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Biomedical subjects

K Donaldson

Publications and source records attributed to K Donaldson.

At least 181 records · Page 10Linked to original sources

Inhalation and injection studies in rats using dust samples from chrysotile asbestos prepared by a wet dispersion process.

Long term inhalation studies and intraperitoneal injection studies in rats were undertaken with a series of chrysotile asbestos dusts. Three dust samples were generated from chrysotile modified by the wet dispersion process (WDC) and one was from unmodified chrysotile. Following a 1 year inhalation period, all the chrysotile samples proved extremely fibrogenic and carcinogenic and there were no significant differences between the WDC dusts and normal chrysotile. In all experimental groups approximately 25% of animals developed pulmonary carcinomas and in the oldest rats advanced interstitial fibrosis occupied on average 10% of all lung tissue. In the injection studies all the dust samples produced mesotheliomas in over 90% of animals. Very little chrysotile remained in the lungs of the animals that survived longest following dust inhalation and what there was was present as individual chrysotile fibrils. It is suggested that chrysotile is potentially the most harmful variety of asbestos as shown in these and other animal studies but that it is removed from lung tissue quite rapidly. In the long lived human species this may mean that except where exposure levels are very high and of long duration, chrysotile should be less hazardous than other asbestos types.

Animals↗

Fluoresceinated alpha 2-macroglobulin as a probe for studying macrophages.

The potential value of fluorescein-conjugated human alpha 2-macroglobulin as a probe for studying macrophages in murine peritoneal exudate and spleen cell suspensions has been investigated using a fluorescence-activated cell sorter. These studies revealed that the number of alpha 2-macroglobulin-positive cells in the mixtures examined correlated closely with their macrophage content as determined morphologically. Furthermore cells separated on the FACS on the basis of their strong alpha 2-macroglobulin binding exhibited macrophage morphology and expressed Fc receptors on their surface. Conversely the alpha 2-macroglobulin-negative population contained few macrophages or Fc rosette-forming cells. Extensive blocking and comparative binding studies with a number of purified human alpha 2-macroglobulin preparations and derivatives thereof, and a variety of purified proteins, confirmed that the binding of fluorescein-conjugated alpha 2-macroglobulin to peritoneal exudate cells was specific. Furthermore the binding occurs in a highly reproducible manner. These observations suggest that fluorescein-conjugated alpha 2-macroglobulin in conjugation with flow cytometry is a sensitive and reliable method for both identifying and isolating subpopulations of macrophages.

Animals↗

Increased release of hydrogen peroxide and superoxide anion from asbestos-primed macrophages. Effect of hydrogen peroxide on the functional activity of alpha 1-protease inhibitor.

The ability of asbestos-elicited murine peritoneal macrophages to release superoxide anion and hydrogen peroxide, following in vitro triggering, has been investigated. The asbestos-elicited macrophages produced increased levels of superoxide and hydrogen peroxide compared to control macrophages and similar levels to those produced by Corynebacterium parvum elicited macrophages. The supernatants from asbestos-elicited macrophages which had been triggered in vitro were capable of impairing the ability of alpha 1-protease inhibitor to inhibit elastase function. The catalase sensitivity of this effect showed it to be due to hydrogen peroxide.

Animals↗

Inhalation studies on the effects of tremolite and brucite dust in rats.

Samples of commercially used asbestos, especially chrysotile, are frequently contaminated by small amounts of other fibrous minerals. Among these are tremolite and brucite although pure tremolite is also produced commercially in relatively small quantities. In order to determine how harmful commercially exploited tremolite might be in comparison with other asbestos types and to explore the possibility that small amounts of tremolite and brucite as contaminants could significantly affect the pathogenicity of industrially used chrysotile, long-term animal inhalation and injection studies using rats were undertaken with what were considered to be mineralogically pure samples of these minerals. Rats treated with tremolite developed very high levels of pulmonary fibrosis as well as 16 carcinomas and two mesotheliomas in a group of 39 animals. Tremolite thus proved to be the most dangerous mineral that we have studied. Animals treated with 'brucite' developed moderate levels of pulmonary fibrosis and two carcinomas. Both tremolite and brucite produced mesotheliomas in greater than 90% of animals following i.p. injection. However, it was found that the supposedly pure brucite in fact contained 10% chrysotile, a level of contamination that could well have been responsible for the pathological changes found in both inhalation and intraperitoneal injection studies. The greatest care should be exercised by industry in handling tremolite or materials contaminated with it.

Air Pollutants, Occupational↗

Immunomodulatory effects of mineral dust. I. Effects of intraperitoneal dust inoculation on splenic lymphocyte function and humoral immune responses in vivo.

Syngeneic PVG rats and C57BL6 mice were inoculated with liquid suspensions of UICC chrysotile A asbestos, DQ12 quartz or the inert particulate TiO2. Rats received 10 mg and mice received 2.5 mg dust intraperitoneally on day 0. Injection of the pathogenic dusts asbestos and quartz was associated with a significant reduction in mitogenic response to PHA and Con A detected in splenocytes removed from animals 14 days after inoculation. Injection of TiO2 had no significant effect on splenocyte mitogenesis. Pretreatment of C57BL6 mice with asbestos intraperitoneally also impaired subsequent production of antibody to the protein antigen KLH. These data show that intraperitoneal injection of chrysotile A asbestos and DQ12 quartz had a marked effect on generalized T lymphocyte function. An additional immunomodulatory effect of asbestos injection was shown in C57BL6 mice where in vivo humoral immune responses were suppressed.

Animals↗

Asbestos-activated peritoneal macrophages release a factor(s) which inhibits lymphocyte mitogenesis.

Intraperitoneal asbestos injection in mice has previously been reported to elicit an activated macrophage population. In the present study supernatants from such macrophages were tested for their effect on thymocyte mitogenesis in response to concanavalin A; control supernatants were obtained from saline- and latex-elicited macrophages. Supernatants from asbestos-elicited macrophages were significantly inhibitory to thymocyte mitogenesis while saline- and latex-elicited macrophages did not release significant amounts of such activity. Asbestos-activated macrophage supernatants were inhibitory in a dose-dependent way and the activity was not secreted by macrophages from mice which had received asbestos in the long term. The inhibitory activity was partially dialysable. Supernatants prepared by treating macrophages in vitro with a lethal dose of asbestos were not inhibitory suggesting that the inhibitory activity in the supernatants of asbestos-activated macrophages did not leak from dead or dying cells. The asbestos macrophage supernatant was also significantly inhibitory to mature T-cell-enriched spleen cells but had no effect on fibroblasts, suggesting that the inhibitory effect could be lymphoid cell specific.

Animals↗

An improved macrophage spreading assay--a simple and effective measure of activation.

The development of a quantitative spreading assay of macrophage activation is described. The assay involved incubation of macrophages on glass coverslips for 1 hour and assessment of cell size using a microscope attached to a microcomputer-assisted digitising system which allowed the diameter of 200 cells to be assessed within 10 minutes. Mouse peritoneal macrophages were used in the development of the assay. Internal consistency of the assay was shown by minimal inter-observer, intra-observer and inter-animal variation. Validation of the assay as a measure of macrophage activation was confirmed by the use of in vivo and in vitro activating agents. Once validated the assay was used to detect activation in alveolar macrophages from rats exposed to airborne asbestos. The macrophage spreading assay described here is quick, reliable, consistent and easy to perform and has a potentially wide application in studies of macrophage function and dysfunction.

Animals↗

Chemiluminescence of asbestos-activated macrophages.

Chemiluminescence, a measure of reactive oxygen release by phagocytes, was compared in peritoneal exudate macrophages elicited with chrysotile asbestos, Corynebacterium parvum and saline. Chrysotile asbestos- and C. parvum-activated macrophages produced significantly more chemiluminescence than saline-elicited macrophages. In a second series of experiments the ability of opsonized chrysotile asbestos to act as a trigger for the release of chemiluminescence was tested. Opsonized chrysotile asbestos produced a dose-related release of chemiluminescence from activated macrophages except at the highest dose where chemiluminescence was reduced due, possibly, to a toxic effect of chrysotile during the assay. Opsonized latex also triggered a dose-related chemiluminescent response from activated macrophages. The potential role of toxic reactive oxygen species, released from macrophages, in the development of asbestos-related pulmonary inflammation and fibrosis are discussed.

Animals↗

Concanavalin A receptors and capping in control and activated macrophages.

Macrophages activated in vivo with Corynebacterium parvum and asbestos showed an increased sub-population of cells which capped spontaneously on incubation with fluoresceinated Concanavalin A compared to saline-induced control macrophages. This capping was unaffected by colchicine but was inhibited by cytochalasin B. The spontaneous capping of activated macrophages did not appear to be directly related to the total number of Concanavalin A receptors as measured by specific 125I-labelled Concanavalin A binding to activated and control macrophages. Capping also did not appear to be related to the position of Concanavalin A receptors as judged by trypsin accessibility of bound 125I-labelled Concanavalin A.

Animals↗

Water balance and protein intake in the Mongolian gerbil (Meriones unguiculatus).

During water deprivation the gerbil increasingly reduced its intake of protein as the protein content of the diet increased thus minimising water stress. When given a choice of diet, increased preference was shown for that with the lower protein content. Despite the reduction in intake, the gerbils became dehydrated, the extent being least on the low protein diet. Gerbils remained in water balance on a high protein diet with 2% saline to drink. The predicted maximum urine osmolality, from renal structural indices, was less than the actual measured value, possibly reflecting the well-developed pelvic anatomy.

Animals↗

Documentation of occupational therapy services: a process model.

This paper describes a process model of occupational therapy and the Occupational Therapy Sequential Client Care Record (OTSCCR). The OTSCCR incorporates the model and provides comprehensive documentation of occupational therapy services from referral through outcome of intervention.

Hospital Departments↗

Cytotoxic effect of asbestos on macrophages in different activation states.

The in vitro effects due to phagocytosis of asbestos by mouse peritoneal macrophages in various stages of activation have been compared. The amphiboles proved relatively inert; chrysotile, however, expressed a greater degree of cytotoxicity toward those populations of macrophages induced in vivo with asbestos, than toward any of the other populations of cells. These results are compared with data concerning the enzyme release from the different populations of macrophages following phagocytosis of asbestos. The results indicate that those macrophages that have been exposed to a prior stimulation of either amphibole or serpentine asbestos in vivo are particularly sensitive to exposure to a second dose of a toxic fiber.

Animals↗

Interactions of asbestos-activated macrophages with an experimental fibrosarcoma.

Supernatants from in vivo asbestos-activated macrophages failed to show any cytostatic activity against a syngeneic fibrosarcoma cell line in vitro. UICC chrysotile-induced peritoneal exudate cells also failed to demonstrate any growth inhibitory effect on the same cells in Winn assays of tumor growth. Mixing UICC crocidolite with inoculated tumor cells resulted in a dose-dependent inhibition of tumor growth; this could, however, be explained by a direct cytostatic effect on the tumor cells of high doses of crocidolite, which was observed in vitro.

5'-Nucleotidase↗

A comparison of the pathological effects in rats of the UICC reference samples of amosite and chrysotile with those of amosite and chrysotile collected from the factory environment.

Inhalation studies were undertaken in rats to compare the pathogenic effects of samples of UICC amosite and chrysotile with those of amosite and chrysotile samples collected from the factory environment. Fibre length and diameter studies on the four dust samples showed that the UICC chrysotile cloud contained more long fibres than factory chrysotile, although the factory fibres tended to be thicker. The factory amosite cloud contained more long fibres than UICC amosite, but again the factory fibres were thicker. The factory dusts contained considerable amounts of impurities; in spite of this, the lung dust content of asbestos at the end of 12 months' dusting was higher with factory dusts than with UICC material. The UICC chrysotile and factory chrysotile produced similar levels of interstitial fibrosis, but the factory dust produced fewer malignant lung tumours. The factory amosite produced much more interstitial fibrosis than UICC amosite, but neither amosite cloud produced any malignant pulmonary tumours. These results are discussed in relation to current theories on the importance of fibre dimensions in the pathogenesis of lung disease.

Animals↗