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Biomedical subjects

M Lekka

Publications and source records attributed to M Lekka.

17 recordsLinked to original sources
PubMed2004

Probing molecular interaction between concanavalin A and mannose ligands by means of SFM.

Recently, the scanning force microscope (SFM) has been widely used for direct monitoring of specific interactions between biologically active molecules. Such studies have employed the SFM liquid-cell setup, which allows measurements to be made in the native environment with force resolution down to a tenth of a picoNewton. In this study, the ligand-receptor strength of monoclonal anti-human prostatic acid phosphatase and prostatic acid phosphatase, representing an antigen-antibody system with a single type of interaction, was determined. Then, the interaction force occurring between concanavalin A and the carbohydrate component of the glycoproteins arylsulfatase A and carboxypeptidase Y was measured. High mannose-type glycans were sought on the human prostate carcinoma cell surface. Application of an analysis based on the Poisson distribution of the number of bonds formed in all these measured systems allowed the strength of the molecular interaction to be calculated. The values of the force acting between two single molecules were 530+/-25, 790+/-32, and 940+/-39 pN between prostatic acid phosphatase and monoclonal anti-human prostatic acid phosphatase, between concanavalin A and arylsulfatase A, and between concanavalin A and carboxypeptidase Y, respectively. The value calculated from data collected for the force between concanavalin A and mannose-containing ligands present on the surface of human prostate carcinoma cells was smaller, 116+/-17 pN. The different values of the binding force between concanavalin A and mannose-containing ligands were attributed to the structural changes of the carbohydrate components.

Binding Sites↗
PubMed2001

The effect of chitosan on stiffness and glycolytic activity of human bladder cells.

The cell's cytoskeleton together with the cell membrane and numerous accessory proteins determines the mechanical properties of cell. Any factors influencing cell organization and structure can cause alterations in mechanical properties of cell (its ability for deformation and adhesion). The determination of the local elastic properties of cells in their culture conditions has opened the possibility for the measurement of the influence of different factors on the mechanical properties of the living cells. The effect of the chitosan on the stiffness of the non-malignant transitional epithelial cells of ureter (HCV 29) and the transitional cell cancer of urine bladder (T24) was determined using scanning force microscopy. The investigations were performed in the culture medium (RPMI 1640) containing 10% fetal calf serum in the presence of the microcrystalline chitosan of the three different deacetylation degrees. In parallel, the effect of chitosan on production of lactate and ATP level was determined. The results showed the strong correlation between the decrease of the energy production and the increase in Young's modulus values obtained for the cancer cells treated with chitosan.

Cell Membrane↗
PubMed2001

The characteristics of bronchoalveolar lavage from a patient with antiphospholipid syndrome who developed acute respiratory distress syndrome.

The purpose of this study was to investigate the biochemical characteristics as well as the occurrence and specificity of antiphospholipid antibodies in the bronchoalveolar lavage (BAL) fluid from a patient with both antiphospholipid antibodies syndrome (APS) and acute respiratory distress syndrome (ARDS). Proteins, lipids, cells and autoantibodies were determined. Immunoglobulins were purified with affinity chromatography. Autoantibody identification was assessed with enzyme-linked immunosorbent assay (ELISA) and with electrophoresis, followed by immunoblotting and revelation with antihuman IgG-peroxidase conjugate. Antiphospholipid antibodies were found to be present in the BAL fluid as well as in the serum from a patient with APS. Specifically, antiphosphatidylserine and antiphosphatidic acid IgG antibodies in the BAL fluid and antiphosphatidylcholine and anticardiolipin IgG antibodies in the serum were detected at high levels. BAL fluid protein and the percentage of neutrophils were found to be increased. A quantitative as well as qualitative deficiency of surfactant phospholipids was also observed. Antibodies directed against surfactant phospholipids could cause surfactant abnormalities and an inflammatory reaction. These disorders may be one of the causes of the ARDS or a factor in the perpetuation of the inflammation.

Adult↗
PubMed2001

Autoantibodies to lipids in bronchoalveolar lavage fluid of patients with acute respiratory distress syndrome.

OBJECTIVE: To investigate the presence of autoantibodies to lipids in the bronchoalveolar lavage (BAL) fluid from adult patients with acute respiratory distress syndrome (ARDS). DESIGN: Analysis of immunoglobulin G (IgG) in BAL fluid by electrophoresis followed by immunoblotting and characterization of immunoglobulins as antilipid autoantibodies. SETTING: Intensive care unit of a university hospital and two research university laboratories. SUBJECTS: Twenty-seven mechanically ventilated patients in total, including nine patients with ARDS and two control groups. INTERVENTIONS: Patients were ventilated with a mechanical ventilation mode. Six aliquots of 20-mL sterile normal saline at 37 degrees C were infused through the working channel of the bronchoscope. MEASUREMENTS: Total protein, detection of IgG by electrophoresis followed by immunoblotting, and characterization of IgG by enzyme-linked immunosorbent assay using different lipids as target antigens. MAIN RESULTS: Antiphospholipid autoantibodies are present in BAL fluid of ARDS patients. Among the phospholipids tested, phosphatidic acid and phosphatidylserine gave the most significant activity. The IgG fraction, purified from BAL fluids by affinity chromatography, gave the same pattern of binding as that of the BAL fluid. CONCLUSION: The presence of antiphospholipid autoantibodies in BAL fluid suggests involvement of autoimmune mechanisms in the pathogenesis of ARDS.

Adult↗
PubMed1999

Elasticity of normal and cancerous human bladder cells studied by scanning force microscopy.

Scanning force microscopy was used for the determination of the elastic properties of living cells in their culture conditions. The studies were carried out on human epithelial cells. Two similar lines of normal cells (Hu609 and HCV29) and three cancerous ones (Hu456, T24, BC3726) were measured using the scanning force microscope in order to collect the force versus indentation curves. The BC3726 line originates from the HCV29 cell line which was transformed by the v-ras oncogene. To evaluate their elastic properties, Young's modulus values were determined. The present study has shown that normal cells have a Young's modulus of about one order of magnitude higher than cancerous ones. Such a change might be attributed to a difference in the organisation of cell cytoskeletons and requires further studies.

Biophysical Phenomena↗
PubMed1997

Proteins and phospholipids in BAL from patients with hydrostatic pulmonary edema.

The purpose of the present study is twofold: to evaluate alterations in total phospholipid content and individual phospholipid classes of the surfactant, and to detect markers of inflammatory reaction in bronchoalveolar lavage (BAL) from patients with hydrostatic pulmonary edema (HPE). Mechanically ventilated patients with HPE (Group 1) were compared with mechanically ventilated patients without cardiopulmonary disease (Group 2), considered as the control group. Group 3, including patients with high-permeability pulmonary edema, was used for further comparison. BAL was obtained and immediately cooled at 4 degrees C. Total proteins, albumin, and platelet-activating factor--acetylhydrolase (PAF-AcH) were measured. Total lipids were extracted and analyzed after thin-layer chromatographic separation. PAF was determined with bioassay. Total BAL proteins and albumin were found significantly higher in patients with HPE compared with control, but were lower compared with adult respiratory distress syndrome (ARDS). PAF was elevated in patients with HPE and ARDS, whereas in the control group it was actually in nondetectable levels. PAF was significantly higher in ARDS than in HPE patients. BAL neutrophils concentration was higher in HPE compared with control, but lower compared with ARDS. There was an inverse correlation between PAF-AcH and PAF. Quantitative reduction of total BAL phospholipids (PL) and qualititative deficiency was observed in both patients with HPE and ARDS. The findings of this study suggest that there is evidence of inflammation in the airspaces of patients with HPE.

1-Alkyl-2-acetylglycerophosphocholine Esterase↗
PubMed1996

Rapid tracheal infusion of surfactant versus bolus instillation in rabbits: effects on oxygenation, blood pressure and surfactant distribution.

UNLABELLED: Surfactant bolus instillation may be associated with a drop in blood pressure. Platelet-activating factor (PAF) has been found in surfactant preparations. The aim of this study was to evaluate rapid tracheal infusion of surfactant during 5 min as an alternative to bolus instillation and to examine whether a PAF receptor antagonist is able to prevent the decrease in blood pressure. METHODS: Surfactant deficiency was induced in 16 adult rabbits by lung lavages with saline. Six animals received a bolus of a porcine surfactant preparation (Curosurf (CS); 200 mg/kg), labeled with red microspheres to assess pulmonary distribution. In another 5 rabbits, the same amount of labelled CS was instilled by tracheal infusion within 5 min. A third group of 5 animals received 3 mg/kg body weight of the PAF antagonist WEB 2170 before CS bolus instillation. RESULTS: After CS bolus administration, mean PaO2 increased by 44.7 +/- 8.3 kPa (mean +/- SD) within 2 min and remained at this level. Mean arterial blood pressure dropped transiently by 2.3 +/- 2 kPa within 5 min. Pulmonary distribution of surfactant was even. After infusion, mean PaO2 rose by 22.4 +/- 16.3 kPa within 15 min. Blood pressure dropped by 1.8 +/- 1.1 kPa within 15 min. The distribution was extremely uneven. Blood pressure decreases also occurred after pretreatment with PAF receptor antagonist. CONCLUSION: Rapid tracheal infusion of surfactant results in poorer oxygenation, an inhomogeneous distribution and a similar decrease in blood pressure compared to the bolus instillation method. Blood pressure changes could not be prevented by a PAF receptor-specific antagonist.

Animals↗
PubMed1996

Scanning force microscopy of biological samples.

The SFM setup has been used to image the three-dimensional surface structures of various biological specimens in air and in liquid cell. The samples like red blood cell (RBC), lymphocytes, aggregates of pronase, and surface of bone measured in air, and macrophage surface measured in PBS solution are presented.

Bone and Bones↗
PubMed1994

Surface characterization of ceramic brackets: a multitechnique approach.

The purpose of this study was to investigate the microstructure and bonding mechanisms for four types of ceramic brackets by using information from several methods. Two of the bracket types provided exclusively micromechanical retention as a bonding mechanism, with the use of microspheres or microcrystals to achieve a rough structure for the bracket base. A silane layer that provided chemical adhesion was found to cover the bases of the two other types of brackets; one type also used central regions of increased roughness to provide additional micromechanical retention. Polarized-light optical microscopy showed that the silane layer was not continuous. Small area x-ray photoelectron spectroscopy analysis (SAXPS) of the silane layer was consistent with the presence of gamma-methacryloxypropyl trimethoxysilane. The study of the ceramic bracket bases revealed a wide variety in composition, structure, morphologic condition, and coating treatment that implies different bonding mechanisms to orthodontic adhesives.

Aluminum Oxide↗
PubMed1993

Isolation of a phospholipid inhibitor of platelet activating factor-induced activity from perfused rat liver: identification as phosphatidylglycerol.

An endogenous inhibitor of platelet activating factor action was isolated from perfused rat liver. It was purified by thin-layer chromatography and high-performance liquid chromatography and subjected to chemical modifications in order to identify its structure. On the basis of its fast atom bombardment-mass spectrum it was characterized as phosphatidylglycerol composed mainly of 16:0/18:1 and 16:0/20:2 fatty acyl chains ([M+H]+ at m/z 749 and 775, respectively) and very minor levels of 18:0/18:1 and 18:0/20:2. The purified compound exhibited inhibition on rabbit platelet aggregation induced by 5 x 10(-10) M platelet activating factor (PAF) at an EC50 value near 2.5 x 10(-6) M and on the serotonin secretion at an EC50 7 x 10(-6) M. Other phospholipids isolated from the liver preparations, such as phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, cardiolipin (diphosphatidylglycerol), and phosphatidic acid, exhibited no inhibitory activity in the concentration range from 1 x 10(-4) to 1 x 10(-7) M nor did they induce any aggregation, or lysis, of the platelets. Of importance, phosphatidylglycerol could inhibit thrombin- and ADP-induced aggregation of rabbit platelets. These results suggested a possible site of inhibition common to the signal transduction pathway of these agonists. Preliminary binding experiments showed a noncompetitive type of inhibition on PAF binding to intact rabbit platelets.

Adenosine Diphosphate↗
PubMed1993

Failure mode analysis of ceramic brackets bonded to enamel.

The purpose of this study was to evaluate in vitro the failure pattern of ceramic brackets bonded to enamel with a light-cured orthodontic adhesive. Five types of ceramic brackets and 125 incisors were used in the study. The brackets were bonded onto enamel with a light-cured orthodontic adhesive. After 1 week storage and thermal cycling, the samples were debonded by one operator according to the individual technique for each bracket group proposed by each manufacturer. The fracture surfaces were examined under a stereomicroscope to reveal the type of failures. The effect of the debonding procedure on enamel structure was significantly affected by the various bonding mechanisms of the bracket bases. Cohesive enamel fractures were detected from brackets that provided a bonding mechanism of micromechanical retention and chemical adhesion. The brackets that combined mechanical retention and chemical adhesion, presented both cohesive resin fractures and fractures located at the bracket resin or the resin enamel interface. The higher frequency of cohesive bracket fractures was obtained from a monocrystalline bracket.

Ceramics↗
PubMed1991

Porosity of pit and fissure sealants.

The aim of this study was to investigate the bulk porosity of a range of pit and fissure sealants. The following materials were included in the study: Concise White Sealant and Delton (chemically cured); Delton (visible light-cured); Helioseal, Visioseal and Prolite (visible light-cured with delivery from the bottle via cannule); Delton, Luma Seal and Prisma Shield (visible light-cured with syringe-type direct delivery systems). Five cylindrical samples, 7 mm in diameter and 1 mm in thickness, were prepared from each sealant and the following parameters were analysed in a computer-controlled mercury porosimeter: (i) total cumulative pore volume; (ii) specific surface area of pores; (iii) percentage total porosity (iv) bulk density; and (v) pore radius distribution. According to the results, the chemically cured sealants exhibited total cumulative pore volume, specific surface area of pores and total porosity. Significant differences were detected within the group of visible light-cured materials. The visible light-cured sealants with syringe-type delivery exhibited the lowest porosity.

Analysis of Variance↗
PubMed1990

Etched glass ionomer liners: surface properties and interfacial profile with composite resins.

The purpose of this study was to assess the surface alterations induced by acid etching on two glass ionomer lining cements (Ketac Bond, G-C lining cement) and to evaluate their interface with a composite resin (Herculite XR) following various surface treatments. The changes in the surface composition and topography of the etched liners were studied by electron microprobe and ESCA analyses. The interfacial treatments performed on the ionomer surfaces were: (i) application of a bonding resin; (ii) 20-s acid etching and application of a bonding resin; (iii) application of a dental adhesive based on a chlorophosphate ester of BisGMA (Scotch-bond LC), without acid etching. The interfacial sealing efficiency of these treatments was studied by the silver nitrate microleakage technique combined with optical and electron microscopy. According to the results the etched surfaces of both the liners present excessive porosity with glass and matrix dissolution. Significant changes in the surface chemistry of the liners were detected indicating severe degradation. The microleakage study revealed interfacial gaps and fractures in the etched samples. The best results were obtained from the non-etched ionomer liners which were subjected to the adhesive treatment. The efficiency of acid etching as a necessary step in the 'layered' technique is seriously questioned.

Acid Etching, Dental↗
PubMed1990

[Interfacial strength and microleakage pattern of paired visible light cured composites].

The clinical build up of visible light cured restorations in many cases is performed by layering different types of composites to obtain enhanced characteristics. The purpose of this study was to evaluate the interfacial strength and microleakage pattern of paired light-cured restorations. Two small particle (SP), two pure hybrid (HB) and two microfilled materials (MF) were selected. Cylindrical molds were used to prepare double layer specimens for the groups SP-HB, SP-MF, HB-MF and the controls. The last group was comprised of double layer specimens respectively. All the samples were immersed in normal saline at 37 degrees C and divided in two series. The first series included samples immersed in normal saline for 24 h and accelerated aged for 1 h in saline at 37 degrees C plus 1000 times thermal cycling between 5 degrees-60 degrees C. These samples were sheared off at 0.005 inch/min cross-head speed. The second series was composed of the samples subjected to the some experimental conditions but tested for their interfacial microleakage pattern utilizing the silver nitrate technique. According to the results pairing SP-MF, SP-HB increases interfacial strength compared to MF-MF, and HB-HB. The combination of HB-MF results in no statistical significance from MF-MF or HB-HB. Minimal leakage was detected in some cases but did not correlate to bond strength values.

Composite Resins↗