Rat lung reactivity to natural and man-made fibrous silicates following short-term exposure.

The inflammatory and fibrogenic potential of three naturally occurring and two man-made industrial minerals were compared. Groups of five rats each received respectively a single intratracheal instillation of saline (control), UICC chrysotile B asbestos, short chrysotile 4T30, attapulgite, xonotlite (a calcium silicate), and Fiberfrax (an aluminum silicate) at doses of 1, 5, and 10 mg. One month after the treatment, assessment of lung morphology and bronchoalveolar lavage were performed on each animal. Under these conditions, UICC chrysotile B at all doses tested (1, 5, and 10 mg) induced fibrotic lesions in bronchiolar tissues while short chrysotile 4T30 (1, 5, and 10 mg) caused focal accumulation of inflammatory cells in the alveolar structures but no apparent fibrosis. Compared to these positive reactions with different fibrogenicity, xonotlite caused minimal inflammatory reactions detectable only at high dose (10 mg) and by bronchoalveolar analysis. By contrast, the rat lung reacted more significantly to attapulgite and Fiberfrax although the tissue reaction differed considerably for these two materials. While attapulgite, at doses up to 10 mg caused minimal reactions characterized by mononuclear cell infiltration mainly in the alveolar structures, Fiberfrax at 1 mg and higher caused significant granulomatous reactions and the appearance of early fibrosis. Overall the order of lung biological reactivity observed for the various silicates was xonotlite much less than attapulgite less than short chrysotile 4T30 less than Fiberfrax less than UICC chrysotile B. These observations indicate that Fiberfrax, attapulgite, and, to a lesser extent, xonotlite are biologically active within the time span studied and potentially deleterious for lung tissue.

Cytotoxicity of respirable dusts from industrial minerals: comparison of two naturally occurring and two man-made silicates.

The membranolytic and cytotoxic properties of two naturally occurring (chrysotile asbestos; attapulgite clay) and two man-made (Fiberfrax, an aluminium-silicate, and xonotlite, a calcium silicate) industrial minerals were compared. "Short" fiber fractions of chrysotile and Fiberfrax were obtained by sedimentation in demineralized water, while the attapulgite and xonotlite samples were used as obtained. The aluminium silicate fibers were found to be non- hemolytic, while for the other three silicates, chrysotile had the strongest hemolysis potential, followed very closely by xonotlite; attapulgite was less hemolytic than the former two silicates, but was nevertheless highly hemolytic to the rat erythrocytes. Using rat pulmonary alveolar macrophages, the in vitro cytotoxicity assays showed that with fresh cell monolayers, all four silicates were equivalent in causing cell damages at a dose of 250 micrograms; at a lower dose (50 micrograms), the intensity of the cytotoxic effect was in the decreasing order: Fiberfrax greater than attapulgite greater than chrysotile greater than xonotlite. With one day-old cultured cell monolayers, a dose of 250 micrograms of the silicates fibers was less cytotoxic, with the exception of the attapulgite fibers which remained essentially as cytotoxic as with the fresh cell monolayers. The reduced cytotoxic response was especially noticeable with the chrysotile fibers. At 50 micrograms, the cytotoxicity scale of the mineral dusts with one day-old cell monolayers was essentially the same as the one obtained with the fresh cell monolayers, that is: Fiberfrax approximately equal to attapulgite greater than chrysotile greater than or equal to xonotlite. Overall, these in vitro tests imply: 1) that all four industrial silicates tested can be considered to be "biologically active"; 2) that on the basis of their different reactivities with the two types of cell culture conditions used, their biological reactivity in vivo might be quite distinct. This might be especially true for at least the chrysotile, attapulgite and xonotlite short fibers, considering that these three types of silicate dusts have very similar dimensions. Moreover, for the chrysotile and attapulgite samples, fiber numbers is probably not an important factor, since the density of the two silicates is roughly the same. The unaltered cytotoxic responses of the American attapulgite fibers in the two macrophage assays correlate well with the fact that short attapulgite fibers seems to have a stronger in vivo reactivity than short chrysotile fibers. In fact, it might be the strongest of the four industrial silicates tested.(ABSTRACT TRUNCATED AT 400 WORDS)

The adsorption of polyaromatic hydrocarbons on natural and chemically modified asbestos fibers.

Many reports indicate that the carcinogenic (genotoxic) potential of benzo[a]pyrene (B[a]P) may be enhanced several-fold by the promoter (epigenetic) effect of asbestos particles. This promoting effect could be related to the fact that when B[a]P is adsorbed onto the particles, there is a resulting enhanced transport and uptake of the carcinogen into microsomial membranes. These in vitro data bear relevance to the epidemiological studies which indicate an association between exposure to inhaled asbestos dusts and the high incidence of pulmonary cancers in smokers. Using HPLC, it has been observed that B[a]P has great affinity for natural asbestos fibers, and that chemical modification of natural chrysotile with POCl3 results in the complete loss of this adsorption potential of chrysotile for benzo[a]pyrene.

An assessment of the fibrogenic potential of very short 4T30 chrysotile by intratracheal instillation in rats.

Three groups of five rats each received, respectively, a single intratracheal instillation of saline (control), 5 mg of UICC chrysotile B asbestos, and 5 mg of a preparation of very short chrysotile fibers (4T30, 100% less than 8 micron) isolated by a sedimentation procedure. At various intervals after the treatment (1 to 60 days), assessment of lung morphology was performed on each animal. Although the two types of chrysotile fibers have similar chemical composition, structure, and surface charge, the lung tissue reaction differed considerably. Lungs of animals exposed to UICC chrysotile B showed significant pathological alterations as early as 7 days following treatment. The lesions were localized in and around terminal bronchioles and consisted of inflammatory cells, fibroblasts and collagen deposition which distorted and obstructed small airways. Reaction to very short 4T30 chrysotile fibers was quite distinct. Seven days after treatment, lungs of these animals showed alveolar and interstitial accumulation of inflammatory cells. The alveolitis persisted 60 days after treatment and no fibrosis was apparent. It appears that very short 4T30 chrysotile fibers are much less fibrogenic than UICC chrysotile B and that intratracheal instillations in rats may represent a useful mean of rapidly assessing the fibrogenic potential of various dusts. These observations support the concept that fiber length is an important factor for fibrogenicity of asbestos.

Cytotoxic effects of aramid fibres on rat pulmonary macrophages: comparison with chrysotile asbestos.

Aramid fibres have been proposed as a substitute for many asbestos uses. Although dimensional characteristics of aramid fibres vary with the type of application, some of the commercial grades proposed contain fibres whose geometry is clearly in a range where biological reactivities have been reported for other natural or man-made fibres. We wish to report that short aramid fibres, when tested on cultures of rat pulmonary alveolar macrophages (PAMs), induce the commonly recognized signs of a cytotoxic effect, that is: leakage of cytoplasmic and lysosomal marker enzymes, concomittant with a decreased ATP cell content.

Effect of age and glucocorticoid administration on the proteolytic activity of gastric mucosa: a comparative study in the young rat, calf and piglet.

The proteolytic activity of gastric mucosa markedly increased from 2 to 4 wk of age in rats and from birth to 8 wk in pigs. In calves it was at adult level at birth, then decreased until d 14 and remained at a relatively low level up to d 56 of age. Early postnatal glucocorticoid injections increased (P less than .05) the proteolytic activity of gastric mucosa in rats, decreased (P less than .05) the abomasal proteolytic activity in calves and had no significant effect in pigs. The high levels of proteolytic activity observed in the rat at earlier ages after glucocorticoid treatment suggested a precocious maturation of the gastric mucosa. Injections of different doses of hydrocortisone acetate and dexamethasone to calves and rats indicated that proteolytic activity of gastric mucosa is only sensitive to high doses (i.e., nonphysiological) of glucocorticoids. The response of gastric mucosa to glucocorticoid injections depended on the animal's age; maximum response was obtained in the early postnatal period while minimal response was observed after weaning.

Effects of administration of glucocorticoids on pancreas and growth of dairy calves.

The possibility of reducing calf weaning age by accelerating development of the pancreas of dairy calves with hormone administration was studied. Hydrocortisone acetate or dexamethasone was injected at 3 or 17 days of age to determine their effects on the pancreas. Hydrocortisone acetate injections (50 mg/kg body weight) to 3-day-old calves increased within 4 days the pancreas weight, amylase and chymotrypsin activities, and proteins, ribonucleic acid, and deoxyribonucleic acid contents. However, the glucocorticoid injection had no effect on activities of digestive enzymes of the 21-day-old calves. The pancreas hypertrophied in the 7-day-old calves following hormonal treatment. In the 21-day-old calves administration of glucocorticoids brought about hyperplasia of the pancreas but no hypertrophy of the tissue. This response of the pancreas to injections of glucocorticoids in the 7-day-old calves was interpreted as an acceleration of normal development of the tissue. Hydrocortisone acetate injection to 3-day-old calves had no effect on feed consumption but had a small depressive effect on growth of calves at 14 and 21 days of age.

The hemolytic activity of chrysotile asbestos fibers: a freeze-fracture study.

Short asbestos fibers isolated by a sedimentation procedure have a strong hemolytic activity. In the presence of ferritin particles, hemolysis by chrysotile fibers is inhibited at least during the first 10 min. Freeze-fracture studies show that after 20 sec or 2 min of contact between the fibers and the RBC membrane, the intramembranous particles remain randomly distributed over the whole surface of the P-face. On the E-face of the asbestos-treated red blood cell membranes, the number of intramembranous particles is significantly reduced. With the transmission electron microscopy, it is not possible to resolve the trilaminar structure of the ghost membrane around the deeply buried asbestos fibers. It is postulated that the membrane defects brought about by asbestos are caused by the adsorption of one membrane constituent, possibly phospholipids, on the chrysotile fibers.

Changes in rat gastric proteases with age: effect of hydrocortisone.

Gel filtration on Sephadex G-75 of rat gastric mucosal preparation shows two peaks of proteolytic activity. These peaks differ at different ages of animals. The proteolytic activity of the gastric mucosa is spread over a wider range of pH in younger animals than older ones with a shift from higher pH towards lower pH values with increasing age. Hydrocortisone injection to nine day-old rats results in qualitative changes of gastric mucosal proteases, the proteolytic activity pattern of young treated animals being similar to that of untreated adult rat.

Isoelectric focusing of pure rat pancreatic juice in polyacrylamide gel : possibility of concomitant estimation of radioactivity incorporated into single identified protein constituents.

A method is described for the isoelectric focusing of rat pancreatic juice on columns of polyacrylamide gel containing urea. Twenty-six bands were separated, and 6 were identified. Application of this method of separation to in vivo radioactively labelled pancreatic juice resulted in bands which could be counted with precision. This procedure allowed also the expression of radioactivity of one constituent relative to any single band, or relative to total radioactivity, concomitantly.