characteristics, the effects of these parameters on the required cutting force, specific energy consumption, acoustic emission generation and bit temperature rise were also studied. Also, the preliminary experiments were performed to compare the bit wear between PCD and WC tipped bits."

Dec 1, 1995

By T. R. Taubert, D. H. Carlson, J. H. Johnson

"ObjectiveDevelop a portable emissions measurement apparatus to sample and measure tailpipe concentrations of diesel particulate matter (DPM), carbon monoxide (CO), carbon dioxide (COJ, nitric oxide (NO), and nitrogen dioxide (NOJ under steady-state engine load and speed conditions during a 1-minute test.BackgroundThe U.S. Bureau of Mines (USBM) has a comprehensive research program to develop ways to reduce exposure of miners to diesel exhaust pollutants. Engine maintenance has been shown to have important effects on such exposure.Accurate measurements of tailpipe emissions are becoming an essential part of a good maintenance program for underground mines that use diesel-powered vehicles. Past studies of particulates in U.S. underground coal mines have shown that DPM alone may be present at concentrations greater than 1 milligram per cubic meter (mg/m3), half the 2-mg/m3 limit for respirable coal mine dust. A poorly tuned (smoking) diesel engine may be the sole cause for respirable dust concentrations exceeding the limit. Although there is currently no standard specifically for DPM in U.S. mines, regulatory steps are underway, and significant reductions of DPM levels in both coal and metal mines will most likely be required.Tailpipe emission measurements can provide diagnostic information to identify engine malfunctions such as clogged air cleaners, faulty injectors, or incorrect injection timing. In some cases, mine vehicle tailpipe DPM concentrations have been reduced up to 50 percent by engine maintenance and adjustments.An accurate measuring instrument would also be useful for determining the effects of exhaust control devices on tailpipe emissions. Little information is currently available about the overall effectiveness of exhaust controls after long-term use. Raw exhaust concentrations with and without the control device can be measured in underground mines, and from these data a control efficiency can be calculated. Knowledge of the effectiveness of exhaust control devices over time is needed to make intelligent decisions on bow to use and maintain these devices, and when to replace them."

Dec 1, 1995

By A. W. Kbair, L. D. Gehl

"INTRODUCTION The rock-bit interaction during the cutting process in underground mines has been a matter of great concern to mining operators, researchers and manufacturers, particularly after the passage of strict environmental laws. Coal cutting bears directly on airborne coal dust generation and rock cutting generates frictional sparks which may cause face ignitions, jeopardizing the productivity, safety and economy of the industry. It is imperative to investigate into the factors which have influence on bit we.ar. Hurt and Evans (1981) concluded that the point attack bits have greater ignition potential than the conventional wedge shaped bits, and also worn bits were more responsible for ignition than the new bits. The results of a study (Khair et al., 1992) suggested that after 7% weight loss, the bit should be replaced. The polycrystalline compact diamond (PDC) tipped bits are reported to be cost effective in a study done by USBM (Twin Cities Research Center), and it was stated that the PDC tipped drill bits had 80 to 100 times longer life than the ordinary tungsten carbide (WC) tipped bits.This paper presents the results of the experiments which were conducted utilizing a coal cutting simulator at the laboratory of the Department of Mining Engineering, West Virginia University, USA. The description of the machine and the experimental set-up have been described elsewhere (Khair et al., 1989; Gehi & Khair, 1994), and therefore, will not be discussed here. The rock types which were used in experimen~ are Tennessee sandstone, Berea sandstone and Indiana limestone. The operating parameters include (1) drum speed, (2) water spray pressure and (3) bit type. Besides studying the wear characteristics influenced by the above mentioned variables, the effect of these parameters on the required cutting force, specific energy consumption, acoustic emissions rate and rise in temperature of the bit were also studied. A preliminary study was also done to compare we.ar characteristics of the WC bits with those of the poly-crystalline compact diamond tipped bits."

Dec 1, 1995

By S. L. Weber, V. Castranova, D. E. Banks, J. K. H. Ma, Y. Rojanasakul, J. Y. C. Ma, M. Bhat

"The mechanism of bleomycin-induced pulmonary fibrosis is not ye: clear. Recent studies have shown that alveolar macrophages (AM) can be stimulated by bleomycin in vitro releasing inflammatory cytokines, suggesting that the interaction of bleomycin with AM is an important step in the drug-induced fibrotic process. Bleomycin is known to bind DNA and generate oxygen radicals through complexation with Fe2+ and oxygen. To provide,more insight into the cellular oxidative property of bleomycin, v.s have developed a fluoromicroscopic method using 2',7'-dichlorofluorescin diacetate (DCFHDA) as an oxidative fluorescence probe to study the deomycin-induced intracellular oxidation in rat AM and the inhibition of the oxida¬tion by taurine, a compound known to inhibit the bleomycin-induced fibrosis. Bleomycin at 5 to 20 ug/ml has a moderate stimulatory effect (1.87- to 2.66-fold) on the secretion of superoxide anion. A high concentration of bleomycin (20 ug/ml), however, inhibits cell response to zymosan-induced secretion of superoxide anion. At 4 ug/ml, bleomycin has no effect on cell membrane integrity or morphology but results in a significant increase in intracellular oxidation. This oxidative process is Fe2-,-dependent and is accompanied by an increase in intracellular calcium (35 nM). Both the intracellular oxidation and calcium rise induced by internalized bleomycin are inhibited by pretreatment of cells with varying concentrations of taurine (25, 125, and 187.5 uM). The inhibitory effect on intracellular oxidation .vas found to be 36, 57, and 60%, respectively. These results demonstrate a stimulation-inhibition relationship between bleomycin and taurine on re cellular oxidation at a subcytotoxic dose of bleomycin, suggesting that the oxidative effect of the intracellular bleomycin-Fe2 complex is important in the initiation of the fibrotic process. — Environ Health Perspect 102 (Suppl10):91-96 (1994) IntroductionThe pulmonary system is most susceptible to oxygen radical-induced diseases due to its constant exposure to the highest oxygen tension, and the potential encounters with oxidant gases such as ozone, nitrogen oxides, and toxic doses of dust particles. The lung is also a well-perfused organ and thereby vulnerable to the effect of circulating xenobiotics such as paraquat, and, bleomycin, which is known to cause pulmonary injuries (1). Alveolar macrophages (AM), which act as a means of defense by phagocytizing inhaled dust particles and microorganisms while increasing production of reactive oxidative species, have been shown to play an important role in mechanisms leading to oxidative lung injury and fibrosis (2-4)."

Dec 1, 1995

By A. W. Khair, L. D. Gehl

"This paper describes the mechanism of coal and rock cutting from quasi-static indentation through dynamic indentation to rotary cutting. The results indicate that during quas~static indentation, a crushed zone, and subsurface fractures develop. The crushed zone is a potential source of dust generation. Dynamic indentation produces a crushed zone due to the frictional resistance offered by the coal or rock during penetration of the bit. Confining pressure changes the mode of failure from tension to mixed tension and shear. The wider bit causes larger crushed zone and more dust generation. The rotary cutting is a combination of quasi-static and dynamic indentation which results in chip formation. 1. INTRODUCTIONThe trend from conventional mining to continuous machine-cutting of coal, using more productive machinery is inexorable. Intolerable levels of dust generated at the face is an indication of inefficient coal or rock cutting by these machines. The Federal Government of the United States paid about 11.7 billion dollars workman compensation to more than 470,000 miners with Coal Workers Pneumoconiosis (Newmeyer, 1981). Bieniawski and Zipf (1985) reported that annual benefit payments were about 2 billion dollars. The need for research m improving coal cutting 1s therefore paramount. The enactment of Health and Safety Act in 1969 in the United States triggered the initial investigation into coal cutting mechanism and cutting tool design. The European countries were also engaged in the same area of research, however, their emphasis was in the use of wedge shaped bits. In 1981, the United States Bureau of Mines commissioned the South West Research Institute to investigate the effect of operating parameters mainly on airborne respirable dust in order to provide a healthier and safer atmosphere for coal mine workers.A research program by the Department of Mining Engineering, West Virginia University, USA was initiated to study the fragmentation mechanism during coal/ rock cutting. The research program was divided into three areas of research, namely quasi-static indentation, dynamic indentation and rotary cutting."

Dec 1, 1995

By B. Z. Zhong, T. Ong, W. E. Wallace, W. Z. Whong, Z. W. Gu, M. J. Keane

"Abstract-Diesel emission particles (DEP) from an engine operating on the Federal Test Procedure VDC were dispersed in an aqueous mixture of dipalmitoyl lecithin (DPL). a major component of pulmonary surfactant and saline and were tested for genotoxicity using the unscheduled DNA synthesis (UDS) assay with autoradiography techniques in cultured Chinese hamster lung cells. A DEP sample from the same source was dispersed in dimethyl sulphoxide (DMSO) and subjected to the same assay. Both dispersions (in DPL and DMSO) were found to induce UDS in a dose-related manner. After separation of the sample into supernatant and sediment fractions. the activity of the DEP sample was found to reside in the supernatant fraction for the DMSO-dispersed sample; and in the sedimentary fraction for the surfactant-dispersed sample. These findings further indicate that genotoxic activity associated with DEP inhaled into the lung may become bioavailable through the dispersion properties of pulmonary surfactant components.INTRODUCTIONTHE mutagenicity of diesel emission particles (DEP) has been studied intensively (LEWTAS and WILLIAMS, 1986). Carcinogenic effects of diesel exhaust in exposed animals have been reviewed (NIOSH, 1988) and an evaluation of carcinogenic risks to humans from diesel engine exhausts has been published (IARC, 1989). Using components of pulmonary surfactant such as dipalmitoyl lecithin (D PL), dipalmitoyl phosphatidylethanolamine (DPPE) or dipalmitoyl phosphatidic acid (DPPA) dispersed in physiological saline, we have previously shown that diesel soots express mutagenic activities in the Salmonella histidine reversion assay system and in the sister chromatid exchange assay using the cultured Chinese hamster lung (V79) cells. Mutagenic activity of diesel soots dispersed in DPL, DPPE and DPPA was quantitatively comparable to and in some cases greater than, the activity expressed by an equal mass of soot extracted by dichloromethane-dimethyl sulphoxide (DCM-DMSO) (WALLACE et al., 1989).Unscheduled DNA synthesis (UDS) is accepted and frequently included, as one of the tests in the battery of assays used to assess carcinogenic-mutagenic potential -of chemicals (MITCHELL et al., 1983). In this study, induction of UDS in V79 cells by DEP, dispersed in the major component of pulmonary surfactant DPL, was carried out. The purpose of this study is to validate the suggestion generated from our previous work that mutagens associated with inhaled particles may be dispersed or solubilized in the phospholipid component of pulmonary surfactant and become active in such a phase (WALLACE er al., 1989) and to test the fractionated DEP sample using UDS assay for a more complete evaluation of its genotoxicity."

Dec 1, 1995

By J. B. Vander Sande, L. C. Rainey, P. Bolsaitis, B. Dirsa

"The structure and composition of silica-rich particles recovered by lavage from the lungs of three active miners with different medical histories were studied using high-resolution electron microscopy and chemical microanalysis. The results arc compared to the similarly determined structure and composition of respirable-size mineral particles obtained from roof-bolter dust-box samples from two coal mines of widely different bulk quartz concentrations. The results show that the lungs of the miners contain silica-based particles with structures not found in the mine samples. Also, the particle structures and compositions found in the macrophages were different in each of the miners. The rau.lu suggest the possibility that intracellular processes may affect the susceptibility of individuals to silica-induced pneumoconiosis. The cytotoxic and fibrogenic potentials of mine dusts result from the activity of their individual components. Whether this effect is additive or even mass (i.e., dose) related is questionable and has been the subject of many investigations. Particle size and size distribution of various constituent minerals, the surface structure and properties of individual particles, and the self-cleaning capability of the lung (i.e., the capacity of the macrophage system) may, among other variables, affect the pathogenic changes induced by dusts lodged inside lungs. Extensive epidemiological investigations conducted on miners from different coalmining districts, where equivalent dust exposures have been shown to result in vastly different frequencies of lung disease incidence, also confirm this hypothesis (1) In a recent study, Ferrer et al. (2) used energy-dispersive X-ray analysis (EDXA) to investigate the inorganic element content in pleura and lung in reference, silica exposed non-pneumoconiosis, and silicotic populations. Constant depositions of silicon and calcium were detected in visceral pleura, parietal pleura, and lung of the refence group and in visceral pleura and lung of the exposed non-pneumoconiosis and silicotic groups. Comparison of the silicon content in pleura between silicotic and exposed non-pneumoconiosis subjects showed a nonsignificant probability of difference, whereas there were no differences with respect to the silicon content in lungs."

Dec 1, 1995

By M. J. Reasor, J. M. Antonini, L. Wu, K. Van Dyke, Z. Ye

The inhalation of silica has been shown to produce a dramatic inflammatory and toxic response within the lungs of humans and· laboratory animals. Currently, no effective treatment exists for workers who may have been exposed to the inhalation of silica. The objective of this study was to develop an animal model in which we could evaluate the effect that anti-inflammatory steroids have on the acute silica-induced pulmonary inflammatory response. Male Fischer 344 rats were pretreated with either dexamethasone (2 mg/kg) or saline vehicle (i.p.) on days 1, 3, and 5. On day 6, the animals from the two groups were then intratracheally instilled with either silica (20 mg/0.5 ml saline vehicle) or saline vehicle (0.5 ml). Twenty-four hours after the instillations in the non-steroid group, significant increases occurred in total protein, total number of cells, neutrophils, and lymphocytes recovered from the lungs of animals treated with silica compared to saline controls. Silica also caused dramatic increases in the luminol-dependent chemiluminescence (LDCL) of lung tissue and bronchoalveolar lavage (BAL) cells. The LDCL reaction was markedly decreased by either superoxide dismutase (SOD) or N-nitro-L-arginine methyl ester hydrochloride (L-NAME). SOD is involved in the enzymatic breakdown of superoxide anion, while L-NAME, a nitric oxide (NO) synthase inhibitor, prevents the formation of NO. When the superoxide anion and NO react, they form the highly oxidizing substance peroxynitrite. This study then implicates peroxynitrite as an agent which may be involved in the silica-induced oxidant lung injury. When the animals were pretreated with the steroid dexamethasone, there was a complete protection against the biochemical, cellular, and chemiluminescent indices of damage caused by silica. The mechanism in which the steroid protects the lung from damage may be due to the ability of dexamethasone to block the induction of NO synthase. With further study in animals, the anti-inflammatory steroids may be useful in the treatment of silicainduced lung injury.

Dec 1, 1995

By H. Netuka, J. V. Horak, A. W. Khair

"A general review of major problems concerning mathematical modelling of roof bolting problem is given. A mathematical model (MM) of interactive set Bolt - Resin - Rock and its discretization based on the Finite Element Method (FEM) is developed to calculate numerically course of displacement, stress and strain fields in this set. This new MM enables to get also course of the normal and tangential components of contact stress vector, normal displacement and then mutual tangential slip at the bolt - resin and resin - rock interfaces, respectively.Behavior of system on all interfaces is modelled in the MM as a ""unilateral contact"" governed by so called non-classical boundary condition of Signorini's type including Coulomb's law of friction between resin and rock, and ""a given friction"" law between bolt and resin (see Duvaut, Lions (1972) for mathematical formulation).Numerical example and some values of resulting stresses are shown and remarks on necessary and sufficient conditions for solvability of the problem are also given.IntroductionAs is well known roof bolting has become the primary support system in almost all underground coal mines. But the ""theoretical"" design criteria for roof bolt patterns are rather problematic because: "" ... real mechanisms by which the roof bolts reinforce the immediate roof of underground coal mine entry are still relatively unknown"", see Peng (1986).The aim of this contribution is to present brief description of fundamentals of mathematical theory (see Hlavacek et al., 1988) which could be, according to our opinion and knowledge, very helpful for numerical modelling of a such type of problems. For the sake of brevity we restrict ourselves here to just one of the simplest possible problem: analysis of the behavior of interactive set composed by only single full - length - grouted bolt, resin (or cement) and suitable part of domain of a ""immediate"" neighborhood of bolt which should represent physical action of real rock material."

Dec 1, 1995

By R. L. Grayson, W. E. Wallace, J. W. Stephens, M. D. Attfield, P. S. Brower, V. Vallyathan, M. J. Keane, J. C. Harrison

Pure respirable quartz is a known pathogenic agent for lung disease. However, for mixed dusts, neither mineralogical nor in vitro cytotoxicity assays unequivocally predict the contribution of quartz to their pathogenic potential.First, although Coal Workers' Pneumoconiosis (CWP) and Progressive Massive Fibrosis (PMF) prevalence have been found to correlate with total respirable coal mine dust exposure, and with coal rank (Attfield and Morring, 1992), epidemiological studies generally have not found the quartz component of coal mine dust as measured by X-ray diffraction or infrared spectroscopy (each of which essentially measures the mass-percent quartz in a bulk sample) to be clear predictors of CWP prevalence or incidence (Walton et al., 1971). Second, the properties which distinguish the extreme fibrogenic potential of quartz are not revealed by conventional in vitro bioassay comparison with other silicate mineral dusts. As an example, quartz and kaolin (aluminosilicate) clay in vitro cytotoxicities are comparable for lavaged primary pulmonary macrophages.The studies discussed here address these two anomalies from a general hypothesis that the pathogenic activity of respired quartz particles is associated with properties of, and interactions with, the particle surfaces. One factor in the expression of toxic activity by a particle surface is the biological availability of that surface. Anomalous lack of correlation of disease prevalence with mass percent quartz in exposures to some mixed composition respirable-sized dusts suggests that individual particle surface features, invisible by bulk sample mass analysis, may alter the biological availability of a fraction of quartz particles in a dust. For pure-surfaced mineral particles, the comparably strong prompt membranolytic activities of quartz and kaolin in vitro suggest that upon deposition of such particles in the lung there must be other interactions with particle surfaces which distinguish quartz. This might be due to toxic surface-associated sites specific to silica, or to interactions of other sites on other mineral particles which affect the biological availability of their toxic functional groups. For example, alurninol groups on clays affect the availability or activity of adjacent silanol groups. Surface activities which distinguish quartz pathogenici ty might be manifest in one or more of the sequential interactions upon deposition in the lung; first with the pulmonary surfactant system, then with extracellular and phagolysosomal enzymatic digestive processes, or with subsequent cytotoxic processes.

Dec 1, 1995

By D. L. Nussear, W. E. Wallace, N. N. Clark, M. Gautam, G. Hong-Guang

"Results from a study directed towards identifying and measuring the mutagenicity of diesel exhaust particulate matter involved in mining exposures, especially those which may synergistically affect genotoxic hazard, are presented in this paper. Particulate matter emissions from a diesel engine, representative of the ones found in underground mines, were sampled and assayed to determine the genotoxic potential as a function of engine operating conditions. Diesel exhaust was diluted in a multi-tube mini-dilution tunnel and the particulate matter was collected on 70 mm fluorocarbon coated glass fiber filters as well as on 8"" x 10"" hi-volume filters. A six mode steady state duty cycle was used to relate engine operating conditions to the mutagenicity potential.Protocols developed for using a primary component of pulmonary surfactant as a medium for collecting and applying respirable particulate material to cells for mutagenicity testing offer a true representation of in-vivo delivery of such respired aerosols to pulmonary cells and the genetic material within . The surfactant dispersion technique is discussed in this paper. Methods have been developed to sample and assay the diesel soot and diesel soot/mineral dust complexes which would not compromise the surface and physical properties of the particulate material."

Nov 1, 1995

By D. E. Banks, J. K. H. Ma, Y. Rojanasakul, J. Y. C. Ma, C. J. Malanga

There is evidence to suggest that cell injury induced in alveolar macrophages (AM) following phagocytic activation by silica particles may be mediated through changes in intracellular free calcium [Ca 2+}. However, the mechanism of silicainduced cytotoxicity relative to [Ca2+], overloading is not yet clear. To provide a better insight into this mechanism, isolated rat AMs were exposed to varying concentrations of crystalline silica (particle size< 5 µm in diameter) and the fluctuation in their [Ca2+]; and cell integrity were quantitatively monitored with the fluorescent calcium probe, Fura-2 AM, and the membrane integrity indicator, propidium iodide (Pl). Results from this study indicate that silica can rapidly increase [Ca2+]; in a dose-dependent manner with a characteristic transient calcium rise at low doses (<0.1 mg/ml) and an elevated and sustained rise at high doses (>0.1 mg/ml). Depletion of extracellular calcium [Ca 2+]0 markedly inhibited the [Ca2+]; rise (˜90%), suggesting that Ca2+ influx from extracellular source is a major mechanism for silica-induced [Ca2+]; rise. When used at low doses but sufficient to cause a transient [Ca2+]; rise, silica did not cause significant increase in cellular Pl uptake during the time of study, suggesting the presevation of membrane integrity of AMs under these conditions. At high doses of silica, however, a marked increase in Pl nuclear fluorescence was observed. Depletion of [Ca 2+] 0 greatly inhibited cellular Pl uptake, induced by 0. 1 mg/ml or higher doses of silica. This suggests that Ca 2 + influx, as a result of silica activation, is associated with cell injury. Indeed, our results further demonstrated that the low dose effect of silica on Ca 2+ influx is inhibited by the Ca2+ channel blocker nifedipine. At high doses of silica (>0.1 mg/ml), cell injury was not prevented by nifedipine or extracellular Ca2+ depletion, suggesting that other cytotoxic mechanisms, i.e., nonspecific membrane damage due to lipid peroxidation, are also responsible for the silica-induced cell injury. Silica had no significant effect on cellular ATP content during the time course of the study, indicating that the observed silicainduced (Ca2 ... ); rise was not due to the impairment of Ca2 + -ATPase pumps, which restricts Ca2+ efflux. Pretreatment of the cells with cytochalasin B to block phagocytosis failed to prevent the effect of silica on [Ca2+]; rise. Taken together, these results suggest that the elevation of [Ca2+]; caused by silica is due mainly to Ca2+ influx through plasma membrane Ca2+,channels and nonspecific membrane damage (at high doses). Neither ATP depletion nor Ca2+ leakage during phagocytosis was attributed to the silica-induced [Ca2+]; rise.

Nov 1, 1995

By P. R. Miles, J. K. H. Ma, J. Y. C. Ma, L. Bowman

"The bisbenzylisoquinoline alkaloids are compounds which can be isolated from plants and which exhibit a broad range of pharmacological activities. Perhaps the best known alkaloid of this group is tetrandrine (TT), a compound isolated from the root of a traditional Chinese medicinal herb, Stephania tetrandra. It has been tested for its effects on the cardiovascular system, tumors, and the lungs. For example, tetrandrine has been shown to act as a calcium entry blocker in cardiac sarcolemmal vesicles (King et a/., 1988) and has been used in China as a treatment for angina and hypertension. Its possible effects as a tumor-inhibitory substance have also been investigated (Kupchan and Atland, 1973). In addition, there are several lines of evidence to suggest that it is a possible therapeutic agent for the treatment of silicosis. Yu et a/. (1983) have shown that TT inhibits the development of experimental silicosis in rats when it is administered orally after intratracheal instillation of quartz. Other studies have shown that the alkaloid inhibits the synthesis of collagen by fibroblasts and the formation of silicotic nodules in rats exposed to silica (Huang et al., 1981; Liu et al., 1983). In fact, tetrandrine has been used clinically in China as an antifibrotic agent and has been shown to improve both the appearance of chest radiographs and the measurements of diffusing capacity in patients with silicosis (Li et al., 1981).Since tetrandrine may have a potential use as a therapeutic agent for fibrotic lung diseases, its effects on various pulmonary cells and tissues should be studied. Alveolar type II cells are located in the alveolar regions of the lungs and are important because they are responsible for the synthesis and secretion of pulmonary surfactant (Dobbs et al., 1982; King, 1984), regeneration of the alveolar epithelium after lung injury (Kauffman et al., 1974), and metabolism of foreign compounds (Bend et al., 1985). Furthermore, in silicosis, for which TT has been suggested as a treatment, there are increased amounts of pulmonary surfactant produced (Dethloff et al., 1986a,b) as a result of increased rates of surfactant synthesis by type II cells (Dethloff et a/., 1989; Miller et al., 1988). Because type II cells play a critical role in maintaining normal lung function and because type II cells are affected in silicosis, for which 'TT has been used as a treatment, we studied the effects of tetrandrine on energy metabolism in these cells in vitro. Energy metabolism was chosen because it is a basic cellular function which is neces-sary for other cellular processes to occur. Since type II cells depend on mitochondrial metabolism for energy production (Fisher et al., 1980), we examined the effects of tetrandrine on cellular and mitochondrial ATP production and oxygen consumption. Additional experiments were performed in an effort to determine if intracellular calcium levels may be involved in the TT-induced effects. A preliminary report of those results has appeared previously (Ma et al., 1991)."

Nov 1, 1995

By K. Clifton, K. Kar, M. Gautam, J. T. Jurewicz

"This paper describes the history of LDV measurements employed in research involving gas-solid flows. Much of the discussion deals with measurement techniques employed to obtain separate and simultaneous solid and gas phase velocities. Also, LDV based measurement techniques that arc used to obtain particle size and concentration information in conjunction with the velocity measurements arc presented.INTRODUCTIONThere are numerous examples of industrial processes in which the motion of a solid or droplet dispersed phase in a suspending gas phase is critically important. The applications range from spray droplets injected into combustion chambers to the understanding of the time release mechanisms of particles generated by dissolving pharmaceutical tablets. In many of these processes, the heat, mass, and/or momentum transfer between the dispersed phase and the suspending fluid arc the items of interest. In the past, much of the phenomena were characterized by empirical models derived from flow averaged measurements. Gradual developments in laser Doppler velocimetry (LDV), have provided researchers with an unobtrusive measurement technique capable of obtaining the particle size velocity and concentration with good spatial resolution.In the two decades that LDV instrumentation has been available, most of the uses have concentrated on single phase fluid flow measurements. The light scattered from seed particles small enough to follow complex fluid motions, including turbulence, is converted into velocity information. While the utility of this instrument in single phase flows has been demonstrated beyond question, the use of LDV in gas-solid multiphase flows has had a more gradual development. The measurements ·of interest in these flow regimes are usually the velocity of the suspending fluid as well as that of the dispersed particulate phase. Early investigators were quick to see the potential of this instrument for measurements in gas-solid flows. The interest soon expanded to the development of instruments that could also simultaneously measure particle size and concentration within a two-phase flow field.This paper tracks the historical development of LDV technology as applied to gas-solid flows. The techniques to measure particle velocity, size, and concentration arc treated in separate sections although much of the development occurred simultaneously. Finally, some unique applications conducted at West Virginia University and other locations will be discussed in the application section."

Nov 1, 1995

By R. V. Ramani, R. L. Grayson, R. L. Frantz, V. A. Marple, W. R. Myers, K. L. Rubow, Mutmansky. J. M., T. J. Stobbe

"An investigation of protocols for research performed from 1953 to 1988 on pneumoconioses (with emphasis on diseases related to coal and silica) was conducted in the Generic Mineral Technology Center for Respirable Dust. In addition, interviews were conducted with prominent U.S. medical researchers, aerosol scientists and engineers in government, industry, and universities to determine dust sources, dust preparation methods, laboratory protocols, and determinant variables related to in vitro and in vivo research and assess the comparability of results.Since the original report on the protocol project, additional research regarding free radical existence in freshly produced dust particles and the shape and contamination of particles generated for research purposes has further heightened scientists' concerns. As indicated by current research, fresher dust particles and those with more angular shapes may have a different level of reactivity in biological systems than do older or more rounded particles. Therefore, research on dust preparation methods for biomedical studies was undertaken and will be outlined here.Results of protocol review and interviews are summarized. Research on size reduction methods and their particle characteristics plus some recommendations concerning dust preparation methods and reporting in biomedical publications are also presented. Recent evidence indicates that these recommendations may be of importance in comparing and interpreting research results."

Nov 1, 1995

By V. A. Marple, K. L. Rubow, B. A. Olson

"INTRODUCTION Inertial classification, gravitational sedimentation centrifugation, and thermal precipitation arc techniques that can be used to collect particles for subsequent analysis or for particle classification. The inertial classifiers, which include impactors, virtual impactors, and cyclones, are widely used in the sampling or particles. Settling chambers, which include centrifuges as well as gravitational settling devices, are less widely used. Thermal precipitators arc seldom used.Impactors, in particular, have been used extensively, studied both theoretically and experimentally, and arc the instrument or choice for the determination or aerosol mass size distributions. Many versions or impactors arc available commercially and many more have been designed, built, and used in special studies.Virtual impactors are a more recently developed inertial classifier and possess a feature not round in conventional inertial impactors; namely, that the particles remain airborne after classification compared to the conventional impactor, where the particles arc collected on a solid surface. The feature or keeping particles airborne is important for transporting the particles to other particle-analyzing instruments, or to filters, and in concentrating particles larger than the cutsize in a small fraction of the total now."

Nov 1, 1995

By R. H. Reist, R. D. Dey, V. Castranova, J. P. Durham, Y. Rojanasakul

"Silicosis is a fibrotic pulmonary disease produced by inhalation of silica-containing dusts (Peters, 1986). Chronic silicosis may develop 20-40 years after initial exposure to crystalline silica, and the disease may progress from a simple to a complicated form. Few symptoms are evident in simple silicosis with pulmonary function being relatively normal. However, chest radiographs reveal small rounded ·opacities in the upper lobes of the lungs. These silicotic nodules consist of collagen arranged in a distinctive circular pattern. As fibrotic changes become more severe, the disease progresses to complicated silicosis (progressive massive fibrosis). Complicated silicosis is associated with shortness of breath and decreases in pulmonary function characteristic ofrestrictive lung disease. Chest x rays reveal numerous large silicotic nodules which may be associated with scar emphysema. Several mechanisms have been proposed to explain the cellular events which result in silica-induced lung damage and fibrosis (Lapp and Castranova, 1993). They include: ( l) direct cytotoxicity of silica, (2) activation of oxidant and enzyme release from alveolar macrophages, (3) recruitment and activation of polymorphonuclear leukocytes, and (4) stimulation of fibroblast proliferation and collagen synthesis.Tetrandrine is a bisbenzylisoquinoline alkaloid derived from the Chinese herb Radix Stephania tetrandra. It has a molecular weight of 622.73 and an empirical formula of C38H420 6N2. The chemical structure of tetrandrine has been published previously by our laboratory (Castranova et al., 1991 a). Its structure is characterized by methoxy groups at C7 and C12 , uncharged nitrogens at N2 and N’2 and two 17-carbon ring members connected by a double oxygen bridge between Ca-C7 and C11-C12. Tetrandrine has been shown to reduce both silica-induced fibrosis and the elevation of lung collagen content in rats (Huang et al .. 1981; Yu et al., 1983; ldel, 1987). Furthermore, tetrandrine has been used as an antifibrotic agent in Chinese clinical trials (Li et al.. 1981 ). Preliminary results suggest that administration of tetrandrine to patients with silicosis results in substantial improvement of symptoms, increases in diffusion capacity, and decreases in the size of shadows on lung x rays."

Nov 1, 1995

By J. L. Stauffer, S. L. Lacey, L. J. Gaydos, L. M. Demers, D. C. Kuhn

"Tumor necrosis factor alpha (TNFa) and thromboxane A2 (TXA2) are major products of the activated alveolar macrophage and serve as key mediators of lung injury. In order to determine if the synthesis ofTXA2 and the release ofTNFa are associated, the production of these inflammatory agents by the human alveolar macrophage (AM), as a result of activation by lipopolysaccharide (LPS), was assessed in the absence and presence of the thromboxane synthase inhibitors UK 38,485 (Dazmegrel) and OKY 046. UK 38,485 and OKY 046 inhibited both LPS-stimulated TXA2 production and TNFa release in a dose-dependent manner. Prostaglandin E2 (PGE2) production was not increased by UK 38,485 or OKY 046. Neither LPS nor UK 38,485 had any effect on LT84 production by AM. Neither UK 38,485 or OKY 046 had any effect on LPS-stimulated interleukin-I beta release. However, the TXA2 mimetic, U46619, did not stimulate TNFa release by AM either in the absence or presence of UK 38,485. These findings suggest that 1) UK 38,485 and OKY 046 are inhibitors of both TXA2 production and TNFa release by activated human AM, 2) UK 38,485 probably does not exert its inhibitory action on TNFa release through effects on eicosanoid production and 3) the possibility that TNFa- and TXA2-induced lung injury may be subject to amelioration by imidazole-based compounds should be further evaluated.IntroductionTumor necrosis factor alpha (TNFa), produced by monocytes and macrophages, is a potent peptide mediator of lung injury (1,2). The exaggerated release of TNFa has been implicated in the etiology of acute lung reactions such as septic shock of bacterial or viral origin (3,4). The role of TNFa is expressed primarily through its ability to activate a variety of cell types within the lung environment including polymorphonuclear leukocytes (PMNs, ref. 5), type II epithelial cells (6) and fibroblasts (7). The role of lipid mediators in lung disease has also been extensively characterized (1). In particular, the relationship between pulmonary inflammation and fibrosis and the production of oxygenated derivatives of arachidonic acid (here termed ""eicosanoids"") appears to be a key factor in a variety of disease processes in the lung (8). The alveolar macrophage (AM) synthesizes primarily prostaglandin E2 (PGE2). thromboxane A2 (TXA2) and leukotriene 84 (LT84, ref. 9). These eicosanoids have been shown to produce a variety of effects in the lung including bronchial smooth muscle dilatation (PGE2, ref. 10), vascular smooth muscle contraction and platelet aggregation (TXA2, ref. 11) and oedema and immune cell extravasation (LT84, ref. 12)."

Nov 1, 1995

By N. S. Dalal, A. H. Hoffman, X. Shi, D. E. Banks, J. K. H. Ma, Y. Rojanasakul

Excessive production of reactive oxygen radicals by alveolar macrophages is proposed to play an important role in oxidative lung injury. A major product of oxygen radical formation is the highly reactive hydroxyl radical (•OH) generated via a biologic Fenton reaction. In addition to its known ability to induce lipid peroxidation, recent studies have suggested that the •OH may exert its cytotoxic effect through the alteration of [Ca2+], homeostasis. To test this potential mechanism as well as to investigate the relationship between •OH and Ca" overloading in cytotoxic injury, isolated rat alveolar macrophages were exposed to externally generated radical system, H2O2 (0.01 to 1 mM) and Fe (1 mM) and their [Ca2+] levels and cell injury were monitored using quantitative fluorescence microscopy with the aid of the specific Ca" indicator, Fura-2, and membrane integrity indicator, propidium iodide. Electron spin resonance measurements using the spin trap 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) confirmed the production of the .OH radical by this system. Upon the addition of the radicals, the macrophages displayed a rapid initial rise in [Ca2+] which was followed by a slower but more pronounced [Ca2+], elevation that reached a level 3 to 5 times higher than the basal level. This process preceded cell death as evident by nuclear propidium iodide fluorescence. Depletion of extracellular Ca" inhibited both the [Ca2+], response and cell injury. Preincubation of the cells with the Ca" channel blocker verapamil or .0H radical scavenger mannitol similarly inhibited the [Ca2+], rise and loss of viability. Firefly luciferase assay of cellular ATP content demonstrated that the alterations in [Ca2+], following .0H treatment preceded the depletion of ATP. Changes in membrane potential caused by Na* substitution with K* had no significant effect on either the unstimulated or stimulated [Ca"], levels. Taken together, these measurements indicate that .OH-induced injury in alveolar macrophages is associated with the alteration in [Ca2+], homeostasis. The mechanism of •OH-induced [Ca2+] loading is not due to nonspecific membrane damage or energy depletion, but mainly due to an increase in Ca2+ influx through Ca2+ selective channels. This Ca2+ over-load state as well as energy depletion caused by the oxidant may activate certain cellular degradative processes leading to cell injury and death.

Nov 1, 1995