Identification of potential erythrocyte phospholipid fatty acid biomarkers of advanced lung adenocarcinoma, squamous cell lung carcinoma, and small cell lung cancer.

New biomarkers for lung cancer would be valuable. Our aim was to analyze the fatty acid profiles of the main phospholipid species in erythrocytes from patients with advanced squamous cell lung carcinoma (SCC), lung adenocarcinoma (ADC), and small cell lung cancer (SCLC) and benign lung diseases (chronic obstructive pulmonary disease (COPD) and asthma) to determine the fatty acids that could be use as lung cancer markers. Twenty-eight, 18, 14, 16, and 15 patients with, respectively, SCC, ADC, SCLC, asthma, and COPD and 50 healthy subjects were enrolled in the study. Fatty acid profiles were investigated using gas chromatography/mass spectrometry followed by receiver operating characteristic (ROC) curve analysis. The fatty acid profiles changed significantly in the different pathologies analyzed. Based on the diagnostic yields and operating characteristics, the most significant fatty acids that might be used as biomarkers were as follows: ADC--arachidonic acid (20:4n6) in phosphatidylcholine and oleic acid (18:1n9) in phosphatidylethanolamine (PE); SCC--eicosapentaenoic acid (20:5n3) in PE and palmitic acid (16:0) in phosphatidylserine + phosphatidylinositol (PS+PI); SCLC--eicosadienoic acid (20:2n6) in PS+PI and lignoceric acid (24:0) in sphingomyelin. In conclusion, fatty acids from erythrocyte phospholipid species might serve as biomarkers in the diagnosis, and probably in other aspects related to clinical disease management, of ADC, SCC, and SCLC.

Erythrocyte fatty acids as potential biomarkers in the diagnosis of advanced lung adenocarcinoma, lung squamous cell carcinoma, and small cell lung cancer.

OBJECTIVES: To analyze the fatty acid profiles of erythrocyte total lipids from patients with advanced squamous cell lung carcinoma (SCC), lung adenocarcinoma (ADC), and small cell lung cancer (SCLC) and benign lung diseases (chronic obstructive pulmonary disease [COPD] and asthma) to reveal the fatty acids that could be used as lung cancer biomarkers. METHODS: Thirty, 20, 15, 17, and 19 patients with SCC, ADC, SCLC, COPD, and asthma, respectively, and 55 healthy participants were enrolled in our study. Fatty acid profiles were investigated using gas chromatography/mass spectrometry followed by receiver operating characteristic (ROC) curve analysis. Sialic acid (SA) and cytokeratins were measured by the thiobarbituric acid and immunoradiometric methods, respectively. RESULTS: At least one of the main fatty acids might be used as a biomarker for every type of lung cancer: arachidonic (20:4n6), linoleic (18:2n6), and stearic (18:0) acids for ADC, SCC, and SCLC, respectively. These fatty acids showed diagnostic yields and operating characteristics similar to or higher than the commonly used SA or cytokeratin markers. CONCLUSIONS: Fatty acids from erythrocyte total lipids might be used as diagnostic biomarkers of lung ADC, SCC, and SCLC. Their use in different aspects of the disease process needs to be explored.

Platelet linoleic acid is a potential biomarker of advanced non-small cell lung cancer.

New parameters that could be used as tumor markers for lung cancer would be valuable. Our aim was to analyze the fatty acid profiles of total lipids from erythrocytes and platelets from patients with advanced non-small cell lung cancer (NSCLC), chronic obstructive pulmonary disease (COPD) and asthma to reveal the fatty acids that could be used as NSCLC biomarkers. In our study, 50, 15 and 15 patients with advanced NSCLC, COPD and asthma and 50 healthy subjects were enrolled. Fatty acid profiles were investigated using gas chromatography/mass spectrometry followed by ROC (receiver operating characteristics) curves analysis to gain information about biomarkers. Sialic acid (SA) and cytokeratins were measured by the thiobarbituric acid and immunoradiometric methods respectively. Useful fatty acid markers were as follows: erythrocytes, 22:0 and linoleic acid (LA, 18:2n6); platelets, 16:0, 18:0, and LA. At the cutoff value to obtain maximum accuracy, the best biomarker was platelet LA, with higher diagnostic yields than the commonly used markers SA or cytokeratins (100%, 76%, 75% and 86% sensitivity, specificity, positive predictive value and accuracy, respectively). These findings suggest that platelet LA might be used as a biomarker of NSCLC in relation to different aspects of the disease process that now needs to be explored.

Erythrocyte and platelet phospholipid fatty acids as markers of advanced non-small cell lung cancer: comparison with serum levels of sialic acid, TPS and Cyfra 21-1.

The phospholipid fatty acid profiles of erythrocytes and platelets from fifty patients with advanced non-small cell lung cancer were investigated using gas chromatography/mass spectrometry, followed by "ROC" curves analysis to gain novel biomarker information. Sialic acid and cytokeratins were also examined. Potentially useful fatty acid markers: Erythrocytes: phosphatidylcholine, 18:2n6 and 20:4n6; phosphatidylethanolamine, 22:4n6 and 22:6n3 + 24:1n9. Platelets: phosphatidylcholine, 22.0; phosphatidylethanolamine, 22:5n3 + 24:0. At the cut-off value to obtain maximum accuracy, the best biomarkers were found in platelets: phosphatidylserine + phosphatidylinositol (PS + PI), 21:0; sphyngomyelin: 20:1n9 and 22:1n9. All these fatty acids showed similar/higher diagnostic yields than the commonly used markers sialic acid or cytokeratins.

Comparison of changes in erythrocyte and platelet phospholipid and fatty acid composition and protein oxidation in chronic obstructive pulmonary disease and asthma.

OBJECTIVE: To analyse and compare the phospholipid and fatty acid composition of total lipids and the occurrence of lipid peroxidation and protein oxidation directly in erythrocytes or platelets from chronic obstructive pulmonary disease (COPD) and asthma patients. PATIENTS: Fifteen consecutive outpatients with COPD (all smokers) and asthma (non-smokers) recruited during a moderate-to-severe (COPD) or moderate (asthma) exacerbation. Fifteen subjects with smoking habits similar to those of COPD patients were studied as a control group. METHODS: Phospholipid and total fatty acid compositions were analysed by two-dimensional thin layer chromatography or gas chromatography-mass spectrometry, respectively. The lipid fluorescence of lipid extracts was measured by spectrofluorimetry. Protein carbonyl contents and profiles were measured by immunoblot detection. RESULTS: No differences were found either in erythrocyte or platelet cholesterol or phospholipid levels. Only a decrease in the content of phosphatidylserine + phosphatidylinositol (P<0.003) was detected in platelets from the asthma patients. In erythrocytes, the fatty acid profile changed in both lung pathologies, especially as regards polyunsaturated fatty acids (decreases in arachidonic and 22:4 fatty acid contents). Other observed changes were: COPD, an increase in palmitic fatty acid; asthma, an increase in oleic and decreases in eicosapentaenoic and 22:6 + 24:1 fatty acids. In platelets, the fatty acid profiles revealed many differences between both lung pathologies: COPD, a decrease in 18:1 and increases in 20:5 and 22:5 + 24:0; asthma, a decrease in 20:4 and increase in 22:6 + 24:1. In COPD vs. asthma patients, fatty acid changes were mainly detected in platelets, especially in 18-carbon species, with decreases in stearic and 18:1 fatty acids in the COPD patients. Protein oxidation levels were increased in both lung pathologies in both erythrocytes and platelets. CONCLUSIONS: COPD and asthma are associated with common or specific changes in the lipid composition of erythrocytes and/or platelets. The data point to lipid peroxidation and protein oxidation phenomena in both types of blood cell, although platelets would be more susceptible to stress.

Comparison of changes in erythrocyte and platelet fatty acid composition and protein oxidation in advanced non-small cell lung cancer.

The formation of free radicals and lipid peroxidation products is linked both to carcinogenesis and tumor behavior. Blood samples from 50 patients with advanced (Stages III-IV) non-small cell lung cancer (NSCLC), and from 50 healthy volunteers were used for plasma beta-thromboglobulin (beta-TG) measurements, red blood cell (RBC) and platelet lipid analyses, and lipid fluorescence determinations. Samples from 15 randomly selected patients and 15 controls also were used for analysis of the expression of oxidized proteins. We observed: (a) higher levels of plasma beta-TG in patients, (b) alterations in membrane fatty acids. The RBC fatty acid profile changed especially in the 18-carbon species (increases in stearic and oleic and a decrease in linoleic fatty acids), and in arachidonic acid, which also decreased significantly. The platelet fatty acid profile mainly showed a decrease in arachidonic acid and a parallel increase in palmitic fatty acid; (c) the loss of polyunsaturated fatty acids (PUFA) in RBC and platelets could be correlated with changes in lipid extract fluorescence only for platelets; (d) protein oxidation levels were increased also only in the case of platelets. The changes detected point to platelet activation and lipid peroxidation processes associated with NSCLC. The oxidative stress affected RBC and platelets differently, although changes in PUFA might still have important physiological consequences in both types of cells.

Alterations in erythrocyte membrane protein composition in advanced non-small cell lung cancer.

Cancer can be associated with hematological complications related to red blood cell (RBC) function, whose physiological roles have now been expanded since it is now known that RBC are also signalling cells. The aim of this study was to explore the alterations occurring in the protein composition of RBC in advanced non-small cell lung cancer (NSCLC). Blood samples from 21 patients with advanced (stages III-IV) NSCLC (16 squamous cell carcinomas and 5 adenocarcinomas), and from 21 healthy volunteers were used. Samples from 6 randomly selected patients and 6 controls were used for the screening of erythrocyte ghost alterations by Differential Scanning Calorimetry (DSC). Samples from 15 patients and 15 controls, different from those used in the DSC measurements, were randomly selected for analysis of the expression of glycophorin (GP) species, band 3, and glycoproteins by SDS-PAGE and Western blotting or lectin enzyme immunoassays. Additionally, 5 patients with chronic obstructive pulmonary disease (COPD) were used as a control group representative of a benign inflammatory disease. Blood samples from the COPD patients were used to analyze the expression of GPs, band 3 and syaloglycoproteins. We observed the following in NSCLC: (a) changes in GP expression levels, mainly decreases in the GPA and GPC monomers, and in the GPAB dimers; (b) a decrease in the band 3 protein level, and (c) alterations in the expression of different sialoglycoproteins. RBC from the COPD patients also showed protein abnormalities, some of them, especially at the level of band 3 and the syaloglycoproteins, being similar to those in NSCLC.

Structural characteristics of a lipid peroxidation product, trans-2-nonenal, that favour inhibition of membrane-associated phosphotyrosine phosphatase activity.

Protein-tyrosine phosphatases (PTPs) are very susceptible to oxidation by reactive oxygen species (ROS), which induce the oxidation of catalytic cysteines, thereby inactivating these PTPs. PTPs are also inactivated by treatment with different aldehydes (such as trans-2-nonenal), produced after tissue damage by ROS. However, the molecular mechanisms behind such aldehyde-due inactivation remain unknown. Using commercially available compounds, we examined the structural characteristics of trans-2-nonenal that allow the inhibition of platelet membrane-associated PTP activity, as well as how these compounds affect the dynamics of SH-, CO- and NH2- protein groups on the membranes. PTP was effectively inhibited by physiological amounts of trans-2-nonenal (1-10 microM). Incubation with trans-2-nonene (10 microM) also decreased PTP activity, although to a lower extent. Treatment with nonyl aldehyde almost eliminated PTP inhibition. Decreases in protein thiols were visible after trans-2-nonenal and trans-2-nonene treatments. Both the latter compounds also increased protein carbonyls (although trans-2-nonenal was more effective) and decreased protein amino groups to an equal extent. Collectively, our data indicate that alpha,beta unsaturation (and not a double bond in another position) is the most important structural determinant for PTP inhibition, the alkenal with 9-carbon atoms being the most effective in eliciting such inhibition. The data allow us to predict the modification of sulfhydryls and/or the formation of addition products with lysyl or histidyl residues, and hence the kind of specific antibodies that it would be necessary to generate in order to test such modifications directly.

Effect of photoperiod on testicular function in blinded and non-blinded marbled newts, Triturus marmoratus (caudata, salamandridae).

Adult male marbled newts (Triturus marmoratus) were collected at the beginning of the spermatogenetic period and exposed to different photoperiods (natural photoperiod with progressively increasing daylengths, total darkness, 8L:16D, 12L:12D, 16L:8D, and continuous light) for 3 months at 20°C. To evaluate the effect of photoperiodic input via pineal gland photoreceptors, two additional groups of newts were blinded by a non-aggressive method (an elastic rubber cap was adjusted to the head to cover the eyes but not the pineal photoreceptors). These animals were exposed either to the natural photoperiod or to 12 hr of light per day. Quantitative histologic studies on testicular development and germ-cell volume revealed no significant differences between non-blinded and blinded animals. Testicular size and germ-cell development increased in the following order: total darkness, constant light, 8L:12D, natural photoperiod, 12L:12D, and 16L:8D. These results suggest that (1) long photoperiods enhance testicular development, whereas short photoperiods or an environment of continuous light have the opposite effects and (2) the effect of photoperiods on testicular function in newts is independent of the ocular photoreceptors.