A comparative study of intratracheal and aerosolization instillations of bleomycin inducing experimental lung fibrosis in rat.

We aimed to investigate in the present work, using metabonomics approaches, the scalability of lung fibrosis-biomarkers, in bleomycin (BLM) model of pulmonary fibrosis in rats. Sixty male Wistar rats, weighing 250 ± 10 g, were randomly divided into three groups: a negative control group receiving normal saline treatment (G1), an intratracheal BLM instilled group (G2), and an aerosol BLM instilled group (G3). Rats were investigated at various times after BLM instillation. Metabolic changes observed in different biofluids have been integrated into the results of the histological examination (increase in inflammation, fibrosis score, and TGF-ß immunostaining) which provide a novel pathway of biomarkers in pulmonary fibrosis. These two BLM-models showed an efficacy in the production of pulmonary fibrosis in rats, accompanied by an oxidative stress in lung tissue as assessed by the increase of lipid peroxidation and the depletion in the level of antioxidant enzymes such as superoxide dismutase and catalase. The aerosol model was more advantageous showing fibrotic foci occupying the majority of the lung in contrast to intratracheal instillation characterized by a non-homogeneous distribution of the fibroblastic foci.

Early Detection of Urinary Proteome Biomarkers for Effective Early Treatment of Pulmonary Fibrosis in a Rat Model.

SCOPE: Pulmonary fibrosis (PF) is a progressive and devastating lung disease. With limited effective treatments available in the late stage, PF has a very poor prognosis. Molecular biomarkers are highly desired for PF, especially for its early phase. MATERIALS AND METHODS: Urine is a good biomarker source, and accumulates systemic changes in the body especially in the early-stage of diseases. In this study, a bleomycin (BLM)-induced rat model is used to mimic PF. Using labeled proteome quantitation, some urinary proteins are identified as candidate biomarkers of PF for early detection and disease monitoring. Then, prednisone treatment is administered at different phases of fibrosis. RESULTS: Our results suggested that urine proteins could enable early detection and monitoring of both disease progression and treatment efficacy in the BLM-induced PF model. Early prednisone treatment effectively inhibited pulmonary fibrosis, whereas the same treatment at a later phase had very limited effects. Meanwhile, five proteins showed the potential for monitoring therapeutic response. CONCLUSION: Urinary proteomics has been underutilized in respiratory diseases. These findings will improve our understanding of the pathogenesis of PF and accelerate biomarker discovery in respiratory diseases.

Cigarette smoke toxins deposited on surfaces: implications for human health.

Cigarette smoking remains a significant health threat for smokers and nonsmokers alike. Secondhand smoke (SHS) is intrinsically more toxic than directly inhaled smoke. Recently, a new threat has been discovered - Thirdhand smoke (THS) - the accumulation of SHS on surfaces that ages with time, becoming progressively more toxic. THS is a potential health threat to children, spouses of smokers and workers in environments where smoking is or has been allowed. The goal of this study is to investigate the effects of THS on liver, lung, skin healing, and behavior, using an animal model exposed to THS under conditions that mimic exposure of humans. THS-exposed mice show alterations in multiple organ systems and excrete levels of NNAL (a tobacco-specific carcinogen biomarker) similar to those found in children exposed to SHS (and consequently to THS). In liver, THS leads to increased lipid levels and non-alcoholic fatty liver disease, a precursor to cirrhosis and cancer and a potential contributor to cardiovascular disease. In lung, THS stimulates excess collagen production and high levels of inflammatory cytokines, suggesting propensity for fibrosis with implications for inflammation-induced diseases such as chronic obstructive pulmonary disease and asthma. In wounded skin, healing in THS-exposed mice has many characteristics of the poor healing of surgical incisions observed in human smokers. Lastly, behavioral tests show that THS-exposed mice become hyperactive. The latter data, combined with emerging associated behavioral problems in children exposed to SHS/THS, suggest that, with prolonged exposure, they may be at significant risk for developing more severe neurological disorders. These results provide a basis for studies on the toxic effects of THS in humans and inform potential regulatory policies to prevent involuntary exposure to THS.

Urinary excretion of protein and non-protein hydroxyproline in patients with sarcoidosis and interstitial pulmonary fibrosis.

The determination of total hydroxyproline (Hyp) with separation into protein and non-protein fractions was carried out in 24-hour urine samples of healthy persons, patients with sarcoidosis and patients with interstitial pulmonary fibrosis (i.p.f.). In the group of sarcoidosis Hyp excretion was evaluated taking into account progression of pulmonary changes and disease activity. It was found that total and non-protein Hyp excretion correlated positively in healthy persons as well as in those with sarcoidosis, and interstitial pulmonary fibrosis. An increased excretion of total and non-protein Hyp occurred in 24% of the patients with sarcoidosis while excretion of protein Hyp remained within the normal range. In 30% of the patients with interstitial pulmonary fibrosis increased excretion of total and non-protein Hyp was demonstrated. The excretion of protein Hyp in these patients did not deviate from normal values. The results of our study indicate that determination of total and nonprotein Hyp in urine may be useful in the evaluation of fibrotic changes in the pulmonary diseases mentioned above. However, these parameters are not usefull in differentiation between sarcoidosis and interstitial pulmonary fibrosis.

[Behavior of urinary hydroxyproline and the effect of cigarette smoking in silicosis].

This study was conducted through regular pneumonoconiosis examination according to the law on 1,096 employees of medium and small-sized ceramic enterprises in Tokai district in 1981-82. Interview examination with BMRC questionnaire, X-ray examination and measurement of urinary hydroxyproline to creatinine ratio (HOP ratio) were carried out in order to elucidate the relationship between silicosis and urinary HOP ratio and to demonstrate the effect of smoking on pneumofibrosis. Grade of silicosis was classified into five types (0 to 4) based on the Japanese Classification of Radiographs of Pneumoconioses. In evaluating the behavior of urinary HOP ratio, when smoking factor is added in the early grade of pneumofibrosis (type 1 and type 2), collagen decomposition rate is rapidly repressed and fibroplastic conditions develop to the final grade as type 3 and 4, although smoking itself does not seem to induce pneumofibrosis. To exclude the effects of smoking, nonsmoking group was used for measurement of HOP ratio by grade. The HOP ratio in type 0 was lowest and HOP ratio increased in the order of type 1 and type 2. The turning point was found in type 2 and their HOP ratio decreased one after another. The turning point shifted from type 2 to type 3 in the case of non-smokers without any index symptoms by BMRC questionnaire and also shifted to type 1, in the case of non-smokers with them. Shifting of turning point suggests that index symptoms also promote fibroplastic activities.

Urinary excretion of desmosine (elastin cross-links) in subjects with PiZZ alpha-1-antitrypsin deficiency, a phenotype associated with hereditary predisposition to pulmonary emphysema.

To evaluate the concept that lung elastin degradation is accelerated in homozygous alpha-1-antitrypsin (AAT) deficient persons, we prepared acid hydrolysates of urine and used a radioimmunoassay for desmosine to measure urine concentrations of this elastin-specific cross-link in such persons and in control subjects. Excretion of desmosine in 17 homozygous AAT-deficient (PiZZ) patients with emphysema was compared with that in 27 patients with interstitial lung diseases (16 sarcoid, 5 idiopathic pulmonary fibrosis, 6 other interstitial lung diseases) and 26 healthy subjects. Both smokers and nonsmokers were present in all groups. Urinary desmosine concentration (microgram/100 mg creatinine) was 2.35 +/- 0.93 in the PiZZ patients, 2.49 +/- 1.01 in those with interstitial lung disease, and 2.05 +/- 0.54 in the healthy control subjects (p greater than 0.1, all comparisons). Because abnormal pulmonary elastolysis may be largely completed before symptoms of emphysema develop in AAT-deficient persons, we also tested 6 asymptomatic adults with homozygous AAT deficiency (PiZZ) and 5 PiZZ children. Urine desmosine (microgram/100 mg creatinine) was not significantly elevated in either group compared with that in the age-matched control subjects, although children (PiZZ and age-matched controls) showed higher excretions than did adults (6 asymptomatic PiZZ adults, 2.60 +/- 0.91; 5 PiZZ children, 3.27 +/- 0.62; 10 control children, 3.61 +/- 0.62). These data suggest that pathologic lung elastolysis in the PiZZ subject may constitute too small a fraction of total-body elastin turnover to be detected by this method.(ABSTRACT TRUNCATED AT 250 WORDS)

Radioimmunoassay of main urinary metabolite of prostaglandin F2alpha in patients with diffuse interstitial fibrosing pneumonitis (DIFP) and other respiratory diseases.

Radioimmunoassay of 5alpha, 7alpha-dihydroxy-11-keto-tetra-norprosta-1,16-dioic acid, main urinary metabolite of prostaglandin F2alpha (PGF2alpha-MUM), was performed in patients with various respiratory diseases including diffuse interstitial fibrosing pneumonitis (DIFP, fibrosing alveolitis). Twenty-four hr excretion of PGF2alpha-MUM in patients with primary lung cancer, pulmonary fibrosis secondary to collagen diseases and stationary DIFP was normal. On the other hand, 24 hr excretion of PGF2alpha-MUM in patients with carcinomatous pleuritis was high and that in patients with aggravating DIFP was markedly high. There was no correlation between serum LDH levels and 24 hr excretion of PGF2alpha-MUM.