3-nitrotyrosine and Acute-Lung-Injury

Acute lung injury (ALI)/acute respiratory distress syndrome (ARDS) are serious clinical complications with a high frequency of morbidity and mortality. The initiation and amplification of inflammation is a well-known aspect in the pathogenesis of ALI and related disorders. Therefore, inhibition of the inflammatory mediators could be an ideal approach to prevent ALI. Epigallocatechin-3-gallate (EGCG), a major constituent of green tea, has been shown to have protective effects on oxidative damage and anti-inflammation. The goal of the present study was to determine whether EGCG improves phenotype and macrophage polarisation in LPS-induced ALI. C57BL/6 mice were given two doses of EGCG (15 mg/kg) intraperitoneally (IP) 1 h before and 3 h after LPS instillation (2 mg/kg). EGCG treatment improved histopathological lesions, Total Leucocyte count (TLC), neutrophils infiltration, wet/dry ratio, total proteins and myeloperoxidase (MPO) activity in LPS-induced lung injury. The results displayed that EGCG reduced LPS-induced ALI as it modulates macrophage polarisation towards M2 status. Furthermore, EGCG also reduced the expression of proinflammatory M1 mediators iNOS TNF-α, IL-1β and IL-6 in the LPS administered lung microenvironment. In addition, it increased the expression of KLF4, Arg1 and ym1, known to augment the M2 phenotype of macrophages. EGCG also alleviated the expression of 8-OHdG, nitrotyrosine, showing its ability to inhibit oxidative damage. TREM1 in the lung tissue and improved lung regenerative capacity by enhancing Ki67, PCNA and Ang-1 protein expression. Together, these results proposed the protective properties of EGCG against LPS-induced ALI in may be attributed to the suppression of M1/M2 macrophages subtype ratio, KLF4 augmentation, lung cell regeneration and regulating oxidative damage in the LPS-induced murine ALI.

Topics: 8-Hydroxy-2'-Deoxyguanosine; Acute Lung Injury; Animals; Anti-Inflammatory Agents; Arginase; beta-N-Acetylhexosaminidases; Catechin; Cell Proliferation; Interleukin-1beta; Interleukin-6; Ki-67 Antigen; Kruppel-Like Factor 4; Kruppel-Like Transcription Factors; Lectins; Lipopolysaccharides; Macrophages; Mice; Mice, Inbred C57BL; Nitric Oxide Synthase Type II; Peroxidase; Proliferating Cell Nuclear Antigen; Tea; Triggering Receptor Expressed on Myeloid Cells-1; Tumor Necrosis Factor-alpha; Tyrosine

To explore the effects of artesunate on cigarette smoke-induced lung oxidative damage in mice and the expression of Nuclear factor-E2-related factor 2 (Nrf2).. In vivo: A total of 40 female specific pathogen free BALB/c mice were divided randomly into four groups: normal group, cigarette smoke group, vehicle group and artesunate group. The latter three groups were exposed on cigarette smoke for 40 days. Vehicle (5% NaHCO3 containing 5% dimethyl sulfoxide, 0.1 ml of each mice) or artesunate (30 mg/kg, dissolved in the 0.1 ml vehicle) was given by intraperitoneal injections before each passive smoking of the vehicle or artesunate group. Saline of 0.1ml was given to the normal and cigarette smoke groups as negative controls. Cells in bronchoalveolar lavage fluid (BALF) were collected and analyzed by absolute different cell counts. Interleukin (IL)-8 levels in BALF and 3-nitrotyrosine (NT) levels in lung tissue were tested by emzyme linked immunosorbent assay (ELISA). Malondialdehyde levels in serum, total superoxide dismutase (SOD) activity and total glutathione peroxidase (GPx) activity in lung tissue were detected. The pathological changes of lung tissues were observed by HE staining. And the expression levels of Nrf2 were measured by Westernblotting. In vitro: 16HBE cells were cultured and transfected with Nrf2 siRNA. Cigarette smoke extract (CSE) were used to stimulate the secretion of IL-8 in cells. Cells were divided into five groups: blank group, non-transfected non-artesunate group, non-transfected artesunate group, transfected non-artesunate group and transfected artesunate group. The latter four groups were incubated with CSE, and non-transfected artesunate and transfected artesunate groups were intervened with artesunate (30 μmol/L) before CSE incubation. The IL-8 levels of each group were measured using ELISA kit.. In vivo: The total cell counts of BALF in artesunate group were significantly lower than the vehicle group [21.00(2.50)×10(4)/ml vs 35.50(2.50)×10(4)/ml, P<0.001], especially neutrophil counts [6.00(5.12)×10(4)/ml vs 13.60(5.25)×10(4)/ml, P<0.001]. The IL-8 levels in BALF, malondialdehyde levels in serum, 3-NT levels and total SOD activity in lung tissue of artesunate group were all drastically lower than those in the vehicle group [(508±55) vs (912±68) ng/L, (38.2±8.8) vs (48.7±10.6) μmol/L, (28.5±5.8) vs (50.0±9.7) μg/L and (11.8±1.8) vs (18.0±5.3) U/mg protein, respectively, all P<0.05]. No significant difference of total GPx activity existed in these four groups. And the expression level of Nrf2 in artesunate group significantly increased than that in vehicle group (P=0.008). In vitro: The IL-8 level of the non-transfected artesunate group was significantly lower than the non-transfected non-artesunate group [(352±26) vs (765±22) ng/L, P<0.001], while the IL-8 levels between the transfected artesunate and transfected non-artesunate groups had no significant difference.. Arteunate attenuates cigarette smoke-induced lung oxidative damage in mice and increases the expression level of Nrf2, and its effects might be mediated by the actions of nuclear Nrf2.

Topics: Acute Lung Injury; Animals; Artemisinins; Artesunate; Bronchoalveolar Lavage Fluid; Female; Interleukin-8; Lung; Malondialdehyde; Mice; Mice, Inbred BALB C; NF-E2-Related Factor 2; Nicotiana; Oxidation-Reduction; Random Allocation; Smoking; Tobacco Smoke Pollution; Tyrosine

Hydrogen sulfide (H2S), known as the third endogenous gaseous transmitter, has received increasing attention because of its diverse effects, including angiogenesis, vascular relaxation and myocardial protection.We aimed to investigate the role of H2S in oxidative/nitrative stress and inflammation in acute lung injury (ALI) induced by endotoxemia.. Male ICR mice were divided in six groups: (1) Control group; (2) GYY4137treatment group; (3) L-NAME treatment group; (4) lipopolysaccharide (LPS) treatment group; (5) LPS with GYY4137 treatment group; and (6) LPS with L-NAME treatment group. The lungs were analysed by histology, NO production in the mouse lungs determined by modified Griess (Sigma-Aldrich) reaction, cytokine levels utilizing commercialkits, and protein abundance by Western blotting.. GYY4137, a slowly-releasing H2S donor, improved the histopathological changes in the lungs of endotoxemic mice. Treatment with NG-nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase (NOS) inhibitor, increased anti-oxidant biomarkers such as thetotal antioxidant capacity (T-AOC) and theactivities of catalase (CAT) and superoxide dismutase (SOD) but decreased a marker of peroxynitrite (ONOO-) action and 3-nitrotyrosine (3-NT) in endotoxemic lung. L-NAME administration also suppressed inflammation in endotoxemic lung, as evidenced by the decreased pulmonary levels of interleukin (IL)-6, IL-8, and myeloperoxidase (MPO) and the increased level of anti-inflammatory cytokine IL-10. GYY4137 treatment reversed endotoxin-induced oxidative/nitrative stress, as evidenced by a decrease in malondialdehyde (MDA), hydrogenperoxide (H2O2) and 3-NT and an increase in the antioxidant biomarker ratio of reduced/oxidized glutathione(GSH/GSSG ratio) and T-AOC, CAT and SOD activity. GYY4137 also attenuated endotoxin-induced lung inflammation. Moreover, treatment with GYY4137 inhibited inducible NOS (iNOS) expression and nitric oxide (NO) production in the endotoxemia lung.. GYY4137 conferred protection against acute endotoxemia-associated lung injury, which may have beendue to the anti-oxidant, anti-nitrative and anti-inflammatory properties of GYY4137. The present findings warrant further exploration of the clinical applicability of H2S in the prevention and treatment of ALI.

Topics: Acute Lung Injury; Animals; Antioxidants; Endotoxemia; Hydrogen Peroxide; Hydrogen Sulfide; Inflammation; Inflammation Mediators; Lipopolysaccharides; Lung; Male; Malondialdehyde; Mice, Inbred ICR; Morpholines; NG-Nitroarginine Methyl Ester; Nitric Oxide; Nitric Oxide Synthase Type II; Nitrosation; Organothiophosphorus Compounds; Oxidative Stress; Tyrosine

The aim of this study was to investigate the effect of costunolide (CS) and dehydrocostuslactone (DCE) a well-known sesquiterpene lactones contained in many plants, in a model of lung injury induced by carrageenan administration in the mice. Injection of carrageenan into the pleural cavity of mice elicited an acute inflammatory response characterized by fluid accumulation in the pleural cavity which contained a large number of polymorphonuclear cells (PMNs) as well as an infiltration of PMNs in lung tissues and increased production of tumour necrosis factor α (TNF-α). All parameters of inflammation were attenuated by CS and DCE (15mg/kg 10% DMSO i.p.) administered 1h before carrageenan. Carrageenan induced an up regulation of the intracellular adhesion molecules-1 (ICAM-1) and P-selectin, as well as nitrotyrosine and poly (ADP-ribose) (PAR) as determined by immunohistochemical analysis of lung tissues. The degree of staining for the ICAM-1, P-selectin, nitrotyrosine and PAR was reduced by CS and DCE. Additionally we show that this inflammatory events were associated with NF-κB and STAT3 activation and these sesquiterpenes down-regulated it. Taken together, ours results clearly shown that CS and DCE may offer a novel therapeutic approach for the management of inflammatory diseases.

Topics: Acute Lung Injury; Animals; Anti-Inflammatory Agents; Carrageenan; Disease Models, Animal; Lactones; Male; Mice; NF-kappa B; Pleurisy; Poly (ADP-Ribose) Polymerase-1; Poly(ADP-ribose) Polymerases; Sesquiterpenes; STAT3 Transcription Factor; Tyrosine

Acute lung injury (ALI) and sepsis are major contributors to the morbidity and mortality of critically ill patients. The current study was designed further evaluate the mechanism of pulmonary vascular hyperpermeability in sheep with these injuries.. Sheep were randomized to a sham-injured control group (n=6) or ALI/sepsis group (n=7). The sheep in the ALI/sepsis group received inhalation injury followed by instillation of Pseudomonas aeruginosa into the lungs. These groups were monitored for 24 h. Additional sheep (n=16) received the injury and lung tissue was harvested at different time points to measure lung wet/dry weight ratio, vascular endothelial growth factor (VEGF) mRNA and protein expression as well as 3-nitrotyrosine protein expression in lung homogenates.. The injury induced severe deterioration in pulmonary gas exchange, increases in lung lymph flow and protein content, and lung water content (P<0.01 each). These alterations were associated with elevated lung and plasma nitrite/nitrate concentrations, increased tracheal blood flow, and enhanced VEGF mRNA and protein expression in lung tissue as well as enhanced 3-nitrotyrosine protein expression (P<0.05 each).. This study describes the time course of pulmonary microvascular hyperpermeability in a clinical relevant large animal model and may improve the experimental design of future studies.

Topics: Acute Lung Injury; Animals; Capillary Permeability; Disease Models, Animal; Female; Lung; Microcirculation; Nitric Oxide; Pneumonia; Pseudomonas aeruginosa; Pseudomonas Infections; Pulmonary Circulation; Pulmonary Edema; Pulmonary Gas Exchange; RNA, Messenger; Sepsis; Sheep; Smoke Inhalation Injury; Time Factors; Tyrosine; Vascular Endothelial Growth Factor A

The inflamed lung exhibits oxidative and nitrative modifications of multiple target proteins, potentially reflecting disease severity and progression. We identified sphingosine-1-phosphate receptor-3 (S1PR3), a critical signaling molecule mediating cell proliferation and vascular permeability, as a nitrated plasma protein in mice with acute lung injury (ALI). We explored S1PR3 as a potential biomarker in murine and human ALI. In vivo nitrated and total S1PR3 concentrations were determined by immunoprecipitation and microarray studies in mice, and by ELISA in human plasma. In vitro nitrated S1PR3 concentrations were evaluated in human lung vascular endothelial cells (ECs) or within microparticles shed from ECs after exposure to barrier-disrupting agonists (LPS, low-molecular-weight hyaluronan, and thrombin). The effects of S1PR3-containing microparticles on EC barrier function were assessed by transendothelial electrical resistance (TER). Nitrated S1PR3 was identified in the plasma of murine ALI and in humans with severe sepsis-induced ALI. Elevated total S1PR3 plasma concentrations (> 251 pg/ml) were linked to sepsis and ALI mortality. In vitro EC exposure to barrier-disrupting agents induced S1PR3 nitration and the shedding of S1PR3-containing microparticles, which significantly reduced TER, consistent with increased permeability. These changes were attenuated by reduced S1PR3 expression (small interfering RNAs). These results suggest that microparticles containing nitrated S1PR3 shed into the circulation during inflammatory lung states, and represent a novel ALI biomarker linked to disease severity and outcome.

Topics: Acute Lung Injury; Adult; Aged; Animals; Biomarkers; Capillary Permeability; Case-Control Studies; Cell-Derived Microparticles; Cells, Cultured; Electric Impedance; Endothelial Cells; Endothelium, Vascular; Female; Gene Knockdown Techniques; Humans; Kaplan-Meier Estimate; Lipopolysaccharides; Lung; Male; Mice; Middle Aged; Pulmonary Artery; Receptors, Lysosphingolipid; RNA Interference; Sphingosine-1-Phosphate Receptors; Tyrosine; Ventilator-Induced Lung Injury

To elucidate the effects of low-dose arginine vasopressin on cardiopulmonary functions and nitrosative stress using an established model of acute lung injury.. Prospective, randomized, controlled laboratory experiment.. Investigational intensive care unit.. Eighteen chronically instrumented sheep.. Sheep were randomly assigned to a sham group without injury or treatment, an injury group without treatment (40% total body surface area third-degree burn and 48 breaths of cold cotton smoke), or an injured group treated with arginine vasopressin (0.02 IU·min⁻¹) from 1 hr after injury until the end of the 24-hr study period (each n = 6). All sheep were mechanically ventilated and fluid resuscitated using an established protocol.. There were no differences among groups at baseline. The injury was characterized by a severe deterioration of cardiopulmonary function (left ventricular stroke work indexes and Pao2/Fio2 ratio; p < .01 each vs. sham). Compared with controls, arginine vasopressin infusion improved myocardial function, as suggested by higher stroke volume indexes and left ventricular stroke work indexes (18-24 hrs and 6-24 hrs, respectively; p < .05 each). In addition to an improved gas exchange (higher Pao2/Fio2 ratios from 6 to 24 hrs, p < .01 each), pulmonary edema (bloodless wet-to-dry-weight ratio; p = .018), bronchial obstruction (p = .01), and pulmonary shunt fraction (12-24 hrs; p ≤ .001 each) were attenuated in arginine vasopressin-treated animals compared with controls. These changes occurred along with reduced nitrosative stress, as indicated by lower plasma levels of nitrate/nitrite (12-24 hrs, p < .01 each), as well as lower myocardial and pulmonary tissue concentrations of 3-nitrotyrosine (p = .041 and p = .042 vs. controls, respectively). At 24 hrs, pulmonary 3-nitrotyrosine concentrations were negatively correlated with Pao2/Fio2 ratio (r = -.882; p < .001) and myocardial 3-nitrotyrosine content with stroke volume indexes (r = -.701; p = .004).. Low-dose arginine vasopressin reduced nitrosative stress and improved cardiopulmonary functions in sheep with acute lung injury secondary to combined burn and smoke inhalation injury.

Topics: Acute Lung Injury; Animals; Arginine Vasopressin; Cardiac Output; Central Venous Pressure; Disease Models, Animal; Dose-Response Relationship, Drug; Female; Hemodynamics; Oxygen Consumption; Pulmonary Gas Exchange; Random Allocation; Reference Values; Sheep; Sheep, Domestic; Smoke Inhalation Injury; Stroke Volume; Tyrosine; Vascular Resistance; Vasoconstrictor Agents

Acute lung injury (ALI) is a serious illness, the incidence and mortality of which are very high. Free radicals, such as hydroxyl radicals (OH) and peroxynitrite (ONOO(-)), are considered to be the final causative molecules in the pathogenesis of ALI. Hydrogen, a new antioxidant, can selectively reduce OH and ONOO(-). In the present study, we investigated the hypothesis that hydrogen inhalation could ameliorate ALI induced by intra-tracheal lipopolysaccharide (LPS, 5mg/kg body weight). Mice were randomized into three groups: sham group (physiological saline+2% hydrogen mixed gas), control group (LPS+normal air) and experiment group (LPS+2% hydrogen mixed gas). Bronchoalveolar lavage fluid (BALF) was performed to determine the total protein concentrations and pro-inflammatory cytokines. Lung tissues were assayed for oxidative stress variables, wet/dry (W/D) ratio, histological, immunohistochemistry and Western blotting examinations. Our experiments exhibited that hydrogen improved the survival rate of mice and induced a decrease in lung W/D ratio. In addition, hydrogen decreased malonaldehyde and nitrotyrosine content, inhibited myeloperoxidase and maintained superoxide dismutase activity in lung tissues and associated with a decrease in the expression of TNF-α, IL-1β, IL-6 and total protein concentrations in the BALF. Hydrogen further attenuated histopathological alterations and mitigated lung cell apoptosis. Importantly, hydrogen inhibited the activation of P-JNK, and also reversed changes in Bax, Bcl-xl and caspase-3. In conclusion, our data demonstrated that hydrogen inhalation ameliorated LPS-induced ALI and it may be exerting its protective role by preventing the activation of ROS-JNK-caspase-3 pathway.

Topics: Acute Lung Injury; Administration, Inhalation; Animals; Antioxidants; Apoptosis; bcl-2-Associated X Protein; bcl-X Protein; Bronchoalveolar Lavage Fluid; Caspase 3; Cytokines; Hydrogen; Lipopolysaccharides; Male; Malondialdehyde; MAP Kinase Signaling System; Mice; Mice, Inbred C57BL; Peroxidase; Pulmonary Edema; Superoxide Dismutase; Treatment Outcome; Tyrosine

Prematurely born infants are often treated with supraphysiologic amounts of oxygen, which is associated with lung injury and the development of diseases such as bronchopulmonary dysplasia. Complimentary responses between the lung and liver during the course of hyperoxic lung injury have been studied in adult animals, but little is known about this relationship in neonates. These studies tested the hypothesis that oxidant stress occurs in the livers of newborn mice in response to continuous hyperoxia exposure. Greater levels of glutathione disulfide and nitrotyrosine were detected in lung tissues but not liver tissues from newborn mice exposed to hyperoxia than in room air-exposed controls. However, early increases in 5-lipoxygenase and cyclooxygenases-2 protein levels and increases in total hydroxyeicosatetraenoic acid and prostaglandin levels were observed in the liver tissues of hyperoxia-exposed pups. These studies indicate that free radical oxidation occurs in the lungs of newborn pups exposed to hyperoxia, and alterations in lipid metabolism could be a primary response in the liver tissues. The findings of this study identify possible new mechanisms associated with hyperoxic lung injury in a newborn model of bronchopulmonary dysplasia and thus open opportunities for research.

Topics: Acute Lung Injury; Animals; Animals, Newborn; Arachidonate 5-Lipoxygenase; Coenzyme A; Cyclooxygenase 2; Disease Models, Animal; Glutathione Disulfide; Hydroxyeicosatetraenoic Acids; Hyperoxia; Liver; Lung; Mice; Mice, Inbred C3H; Oxidative Stress; Prostaglandins; Time Factors; Tyrosine

Several lines of evidence suggest a biological role for peroxisome proliferator-activated receptor (PPAR) beta/delta in the pathogenesis of a number of diseases. The aim of this study was to investigate the effects of a high-affinity PPAR-beta/delta agonist, GW0742, in a mouse model of carrageenan (CAR)-induced pleurisy. Injection of CAR into the pleural cavity of mice elicited an acute inflammatory response characterized by accumulation of fluid containing a large number of neutrophils (polymorphonuclear leukocytes) in the pleural cavity, infiltration of polymorphonuclear leukocytes in lung tissues and subsequent lipid peroxidation, and increased production of nitrite/nitrate, TNF-alpha, and IL-1beta. Furthermore, CAR induced lung apoptosis (Bax and Bcl-2 expression), and nitrotyrosine formation was determined by immunohistochemical analysis of lung tissues. Administration of GW0742 (0.3 mg/kg, i.p. bolus) 30 min before and 30 min after a challenge with CAR caused a reduction in all the parameters of inflammation measured. Thus, based on these findings, we propose that a PPAR-beta/delta agonist such as GW0742 may be useful in the treatment of various inflammatory diseases.

Topics: Acute Lung Injury; Animals; Apoptosis; bcl-2-Associated X Protein; Carrageenan; I-kappa B Proteins; Lipid Peroxidation; Lung; Male; Mice; Neutrophils; NF-KappaB Inhibitor alpha; Nitric Oxide Synthase Type II; Pleurisy; PPAR delta; PPAR-beta; Proto-Oncogene Proteins c-bcl-2; Thiazoles; Transcription Factor RelA; Tumor Necrosis Factor-alpha; Tyrosine

Different isoforms of nitric oxide synthases (NOS) and determinants of oxidative/nitrosative stress play important roles in the pathophysiology of pulmonary dysfunction induced by acute lung injury (ALI) and sepsis. However, the time changes of these pathogenic factors are largely undetermined.. Twenty-four chronically instrumented sheep were subjected to inhalation of 48 breaths of cotton smoke and instillation of live Pseudomonas aeruginosa into both lungs and were euthanized at 4, 8, 12, 18, and 24 hours post-injury. Additional sheep received sham injury and were euthanized after 24 hrs (control). All animals were mechanically ventilated and fluid resuscitated. Lung tissue was obtained at the respective time points for the measurement of neuronal, endothelial, and inducible NOS (nNOS, eNOS, iNOS) mRNA and their protein expression, calcium-dependent and -independent NOS activity, 3-nitrotyrosine (3-NT), and poly(ADP-ribose) (PAR) protein expression.. The injury induced severe pulmonary dysfunction as indicated by a progressive decline in oxygenation index and concomitant increase in pulmonary shunt fraction. These changes were associated with an early and transient increase in eNOS and an early and profound increase in iNOS expression, while expression of nNOS remained unchanged. Both 3-NT, a marker of protein nitration, and PAR, an indicator of DNA damage, increased early but only transiently.. Identification of the time course of the described pathogenetic factors provides important additional information on the pulmonary response to ALI and sepsis in the ovine model. This information may be crucial for future studies, especially when considering the timing of novel treatment strategies including selective inhibition of NOS isoforms, modulation of peroxynitrite, and PARP.

Topics: Acute Lung Injury; Animals; Disease Models, Animal; Interleukin-8; Lung; Nitrates; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Nitric Oxide Synthase Type III; Nitrites; Poly Adenosine Diphosphate Ribose; Reverse Transcriptase Polymerase Chain Reaction; Sepsis; Sheep; Time Factors; Tyrosine

Excessive nitric oxide (NO) generated by inducible nitric oxide synthase (iNOS) aggravates acute lung injury (ALI) by producing peroxinitrite. We previously showed that the expression of iNOS and lung injury were suppressed by inhalation of a novel iNOS inhibitor, ONO-1714, in mice with Candida-induced ALI, and that nitric oxide produced by iNOS and apoptosis of epithelial cells were found to have a crucial role in Candida-induced ALI. In the present study, we investigated the effect of NO on the apoptosis of alveolar epithelial cells in Candida-induced ALI. Mice were pretreated by inhalation of ONO-1714 or saline (vehicle control of ONO-1714), and were given an intravenous injection of Candida albicans to induce ALI. After 24 h from injection of Candida albicans, we performed bronchoalveolar lavage and removed lung tissues. We assessed apoptosis on the basis of TUNEL staining and caspase 3 activity. Our results showed that apoptosis was suppressed by inhibition of iNOS-derived NO production by ONO-1714 inhalation. The augmented production of NO increased FasL, TNF-alpha, and mRNA production of Bax of lung that induced apoptosis of alveolar epithelial cells. Inhibition of iNOS-derived NO production by ONO-1714 inhalation ameliorated Candida-induced ALI and improved survival by suppressing apoptosis of alveolar epithelial cells.

Topics: Acute Lung Injury; Amidines; Animals; Apoptosis; bcl-2-Associated X Protein; Bronchoalveolar Lavage Fluid; Candida; Candidiasis; Caspase 3; Fas Ligand Protein; Heterocyclic Compounds, 2-Ring; Humans; In Situ Nick-End Labeling; Lung; Mice; Mice, Inbred BALB C; Nitric Oxide; Nitric Oxide Synthase Type II; Survival Rate; Tumor Necrosis Factor-alpha; Tyrosine