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The effects of age on immune responses in the antigen-instilled dog lung. Antibody responses in the lung and lymphoid tissues following primary and secondary antigen instillation.

To evaluate the effects of age on immunity induced by lung immunization, 11 aged (12-17 years; median age = 14) and 12 young (2-5 years) male Beagle dogs were instilled with 10 mg of keyhole limpet hemocyanin (KLH) in the right cardiac lung lobe and 10(10) sheep red blood cells (SRBC) in the left cardiac lung lobe. Five aged and six young dogs were sacrificed at day 9 after primary antigen instillation. The remainder were given challenge antigen instillations of KLH and SRBC at day 21 and sacrificed 7 days later. Serum, bronchoalveolar lavage fluid and lung tissue from immunized and control lobes, tracheobronchial, mesenteric and popliteal lymph nodes, spleen, and blood were taken at sacrifice. Anti-KLH IgA, IgG and IgM antibody production by cells in lung tissue and lavage fluid from the KLH-exposed lobe was lower at primary immunization and challenge in aged than young dogs. Lavage fluid IgA and IgG levels from the KLH exposed lobe at primary immunization and challenge were lower in aged versus young dogs, while IgM levels were lower only after primary immunization. Localized lung immune memory responses were also markedly lower in aged dogs when compared with young dogs. Anti-SRBC responses were similar to the anti-KLH responses. Our data show that systemic immune responses are significantly lower in aged dogs following primary antigen instillation, but not after antigen challenge in the lung. This was not the case for localized lung immune responses, which were significantly lower in aged dogs even following antigen challenge. The data also show that antibody production by lavage cells is a good index of interstitial lung cell antibody production.

Aging↗

Isolated lung perfusion with melphalan for resectable lung metastases: a phase I clinical trial.

BACKGROUND: Current 5-year survival after complete resection of pulmonary metastases is 20% to 40%, and many patients develop intrathoracic recurrences. Isolated lung perfusion is an experimental technique to deliver high-dose chemotherapy to the lung without systemic exposure. A phase I trial of isolated lung perfusion with melphalan (MN) combined with pulmonary metastasectomy for resectable lung metastases was conducted to define the dose-limiting toxicity and maximum tolerated dose. METHODS: From May 2001 to August 2003, 16 patients underwent isolated lung perfusion with MN, followed by surgical resection of lung metastases. Patients were treated with increasing MN doses (15, 30, 45, and 60 mg). For each dose level, normothermia (37 degrees C) and hyperthermia (42 degrees C) were evaluated (n = 3 per level). Serum samples were obtained during the procedure. Pulmonary, hematologic, and nonhematologic toxicities were recorded. The primary tumor was colorectal in 7 patients, renal in 5, sarcoma in 3, and salivary gland in 1. Isolated lung perfusion was performed unilaterally in 11 patients, and staged bilaterally in 5. RESULTS: In total, 21 procedures of isolated lung perfusion with complete metastasectomy were performed without technical difficulties. Operative mortality was 0%, and no systemic toxicity was encountered. Grade 3 pulmonary toxicity developed at a dose of 60 mg of MN at 37 degrees C in 2 of 3 patients at this dose, terminating the trial. CONCLUSIONS: Isolated lung perfusion with MN combined with pulmonary metastasectomy is feasible. Dose-limiting toxicity occurred at a dose of 60 mg of MN at 37 degrees C, and the maximum tolerated dose was set at 45 mg of MN at 42 degrees C.

Adult↗

Cytotoxic concentrations of alpha-defensins in the lungs of individuals with alpha 1-antitrypsin deficiency and moderate to severe lung disease.

The lungs of individuals with alpha(1)-antitrypsin (alpha(1)-AT) deficiency have a reduced antiprotease protective shield. Typically, numbers of neutrophils are elevated in the lungs of affected individuals, carrying large amounts of neutrophil elastase (NE) and alpha-defensins that are released upon activation. We hypothesized that in individuals with advanced lung disease associated with alpha(1)-AT deficiency cytotoxic concentrations of alpha-defensins might be present and unopposed, thus further aggravating lung disease. To evaluate this hypothesis, bronchoalveolar lavage was performed in 20 alpha(1)-AT deficient individuals with moderate to severe lung function impairment and 13 healthy volunteers. Cell counts as well as concentrations of NE and alpha-defensins were determined. While concentrations of NE were low (23+/-14nM) and alpha-defensins were undetectable in volunteers, they were significantly elevated in individuals with alpha(1)-AT deficiency (1313+/-723nM and 6605+/-2856nM, both p<0.0001). These concentrations were significantly higher than those found in a historical control of alpha(1)-AT deficient individuals with mild lung disease. Neutrophil numbers and NE concentration correlated with alpha-defensins concentration (r=0.612 and r=0.758, p=0.005 and p=0.0001, respectively). Individuals with alpha(1)-AT deficiency and moderate to severe lung function impairment have lung alpha-defensins concentrations in a range known to induce cytotoxicity in vitro in the absence of normal amounts alpha(1)-AT and thus may contribute to the development of lung disease in this population.

Adult↗

Lung cancer resection combined with lung volume reduction in patients with severe emphysema.

OBJECTIVE: Certain patients with resectable lung cancer and severe respiratory limitation due to emphysema may have a suitable operative risk by combining cancer resection with lung volume reduction surgery. The purpose of this study is to review our experience with such patients. METHODS: A review was conducted on 21 patients with lung cancer in the setting of severe emphysema who underwent an operation designed to provide complete cancer resection and volume reduction effect. RESULTS: In the 21 patients, the mean preoperative forced expiratory volume in 1 second was 0.7 +/- 0.2 L (29% predicted), residual volume was 5.5 +/- 1.0 L (271%), and diffusing capacity for carbon monoxide was 8.0 +/- 2.2 mL/min/mm Hg (34% predicted). In 9 patients, the cancer was located in a severely emphysematous lobe and the lung volume reduction surgery component of the procedure was accomplished with lobectomy alone. In the remaining 12 patients, the cancer resection lobectomy (n = 9) and wedge resection (n = 3) were supplemented with lung volume reduction surgery. Final pathologic staging was stage I in 16 patients, stage II in 2 patients, and stage III in 2 patients. One patient was found to have stage IV disease due to multifocal tumors in separate lobes. There were no hospital deaths. Postoperative complications included prolonged air leak in 11 patients, atrial fibrillation in 6 patients, and reintubation for ventilatory assistance in 2 patients. All patients showed improved lung function postoperatively. Survival was 100% and 62.7% at 1 and 5 years, respectively. CONCLUSIONS: Patients with severe emphysema and resectable lung cancer who have a favorable anatomy for lung volume reduction surgery may undergo a combined cancer resection and lung volume reduction surgery with an acceptable risk and good long-term survival.

Aged↗

Adenovirus mediated IL-10 gene transfer to the airway of the rat lung for prevention of lung allograft rejection.

BACKGROUND: The ability to express genes with potential immunoregulatory capacity could reduce allograft rejection (AR). We examined the feasibility of transferring the viral interleukin-10 (vIL-10) gene into rat lungs by intra-bronchial instillation and the subsequent effects of delivered vIL-10 on acute lung AR. METHODS: First, the adenoviral beta-galactosidase vector (adv-beta-gal) particles were instilled into the airway of the rat lung and protein synthesis of beta-gal was examined by histochemical staining. Next, the ability of the adenoviral vIL-10 vector (adv-vIL-10) transfection to modify AR was examined in a highly histoincompatible rat lung transplant model (BN-->Lew). Donor left lungs were transfected with 3 x 10(8) pfu/0.3 mL of adv-vIL-10 (vIL-10 group) or adv-beta-gal (control group) 3 days before transplantation. On day 6 post-transplant, lung allografts were harvested and AR was graded histologically (stage 0-4). Several pathological categories of inflammation (perivascular, peribronchial, or peribronchiolar mononuclear infiltrates, edema, vasculitis, intraalveolar hemorrhage, and necrosis) were also examined and scored on a scale of 0-4 as previously described. RESULTS: A successful transgene protein synthesis by adv-beta-gal in alveolar epithelial cells and alveolar macrophages was confirmed by histochemical staining with X-gal. The vIL-10 group showed a trend toward an improved stage of AR (3.75 +/- 0.5 vs. 4.0 +/- 0), and also a decreased pathological scores for edema (3.5 +/- 0.6 vs. 4.0 +/- 0), intraalveolar hemorrhage (2.3 +/- 1.0 vs. 2.5 +/- 0.6) and necrosis (1.5 +/- 0.5 vs. 1.75 +/- 1.3) compared with the control group, however, the differences in any pathological scores between the two groups did not reach a statistical significance. CONCLUSIONS: 1. A successful transgene protein synthesis in alveolar epithelial cells was ensured by intra-bronchial instillation of an adenoviral vector encoding beta-galactosidase gene. 2. Transferring the vIL-10 gene into rat lungs by intra-bronchial instillation did not seem to reduce lung AR significantly, as opposed to the results of our previous experiments in a rat cardiac allograft model. This discrepancy may be explained by several potential factors including the immunogenecity of adenoviral vectors in conjunction with the nature of the lung more susceptible to immune response and inflammation.

Adenoviridae↗

Lung cancer rates as an index of tobacco smoke exposures: validation against black male approximate non-lung cancer death rates, 1969-2000.

BACKGROUND: Researchers use lung cancer death rates (rates) as an index of the cumulative burdens of smoking. That index lacks direct validation and calibration. So this study directly validates and calibrates that index against annual approximately non-lung (all-sites minus lung and stomach) rates from 1969 to 2000 in United States black men, then estimates their cancer death rate smoking-attributable fractions (SAFs). METHODS: This study uses linear regression, age-adjusted rates from http://www.seer.cancer.gov/canques, and the formula SAF = (1- ((rate in the unexposed) / (rate in the exposed))). Estimated rates in the unexposed range between the 1969 rate and the rate predicted for a population with no smoking-attributable lung cancers. Stomach and lung cancer rate SAFs were based on published cohort studies. RESULTS: Lung cancer death rates predicted 98% and 97% of the variances in approximately non-lung cancer death rates throughout their 1969-1990 34% rise and subsequent declines, respectively (each P < 0.0001). The findings suggest that the SAF of the all-sites cancer death rate in black men peaked at 66% in 1990. CONCLUSIONS: Lung cancer death rates were a good index of smoke exposure for predicting approximately non-lung cancer death rates in black men. Smoking may cause most premature cancer deaths in black men.

Black or African American↗

Does human leukocyte antigen matching influence the outcome of lung transplantation? An analysis of 3,549 lung transplantations.

BACKGROUND AND OBJECTIVE: Human leukocyte antigen (HLA) compatibility has been shown to improve the outcome of renal and cardiac transplantation. However, its impact on outcome following lung transplantation is not clear, with several single-center studies reporting inconsistent results. We studied the influence of HLA matching on survival and the development of rejection and obliterative bronchiolitis after lung transplantation, using data from the United Network for Organ Sharing/International Society for Heart and Lung Transplantation registry. METHODS: The study population included adult patients who received cadaveric lung transplants between October 1987 and June 1997 for whom HLA data were available. Two cohorts were examined, depending on the era of transplantation: (1) October 1987 to June 1997 (n = 3,549): Differences in actuarial survival as stratified by either the total number of HLA mismatches or the number of mismatches at each HLA locus were determined using a log-rank test. Multivariate logistic regression models were developed to determine independent predictors of survival at 1, 3, and 5 years following lung transplantation. (2) April 1994 to June 1997 (n = 1,796): The association of HLA mismatching with acute rejection and obliterative bronchiolitis was determined using a chi-squared analysis. RESULTS: Only 164 patients (4.6%) received lung grafts with 2 or fewer HLA mismatches. Univariate analyses demonstrated a significant difference in post-transplant survival by mismatch level, with the total number of HLA mismatches (p = 0.0008) and mismatching at the HLA-A locus (p = 0.002) associated with worse survival. Multivariate logistic regression demonstrated that the number of mismatches at the HLA-A and HLA-DR loci predicted 1-year mortality (incremental odds ratios 1.18, p = 0.01, and 1.15, p = 0. 03, respectively). The total number of HLA mismatches predicted 3- and 5-year mortality (incremental odds ratios 1.13 at 3 years, p = 0. 0004, and 1.14 at 5 years, p = 0.0002). However, other covariates such as repeat transplantation, transplantation for congenital heart disease, advanced recipient age, and an early era of transplantation were stronger predictors of mortality. We found no significant association between HLA mismatching and the development of obliterative bronchiolitis, although there was an association between mismatching at the HLA-A locus and acute rejection episodes requiring hospital admission (p = 0.008). We also found no association between mismatching at the HLA-B locus and rejection episodes requiring either hospitalization or the alteration of anti-rejection medications (p = 0.034). CONCLUSION: Although the number of HLA mismatches at the HLA-A and HLA-DR loci predicted 1-year mortality and the total number of mismatches predicted 3- and 5-year mortality following lung transplantation, the effect of each covariate was small in this multicenter study of 3,549 patients. Further close follow-up of registry patients is necessary to determine the effect of HLA matching on long-term survival and freedom from obliterative bronchiolitis and rejection following lung transplantation. A prospective study of HLA matching for lung transplantation should not yet be considered in view of the small number of grafts with 2 or fewer mismatches and the modest effect of HLA matching on outcome.

Adult↗

Preferential elevation of Prx I and Trx expression in lung cancer cells following hypoxia and in human lung cancer tissues.

Transient/chronic microenvironmental hypoxia that exists within a majority of solid tumors has been suggested to have a profound influence on tumor growth and therapeutic outcome. Since the functions of novel antioxidant proteins, peroxiredoxin I (Prx I) and II, have been implicated in regulating cell proliferation, differentiation, and apoptosis, it was of our special interest to probe a possible role of Prx I and II in the context of hypoxic tumor microenvironment. Since both Prx I and II use thioredoxin (Trx) as an electron donor and Trx is a substrate for thioredoxin reductase (TrxR), we investigated the regulation of Trx and TrxR as well as Prx expression following hypoxia. Here we show a dynamic change of glutathione homeostasis in lung cancer A549 cells and an up-regulation of Prx I and Trx following hypoxia. Western blot analysis of 10 human lung cancer and paired normal lung tissues also revealed an elevated expression of Prx I and Trx proteins in lung cancer tissues. Immunohistochemical analysis of the lung cancer tissues confirmed an augmented Prx I and Trx expression in cancer cells with respect to the parenchymal cells in adjacent normal lung tissue. Based on these results, we suggest that the redox changes in lung tumor microenvironment could have acted as a trigger for the up-regulation of Prx I and Trx in lung cancer cells. Although the clinical significance of our finding awaits more rigorous future study, preferential augmentation of the Prx I and Trx in lung cancer cells may well represent an attempt of cancer cells to manipulate a dynamic redox change in tumor microenvironment in a manner that is beneficial for their proliferation and malignant progression.

Antioxidants↗

Lipid metabolism by rat lung in vitro. Effect of starvation and re-feeding on utilization of (U- 14 C)glucose by lung slices.

1. The incorporation of [U-(14)C]glucose into several lipid components of lung and liver slices, and the activities of glucose 6-phosphate dehydrogenase (EC 1.1.1.49), 6-phosphogluconate dehydrogenase (EC 1.1.1.44), ;malic' enzyme (EC 1.1.1.40) and NADP-isocitrate dehydrogenase (EC 1.1.1.42) of the cell cytosol were examined in normal, starved and re-fed rats. 2. Lipogenesis and the activities of these enzymes in liver were decreased markedly in rats starved for 72h. Re-feeding starved rats on a fat-free diet for 72h resulted in the well documented hyperlipogenic response in liver, particularly in its ability to convert glucose into neutral lipid, and increased activities of glucose 6-phosphate dehydrogenase, ;malic' enzyme and 6-phosphogluconate dehydrogenase to values approx. 700, 470 and 250% of controls respectively. 3. Approx. 70% of the total label in lung lipids was present in the phospholipid fraction. Hydrolysis of lung phospholipids revealed that lipogenesis from glucose was considerable, with approx. 40% of the total phospholipid radioactivity present in the fatty acid fraction. 4. Incorporation of glucose into total lung lipids was decreased by approx. 40% in lung slices of starved rats and was returned to control values on re-feeding. Although phospholipid synthesis from glucose was decreased in lung slices of starved rats, the decrease proportionally was greater for the fatty acid fraction (approx. 50%) as compared with the glycerol fraction (approx. 25%). 5. The activities of lung glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and NADP-isocitrate dehydrogenase were not affected by the dietary alterations. ;Malic' enzyme activity was not detected in lung cytosol preparations. 6. The results are discussed in relation to the surface-active lining layer (surfactant) of the lung.

Animals↗

Predictors of renal function following lung or heart-lung transplantation.

BACKGROUND: Renal failure is a common complication following non-renal solid organ transplantation. The purpose of our study was to define the rate of decline in renal function and to identify independent risk factors associated with renal failure following lung or heart-lung transplantation. METHODS: Between May 1986 and December 1998, 219 patients underwent lung or heart-lung transplantation at the University of Minnesota and survived at least six months (33 heart-lung, 66 bilateral single lung, and 120 unilateral single lung transplants). The mean age at the time of transplant was 45.9 +/- 11.6 years (mean +/- SD; range, 15 to 65 years), and the mean pre-transplant serum creatinine level was 0.88 +/- 0.19 mg/dL. All patients were treated with a calcineurin inhibitor (164 cyclosporine, 55 tacrolimus). RESULTS: During the follow-up period (median 44 months, range 6.8 to 163 months), 16 patients (7.3%) developed end-stage renal disease. The cumulative incidence of doubling of serum creatinine was 34% at one year, 43% at two years and 53% by five years. Factors associated with the primary end point of the time to doubling of the baseline serum creatinine by proportional hazards regression were cumulative periods with diastolic blood pressure greater than 90 mm Hg [relative risk (RR) 1.30, P = 0.02] and the serum creatinine value at one month post-transplantation (RR 1.28, P = 0.03). Use of tacrolimus during the first six months after transplantation was associated with a significant decrease in the risk for time to doubling of serum creatinine (RR 0.38, P = 0.009) and a lower rate of acute rejection. CONCLUSIONS: These results suggest that potential renoprotective strategies following lung or heart-lung transplantation include avoidance of peri-transplant renal injury, diligent blood pressure control, and preferential use of tacrolimus over cyclosporine.

Acute Disease↗

Short-term tracheal occlusion in fetal lambs with diaphragmatic hernia improves lung function, even in the absence of lung growth.

BACKGROUND/PURPOSE: Prolonged tracheal occlusion (TO) accelerates lung growth but impairs surfactant production. Short-term TO results in less lung growth but preserves type II cell function. The authors studied the effects of short-term TO on lung physiology in diaphragmatic hernia. METHODS: Diaphragmatic hernia was created in 9 fetal lambs at 90 to 95 days. Five were left uncorrected (CDH), 4 underwent 2-week TO (108 to 122 days; CDH + TO). Five unoperated lambs served as controls. Near-term (136 days) fetuses were ventilated for 90 to 150 minutes. Pulmonary arterial pressure, postductal blood gases, quasistatic compliance, total lung capacity (TLC), and lung weight to body weight (LW/BW) were measured. RESULTS: There was an overall survival rate of 89% at full term. Short-term occlusion did not induce lung growth (TLC and LW/BW, 6.07 +/- 2.92 mL/kg and 0.022 +/- 0.008 in CDH, 4.86 mL/kg and 0.019 +/- 0.005 in CDH + TO, 10.81 +/- 3.55 mL/kg and 0.036 +/- 0.006 in controls, respectively). However, pulmonary hypertension in CDH (47.4 +/- 12.32/35.8 +/- 12.19 torr) was corrected by short-term occlusion (20.2 +/- 4.0/16.0 +/- 4.8 torr in CDH + TO, P< .05, single-factor analysis of variance [ANOVA]; similar to control). Best pO2 and pCO2 improved after occlusion (CDH, 48.6 +/- 6.7 torr and 107.1 +/- 34.3 torr, respectively; CDH + TO, 101.5 +/- 16.3 torr and 81.9 +/- 2.4 torr; control, 291.4 +/- 4.7 torr and 37.7 +/- 17.3), as did oxygenation index (P < .05, CDH vCDH + TO; CDH, 97.2 +/- 23.0; CDH + TO, 28.7 +/- 3.1; control, 5.6 +/- 0.6). CONCLUSIONS: Short-term TO corrects pulmonary hypertension and improves gas exchange in fetal lambs with diaphragmatic hernia despite failure to produce accelerated lung growth. Inducing lung maturation and correcting the physiological derangement in diaphragmatic hernia may be more important than achieving lung growth alone.

Analysis of Variance↗

Platelet-activating factor induces lung inflammation and leak in rats: hydrogen peroxide production along neutrophil-lung endothelial cell interfaces.

Interleukin-1 (IL-1) levels and neutrophils are increased in the lung-lavage fluid of patients with acute lung injury (ALI), and instilling IL-1 intratracheally into rats causes rapid lung-neutrophil influx and neutrophil-dependent lung leakage. IL-1 insufflation also increases platelet-activating factor (PAF) activity in rat lung, and PAF is increased in the lung-lavage fluid of ALI patients. To assess the direct effects of PAF on the lung, we administered PAF intratracheally in rats. We found that rats given PAF (5 microg) intratracheally had increased lung nuclear factor-kappaB activation, myeloperoxidase activity, numbers of lavage neutrophils, lavage neutrophil nitroblue tetrazolium reduction, and leakage compared with sham-treated rats administered saline solution intratracheally. Electron microscopic examination also indicated that lungs from rats given PAF intratracheally had increased neutrophil infiltration, cell damage, and neutrophil-endothelial cell interface cerium chloride staining - a marker of hydrogen peroxide production - compared with sham-treated rats. Simultaneous treatment with a PAF receptor-antagonist, WEB 2086, decreased the aforementioned changes observed after intratracheal administration of PAF.

Animals↗

Improved lung allograft function after fundoplication in patients with gastroesophageal reflux disease undergoing lung transplantation.

OBJECTIVES: Bronchiolitis obliterans is the greatest limitation to the long-term applicability of lung transplantation. Although alloimmune events are important, nonimmune events, such as gastroesophageal reflux, might contribute to lung injury and the development of bronchiolitis obliterans syndrome. METHODS: We retrospectively studied the 396 patients who underwent lung transplantation at the Duke Lung Transplant Program from April 1992 to April 2002. Reflux was assessed for using an ambulatory 24-hour esophageal pH probe. RESULTS: Reflux assessment with an esophageal pH probe was obtained in 128 patients after lung transplantation. Abnormal pH study results were present in 93 (73%) patients. Forty-three patients underwent a surgical fundoplication. There was no in-hospital or 30-day mortality in the patients undergoing fundoplication. At the time of fundoplication, 26 patients met the criteria for bronchiolitis obliterans syndrome. After fundoplication, 16 patients had improved bronchiolitis obliterans syndrome scores, with 13 of these patients no longer meeting the criteria for bronchiolitis obliterans syndrome. In patients at least 6 months after lung transplantation and 6 months after fundoplication, the forced expiratory volume in 1 second improved by an average of 24% (mean forced expiratory volume in 1 second before fundoplication, 1.87 L; mean forced expiratory volume in 1 second after fundoplication, 2.19 L/sec; P <.0002). Overall actuarial survival was significantly better in patients who had either normal pH studies or who had fundoplication. CONCLUSIONS: Gastroesophageal reflux disease is very common after lung transplantation and appears to contribute to mortality and development of bronchiolitis obliterans syndrome. Fundoplication in lung transplant recipients with gastroesophageal reflux disease is associated with significant improvements in lung function, particularly if performed before the late stages of bronchiolitis obliterans syndrome.

Actuarial Analysis↗

Environmental exposure and lung cancer among nonsmokers: an example of Taiwanese female lung cancer.

Lung cancer is the leading cause of cancer death worldwide and in Taiwan. Cigarette smoking is considered to be the most important risk factor, since about 90% of lung cancer can be related to cigarette smoking. Despite the recent decrease of cigarette smoking, lung cancer is still the leading cause of cancer death in the United States. In Taiwan, only around 50% of lung cancer incidence could be associated with cigarette smoking, particularly less than 10% of Taiwanese women are smokers. Thus, the aetiology of lung cancer for nonsmokers remains unknown. DNA damages including bulky and oxidative damage may be related with mutation of tumor suppressor genes, such as p53 gene. The high DNA adduct levels in female may be associated with frequent exposure to indoor cooking oil fumes (COF) and outdoor heavy air pollution. Oxidative stress induced by COF was also discussed. Different p53 mutation spectra and mutation frequency between genders reflected that different environmental factors may be involved in nonsmoking male and female lung cancer development. Most importantly, our recent report has demonstrated that human papillomavirus (HPV) infection was associated with nonsmoking female lung cancer. Based on our studies with Taiwanese nonsmoking lung cancer as the model, the possible aetiological factors of lung cancer incidence in Taiwanese nonsmokers were elucidated.

Adult↗

Conditional expression of the mutant Ki-rasG12C allele results in formation of benign lung adenomas: development of a novel mouse lung tumor model.

To determine the effects of expression of mutant Ki-ras on lung tumorigenesis, we developed a bitransgenic mouse model that expresses the human Ki-ras(G12C) allele in alveolar type II and/or Clara cells in a tetracycline-inducible, lung-specific manner. Expression of Ki-ras(G12C) caused multiple, small lung tumors over a 12-month time period. Although tumor multiplicity increased upon continued Ki-ras expression, most lung lesions were hyperplasias or well-differentiated adenomas. This is in contrast to the more severe phenotypes observed in other transgenic mouse models in which different mutant Ki-ras alleles were expressed in the lung. Expression of Ki-ras(G12C) was associated with a 2-fold increase in the activation of the Ras and Ral signaling pathways and increased phosphorylation of Ras downstream effectors, including Erk, p90 ribosomal S6 kinase, ribosomal S6 protein, p38 and MAPKAPK-2. In contrast, expression of the transgene had no effect on the activation of the JNK and Akt signaling pathways. Withdrawal of doxycycline for 1 month resulted in almost a complete absence of proliferative pulmonary lesions, suggesting tumor regression in the absence of Ki-ras expression. Mutant Ki-ras(G12C) expression was sufficient for initial lung tumor transformation, required for maintenance of tumor phenotype, and induced transformation of lung epithelial cells by the activation of multiple effector pathways. These results describe a novel mouse lung tumor model demonstrating benign tumor development in the absence of tumor progression, which will provide a new tool for understanding the early stages of lung tumor pathogenesis.

Adenoma↗

How the left lung is perfused after ligating the left pulmonary artery in the pig at birth: clinical implications for the hypoperfused lung.

The left pulmonary artery and ductus arteriosus were ligated in 14 pigs at birth. Animals were sacrificed at intervals from 2 to 24 weeks of age. In the right lung the pulmonary artery and in the left, either the distal pulmonary artery, bronchial arteries or both were injected. The fixed lung specimens were studied by arteriography, dissection and microscopic examination of serial and random sections of lung tissue. The bronchial arterial circulation to, and within the right lung appeared normal and was similar to that described in the human lung. In the left lung, the bronchial arterial circulation hypertrophied rapidly during the first 2 weeks, and large anastomoses between pulmonary and systemic circulations were found at the same sites as in the normal pig lung. The position and structural characteristics of the anastomosing arteries is described in the different types of broncho-pulmonary connection. In most animals aged 16 weeks or more, peripheral bronchial arteries immediately proximal to the anastomotic sites, developed intimal and medial proliferation. The left lung continued to grow although in all animals it was small. The axial pulmonary artery and its branches became smaller with age. These findings help explain how the lung is perfused and grows in children with congenital heart disease and an acquired collateral pulmonary arterial circulation.

Animals↗

High incidence of migration inhibition reactivity to lung tumor-associated antigen by normal donors in close contact with lung cancer patients or materials.

A high incidence of indirect leukocyte migration inhibition reactivity of normal donors to a 3-M KCl extract from a fresh pleural effusion of a patient with a lung adenocarcinoma (designated 7661) was observed. When these normal donors were classified according to contact with lung cancer patients or materials, 22 of 32 (72%) normal donors in contact with lung cancer patients or materials were reactive with the 7661 extract as compared to only 3 of 76 (4%) who had no contact. Of normal donors involved in the direct care of lung cancer patients, 14 of 20 (70%) were positive, whereas only 2 of 10 (20%) hospital personnel who worked with noncancer patients were reactive. Among laboratory personnel who handled blood and tissue specimens from lung cancer patients, 8 of 11 (73%) were positive with the 7661 extract, whereas none of 5 laboratory workers who worked with cancer materials unrelated to lung cancer were positive. Also, none of 13 personnel working in laboratories adjacent to those where lung cancer tests were performed were reactive with 7661. None of the 16 blood bank donors and none of 11 secretarial and clerical staff who worked in biochemical laboratories were positive. Reactivity was no correlated with a smoking history. Thus development of reactivity appeared to require direct contact with lung cancer patients or materials. The results suggested a horizontal transmission of reactivity against an antigen associated with lung cancer.

Adenocarcinoma↗

Quantification of DNA adducts formed in liver, lungs, and isolated lung cells of rats and mice exposed to (14)C-styrene by nose-only inhalation.

Bronchiolo-alveolar tumors were observed in mice exposed chronically to 160 ppm styrene, whereas no tumors were seen in rats up to concentrations of 1000 ppm. Clara cells, which are predominant in the bronchiolo-alveolar region in mouse lungs but less numerous in rat and human lung, contain various cytochrome P450s, which may oxidize styrene to the rodent carcinogen styrene-7,8-oxide (SO) and other reactive metabolites. Reactive metabolites may form specific DNA adducts and induce the tumors observed in mice. To determine DNA adducts in specific tissues and cell types, rats and mice were exposed to 160 ppm [ring-U-(14)C]styrene by nose-only inhalation for 6 h in a recirculating exposure system. Liver and lungs were isolated 0 and 42 h after exposure. Fractions enriched in Type II cells and Clara cells were isolated from rat and mouse lung, respectively. DNA adduct profiles differed quantitatively and qualitatively in liver, total lung, and enriched lung cell fractions. At 0 and 42 h after exposure, the two isomeric N:7-guanine adducts of SO (measured together, HPEG) were present in liver at 3.0 +/- 0.2 and 1.9 +/- 0.3 (rat) and 1.2 +/- 0.2 and 3.2 +/- 0.5 (mouse) per 10(8) bases. Several other, unidentified adducts were present at two to three times higher concentrations in mouse, but not in rat liver. In both rat and mouse lung, HPEG was the major adduct at approximately 1 per 10(8) bases at 0 h, and these levels halved at 42 h. In both rat Type II and non-Type II cells, HPEG was the major adduct and was about three times higher in Type II cells than in total lung. For mice, DNA adduct levels in Clara cells and non-Clara cells were similar to total lung. The hepatic covalent binding index (CBI) at 0 and 42 h was 0.19 +/- 0.06 and 0.14 +/- 0.03 (rat) and 0. 25 +/- 0.11 and 0.44 +/- 0.23 (mouse), respectively. The pulmonary CBIs, based on tissues combined for 0 and 42 h, were 0.17 +/- 0.04 (rat) and 0.24 +/- 0.04 (mouse). Compared with CBIs for other genotoxicants, these values indicate that styrene has only very weak adduct-forming potency. The overall results of this study indicate that DNA adduct formation does not play an important role in styrene tumorigenicity in chronically exposed mice.

Animals↗