Conventional and toxicogenomic assessment of the acute pulmonary damage induced by the instillation of Cardiff PM10 into the rat lung.

There is strong epidemiological evidence of association between PM10 (particulate matter with an aerodynamic diameter less than or equal to 10 microm) and adverse health outcomes including death and increased hospital admissions for cardiopulmonary conditions. Ambient PM10 surrogates such as diesel exhaust particles (DEP), a common component of UK PM10 have been shown to induce lung inflammation in both humans and rodents. To date, few studies have reported on the toxicological response of UK PM10 in experimental animals. This study examines the pulmonary toxicological responses in male Sprague Dawley rats following the intratracheal instillation of Cardiff urban PM10. A mild but significant change in lung permeability was observed in the lung post-instillation of a high (10 mg) dose of the whole PM10 as adjudged by increases in lung to body weight ratio and total acellular lavage protein. Such effects were less marked following instillation of a water-soluble fraction (80% of the total mass) but histological examination showed that lung capillaries were swollen in size with this treatment. In conclusion, conventional toxicological, histological and toxicogenomic studies have indicated that Cardiff PM10 exhibits low bioreactivity in the form of mild permeability changes. Differential gene expression was observed when the lung was treated with whole PM10, containing durable particles, in comparison with the water-soluble fraction of PM10 that was devoid of particles. Such changes were linked to different histopathological events within the lung.

The spatial and temporal variations in PM10 mass from six UK homes.

People spend the majority of their time indoors mostly in the domestic environment, where their health may be effected by significant airborne particulate pollution. The indoor/outdoor air quality at six homes in Wales and Cornwall was investigated, based on different locations (urban, suburban, rural) and household characteristics (smokers, non-smokers). The spatial and temporal variations in PM10 mass were monitored for a calendar year, including ambient weather conditions. The activities of individuals within a household were also recorded. Monitoring of PM10 took place inside (kitchen, living room, bedroom) homes, along with concomitant collections outdoors. Samples were subjected to gravimetric analysis to determine PM10 concentrations and examined by scanning electron microscopy to identify the types of particles present on the filters. The results of the study show there are greater masses of PM10 indoors, and that the composition of the indoor PM10 is controlled by outdoor sources, and to a lesser extent by indoor anthropogenic activities, except in the presence of tobacco smokers. The indoor and outdoor PM10 collected was characterised as being a heterogeneous mixture of particles (soot, fibres, sea salt, smelter, gypsum, pollen and fungal spores).

Leptin, body composition, adrenal and gonadal hormones among captive male baboons.

Morphometric and hormonal measures were collected from 21 captive savanna baboons (Papio cynocephalus) maintained at the Tulane National Primate Research Center in order to determine age-related patterns in leptin levels over the life course as well as their relationships to body composition and adrenal and gonadal steroids. Comparison of leptin levels between peri-pubertal, adolescent, young adult, and fully mature males show lower levels among adolescent as compared with young adult males (P = 0.05 by Kruskal-Wallis ANOVA). In addition, abdominal fat varied among age groups (P = 0.003 by Kruskal-Wallis ANOVA) with the peri-pubertal animals lower than the adolescents, young adults, and prime adults. However leptin was not related to any measure of body composition, including abdominal fat, or to adrenal hormones (dehydroepiandrosterone, dehydroepiandrosterone-sulfate, and cortisol) or gonadal hormones (testosterone and estradiol). Age-related changes in leptin appear similar to those reported for captive rhesus macaques, while the failure to find an association between leptin and abdominal fat is interestingly different. These results confirm elevated levels of leptin in captive baboons compared with their wild counterparts and suggest that they result from changes in fetal development.

Pulmonary antioxidants exert differential protective effects against urban and industrial particulate matter.

This investigation focuses on the application of an in vitro assay in elucidating the role of lung lining fluid antioxidants in the protection against inhaled particles, and to compare the toxicities of different airborne particulate matter (PM), PM10, collections from South Wales, UK. PM collections from both urban and industrial sites caused 50% oxidative degradation of DNA in vitro at concentrations as low as 12.9 +/- 2.1 microg ml(-1) and 4.9 0.9 mg ml-1 respectively. The primary source of this bioreactivity was found to be the soluble fraction of both particle collections. The coarser PM(10-2.5) fraction also showed greater oxidative bioreactivity than the PM(2.5-0.1) in both cases. When repeated in the presence of a low molecular weight fraction of fresh pulmonary lavage fluid, as well as in artificial lung lining fluid (200 microM urate, glutathione and ascorbate), the DNA damage was significantly reduced in all cases (P < 0.05). The antioxidants exerted a greater effect on the industrial samples than on the urban samples, and on the PM(10-2.5) fractions than on the PM(2.5-0.1) fractions, supporting the previous findings that respirable PM and urban samples contain fewer free radical sources than inhalable PM and industrial samples.

Pulmonary epithelial response in the rat lung to instilled Montserrat respirable dusts and their major mineral components.

BACKGROUND: The Soufriere Hills, a stratovolcano on Montserrat, started erupting in July 1995, producing volcanic ash, both from dome collapse pyroclastic flows and phreatic explosions. The eruptions/ash resuspension result in high concentrations of suspended particulate matter in the atmosphere, which includes cristobalite, a mineral implicated in respiratory disorders. AIMS: To conduct toxicological studies on characterised samples of ash, together with major components of the dust mixture (anorthite, cristobalite), and a bioreactive mineral control (DQ12 quartz). METHODS: Rats were challenged with a single mass (1 mg) dose of particles via intratracheal instillation and groups sacrificed at one, three, and nine weeks. Acute bioreactivity of the particles was assessed by increases in lung permeability and inflammation, changes in epithelial cell markers, and increase in the size of bronchothoracic lymph nodes. RESULTS: Data indicated that respirable ash derived from pyroclastic flows (20.1% cristobalite) or phreatic explosion (8.6% cristobalite) had minimal bioreactivity in the lung. Anorthite showed low bioreactivity, in contrast to pure cristobalite, which showed progressive increases in lung damage. CONCLUSION: Results suggests that either the percentage mass of cristobalite particles present in Montserrat ash was not sufficient as a catalyst in the lung environment, or its surface reactivity was masked by the non-reactive volcanic glass components during the process of ash formation.

[Physicochemical characterization of the PM10 in ambient air of northwestern Beijing urban area during heating-period].

Based on monitoring data, the daily and day-to-day changes of PM10 mass concentrations in northwestern Beijing urban area during heating-period were discussed. XRD and SEM were applied to study the mineral compositions and morphology of the PM10, respectively. It was concluded that concentration of PM10 at nighttime was higher than that at daytime generally. By XRD analysis, it was found that the finer the particles, the less the minerals in it. Furthermore, from the SEM micrograph, five types of the particles, mostly from coal burning and diesel exhaust in number concentration, were classified as chain-like aggregate, cluster aggregate, spherical particle, flake-like particle and non-regular particle.

Can toxicogenomics provide information on the bioreactivity of diesel exhaust particles?

Epidemiologists have linked increased cardio-respiratory hospital admissions, morbidity and mortality rates and increases in particulate matter with an aerodynamic diameter less than 10 microns (PM10) concentrations (Anderson et al., 1991). PM10 consist of a heterogeneous mixture of particles that include minerals, metal oxides, sea salt, biological components and soot. In urban locations, soot, especially ultrafine diesel exhaust particles (DEP), accounts for 20-80% by mass of the airborne PM10 arising from vehicular activities. In the experiment described here, control [NaCl] and 1.25 mg of DEP were instilled into rat lung and the responses assessed using changes in lung permeability, inflammation and epithelial cell markers in lavage fluid, with the addition of a new technique of gene expression profiling using macroarrays. The aim of the study was to use these macroarrays as a sensitive measurement of acute up- or down-regulation of genes that were taking place in the rat lung in response to the small instilled mass of DEP. DEP instillation caused a slight oedematous lung with no overt inflammation and ten out of a possible 207 (5%) rat stress genes were repeatedly changed in response to DEP instillation. Therefore, the conclusion from the macroarray analysis is in agreement with the conventional toxicology and suggest that DEP elicits a low bioreactive response in a healthy rat lung.

Regulation of human and mouse procathepsin E gene expression.

Cathepsin E is an intracellular aspartic proteinase that is considered to have a number of physiological roles including antigen processing. Quantitation of procathepsin E mRNA by LightCyclertrade mark technology indicated that the gene was transcribed in lung but not in kidney of both human and mouse origin. In contrast, the transcript was present in mouse spleen and alveolar macrophages but not in the counterpart tissue/cells from humans. Regulation of human and mouse procathepsin E gene expression was shown not to be influenced by the extent of CpG methylation but depended on the recognition of potential binding motifs in each promoter region by transcription factors such as GATA1, PU1 and YY1, as revealed by functional analysis using a series of promoter/luciferase reporter gene fusion constructs. Thus the extent to which the procathepsin E gene is expressed in a particular cell type may depend on the balance between the effects produced by positive-acting, cell-specific transcription factors such as GATA1 and PU1 and the negative influence of the ubiquitous YY1 factor. In this way, the relative abundance and influence of general and cell-specific transcription factors can govern the production of cathepsin E and thereby account for the sporadic cell and tissue distribution of this enzyme in different species.

Effects of dehydroepiandrosterone and quinapril on nephropathy in obese Zucker rats.

BACKGROUND: The obese Zucker rat exhibits insulin resistance, develops nephropathy at an early age, and may be a model of diabetic nephropathy. Dehydroepiandrosterone (DHEA) may ameliorate many of the factors that contribute to diabetic nephropathy, while angiotensin-converting enzyme inhibitors are known to be effective. One marker of nephropathy is the expression of alpha-smooth muscle actin. METHODS: We studied the effect of DHEA on the expression of alpha-smooth muscle actin in obese Zucker rats and compared the changes with those in a control group, a group given quinapril, and a group on a low-calorie diet. DHEA (0.6%) added to plain chow, quinapril (0.3 mg/kg) added to drinking water, and a low-calorie diet based on pair-feeding were administered to obese rats from age 4 to 20 weeks. Immunohistochemical expression of alpha-smooth muscle actin, a marker of interstitial and glomerular fibrosis and an early indicator of nephropathy, was measured semiquantitatively in glomeruli, cortical interstitium, and medullary interstitium on a scale of 0 to 4 and was reported as mean +/- SEM. RESULTS: When compared with the obese control group, quinapril exhibited a marked reduction in alpha-smooth muscle actin staining in glomeruli, cortical interstitium, and medullary interstitium (P < 0.0005); DHEA reduced alpha-smooth muscle actin staining in cortical interstitium and medullary interstitium (P < 0. 005), and a low-calorie diet reduced alpha-smooth muscle actin staining in cortical and medullary interstitium (P < 0.005), which was similar to the effects of DHEA. CONCLUSIONS: DHEA was similar to a low-calorie diet in reducing the immunohistochemical staining of alpha-smooth muscle actin in obese Zucker rats. However, quinapril exerted a marked protective effect on the development of fibrosis, as indicated by alpha-smooth muscle actin staining, which was significantly less than that of DHEA at the doses studied.

Reproductive maturation in a sample of captive male baboons.

Though baboons have been considered an appropriate non-human primate model for studying human reproductive and endocrine development. the overall similarity of reproductive maturation between the two species is unclear. This paper examines the role of testicular and adrenal hormones for pubertal changes in a cross-sectional sample of 21 captive male savanna baboons. Morphometric and hormonal indices demonstrate changes in size and gonadal function, but not adrenal function, during pubertal maturation among baboons. Results also indicate that gonadal, but not adrenal, androgens are related to morphometric variables. We conclude that savanna baboons do not make an appropriate evolutionary model of human pubertal maturation.

Serum leptin, lipids, free fatty acids, and fat pads in long-term dehydroepiandrosterone-treated Zucker rats.

The obese Zucker rat has a genetically flawed leptin system and is a model of hyperphagia, obesity, hyperlipidemia, and markedly elevated leptin levels. Dehydroepiandrosterone (DHEA) administration reduces hyperphagia, hyperlipidemia, and obesity in Zucker rats. Since serum leptin levels are associated with body fat, we wondered what the effects of fat pad weight reduction from DHEA administration would have on leptin levels. This experiment investigated the effects of DHEA on intra-abdominal fat pads, serum lipids, and peripheral leptin in male lean and obese Zucker rats that were administered DHEA in their food from 4 weeks of age to 20 weeks. Lean and obese rats received plain chow or chow containing DHEA. Additional chow-fed groups of lean and obese weight-matched controls and obese pair-fed rats helped to control for the reduced body weight, food intake, and fat pad weights seen with DHEA administration. DHEA administration to lean Zucker rats reduced body weight and fat pad weights, but leptin levels showed a lower trend. Among obese rats, both DHEA treatment and pair-feeding reduced body weight and fat pad weights, but only DHEA lowered leptin levels. The weight-matched controls had reductions in fat pad weights similar to the DHEA-treated group, but with increased leptin levels. Thus, DHEA may exert a small, independent effect on leptin levels in this animal model, but the reduction is less than what would be expected.

Dehydroepiandrosterone-mediated decrease in caloric intake by obese Zucker rats is not due to changes in serum entrostatin-like immunoreactivity.

To understand the mechanism(s) of appetite modulation by DHEA, we have undertaken a series of studies to examine the effects of DHEA on neurotransmitters and neuropeptides known to affect appetitive behavior. Here, we report the effect of DHEA on serum enterostatin-VPDPR or E, a pentapeptide known to cause selective diminution in fat intake. Four-week-old lean (fa/+) and obese (fa/fa) Zucker rats were divided into control and treatment groups. DHEA-treated groups received powdered chow containing 0.6% DHEA ad lib for 16 weeks. Another group of obese rats was pair fed to match the intake of the obese DHEA-treated rats. At the end of this period, trunk blood was collected from fasted rats for assay of E-like immunoreactivity (E-LI) by ELISA. DHEA treatment caused a significant diminution in circulating E-LI in both lean (control: 2030 +/- 226; treated: 752 +/- 145 ng/mL; n = 10, p < 0.0001) and obese (control: 2489 +/- 391, n = 6; treated: 1123 +/- 185 ng/mL, n = 7; p = 0.0003) rats. Because DHEA treatment decreases caloric intake and body weight, we examined the effect of caloric intake and body weight on E-LI levels. Serum ELI levels were lower in the obese DHEA-treated group compared to that of obese pair fed (pair fed: 1589 +/- 313, n = 6; DHEA: 1123 +/- 185 ng/mL, n = 7), but the differences were statistically insignificant (p = 0.185). Also, both weight-matched lean and obese control rats had significantly (p < 0.008) higher E-LI than their DHEA-treated counterparts. To examine whether the decrease in serum E-LI following DHEA treatment could be due to increased peptide metabolism, the rate of disappearance of endogenous E-LI from serum (obese control and DHEA-treated) at 37 degrees C was evaluated. The results show an attenuation of peptide metabolism in serum from DHEA-treated rats, a finding contrary to our expectations. In summary, DHEA treatment lowers serum E-LI levels both in lean and obese Zucker rats. This decrement in peptide level is not secondary to changes in body weight or caloric intake due to DHEA, or due to altered serum peptide metabolism. Although DHEA appears to be a potent modulator of E-LI levels, the relationship between DHEA and E-LI in relation to appetitive behavior remains to be clarified.

Density and substrata are important in lung type II cell transdifferentiation in vitro.

Morphological techniques and metabolic cell marker assays were used to study the transdifferentiation of pulmonary type II epithelial cells to type I-like cells in vitro. In the lung this process is important during remodelling and alveolar repair. Type II cell phenotype was best maintained over eight days when densely packed cells were plated out on a commercially available extracellular matrix. Such cells retained type II cell characteristics (lamellar bodies, high activities of gamma glutamyl transpeptidase and alkaline phosphatase) but expressed low levels of rT1(40) a surface protein marker of type I cells. In contrast, low density cultures, irrespective of substratum, exhibited rapid cell spreading, loss of lamellar bodies, loss of type II cell enzyme markers and expressed high levels or rT1(40). Conditions have been described whereby the same isolate of type II cells can be used to produce differential epithelial phenotypes and use can be made of this for further characterisation or to investigate the effect of toxins on different lung cell types in vitro.

Prevalence of previous appendectomy among patients needing gastrointestinal endoscopy.

BACKGROUND: The appendix may be an immune modulator of the gut, and its absence may lead to an increase in gastrointestinal illnesses. If this is true, we may expect patients needing endoscopy to have a higher prevalence of previous appendectomy. METHODS: We did a case-control study at the University of Oklahoma Hospital for 13 months. Subjects having endoscopic evaluation at the University of Oklahoma Hospital formed the study group. Patients seen at the general medicine clinic of the University of Oklahoma served as controls. We recorded the patient's name, age, sex, race, history of smoking, and history of appendectomy or tonsillectomy. RESULTS: The endoscopy group had 524 patients; 469 patients were in the control group. There were no differences based on race or history of smoking. There was greater prevalence of previous appendectomy in the endoscopy group (33.46% vs. 20.55%). The prevalence of tonsillectomy was 29.28% in the study group vs. 21.61% among the controls. Multiple regression revealed that history of appendectomy and not tonsillectomy was related to the performance of endoscopy. CONCLUSION: History of appendectomy is associated with greater performance of endoscopy.

Bioreactivity of carbon black and diesel exhaust particles to primary Clara and type II epithelial cell cultures.

OBJECTIVES: To begin to elucidate the mechanisms of particle toxicity to the lung, the bioreactivity of four carbon black (CB) and diesel exhaust particles ((DEPs), a surrogate for particulate matter of aerodynamic diameter < 10 microns (PM10), were examined with primary cultures of Clara and type II epithelial cells. METHODS: The particles were extensively characterised by surface chemistry, size, and aggregation properties. Toxicity of the particles was assessed by determining cell attachment to an extracellular matrix substratum. RESULTS: The spherulite size range for the particles ranged from 50, 40, 20, 20, and 30 nm for CB1-4 and DEPs. All particle samples had different surface chemical compositions. CB1 was the least toxic to Clara (170 micrograms) and type II cells (150 micrograms) and CB4 was the most toxic (55 micrograms and 23 micrograms respectively). DEPs stored for 2 weeks were equally toxic to both epithelial cell types (27-28 micrograms). DEPs became progressively less toxic to type II cells with time of storage. Both primary epithelial cell types internalised the particles in culture. CONCLUSIONS: Bioreactivity was found to be related to CB particle spherulite size and hence surface area: the smaller the particle and larger the surface area, the more toxic the particles. Also, CB particles with the most complicated surface chemistry were the most bioreactive. Freshly prepared DEPs were equally toxic to type II and Clara cells and they became progressively less toxic to the type II cells with time. With all CB and DEPs, the primary epithelial cells internalised the particles, although this was noted most in cells of low functional competence.

Long-term oral administration of dehydroepiandrosterone has different effects on energy intake of young lean and obese male Zucker rats when compared to controls of similar metabolic body size.

AIM: The effects of dehydroepiandrosterone (DHEA) on appetite and weight in the Zucker rat have been examined by many investigators who have reported appetite suppression and metabolic effects. However, these studies compared the treated animals to controls of a similar age. Since animals of different sizes consume different amounts of food, perhaps the treated animals should be compared to controls of a similar size. We studied the effects of DHEA on energy intake and weight gain and analysed the effects by age and metabolic body size. METHODS: Lean (n = 21) and obese (n = 16) male Zucker rats were fed plain chow or chow containing 6 g DHEA/kg chow (0.6% wt/wt) from age 4 wk to 20 wk. Daily energy intakes and body weights were determined at least once weekly. RESULTS: As expected, the lean and obese rats given DHEA exhibited less daily energy intake (kJ/d) and less weight gain than their respective controls of the same age. The lean rats given DHEA did not exhibit any difference in daily energy intake when determined relative to body weight (b.w.) (kJ x d-1 x g b.w.-1) compared to lean controls of the same metabolic body size, while the obese rats given DHEA exhibited less daily energy intake relative to b.w. (kJ x d-1 x g b.w.-1) compared to obese control of the same metabolic body size. CONCLUSIONS: Though DHEA reduced total energy intake among the lean and obese Zucker rats, only the obese rats exhibited less energy intake relative to b.w. compared to controls of the same metabolic body size. Thus, DHEA may exert different effects on energy intake relative to b.w. in lean and obese Zucker rats and perhaps the lean Zucker rat is a better model for evaluating the metabolic effects of DHEA since it does not exhibit any effect on energy intake relative to b.w. compared to rats of the same metabolic body size.

A comparative study of the isolation of type II epithelial cells from rat, hamster, pig and human lung tissue.

Data is reported on the reproducibility and purity of alveolar type II cell isolations from 4 species. Human and pig type II cells were isolated using a tissue slice method to remove blood and contaminating cells, whilst rat and hamster cells were isolated using the method of protease instillation. All cells were purified on Percoll gradients and by differential attachment. Cell type purity was assessed by phase contrast microscopy, electron microscopy (EM), percentage of cells alkaline phosphatase (AP) positive and percentage of cells staining strongly for NADPH dependent nitro blue tetrazolium reductase (NBT). These enzymes are considered as markers for type II and Clara cells respectively. The purity of all cell preparations was enhanced following 24 h culture on a biomatrix and whilst plating efficiency was similar for all species, the human tissue consistently yielded the highest purity of type II cells. All cells with lamellar bodies did not contain AP, and activity was variable between species. Further studies are needed to determine if NBT is equally nonspecific as a cell marker enzyme. In summary, sufficient type II cells of high purity can be isolated thus permitting interspecies comparative studies to investigate the effects of selective and non-specific pulmonary toxins, but more specific marker enzymes are required to identify Type II and Clara cells.