How do climate change experiments alter plot-scale climate?

To understand and forecast biological responses to climate change, scientists frequently use field experiments that alter temperature and precipitation. Climate manipulations can manifest in complex ways, however, challenging interpretations of biological responses. We reviewed publications to compile a database of daily plot-scale climate data from 15 active-warming experiments. We find that the common practices of analysing treatments as mean or categorical changes (e.g. warmed vs. unwarmed) masks important variation in treatment effects over space and time. Our synthesis showed that measured mean warming, in plots with the same target warming within a study, differed by up to 1.6  ∘ C (63% of target), on average, across six studies with blocked designs. Variation was high across sites and designs: for example, plots differed by 1.1  ∘ C (47% of target) on average, for infrared studies with feedback control (n = 3) vs. by 2.2  ∘ C (80% of target) on average for infrared with constant wattage designs (n = 2). Warming treatments produce non-temperature effects as well, such as soil drying. The combination of these direct and indirect effects is complex and can have important biological consequences. With a case study of plant phenology across five experiments in our database, we show how accounting for drier soils with warming tripled the estimated sensitivity of budburst to temperature. We provide recommendations for future analyses, experimental design, and data sharing to improve our mechanistic understanding from climate change experiments, and thus their utility to accurately forecast species' responses.

Diatoms response to salinity changes: investigations using single pulse and cross polarisation magic angle spinning 29Si NMR spectra.

Diatoms Thalassiosira pseudonana and Chaetoceros muelleri (Bacillariophyceae) were cultured at three different salinities (26, 36 and 46 practical salinity units (PSU)) and their silica content examined using natural abundance 29Si magic angle spinning (MAS) nuclear magnetic resonance (NMR) spectroscopy. The samples were investigated using both single pulse (SP) and cross-polarisation (CP) MAS experiments. In addition, samples of T. pseudonana were examined using variable contact time CP MAS experiments allowing the dynamics (TSiH and T1rρH) of CP to be determined. Comparison of SP and CP results revealed a change to a less condensed silica state when diatoms were cultured at salinities away from optimal (36 PSU). Further, an increased amount of organic material (but not its composition) was determined to be present for such samples affecting CP experiments. The location of the organic material, on the diatom surface or within the frustule, was unable to be determined.

Tetra-porphyrin molecular tweezers: two binding sites linked via a polycyclic scaffold and rotating phenyl diimide core.

The synthesis of a tetra-porphyrin molecular tweezer with two binding sites is described. The bis-porphyrin binding sites are aligned by a polycyclic scaffold and linked via a freely rotating phenyl diimide core. Synthesis was achieved using a divergent approach employing a novel coupling method for linking two polycyclic units to construct the core, with a copper(ii)-mediated phenyl boronic acid coupling found to extend to our polycyclic imide derivative. We expect this chemistry to be a powerful tool in accessing functional polycyclic supramolecular architectures in applications where north/south reactivity and/or directional interactions between modules are important. Porphyrin receptor functionalisation was undertaken last, by a four-fold ACE coupling reaction on the tetra-epoxide derivative of the core.

Effect of particle size on the lymphatic distribution of 111Indium-aminopolystyrene through intrapleural administration.

The study examined the impact of size on lymphatic particle distribution through intrapleural (ipl.) administration. Aminopolystyrene of three sizes, 0.29 microm, 2.18 microm, and 11.2 microm were radiolabeled with 111Indium and their biodistributions were evaluated in rats after ipl administration. Animals received either particles of three different sizes (4 mg, 200 microCi/animal) or unconjugated 111Indium as control. The percentage of injected dose (%ID) per organ or sample was determined for left (L) and right (R) mediastinal lymph nodes (LN), blood, lung, and pleural wash. The biodistribution of 2.18 microm 111In-aminopolystyrene was further investigated at 6 h, 24 h, 48 h, and 72 h following ipl administration to examine the possible particle retention time. The 2.18 microm particles had significantly higher uptake in both LLN and RLN compared to other sizes. The systemic uptake was minimal. At 72 h, there was still 3.2 +/- 3.2% and 2.1 +/- 1.8% of injected dose retained in the LLN and RLN, respectively. Scintigraphic imaging revealed significant accumulation of the radioactivity in mediastinal nodes. Particle size has significant impact on lymphatic particle distribution through ipl administration. Approximately 2 microm seems to be a suitable size.

Secondary screening system for preclinical testing of human lung cancer therapies.

The National Cancer Institute has instituted a primary screening system for testing new agents against cultured cancer cell lines. The purpose of this study was to determine the feasibility of using a nude rat orthotopic (organ-specific) human lung cancer model system as an in vivo secondary screen for general evaluation of new anticancer agents and therapies active against lung cancer. To make this determination, we tested whether this system allows measurement of uptake and tumoricidal activity of anticancer therapies. Tumor-bearing lungs from 53 Rowett nude rats with orthotopically implanted human large-cell undifferentiated lung carcinoma (NCI-H460) were perfused ex vivo for 1 hour with or without each of two anticancer modalities. Lungs were perfused with blood-free perfusate alone (untreated control), perfusate with 100 micrograms/mL doxorubicin (treated positive control), or perfusate with lymphokine-activated killer cells plus human recombinant interleukin-2 (LAK/rIL-2). Weight gain during perfusion was the criterion used to quantitate lung injury. Treatment efficacy was measured by clonogenic assay after enzymatic disaggregation of the perfused tumors. Doxorubicin levels in the tumor and in the uninvolved lung were measured by high-performance liquid chromatography. Both treatment groups showed only slight increases in lung weight compared with that in the untreated control group, suggesting good lung tolerance of the procedure. Lung and tumor levels of doxorubicin were 320 +/- 21 ng/mg of tissue and 32 +/- 5 ng/mg of tissue (means +/- SE), respectively. Clonogenic assay demonstrated a fivefold to 10-fold reduction in the surviving fraction of tumor cells with doxorubicin but no change with LAK/rIL-2.(ABSTRACT TRUNCATED AT 250 WORDS)

Mucin gel formed by tumorigenic squamous lung carcinoma cells has Le(a)-X oligosaccharides and excludes antibodies from underlying cells.

Cells of cloned lines of human squamous lung carcinomas elaborate large glycoproteins that are associated with their tumorigenic potential. Two groups of clones (called Le(a)-X-positive and Le(a)-X-negative) were studied that either do or do not express the Le(a)-X oligosaccharide associated with large glycoproteins and mucins secreted by these clones. Le(a)-X-positive cells elaborate a mucin gel complex associated with their apical surfaces, which appears as a mosaic of extracellular plates. Clones of this type are tumorigenic in nude rodents when injected s.c. or when introduced into the lungs via intrabronchial aerosol. By contrast, the Le(a)-X-negative clones do not form extracellular plates and are not tumorigenic in the lungs or subcutaneously. We demonstrate that the extracellular plates of Le(a)-X-positive cells exclude antibodies from interacting with the underlying squamous lung carcinoma cells and may therefore exert an immunoprotective effect. In support of this possibility it was found that: (a) There is a substantial inflammatory cell infiltrate associated with regressing nodules of Le(a)-X-negative cells in nude rodent lung and subcutaneous nodules, while there is no observable infiltration associated with progressing Le(a)-X-positive tumors. (b) In the brain (an immunoprivileged site) tumors develop and progress when either Le(a)-X-negative or -positive cells are introduced.

Irradiated nude rat model for orthotopic human lung cancers.

The development of improved animal models for biological and preclinical studies of human lung cancer is important because lung cancer is the leading cause of cancer death in the United States. To determine whether the Rowett nude rat could serve as an orthotopic (organ-specific) model of this disease, nude rats (CR: NIH-RNU), with and without 500 rads of prior gamma-irradiation, were implanted intrabronchially with 10(7) cultured cells from 3 human lung cancer lines. Without irradiation, the NCI-H460 large-cell undifferentiated carcinoma had a 54% take-rate, whereas the NCI-H125 adenosquamous carcinoma and A549 adenocarcinoma had take-rates of 7 and 33%, respectively; irradiation increased the respective take-rates to 100, 83, and 90%. In irradiated rats, tumor age versus weight measurements showed progressive growth for all three tumors, with growth rates in the order: NCI-H460 greater than A549 greater than NCI-H125, requiring approximately 3, 5, and 9 weeks, respectively, for average tumor sizes to exceed 500 mg. The small-cell carcinoma cell line NCI-H345 was implanted only into irradiated rats and resulted in more slowly growing tumors. Histopathological study showed all model tumor types to have histological characteristics consistent with the clinical tumors from which the cell lines were derived. Each tumor type had a different growth pattern, with some of the the A549- and NCI-H125-derived tumors metastasizing to contralateral lung and/or regional lymph nodes. There was no evidence for immunological rejection in irradiated, tumor-bearing rats. Nonirradiated, implanted rats without gross tumor exhibited peribronchiolar mononuclear cell infiltration with or without fibrosis, suggesting prior immunological rejection. The successful orthotopic growth of these 4 human lung cancer cell lines in irradiated nude rats suggests that this model could be useful for biological and preclinical studies of human lung cancer, both in intact rats and via ex vivo perfusion of their tumor-bearing lungs.

Occurrence of the Perfect Stage of Powdery Mildew of Sugar Beets in Southern Montana in 2003.

Wide spread powdery mildew infections on sugar beets were observed at the Southern Agricultural Experiment Station in Huntley, MT during September, 2003. Throughout the area, lower leaves were frequently heavily covered by the vegetative stage of the fungus with plants at the edge of the field having clearly visible abundant mature (black) and immature (brown) globose ascocarps on the leaf surfaces and stems. The fruiting structures had mostly branched appendages and were imbedded in the superficial mycelium. Their diameter ranged from 70 to 100 µm. Each ascocarp contained five to eight asci with one to four ascospores (mostly three) per ascus. Elliptical ascospores were hyaline and measured 20 to 25 µm long and 12 to 20 µm wide. On the basis of the descriptions given for isolates from Idaho and Colorado (1) and the usage of Erysiphe polygoni DC for powdery mildew on sugar beet in the United States, this isolate may be classified as E. polygoni DC. However, measurements taken show that ascocarps, asci, and ascospores also fall within the range of E. betae (Vanha) Weltz. as described by Weltzien (2). We strongly suggest that these species be compared by using rDNA analysis of the ITS region to determine whether they are separate species. If survival of the ascocarps and the viability and pathogenicity of the ascospores can be confirmed, epidemics of sugar beet powdery mildew could be understood as local and regional events that are not dependant on long distance dispersal of conidiospores. The occurrence of the perfect stage also could lead to the more frequent appearance of new races through genetic recombination. References: (1) J. J Gallian and L. E. Hanson. Plant Dis. 87:200, 2003. (2) H. C. Weltzien. Phytopathol. Z. 47:123, 1963.

Characterization of a highly metastatic, orthotopic lung cancer model in the nude rat.

The prevailing subcutaneous nude rodent tumor xenograft models used for biological and preclinical studies do not optimally reflect some important biological properties of cancer, especially invasion and metastasis. Orthotopic models have been developed to address this need. However, for lung cancer none of the available models are optimal, in that none originate from an orthotopic (bronchial) primary site and exhibit extensive extrathoracic metastasis. Our goal was to develop a consistent rodent model of non-small cell lung cancer with both of these properties. Groups of male Rowett nude rats were given 500 rads of gamma radiation and then endobronchially implanted in the right caudal lobe airway with 50 mg of small NCI-H460 tumor fragments taken from an orthotopic donor tumor. They were then sacrificed at selected post-implantation times and evaluated grossly and histologically for animal weight, primary tumor take and size, and metastatic tumor incidence at multiple sites. At a late time point (32-35 days), consistency of primary tumor size and metastasis was estimated by comparing results from four groups of rats implanted on different occasions. The results showed that the primary tumors grew steadily, reaching four grams by days 32-35. Rats gained weight until days 14 to 21, but then began to show cachexia. High metastatic rates (>60%) were seen for mediastinal lymph nodes (by 21 days), and kidney, bone and brain (by 28 days). Mean primary tumor size and the incidences of both regional and systemic metastasis were consistent at 32-35 days in four different groups of six animals. In conclusion, this orthotopic lung cancer model is highly metastatic and consistent in terms of both primary tumor growth and metastatic behavior. It is the only available rodent model of human lung cancer emanating from an endobronchial site and metastasizing to multiple extrapulmonary sites, and should be very useful for both biological and preclinical studies of lung cancer, particularly where studies of antimetastatic activity are of interest, and/or where survival studies are desired.

The biology of some intraerythrocytic parasites of fishes, amphibia and reptiles.

Fishes, amphibia and reptiles, the ectothermic vertebrates, are hosts for a variety of intraerythrocytic parasites including protists, prokaryotes, viruses and structures of uncertain status. These parasites may experience host temperature fluctuations, host reproductive strategies, population genetics, host habitat and migratory behaviour quite unlike those of endothermic hosts. Few blood infections of fishes, amphibia and reptiles have proven pathogenicity, in contrast to the many intraerythrocytic parasites of mammals and some birds which harm their hosts. Although not given the attention afforded to intraerythrocytic parasites of endotherms, those of ectotherms have been studied for more than a century. This review reports on the diversity, general biology and phylogeny of intraerythrocytic parasites of ectotherms. The existence of taxonomic confusion is emphasized and the main taxonomic features of most of the 23 better characterized genera, particularly the kinetoplastid and apicomplexan protists, are summarized. Transmission of protistan infections of aquatic ectotherms is also discussed. Leeches can transfer sporozoties or merozoites to the vertebrate host during feeding. Dormant sporozoites of Lankesterella may permit transmission of species of this genus between vertebrates by predation. The fish haemogregarine, Haemogregarina bigemina, probably has gnathiid isopods, rather than leeches, as its definitive hosts. Hepatozoon spp. in aquatic hosts, and Progarnia of caiman, may also use invertebrate hosts other than leeches. Protistan infections of terrestrial or semi-terrestrial hosts are transmitted by a variety of arthropods, or, in some cases, leeches, contaminated paratenic hosts, or sporocysts free in water. Transfer of protists between vertebrates by predation and congenitally may also occur. The biology of the host cells of these infections, the red blood cells of ectotherm vertebrates, is summarized and compared with that of mammalian erythrocytes. Erythropoiesis, the nature of the surface molecules (especially the possible existence of a major histocompatibility complex), the haemoglobins, and the shape and size of erythrocytes are discussed. The exoerythrocytic sites in which protists, prokaryotes, viruses and structures of uncertain status exist before erythrocyte entry are described. Tissue merogony, tissue cysts and invasion of the white cell series occur in a variety of protistan infections. Intraerythrocytic stages of protistan infections are also discussed, including modes of entry to erythrocytes, survival mechanisms, and multiplication. The impact of infection on host populations is difficult to assess, in part because there is no agreement in the literature on the criteria used to evaluate parasite-induced cost to the host. Almost all studies have been on haemogregarine and Plasmodium infections in, mainly, lizards, but also fishes and snakes. Some infections may be responsible for mortality in their hosts, but hosts themselves may be short-lived, or have a limited ability to recover from infection.

Biochemical analyses on single amphidial glands, excretory-secretory gland cells, pharyngeal glands and their secretions from the avian nematode Syngamus trachea.

The gape nematode, Syngamus trachea, has been used as a model to study nematode secretions. Individual and intact pairs of amphidial glands, pharyngeal glands and pairs of excretory-secretory gland cells have been dissected and their secretory products analysed. The protein profiles of each gland and the total nematode secretions were analysed on 12.5% homogeneous SDS-PAGE minigels. The protein analyses revealed that the structural protein profile of each gland is different. The amphidial gland secretes two major proteins of 36.0 and 41.5 kDa, the excretory-secretory gland cell secretes a protein of 28.2 kDa and a protein of 14.3 kDa, and the pharyngeal gland secretes proteins of 41.5 and 14.6 kDa. Analysis of the total nematode secretions revealed all of the above major secretory proteins and an additional protein of 49.3 kDa. Syngamus trachea secretes acetylcholinesterases and its secretions contain multiple proteases.

A study of smoking, p53 tumor suppressor gene alterations and non-small cell lung cancer.

PURPOSE: The purpose of this study was to investigate the relationship between smoking and p53 tumor suppressor gene alterations, and their association with clinicopathologic features and prognosis in non-small cell lung cancer (NSCLC). METHODS: For 111 of 119 stage I-III NSCLC patients that had been followed prospectively, tumor p53 protein accumulation was measured immunohistochemically (IHC). Staining was evaluated as a score (p53IHCS) combining intensity and percent distribution. RESULTS: Forty-eight of 111 (43%) tumors had p53IHCS > 1. p53 IHC was associated with increasing tumor size (T) (p = 0.035), nodal status (N) (p = 0.091), stage (p = 0.054), and histology: squamous cell carcinoma (70%) and adenocarcinoma (27%) (p = 0.0002). In logistic regression analysis, p53 IHC was associated with squamous cell histology versus other histotypes [adjusted odds ratio (OR)5.90, 95% confidence interval (CI) 2.34-14.90]. p53 IHC was not associated with smoking variables. In multivariate proportional hazards analysis, p53IHCS and pack-years smoked (PY), both as continuous variables, were negative prognostic factors. The adjusted hazard ratios (HR) for the survival outcome recurrence for p53IHCS and PY were 1.20 (95% CI 1.02-1.40) and 1.03 (95% CI 1.01-1.04), and for death due to recurrent disease (DRD) were 1.35 (95% CI 1.11-1.64) and 1.03 (95% CI 1.01-1.04), respectively. Comparing the 75th percentile to the baseline 0, the adjusted HR for p53IHCS (5 vs. 0) was 4.5 and for PY (55 vs. 0) was 5.1 for the outcome DRD. Both variables demonstrated a dose-response relationship with survival. CONCLUSIONS: PY and p53IHCS are significant, independent and important predictors of recurrence and DRD in stage I-III NSCLC.

Median sternotomy for resection of pulmonary metastases.

Thoracic surgeons have been reluctant to adopt the median sternotomy as a suitable approach to pulmonary operations. However, its lesser functional morbidity and the capability to assess both lungs are advantageous in selected patients. Examined herein is a concentrated experience with 53 median sternotomies in 46 patients for the resection of pulmonary metastases. Forty of the 46 patients had soft tissue or osteogenic sarcoma. Full-lung tomograms diagnosed but did not accurately reflect the extent of disease. Fifty-three percent more tumor nodules were found at median sternotomy than were seen on full-lung tomography. Eleven of 18 patients (61%) thought to have unilateral disease by full-lung tomography had bilateral metastases found at median sternotomy. Most median sternotomies involved wedge resections (mean 8.9 range one to 52). Two segmentectomies, six lobectomies, and one pneumonectomy were also performed. Repeat median sternotomy was accomplished seven times; one patient underwent four median sternotomies. Sixty-six percent of the nodules removed proved to be tumor. Complications included one reoperation for bleeding; three patients had air leaks for more than 1 week, including one bronchopleural fistula; four required respirator assistance for more than 3 days; and there were four major infections. There was no operative mortality. Because of its low morbidity, the high incidence of unsuspected bilateral disease, and the elimination of a second operative procedure, median sternotomy is the approach of choice for the surgical treatment of pulmonary metastases.

Lung perfusion with chemotherapy in patients with unresectable metastatic sarcoma to the lung or diffuse bronchioloalveolar carcinoma.

Eight patients with metastatic sarcoma to the lung (n = 4) or diffuse bronchioloalveolar carcinoma of the lung (n = 4) underwent isolated lung perfusion with chemotherapy in a pilot study. Ages ranged from 18 to 60 years and half were female. The left lung was perfused in three patients (single lung perfusion) and both lungs in five patients (total lung perfusion). Perfusions ranged from 45 to 60 minutes at ambient or normothermic temperatures. One patient received perfusion at moderate hyperthermia (40 degrees C). Escalating doses of doxorubicin (1 to 10 micrograms/ml perfusate) was used in six patients, whereas two received cisplatin (14 and 20 micrograms/ml perfusate). There were two major complications and no objective responses. The isolated perfusion systems gave excellent separation between systemic and pulmonary circulations with zero to 15% of the measured peak drug concentration of the pulmonary perfusate detected in the systemic circulation. Drug concentrations in normal lung and tumor generally increased with higher drug dosages and drug was detectable in mediastinal lymph nodes of three out of four patients in whom sampling was done. Isolated lung perfusion with chemotherapy can be done safely in patients with lung malignancies and evidence suggests that higher drug dosages should be well tolerated.

Thoracic mass lesions in immuno-incompetent patients.

An aggressive surgical approach was used in the diagnosis and treatment of seven immuno-incompetent patients who presented with focal thoracic mass lesions. In five of the seven patients, minor diagnostic procedures had failed to provide a diagnosis. All seven patients were subjected to exploratory thoracotomy with resection and/or drainage of the involved area. Two patients had parenchymal masses, two had lung abscesses, two had empyemas with trapped lung, and one had a bronchial fistula. An accurate diagnosis and full resolution of the intrathoracic process was obtained in all patients. There was little morbidity and no operative mortality in this series. Resection of focal thoracic lesions in immuno-incompetent patients combines accurate diagnosis with precise therapy and is well tolerated in this high risk group of patients.

Tolerance of the isolated perfused lung to hyperthermia.

With the use of in vivo isolated lung perfusion for targeting antitumor therapy in the treatment of lung cancer, tolerance of normal lung tissue to the tumoricidal conditions becomes the limiting factor. This study was performed to determine the short-term tolerance of the lung to hyperthermia. Isolated dog lung lobes were perfused with autologous blood or an artificial salt solution at constant flow. Measurements of lung weight, extravascular water, vascular volume, serotonin uptake, urea permeability surface area product, perfusion pressure, and lung compliance were made with the temperature at about 37 degrees C. The temperature was then set at between 37 degrees and 45 degrees C, and at the end of the subsequent 2 hours the measurements were repeated. When the temperature was less than about 44.4 degrees C, hyperthermia had no detectable influence on the measured variables. Thus on the time frame consistent with in vivo perfusion therapy the normal lung appears to tolerate a fairly severe hyperthermia.

Analysis of prognostic factors in patients undergoing resection of pulmonary metastases from soft tissue sarcomas.

The selection of objective criteria that can reliably predict survival in patients undergoing resection of pulmonary metastases remains controversial. Between 1974 and 1982, 487 patients with soft tissue sarcomas presented to the National Cancer Institute. Eighty patients underwent thoracotomy for putative metastases and 67 patients had histologically proved pulmonary metastases. The 3 year tumor-free survival rate was 30% by actuarial analysis. Patients with resectable metastases had significantly prolonged post-thoracotomy survival compared to those patients with unresectable metastases. The most significant preoperative predictors of survival were the tumor doubling time, the number of metastases on preoperative linear chest tomograms, and the disease-free interval. Patients with a tumor doubling time of 20 days or more had a significantly longer post-thoracotomy survival (22 months median) than patients with a tumor doubling time of less than 20 days (6 months median). Those patients with four nodules or less on preoperative tomograms had significantly longer post-thoracotomy survival times (23 months median) than those patients with more than four nodules (6 months median). Patients with a disease-free interval of more than 12 months had a longer post-thoracotomy survival (32 months median) than patients with a disease-free interval of 12 months or less (10 months median). Combining these three prognostic factors significantly increased the predictive ability of this model. These criteria provide an accurate and rapid method to identify preoperatively those patients who will maximally benefit from resection of pulmonary metastases from soft tissue sarcomas.

Interstitial pulmonary infiltrate following combined therapy for esophageal carcinoma.

Seven patients with squamous cell carcinoma of the esophagus received radiation therapy in twice weekly 400 rad fractions over five weeks followed by esophagectomy. Four of these patients developed severe interstitial pulmonary infiltrates and died of pulmonary insufficiency 18-50 days after surgery. In three of these patients the infiltrates were detected within 24 hours of surgery while the infiltrate was not present until the eighth postoperative day in the fourth patient. Postmortem examination revealed widespread dilatation of interstitial and subpleural lymphatics. It is postulated that the combination of large fraction radiation therapy followed by extensive surgery resulted in lymphatic obstruction.

The limits of surgical resection alone for non-small cell lung cancer.

Surgical resection remains the best treatment for Stages I and II non-small cell lung cancer. In Stage IIIA disease the use of induction therapy has become widespread, although evidence supporting this approach is still preliminary. However, in subsets of patients with T3 tumours without mediastinal nodal involvement and those with certain single station, non-bulky N2 disease, surgery alone is still the preferred therapy. Studies show survival rates with surgery alone the same or higher than those achieved by most induction therapy regimens.

Extracellular flush solution that contains blood, mannitol, albumin, and prostacyclin protects rat lungs from six hours of ischemia.

We have used an isolated rat lung model to compare the quality of preservation of different flush techniques with each other and with topical cooling alone. Lung injury was assessed by recording lung weights after reperfusion after 4 and 6 hours of ischemia. The flush solutions studied were intracellular (Collins-Sacks), traditional extracellular, extracellular with low potassium plus dextran, and extracellular containing blood, mannitol, albumin, and prostacyclin (Wallwork's solution). Flushing with Wallwork's solution before both 4 and 6 hours of ischemia gave superior protection from lung edema after reperfusion over all the other methods.