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Membranes constitute a meeting point for lipids and proteins. Not only do they define the entity of cells and cytosolic organelles but they also display a wide variety of important functions previously ascribed to the activity of proteins alone. Indeed, lipids have commonly been considered a mere support for the transient or permanent association of membrane proteins, while acting as a selective cell/organelle barrier. However, mounting evidence demonstrates that lipids themselves regulate the location and activity of many membrane proteins, as well as defining membrane microdomains that serve as spatio-temporal platforms for interacting signalling proteins. Membrane lipids are crucial in the fission and fusion of lipid bilayers and they also act as sensors to control environmental or physiological conditions. Lipids and lipid structures participate directly as messengers or regulators of signal transduction. Moreover, their alteration has been associated with the development of numerous diseases. Proteins can interact with membranes through lipid co-/post-translational modifications, and electrostatic and hydrophobic interactions, van der Waals forces and hydrogen bonding are all involved in the associations among membrane proteins and lipids. The present study reviews these interactions from the molecular and biomedical point of view, and the effects of their modulation on the physiological activity of cells, the aetiology of human diseases and the design of clinical drugs. In fact, the influence of lipids on protein function is reflected in the possibility to use these molecular species as targets for therapies against cancer, obesity, neurodegenerative disorders, cardiovascular pathologies and other diseases, using a new approach called membrane-lipid therapy.

Patients with chronic obstructive pulmonary disease (COPD) often lose weight during the course of their disease. We hypothesized that this may be due to skeletal muscle apoptosis. To investigate this possibility, we obtained quadriceps femoris biopsies in 15 patients with COPD (8 with normal body mass index [BMI] and 7 with low [< 20 kg/m(2)] BMI), 8 healthy volunteers, and 6 sedentary subjects undergoing orthopedic surgery (both groups with normal BMI). Skeletal muscle apoptosis was assessed by the transferase-mediated dUTP nick end labeling (TUNEL) technique and the immunodetection of poly-(ADP-ribose)-polymerase proteolytic fragments. Exercise tolerance on a cycloergometer was also determined in patients with COPD. We found that skeletal muscle apoptosis (by both techniques) was increased in patients with COPD and low BMI as compared with the other three groups (p < 0.005). In patients with COPD, BMI was inversely related to skeletal muscle apoptosis (TUNEL, p = 0.009), and it was better correlated with exercise capacity (p = 0.006) than with the degree of airflow obstruction present (p = 0.02). Markers of skeletal muscle apoptosis were not related to any of the measured lung function variables. This study shows that skeletal muscle apoptosis (1) is increased in patients with COPD having low BMI; and (2) is associated with a lower exercise tolerance despite a similar degree of lung function impairment.

Nowadays we understand cell membranes not as a simple double lipid layer but as a collection of complex and dynamic protein-lipid structures and microdomains that serve as functional platforms for interacting signaling lipids and proteins. Membrane lipids and lipid structures participate directly as messengers or regulators of signal transduction. In addition, protein-lipid interactions participate in the localization of signaling protein partners to specific membrane microdomains. Thus, lipid alterations change cell signaling that are associated with a variety of diseases including cancer, obesity, neurodegenerative disorders, cardiovascular pathologies, etc. This article reviews the newly emerging field of membrane lipid therapy which involves the pharmacological regulation of membrane lipid composition and structure for the treatment of diseases. Membrane lipid therapy proposes the use of new molecules specifically designed to modify membrane lipid structures and microdomains as pharmaceutical disease-modifying agents by reversing the malfunction or altering the expression of disease-specific protein or lipid signal cascades. Here, we provide an in-depth analysis of this emerging field, especially its molecular bases and its relevance to the development of innovative therapeutic approaches.

Background-Neutrophils are likely to play a major role in the inflammatory response seen in chronic obstructive pulmonary disease (COPD). This study sought to address the hypothesis that an enhanced neutrophil response to proinflammatory agents in COPD may contribute to their recruitment and activation in the lungs. Methods-Circulating neutrophils were obtained from 10 patients with COPD, eight long term smokers with normal lung function, and eight healthy never smoking controls. The in vitro production of reactive oxygen species (ROS) was measured by the NADPH oxidase method (respiratory burst) and the surface expression of several adhesion molecules (Mac-1, LFA-1 and L-selectin) was measured by flow cytometry. Measurements were obtained under basal conditions and after stimulation with phorbol myristate acetate (PMA) and tumour necrosis factor alpha (TNF ). mRNA levels of p22-phox (a subunit of NADPH oxidase) and Mac-1 (CD11b) were also determined by reverse transcriptase polymerase chain reaction (RT-PCR). 15) v 263 (11); mean diVerence -77 (95% CI of the diVerence -119 to -34), p=0.001). These diVerences were also apparent when patients with COPD were compared with non-smokers (p<0.05). The mRNA levels of p22-phox and Mac-1 (CD11b) were similar in patients with COPD and smokers with normal lung function, suggesting that the observed differences were due to post-transcriptional regulation. Results-PatientsConclusions-These results demonstrate an enhanced neutrophil response to proinflammatory agents in patients with COPD which may contribute to their enhanced recruitment and activation in the lungs of these patients. These findings support those of other studies which have indicated that the neutrophil is likely to play a major role in the pathogenesis of this disease. (Thorax 2001;56:432-437)

Telomeres are complex DNA-protein structures located at the end of eukaryotic chromosomes. Telomere length shortens with age in all replicating somatic cells. It has been shown that tobacco smoking enhances telomere shortening in circulating lymphocytes. The present study investigated whether this effect was further amplified in smokers who develop chronic obstructive pulmonary disease.Telomere length was determined by fluorescence in situ hybridisation in circulating lymphocytes harvested from 26 never-smokers, 24 smokers with normal lung function and 26 smokers with moderate-to-severe airflow obstruction (forced expiratory flow in one second 48¡4% predicted).In contrast to never-smokers, telomere length significantly decreased with age in smokers. There was also a dose-effect relationship between the cumulative long-life exposure to tobacco smoking (pack-yrs) and telomere length. The presence and/or severity of chronic airflow obstruction did not modify this relationship.The results of the current study confirm that smoking exposure enhances telomere shortening in circulating lymphocytes. It also demonstrates a dose-effect relationship between exposure to tobacco smoking and telomere length, but failed to show that this effect is amplified in smokers who develop chronic obstructive pulmonary disease. An accelerated rate of lung function decline with age is one of the central pathophysiological characteristics of COPD [1]. In the present study, the authors hypothesised that the effects of tobacco smoking upon telomere length shortening would be enhanced in smokers who developed COPD, compared with those whose lung function was preserved despite their habit. To test this hypothesis, the rate of telomere shortening was compared with age in circulating lymphocytes harvested from smokers with and without COPD. KEYWORDS METHODS Population and ethicsThe study included 50 life-long smokers, 24 of whom had normal lung function and 26 with evidence of COPD according to international guidelines [1], and 26 never-smokers. Participants with chronic diseases other than COPD were excluded. The Ethics Committee of Hospital Universitari Son Dureta (Palma De Mallorca, Spain) approved the study and patients gave their informed consent. Lung function measurementsForced spirometry was obtained in all participants according to international recommendations [6]. Reference values were those of a Mediterranean population [7]. Lymphocyte isolationPeripheral blood samples were obtained by venipuncture. Lymphocytes were isolated by mixing 10 mL of venous blood with an equal volume of PBS. This was followed by centrifugation on a 2-mL layer of a Ficoll-Paque research grade gradient (Pharmacia Biotech, Uppsala, Sweden) at 9006g for 30 minutes at 22uC. The . This cell line is characterised by a lack of telomere shortening (and hence, constant telomere length) during cell division due to overexpression of telomerase [4]. Briefly, a suspension containing a mixture of 2610 6 lymphocytes and 2610 6 of 1301 cells was denatured at 82uC...

Despite recent advances in the development of new cancer therapies, the treatment options for glioma remain limited, and the survival rate of patients has changed little over the past three decades. Here, we show that 2-hydroxyoleic acid (2OHOA) induces differentiation and autophagy of human glioma cells. Compared to the current reference drug for this condition, temozolomide (TMZ), 2OHOA combated glioma more efficiently and, unlike TMZ, tumor relapse was not observed following 2OHOA treatment. The novel mechanism of action of 2OHOA is associated with important changes in membrane-lipid composition, primarily a recovery of sphingomyelin (SM) levels, which is markedly low in glioma cells before treatment. Parallel to membrane-lipid regulation, treatment with 2OHOA induced a dramatic translocation of Ras from the membrane to the cytoplasm, which inhibited the MAP kinase pathway, reduced activity of the PI3K/Akt pathway, and downregulated Cyclin D-CDK4/6 proteins followed by hypophosphorylation of the retinoblastoma protein (RB). These regulatory effects were associated with induction of glioma cell differentiation into mature glial cells followed by autophagic cell death. Given its high efficacy, low toxicity, ease of oral administration, and good distribution to the brain, 2OHOA constitutes a new and potentially valuable therapeutic tool for glioma patients.fatty acids | sphingomyelin synthase | cancer drug target | glioma biomarker C ancer cells of undifferentiated phenotype (e.g., glioma) have a poor prognosis and limited treatment options. Primary brain tumors, of which glioma is the most common, are generally associated with very high rates of mortality (ca. 90%), being the median survival of patients about 1 y (1, 2). Chemotherapy provides only modest benefits to radiotherapy and surgery being the alkylating agent temozolomide (TMZ) the reference drug; however, tumor relapse is usually observed, and TMZ only increases the patients' life expectancy about 2.5 m (from 12.1 to 14.6 m: ref.3). The present study was designed to investigate the efficacy of 2OHOA against glioma and its molecular mechanisms of action. 2OHOA exhibited a greater efficacy than TMZ in the treatment of glioma, and there was no relapse after long-term treatment with 2OHOA. This efficacy and lack of toxicity at therapeutic doses has been acknowledged recently by the European Medicines Agency (EMA) to designate 2OHOA orphan drug for the treatment of glioma. In previous studies, we showed that this compound induces cell cycle arrest of lung cancer cells (4-6). Here, we showed that 2OHOA reversed the altered lipid profile of glioma cells and how this modification regulated cell signaling to induce autophagy specifically in glioma but not normal cells.Moreover, in the present study we demonstrated that the changes induced by 2OHOA were specific to cancer cells with no significant effects observed in normal cells and no adverse effects in treated animals, features not shared by most anticancer drugs. The efficacy of this compound in the absence ...

We investigated the expression of adhesion molecules in circulating neutrophils (lymphocyte function-associated antigen-1 [LFA-1], Mac-1, and L-selectin) and endothelial cells (soluble intercellular adhesion molecule-1[sICAM-1]) in 23 patients with stable chronic obstructive pulmonary disease (COPD), 18 subjects with exacerbated COPD, and 23 healthy volunteers. Also, in these circulating neutrophils, we assessed the expression of two G protein subunits (Galphas and Galphai1/2). Compared with control subjects, patients with stable COPD showed increased expression of Mac-1 (p < 0.001) and lower levels of sICAM-1 (p = 0.002); LFA-1 and L-selectin expression was similar in patients and control subjects. During exacerbations, compared with stable patients, the expression of Mac-1 and LFA-1 was reduced (p < 0.001). Finally, the expression of Galphas (but not Galphai1/2) was also reduced (p < 0.001) in circulating neutrophils of patients with COPD, irrespective of the clinical condition of the patient. These results indicate that in patients with COPD: (1) the expression of some neutrophil adhesion molecules (Mac-1) is abnormal, and that this pattern changes during exacerbations; (2) there may be a form of endothelial dysfunction, as suggested by the low sICAM-1 levels; (3) the expression of G protein subunit (Galphas) in circulating neutrophils is downregulated, irrespective of their clinical conditions. Overall, these results indicate the presence of significant systemic abnormalities in COPD.