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1.
Biochim Biophys Acta Mol Basis Dis ; 1864(5 Pt A): 1828-1838, 2018 May.
Article in English | MEDLINE | ID: mdl-29524632

ABSTRACT

Although many studies have been carried out in order to understand the implication of copper (Cu) in the pathogenesis of multiple sclerosis (MS), the exact role that this metal plays in the disease is not still clear. Because of the lack of information in this subject, the present study compared the serum and cerebrospinal (CSF) levels of copper in MS patients in respect to a control group, matched for age and sex, finding a significant increase of metal concentrations, in both biological fluids of MS subjects. To confirm the possible impairment of Cu metabolism, we analyzed ceruloplasmin (Cp) level and activity, seeing as this protein is an established peripheral marker in diseases associated with Cu imbalance. By comparing these two parameters between control and MS subjects, we found an increase of Cp levels, associated with a decrease in Cp activity, in the second group. By analysing these data, free copper levels were calculated, significantly increased in serum of MS subjects; the increase in free copper could be one of the predisposing factors responsible for the Cu altered levels in CSF of MS patients. At the same time, this alteration could be attributable to the inability to incorporate Cu by Cp, probably due to the high oxidative environment found in serum of MS patients. Overall, all these copper alterations may play a role in MS pathogenesis.


Subject(s)
Ceruloplasmin/cerebrospinal fluid , Copper/blood , Copper/cerebrospinal fluid , Multiple Sclerosis/blood , Multiple Sclerosis/cerebrospinal fluid , Adolescent , Adult , Female , Humans , Male , Middle Aged
2.
Leukemia ; 30(3): 640-8, 2016 Mar.
Article in English | MEDLINE | ID: mdl-26487273

ABSTRACT

Bortezomib (bort) has improved overall survival in patients with multiple myeloma (MM), but the majority of them develop drug resistance. In this study, we demonstrate that bone marrow (BM) fibroblasts (cancer-associated fibroblasts; CAFs) from bort-resistant patients are insensitive to bort and protect the RPMI8226 and patients' plasma cells against bort-induced apoptosis. Bort triggers CAFs to produce high levels of interleukin (IL)-6, IL-8, insulin-like growth factor (IGF)-1 and transforming growth factor (TGF) ß. Proteomic studies on CAFs demonstrate that bort resistance parallels activation of oxidative stress and pro-survival autophagy. Indeed, bort induces reactive oxygen species in bort-resistant CAFs and activates autophagy by increasing light chain 3 protein (LC3)-II and inhibiting p62 and phospho-mammalian target of rapamycin. The small-interfering RNA knockdown of Atg7, and treatment with 3-methyladenine, restores bort sensitivity in bort-resistant CAFs and produces cytotoxicity in plasma cells co-cultured with CAFs. In the syngeneic 5T33 MM model, bort-treatment induces the expansion of LC3-II(+) CAFs. TGFß mediates bort-induced autophagy, and its blockade by LY2109761, a selective TßRI/II inhibitor, reduces the expression of p-Smad2/3 and LC3-II and induces apoptosis in bort-resistant CAFs. A combination of bort and LY2109761 synergistically induces apoptosis of RPMI8226 co-cultured with bort-resistant CAFs. These data define a key role for CAFs in bort resistance of plasma cells and provide the basis for a novel targeted therapeutic approach.


Subject(s)
Antineoplastic Agents/pharmacology , Bortezomib/pharmacology , Drug Resistance, Neoplasm/drug effects , Gene Expression Regulation, Neoplastic , Multiple Myeloma/drug therapy , Pyrazoles/pharmacology , Pyrroles/pharmacology , Transforming Growth Factor beta/antagonists & inhibitors , Aged , Aged, 80 and over , Animals , Autophagy/drug effects , Bone Marrow Cells/drug effects , Bone Marrow Cells/metabolism , Bone Marrow Cells/pathology , Drug Combinations , Drug Resistance, Neoplasm/genetics , Female , Fibroblasts/drug effects , Fibroblasts/metabolism , Fibroblasts/pathology , Humans , Male , Mice , Microtubule-Associated Proteins/genetics , Microtubule-Associated Proteins/metabolism , Middle Aged , Multiple Myeloma/genetics , Multiple Myeloma/mortality , Multiple Myeloma/pathology , Plasma Cells/drug effects , Plasma Cells/metabolism , Plasma Cells/pathology , Primary Cell Culture , RNA-Binding Proteins/genetics , RNA-Binding Proteins/metabolism , Signal Transduction , Survival Analysis , TOR Serine-Threonine Kinases/genetics , TOR Serine-Threonine Kinases/metabolism , Transforming Growth Factor beta/genetics , Transforming Growth Factor beta/metabolism , Xenograft Model Antitumor Assays
3.
Mol Biosyst ; 12(1): 219-32, 2016 Jan.
Article in English | MEDLINE | ID: mdl-26588820

ABSTRACT

Classical Hodgkin lymphoma (cHL) is a malignancy with complex pathogenesis. The hallmark of the disease is the presence of large mononucleated Hodgkin and bi- or multinucleated Reed/Sternberg (H/RS) cells. The origin of HRS cells in cHL is controversial as these cells show the coexpression of markers of several lineages. Using a proteomic approach, we compared the protein expression profile of cHL models of T- and B-cell derivation to find proteins differentially expressed in these cell lines. A total of 67 proteins were found differentially expressed between the two cell lines including metabolic proteins and proteins involved in the regulation of the cytoskeleton and/or cell migration, which were further validated by western blotting. Additionally, the expression of selected B- and T-cell antigens was also assessed by flow cytometry to reveal significant differences in the expression of different surface markers. Bioinformatics analysis was then applied to our dataset to find enriched pathways and networks, and to identify possible key regulators. In the present study, a proteomic approach was used to compare the protein expression profiles of two cHL cell lines. The identified proteins and/or networks, many of which not previously related to cHL, may be important to better define the pathogenesis of the disease, to identify novel diagnostic markers, and to design new therapeutic strategies.


Subject(s)
Hodgkin Disease/metabolism , Proteome , Proteomics , Cell Line, Tumor , Flow Cytometry , Gene Expression Regulation, Neoplastic , Hodgkin Disease/genetics , Humans , Models, Biological , Protein Interaction Mapping , Protein Interaction Maps , Proteomics/methods
4.
Curr Cancer Drug Targets ; 12(8): 987-97, 2012 Oct.
Article in English | MEDLINE | ID: mdl-22515523

ABSTRACT

The current therapy for ovarian cancer has advanced from alkylating agents, to a combination of carboplatinum and paclitaxel offering increased survival. Although most patients respond to this first-line therapy, initially, the majority of these patients relapse within 2 years. The mechanisms responsible for acquired drug resistance in ovarian cancer have been elucidated only in part. They include i) enhanced drug export, ii) activation/inhibition of intracellular signalling pathways, iii) molecular alterations in tubulin isotype composition. A better understanding of these mechanisms is needed, in order to develop new approaches, aimed at overcoming resistance to anticancer agents, and to reveal the complexity of causes, which contribute to drug resistance. In this review we offer an updated overview of proteomic studies on the molecular mechanisms of paclitaxel resistance. These proteomic studies also identify potential targets for modulating drug resistance, that could be predictive of response to chemotherapy in ovarian carcinomas.


Subject(s)
Drug Resistance, Neoplasm/drug effects , Ovarian Neoplasms/drug therapy , Ovarian Neoplasms/metabolism , Paclitaxel/therapeutic use , Proteomics/methods , Biomarkers, Tumor/analysis , Female , Humans , Tubulin/metabolism
5.
Nanotechnology ; 21(16): 165502, 2010 Apr 23.
Article in English | MEDLINE | ID: mdl-20348602

ABSTRACT

Surface-enhanced Raman spectroscopy (SERS) allows a new insight into the analysis of cell physiology. In this work, the difficulty of producing suitable substrates that, besides permitting the amplification of the Raman signal, do not interact with the biological material causing alteration, has been overcome by a combined method of hydrothermal green synthesis and thermal annealing. The SERS analysis of the cell membrane has been performed with special attention to the cellular prion protein PrP(C). In addition, SERS has also been used to reveal the prion protein-Cu(II) interaction in four different cell models (B104, SH-SY5Y, GN11, HeLa), expressing PrP(C) at different levels. A significant implication of the current work consists of the intriguing possibility of revealing and quantifying prion protein expression in complex biological samples by a cheap SERS-based method, replacing the expensive and time-consuming immuno-assay systems commonly employed.


Subject(s)
Complex Mixtures/analysis , Gene Expression Profiling/methods , PrPC Proteins/analysis , PrPC Proteins/metabolism , Spectrum Analysis, Raman/methods , HeLa Cells , Humans
6.
J Neuroimmunol ; 210(1-2): 80-6, 2009 May 29.
Article in English | MEDLINE | ID: mdl-19329191

ABSTRACT

This cross-sectional study investigated with two-dimensional gel electrophoresis coupled to MALDI-TOF and MRI the relationship between PBMCs protein expression profile and whole-brain atrophy in 16 unselected RR-MS IFN-treated patients compared with 6 RR IFN-untreated and 12 matched healthy control subjects. Grey/white matter fraction, T1/T2 lesion load and clinical variables were considered too. Twenty six proteins showed significant differential expression among RR IFN-treated patients and control samples. Four of these (IN35, GANAB, PP1B, SEPT2) resulted correlated with clinical and MRI findings in RR IFN-treated MS patients. Future clinical applications remain to be validated by other techniques and confirmed by a larger study.


Subject(s)
Atrophy/pathology , Brain/pathology , Leukocytes, Mononuclear/metabolism , Multiple Sclerosis, Relapsing-Remitting/metabolism , Multiple Sclerosis, Relapsing-Remitting/pathology , Adult , Aged , Atrophy/physiopathology , Brain/physiopathology , Cross-Sectional Studies , Disease Progression , Electrophoresis, Gel, Two-Dimensional , Female , Humans , Leukocytes, Mononuclear/immunology , Magnetic Resonance Imaging , Male , Middle Aged , Multiple Sclerosis, Relapsing-Remitting/immunology , Nerve Fibers, Myelinated/pathology , Phosphoric Monoester Hydrolases/analysis , Phosphoric Monoester Hydrolases/metabolism , Pilot Projects , Predictive Value of Tests , Proteomics/methods , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Young Adult
7.
Biophys J ; 96(4): 1586-96, 2009 Feb 18.
Article in English | MEDLINE | ID: mdl-19217874

ABSTRACT

Molecular flexibility and rigidity are required to determine the function and specificity of protein molecules. Some psychrophilic enzymes demonstrate a higher catalytic efficiency at low temperatures, compared to the efficiency demonstrated by their meso/thermophilic homologous. The emerging picture suggests that such enzymes have an improved flexibility of the structural catalytic components, whereas other protein regions far from functional sites may be even more rigid than those of their mesophilic counterparts. To gain a deeper insight in the analysis of the activity-flexibility/rigidity relationship in protein structure, psychrophilic carbonic anhydrase of the Antarctic teleost Chionodraco hamatus has been compared with carbonic anhydrase II of Bos taurus through fluorescence studies, three-dimensional modeling, and activity analyses. Data demonstrated that the cold-adapted enzyme exhibits an increased catalytic efficiency at low and moderate temperatures and, more interestingly, a local flexibility in the region that controls the correct folding of the catalytic architecture, as well as a rigidity in the hydrophobic core. The opposite result was observed in the mesophilic counterpart. These results suggest a clear relationship between the activity and the presence of flexible and rigid protein substructures that may be useful in rational molecular and drug design of a class of enzymes playing a key role in pathologic processes.


Subject(s)
Carbonic Anhydrases/chemistry , Amino Acid Sequence , Animals , Cattle , Hydrophobic and Hydrophilic Interactions , Kinetics , Light , Models, Molecular , Molecular Sequence Data , Perciformes , Pliability , Protein Conformation , Scattering, Radiation , Sequence Analysis, DNA , Sequence Homology, Amino Acid , Software , Spectrometry, Fluorescence , Temperature , Thermodynamics
8.
Res Vet Sci ; 84(3): 395-408, 2008 Jun.
Article in English | MEDLINE | ID: mdl-18561390

ABSTRACT

Physiological and health related responses to dietary inclusion of genetically modified (GM) full-fat soybean meal (Roundup Ready; GM-soy) and maize (MON810 Bt-maize; GM-maize), as well as non-parental, untransformed lines (nGM-soy and nGM-maize D2), were evaluated in farmed Atlantic salmon (Salmo salar L.) parr during the first 8 months of feeding. Significant effects of dietary GM presence were only found in intestinal Na+-dependent d-glucose uptake and SGLT1 protein level in the region pyloric caeca in which the highest values were found in the GM-soy, intermediate in the nGM-soy, and lowest in the standard FM fed groups. Data from this study confirm that GM soybeans (RRS) and maize (MON810) at inclusion levels of about 6% appear to be as safe as commercially available nGM soy and maize in diets for Atlantic salmon parr. Results from studies with higher inclusion levels and with non-modified, isogenic or near-isogenic parental lines as control groups are pending.


Subject(s)
Animal Feed , Digestion/physiology , Food, Genetically Modified , Glycine max , Immune System/drug effects , Salmo salar/growth & development , Zea mays , Animals , Dietary Carbohydrates , Dietary Fats , Dietary Proteins , Immunoglobulin M/blood , Immunoglobulin M/drug effects , Salmo salar/immunology , Glycine max/genetics , Zea mays/genetics
9.
Biosens Bioelectron ; 21(1): 30-40, 2005 Jul 15.
Article in English | MEDLINE | ID: mdl-15967348

ABSTRACT

In this paper we have tested two different procedures (the "three-step" and the "four-step" procedures) for the covalent immobilization of glutamate dehydrogenase (GDH) onto silicon supports. Atomic force microscopy (AFM), Fourier-transform infrared spectroscopy (FT-IR), fluorescence spectroscopy and an enzymatic assay were used to probe the structure and activity of the immobilized enzyme. Our results demonstrate that coupling through the "three-step" procedure does not significantly affect either the fold pattern or the activity of the enzyme, suggesting that this method could be ideally suited to the development of high quality monolayers for use in enzyme-based planar biosensors.


Subject(s)
Biosensing Techniques/instrumentation , Enzymes, Immobilized , Glutamate Dehydrogenase , Silicon , Microscopy, Atomic Force , NAD/metabolism , Oxidation-Reduction , Spectroscopy, Fourier Transform Infrared
10.
In. Sempé, Carlota; Salcedo, Susana; Maffia, Marta. Azampay: presente y pasado de un pueblito catamarqueño. La Plata, Ediciones Al margen;Universidad Nacional de La Plata, 2005. p.29-61, tab. (126530).
Monography in Spanish | BINACIS | ID: bin-126530
11.
In. Sempé, Carlota; Salcedo, Susana; Maffia, Marta. Azampay: presente y pasado de un pueblito catamarqueño. La Plata, Ediciones Al margen;Universidad Nacional de La Plata, 2005. p.87-108, ilus. (126527).
Monography in Spanish | BINACIS | ID: bin-126527
12.
In. Sempé, Carlota; Salcedo, Susana; Maffia, Marta. Azampay: presente y pasado de un pueblito catamarqueño. La Plata, Ediciones Al margen;Universidad Nacional de La Plata, 2005. p.109-24, ilus. (126526).
Monography in Spanish | BINACIS | ID: bin-126526

Subject(s)
Housing , Cultural Factors
13.
In. Sempé, Carlota; Salcedo, Susana; Maffia, Marta. Azampay: presente y pasado de un pueblito catamarqueño. La Plata, Ediciones Al margen;Universidad Nacional de La Plata, 2005. p.125-42, ilus. (126525).
Monography in Spanish | BINACIS | ID: bin-126525
14.
In. Sempé, Carlota; Salcedo, Susana; Maffia, Marta. Azampay: presente y pasado de un pueblito catamarqueño. La Plata, Ediciones Al margen;Universidad Nacional de La Plata, 2005. p.143-58, ilus. (126524).
Monography in Spanish | BINACIS | ID: bin-126524
15.
In. Sempé, Carlota; Salcedo, Susana; Maffia, Marta. Azampay: presente y pasado de un pueblito catamarqueño. La Plata, Ediciones Al margen;Universidad Nacional de La Plata, 2005. p.159-72, ilus. (126523).
Monography in Spanish | BINACIS | ID: bin-126523
16.
Phys Rev E Stat Nonlin Soft Matter Phys ; 67(4 Pt 1): 041902, 2003 Apr.
Article in English | MEDLINE | ID: mdl-12786391

ABSTRACT

This paper describes the formation of glutamate dehydrogenase monolayers on silicon dioxide, and their characterization by means of physical techniques, i.e., fluorescence spectroscopy and Fourier-transform infrared spectroscopy. Detailed investigations of the intrinsic stability of native proteins in solution were carried out to elucidate the occurrence of conformational changes induced by the immobilization procedure. The enzyme monolayers were deposited on SiO2 after preexposing silicon surfaces to 3-aminopropyltriethoxysilane and reacting the silylated surfaces with glutaric dialdehyde. The optical characterization demonstrates that the immobilization does not interfere with the fold pattern of the native enzyme. In addition, fluorescence spectroscopy, thermal denaturation, and quenching studies performed on the enzyme in solution well describe the folding and unfolding properties of glutamate dehydrogenase. The photophysical studies reported here are relevant for nanobioelectronics applications requiring protein immobilization on a chip.


Subject(s)
Glutamate Dehydrogenase/chemistry , Silicon Dioxide/chemistry , Silicon/chemistry , Biophysical Phenomena , Biophysics , Light , Propylamines , Protein Conformation , Scattering, Radiation , Silanes/chemistry , Spectrometry, Fluorescence , Spectroscopy, Fourier Transform Infrared , Temperature , Time Factors , Tryptophan/chemistry
17.
J Exp Biol ; 206(Pt 4): 705-14, 2003 Feb.
Article in English | MEDLINE | ID: mdl-12517988

ABSTRACT

H(+)/peptide cotransport was studied in brush-border membrane vesicles (BBMV) from the intestine of the haemoglobinless Antarctic teleost Chionodraco hamatus by monitoring peptide-dependent intravesicular acidification with the pH-sensitive dye Acridine Orange. Diethylpyrocarbonate-inhibited intravesicular acidification was specifically achieved in the presence of extravesicular glycyl-L-proline (Gly-L-Pro) as well as of glycyl-L-alanine (Gly-L-Ala) and D-phenylalanyl-L-alanine (D-Phe-L-Ala). H(+)/Gly-L-Pro cotransport displayed saturable kinetics, involving a single carrier system with an apparent substrate affinity (K(m,app)) of 0.806+/-0.161 mmol l(-1). Using degenerated primers from eel and human (PepT1) transporter sequence, a reverse transcription-polymerase chain reaction (RT-PCR) signal was detected in C. hamatus intestine. RT-PCR paralleled kinetic analysis, confirming the hypothesis of the existence of a PepT1-type transport system in the brush-border membranes of icefish intestine. Functional expression of H(+)/peptide cotransport was successfully performed in Xenopus laevis oocytes after injection of poly(A)(+) RNA (mRNA) isolated from icefish intestinal mucosa. Injection of mRNA stimulated D-Phe-L-Ala uptake in a dose-dependent manner and an excess of glycyl-L-glutamine inhibited this transport. H(+)/peptide cotransport in the Antarctic teleost BBMV exhibited a marked difference in temperature optimum with respect to the temperate teleost Anguilla anguilla, the maximal activity rate occurring at approximately 0 degrees C for the former and 25 degrees C for the latter. Temperature dependence of icefish and eel intestinal mRNA-stimulated uptake in the heterologous system (oocytes) was comparable.


Subject(s)
Cadherins , Carrier Proteins/genetics , Carrier Proteins/metabolism , Gastrointestinal Hormones/physiology , Hemoglobins/metabolism , Intestinal Mucosa/physiology , Membrane Transport Proteins , Microvilli/physiology , Oocytes/metabolism , Peptides/metabolism , Perciformes/physiology , Amino Acid Sequence , Amino Acids/metabolism , Animals , Antarctic Regions , Carrier Proteins/chemistry , Cold Climate , Female , Hemoglobins/deficiency , Kinetics , Molecular Sequence Data , RNA, Messenger/genetics , Reverse Transcriptase Polymerase Chain Reaction , Substrate Specificity
18.
J Exp Biol ; 204(Pt 22): 3983-92, 2001 Nov.
Article in English | MEDLINE | ID: mdl-11807116

ABSTRACT

Carbonic anhydrase (CA) activity was measured in blood, intestine, kidney and gill of two Antarctic teleosts, the haemoglobinless Chionodraco hamatus and the red-blooded Trematomus bernacchii, and of the temperate teleost Anguilla anguilla. In all species, the highest CA activity was in the gills, with the greatest activity in C. hamatus. CA activity in the blood was highest in A. anguilla, but none was detected in the blood of C. hamatus despite the presence of plasma CA inhibitors. The enzyme was present but its activity was low in the intestine and kidney of all three species. The existence of very high CA activity in C. hamatus gills compared with the red-blooded species was investigated further by isolating and characterising the branchial cytosolic CA isoforms. The turnover rate of the C. hamatus isoform was significantly higher than that of T. bernacchii and A. anguilla. The isoforms from both the Antarctic species exhibited lower apparent K(m) (K(m,app)) and heat stability than those from A. anguilla. Sensitivity to sulphonamides was similar in all species and was within the range of the mammalian CA II isoform. The branchial CA isoforms of C. hamatus, T. bernacchii and A. anguilla displayed relative molecular masses of 28.9, 29.9 and 31.2 kDa, respectively. The results suggest that the hemoglobinless teleost possesses a different branchial cytosolic CA isoform from that of red-blooded teleosts.


Subject(s)
Anguilla/metabolism , Carbonic Anhydrases/analysis , Fishes/metabolism , Animals , Antarctic Regions , Carbonic Anhydrases/blood , Gills/enzymology , Hemoglobins/analysis , Intestines/enzymology , Kidney/enzymology
19.
J Exp Biol ; 203(Pt 19): 2991-3001, 2000 Oct.
Article in English | MEDLINE | ID: mdl-10976035

ABSTRACT

H(+)/peptide cotransport in brush-border membrane vesicles (BBMVs) from eel (Anguilla anguilla) intestine was studied by measuring d-[(3)H]-phenylalanyl-l-alanine uptake and by monitoring peptide-dependent intravesicular acidification using the pH-sensitive dye Acridine Orange. d-[(3)H]-phenylalanyl-l-alanine influx was greatly stimulated by an inside-negative membrane potential and enhanced by an inwardly directed H(+) gradient. In parallel, vesicular H(+) influx was significantly increased in the presence of extravesicular d-phenylalanyl-l-alanine or a series of glycyl and l-prolyl peptides. H(+)/peptide cotransport displayed saturable kinetics involving a single carrier system with apparent substrate affinities of 0.9-2.6 mmol l(-1) depending on the particular peptide. All substrates tested competed with this system. Pre-incubation of BBMVs with dipeptides prevented diethylpyrocarbonate inhibition of transport activity, suggesting that the substrates mask histidine residues involved in the catalytic function of the transporter. Using human PepT1-specific primers, a reverse transcription-polymerase chain reaction (RT-PCR) signal was detected in eel intestine. Our results suggest that, in eel intestine, a brush-border membrane 'low-affinity'-type H(+)/peptide cotransport system is present that shares kinetic features with the mammalian intestinal PepT1-type transporters.


Subject(s)
Anguilla/metabolism , Cadherins , Carrier Proteins/metabolism , Intestinal Mucosa/metabolism , Membrane Transport Proteins , Symporters , Anguilla/genetics , Animals , Base Sequence , Biological Transport, Active , Carrier Proteins/genetics , DNA Primers/genetics , Dipeptides/metabolism , Humans , Kinetics , Microvilli/metabolism , Peptide Transporter 1 , Peptides/metabolism , Protons
20.
Arterioscler Thromb Vasc Biol ; 19(2): 220-8, 1999 Feb.
Article in English | MEDLINE | ID: mdl-9974401

ABSTRACT

Because oleic acid is implicated in the antiatherogenic effects attributed to the Mediterranean diet, we investigated whether this fatty acid can modulate endothelial activation, ie, the concerted expression of gene products involved in leukocyte recruitment and early atherogenesis. We incubated sodium oleate with human umbilical vein endothelial cells for 0 to 72 hours, followed by coincubation of oleate with human recombinant tumor necrosis factor, interleukin (IL)-1alpha, IL-1beta, IL-4, Escherichia coli lipopolysaccharide (LPS), or phorbol 12-myristate 13-acetate for a further 6 to 24 hours. The endothelial expression of vascular cell adhesion molecule-1 (VCAM-1), E-selectin, and intercellular adhesion molecule-1 was monitored by cell surface enzyme immunoassays or flow cytometry, and steady-state levels of VCAM-1 mRNA were assessed by Northern blot analysis. At 10 to 100 micromol/L for >24 hours, oleate inhibited the expression of all adhesion molecules tested. After a 72-hour incubation with oleate and a further 16-hour incubation with oleate plus 1 microg/mL LPS, VCAM-1 expression was reduced by >40% compared with control. Adhesion of monocytoid U937 cells to LPS-treated endothelial cells was reduced concomitantly. Oleate also produced a quantitatively similar reduction of VCAM-1 mRNA levels on Northern blot analysis and inhibited nuclear factor-kappaB activation on electrophoretic mobility shift assays. Incubation of endothelial cells with oleate for 72 hours decreased the relative proportions of saturated (palmitic and stearic) acids in total cell lipids and increased the proportions of oleate in total cell lipids without significantly changing the relative proportions of polyunsaturated fatty acids. Although less potent than polyunsaturated fatty acids in inhibiting endothelial activation, oleic acid may contribute to the prevention of atherogenesis through selective displacement of saturated fatty acids in cell membrane phospholipids and a consequent modulation of gene expression for molecules involved in monocyte recruitment.


Subject(s)
Endothelium, Vascular/drug effects , Endothelium, Vascular/physiology , Oleic Acid/pharmacology , Arteriosclerosis/prevention & control , Cell Adhesion/drug effects , Cells, Cultured , Diet , Dietary Fats, Unsaturated/pharmacology , Endothelium, Vascular/cytology , Fatty Acids/metabolism , Homeostasis , Humans , Lipid Metabolism , Lipopolysaccharides/pharmacology , Mediterranean Region , Monocytes/physiology , NF-kappa B/drug effects , NF-kappa B/physiology , RNA, Messenger/metabolism , Time Factors , Vascular Cell Adhesion Molecule-1/genetics , Vascular Cell Adhesion Molecule-1/metabolism
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