Detection of human serum amyloid A protein in very low density--and high density lipoproteins of patients after acute myocardial infarction

Two acute-phase proteins have been identified in very low density (VLDL)- and high density lipoproteins (HDL) of patients after acute myocardial infarction. Both proteins have a relative molecular weight of 11,000 and isoelectric points pI 6.08 and 6.27, and do not contain cysteine or sugar residues. Polyclonal antibodies to these acute phase reactants did not cross-react with other serum apolipoproteins. Evidence is given that both proteins are polymorphic forms of the human serum amyloid A protein.     

Two-dimensional electrophoresis of plasma proteins and high density lipoproteins during inflammation

Plasma protein and lipoprotein fractions of five patients were analyzed on day 1, 5, and 15 after severe head injury by combining three types of two-dimensional electrophoresis (2-DE) to obtain information on lipoprotein and apolipoprotein composition. On analysis under nondenaturing conditions in both dimensions on day 5, the samples show modifications of isoelectric point (pI) and molecular weight (Mr) properties of the high density lipoprotein (HDL) fraction in addition to an increase in inflammatory proteins and a return to a normal pattern on day 15. In the second type of 2-DE the samples were analyzed employing isoelectric focusing without denaturant in the first dimension, followed by sodium dodecyl sulfate (SDS) in the second dimension in order to study the protein composition of lipoprotein fractions. On day 5, a decrease of the apolipoproteins apo A-I, apo A-II, and apo C were noted, with simultaneous appearance of an unidentified protein with Mr 12,000 and pI 6.0. In the third type of 2-DE, employing urea and Nonidet P-40 in the first and SDS in the second dimension, the plasma polypeptide composition was studied. The presence of an unidentified polypeptide could be confirmed on day 5, tending to disappear thereafter. This Mr 12,000 component consists of two major spots at pI 5.7 and 6.0 and four minor ones between pI 6.0 and 8.0. These properties suggest that this protein corresponds to serum amyloid A apolipoprotein.     

Interactions of lecithin and pig apolipoproteins of high density lipoproteins at the surface monolayer of reconstituted very small particles

Cosonication of egg yolk lecithin and triolein with apolipoproteins isolated from pig high density lipoprotein (apoHDL) gave us reconstituted high density lipoprotein particles (r-HDLs) of 9 nm in average diameter. They were smaller than microemulsion particles (MEs) composed of the lipids (35 nm). The protein/egg yolk lecithin ratio in the fractionated r-HDLs was higher in the smaller particles. Binding of a hydrophobic probe, 2-p-toluidinylnaphthalene-6-sulfonate (TNS), to MEs, r-HDLs and apoHDL were evaluated on the basis of Halfman and Nishida's method. The reconstitution of apoHDL into MEs led to a 68% reduction in the binding of TNS and a small increase in the alpha-helix content as compared with free apoHDL. The binding experiments also showed the condensation of lecithin molecules at the r-HDL surface. The amphipathic helixes of apoHDL are located in the surface monolayer of egg yolk lecithin surrounding the triolein core. The intercalation of the hydrophobic residues of apoHDL between egg yolk lecithin molecules brings about a pronounced curvature of the surface and a decrease in the particle diameter.     

Cellular uptake and photosensitizing properties of anticancer porphyrins in cell membranes and low and high density lipoproteins

The mechanisms of the phototoxic effect of anticancer porphyrins used in the photodynamic therapy (PDT) of tumours are not yet completely understood. Irradiation of porphyrins gives rise to singlet oxygen which reacts with key residues of proteins, polyunsaturated fatty acids and cholesterol in membranes, leading to inactivation of various enzymes and transporters. Lipoproteins, mainly low density lipoproteins (LDL), are efficient carriers of anticancer porphyrins in blood and can deliver these photosensitizers to tissues through the apolipoprotein (apo) B/E specific LDL receptor pathway. In this review, we discuss some aspects of anticancer porphyrin transport, cellular uptake and photosensitizing properties in cell membranes and lipoproteins.     

Softness of atherogenic lipoproteins: a comparison of very low density lipoprotein (VLDL) and low density lipoprotein (LDL) using elastic incoherent neutron scattering (EINS)

Apolipoprotein B100 (apoB100)-containing plasma lipoproteins (LDL and VLDL) supply tissues and cells with cholesterol and fat. During lipolytic conversion from VLDL to LDL the size and chemical composition of the particles change, but the apoB100 molecule remains bound to the lipids and regulates the receptor mediated uptake. The molecular physical parameters which control lipoprotein remodeling and enable particle stabilization by apoB100 are largely unknown. Here, we have compared the molecular dynamics and elasticities of VLDL and LDL derived by elastic neutron scattering temperature scans. We have determined thermal motions, dynamical transitions, and molecular fluctuations, which reflect the temperature-dependent motional coupling between lipid and protein. Our results revealed that lipoprotein particles are extremely soft and flexible. We found substantial differences in the molecular resiliences of lipoproteins, especially at higher temperatures. These discrepancies not only can be explained in terms of lipid composition and mobility but also suggest that apoB100 displays different dynamics dependent on the lipoprotein it is bound to. Hence, we suppose that the inherent conformational flexibility of apoB100 permits particle stabilization upon lipid exchange, whereas the dynamic coupling between protein and lipids might be a key determinant for lipoprotein conversion and atherogenicity.     

Immunochromatographic rapid analysis of human heart-type fatty acid-binding protein for acute myocardial infarction diagnosis

A method of immunochromatographic assay for rapid detection of human heart-type fatty acid-binding protein (h-FABP) as an early marker of acute myocardial infarction was developed. Gold nanoparticles were used as a visual label. The optimum conditions for assay were determined. The limit of detection in the assay was 1.5 ng/mL, and the variation coefficient did not exceed 8%. Using the developed test system, the clinical diagnosis of acute myocardial infarction (N = 10) was confirmed, and the results of testing the serum of healthy individuals (N = 25) were negative.     

Assessment of role of rosmarinic acid in preventing oxidative process of low density lipoproteins

This study aimed to assess the antioxidant compound effects on oxidisable substrates, using an effective bio-mimetic system based on human low density lipoproteins (LDL). Thermally generated radicals induce LDL oxidative changes to be identified and quantified. The bio-mimetic system thus developed responded linearly to radicals?? concentration over a range of 10^?6-10^?5 mol L^?1. Cu²⁺ accentuates lipoperoxidation but, when rosmarinic acid was present, Cu²⁺ produced an unexpected effect, i.e. increased antioxidant efficiency against lipoperoxidation. Rosmarinic acid inhibits production of lipoperoxides by up to 30 % in the absence of Cu²⁺ and up to 70 % in its presence when the rosmarinic acid-to-Cu molar ratio is 1: 1.     

A simplified hard-sphere equation of state meeting the high and low density limits

A simplified hard-sphere equation of state has been developed, which meets the correct limit for close-packed conditions. It is shown that the proposed equation of state for hard spheres can represent accurately the computer simulation compressibility factor data and virial coefficients over a wide density range. The comparison of the results of the calculations using this equation, the Carnahan-Starling equation, and the two equations proposed by Iglesias-Silva and Hall, shows that the equation proposed here represents the compressibility factor data and the virial coefficients with better accuracy.     

Heterogeneity of serum creatine kinase isoenzyme-MM in acute myocardial infarction

Isoelectric focusing of serum creatine kinase (CK;EC 2.7.3.2) reveals up to 14 CK-MM subbands following acute myocardial infarction (AMI). The "normal" subbands 1 (pI 6.91), 2 (pI 6.65) and 3 (pI 6.35) are faintly present in normal serum and the "abnormal" subbands c (pI 7.25), e (pI 6.85), g (pI 6.50), i (pI 6.28), j (pI 6.20) and k (pI 6.15) are prominently detected in sera with elevated CK. "Abnormal" subbands a (pI 7.55),b(pI7.35),d(pI7.05),f(pI6.72) and h(pI6.40) have only been detected in AMI. The "abnormal" subbands appear, and reach maximum intensity (together with CK-MM 1-3), 3-12 h after infarction, and become faint and anodally convert (as do CK-MM 1-3) within 36 h. Similar changes are detected by nonequilibrium pH gradient electrophoresis which combines CK-MM and CK-MB analysis. In vitro incubation of serum with 0.015 M 2-mercaptoethanol induces conversion of CK-MM 1, 2 and 3 to b and c, d and e, and f and g, respectively. Thus, the complexity of the patterns is explained by a secondary conversion of "normal" to "abnormal" subbands superimposed upon anodal conversion of CK-MM 1----3. The clinical significance of these findings is discussed.     

Unexpected molecular weight dependence of shish-kebab structure in the oriented linear low density polyethylene/high density polyethylene blends

In this study, highly oriented shish-kebab structure was achieved via imposing oscillatory shear on the melts of linear low density polyethylene (LLDPE)/high density polyethylene (HDPE) blends during the packing stage of injection molding. To investigate the effect of molecular weight of HDPE on the formation of shish-kebab structure, two kinds HDPE with large melt flow index (low molecular weight) and small melt flow index (high molecular weight) were added into LLDPE matrix. The structural characteristics of LLDPE/HDPE blends were systematically elucidated through two-dimensional wide-angle x-ray scattering, scanning electron microscopy, and differential scanning calorimetry. Interestingly, an unexpected molecular weight dependence of shish-kebab structure of the prepared samples was found that the addition of HDPE with low molecular weight resulted in an higher degree of orientation, better regularity of lamellar arrangement, thicker lamellar size, and higher crystal melting temperature than that adding HDPE with high molecular weight. Correspondingly, the blend containing low molecular weight HDPE had better tensile strength. A possible mechanism was suggested to elucidate the role of HDPE molecular weight on the formation of shish-kebab structure in the oriented blends, considering the change of chain mobility and entanglement density with change of molecular weight.     

Endothelial progenitor cell transplantation decreases lymphangiogenesis and adverse myocardial remodeling in a mouse model of acute myocardial infarction

Cardiac lymphatic system in the remodeling after acute myocardial infarction (AMI) has been overlooked. We wanted to investigate the role of bone marrow-derived endothelial progenitor cells (EPCs) and their contribution to lymphatic distribution in myocardial remodeling after AMI. Mouse (C57bl/6J) MI models were created by ligation of the left anterior descending coronary artery and were treated with phosphate buffered saline (PBS) or EPCs. Real-time RT-PCR with 2- to 4-week myocardial tissue samples revealed that lymphangiogenetic factors such as vascular endothelial growth factor (VEGF)-C (8.5 fold, P < 0.05), VEGF-D (6.1 fold, P < 0.05), Lyve-1 (15 fold, P < 0.05), and Prox-1 (11 fold, P < 0.05) were expressed at significantly higher levels in the PBS group than the EPC group. The PBS group also showed a significantly higher density of lymphatic vessels in the peri-infarction area. Echocardiography showed that from 2 weeks after the treatment, left ventricle (LV) dimensions at both systole and diastole were significantly smaller in the EPC group than in the PBS group (P < 0.01) and LV fractional shortening was higher in the EPC group accordingly (P < 0.01). Lymphangiogenic markers increased in a mouse MI model. EPC transplantation decreased lymphangiogenesis and adverse ventricular remodeling after AMI. These novel findings suggest that new lymphatic vessels may be formed in severely damaged myocardium, and may be involved in adverse myocardial remodeling after AMI.     

A mycelium with polyelectrolyte complex-bunched hyphae: several important factors affecting on the fermentation performance at a very high cell density

We have reported in the previous paper (Colloids Surf. B (2006) in press) a marked increase in the rate of gluconic acid production at a very high cell concentration (40 g/l) of filamentous fungus (Aspergillus niger IFO 31012) which was immobilized with polyelectrolyte complex consisting of potassium poly(vinyl alcohol) sulfate and trimethylammonium glycol chitosan iodide [6-O-(2-hydroxyethyl)-2-(trimethylammonio)-chitosan iodide]. The present study was carried out to look at what factors play a crucial role in this enhancement. We measured viscosity of broth, mass-transfer coefficient (k(L)a) for oxygen and diffusion coefficient of glucose (substrate). It has become apparent that there is only a difference in the diffusion coefficient of glucose between the free and immobilized cells. Therefore, we believe that the diffusion limitation by substrates as a problem in submerged mycelial processes is improved by immobilization based on polyelectrolyte complexes.     

Erosion‐dependant continuity development in high viscosity ratio blends of very low interfacial tension

This work studies continuity development and cocontinuity in high viscosity ratio EPDM/PP blends. A very low interfacial tension (0.3 mN/m) between the blend components together with high viscosity ratios (11 and 17) result in a variety of unusual morphological features, including isolated nanometer diameter fibers, very large particles, partially coalesced particles, and numerous particles interconnected by fibers. This unique combination of morphologies leads the blend to a novel and stable cocontinuous structure of partially coalesced particles and particles interconnected by fibers. Compared with low to medium viscosity ratio EPDM/PP blends, these cocontinuous networks demonstrate early percolation thresholds, rapid continuity development, and attain cocontinuity at lower compositions of minor phase. The slow surface erosion of the high viscosity EPDM phase during melt blending is shown to be responsible for the generation of these unusual morphological structures. Typically the timescale for erosion phenomena are so small that they have defied study in the mixing environment itself and typical blend morphology studies almost always examine the final steady‐state morphology obtained after several minutes of mixing. The combination of very low interfacial tension and very high viscosity ratios of these EPDM/PP systems provide a unique opportunity to examine erosion phenomena persisting over longer time scales during melt mixing. We propose a new concentration‐dependant erosion mechanism that is based on particle collision–coalescence–separation dynamics. The proposed conceptual mechanism is shown to dramatically accelerate the erosion process and maintain cocontinuity over prolonged periods of mixing. © 2006 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 44: 1919–1929, 2006     

An evaluation of two-dimensional electrophoresis for detection of human serum proteins in acute myocardial infarction

High resolution two-dimensional electrophoresis indicates that serum proteins previously only detected in patients with acute myocardial infarction (AMI) (Gomo et al., Electrophoresis 1983, 4, 298-302) are also present in the serum protein patterns of other patients (AMI negative or demonstrating acute phase response) and are faintly detected even in controls. Thus, these proteins are not specific to AMI and are probably acute phase reactants. However, they do demonstrate a characteristic time course response in sequential samples from AMI patients.     

A procedure for the determination of very low activity levels of tritium in water samples

We describe the procedure developed for the electrolytic concentration of³H in water, together with systematically applied checks to guarantee the reproducibility of the process and to minimize the variability between water samples concentrated simultaneously in the different electrolytic cells. Starting from initial volume of 250 ml water, we obtained enrichment parameters normally greater than 70%, with a dispersion always less than 5%. Measuring the concentrated and the unelectrolyzed samples under the same conditions, we found that the lowest detectable activities had been improved from around 1.5 to 0.3 Bq/l. This allowed us to quantify the³H activity levels of 100% of the rainwater samples collected in Cáceres from 1994 to 1996. The resulting average value for this period was (0.79±0.38) Bq/l.     

Kinetic approach to radiation-induced grafting in the polyethylene-styrene system. IV. Comparison between high density polyethylene and low density polyethylene

The investigation method reported in earlier articles was applied to preirradiation methods of the reaction of low-density polyethylene (LDPE) in liquid and vapor and compared with high-density polyethylene (HDPE). Monomer concentrations during reactions and monomer feed rates were determined gravimetrically. Increasing patterns of the degree of grafting were obtained and compared. Monomer concentration during the reactions was lower in LDPE than HDPE and radical decay was more rapid in LDPE. A model calculation was applied to this experiment and a schematic explanation was attempted. The differences between the reaction mechanisms of HDPE and LDPE are explained.     

The shape of the biodegradation curve for low and high density polyethenes in prolonged series of experiments

Previous work on the ¹⁴CO₂ liberation (measured by liquid scintillation) after biodegradative attempts was extended from two to four years in the case of LDPE ([¹⁴C]-labelled material mixed in the bulk) and was compared with results on HDPE (homogenously labelled by mixing in [¹⁴C]monomer). The trends for LDPE degradation with or without u.v.-sensitizer and its dependence on the surface/volume ratio were found to be similar to HDPE in spite of the fact that LDPE is more inert. The degradation curves were characterized by a straight line progression in the first 100 days of observation before declining parabolically. This is probably due to a rapid exhaust and enzymatic cleavage of the extractible short chain materials from the bulk. The trend was occasionally reversed to a progressive increase of ¹⁴CO₂ liberation in some of the long term runs. Elucidation of this anomaly is being attempted.     

New way of presenting the universal calibration of gel permeation chromatography to include macromolecules with very high segment density

The universal calibration of gel permeation chromatography presents deviations when star‐shaped polymers are used, especially when they present high segment density. Polymers presenting high segment density also have higher values of Flory's parameter Φ than the corresponding flexible, linear polymers. This deviation disappears if we express the hydrodynamic volume of the polymers with the quantity [η]M/Φ (where [η] is the intrinsic viscosity and M is the molecular mass) instead of the commonly used quantity [η]M. © 2005 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 43: 2388–2391, 2005     

Differences in first neighbor orientation behind the anomalies in the low and high density trans-1,2-dichloroethene liquid

Trans-1,2-dichloroethene (HClC=CClH) has several structural and dynamic anomalies between its low- and high-density liquid, previously found through neutron scattering experiments. To explain the microscopic origin of the differences found in those experiments, a series of molecular dynamics simulations were performed. The analysis of molecular short-range order shows that the number of molecules in the first neighbor shell is 12 for the high-density liquid and 11 for the low-density one. It also shows that the angular position of the center of mass of the first neighbor is roughly the same although the molecular orientation is not. In both liquids the first neighbor and its reference molecule arrange mainly in two configurations, each being the most probable in one of the liquids. First neighbors in the configuration that predominates in the high-density liquid tend to locate themselves closer to the reference molecule, an evidence that they are more strongly bonded. This arrangement facilitates a better packing of the rest of molecules in the first neighbor shell so that on average an additional molecule can be included, and is proposed to be the key in the explanation of all the observed anomalies in the characteristics of both liquids.     

Stability of the interphase interaction and changes in the microdestruction mechanisms in low density polyethylene on high energy irradiation

The effect of gamma irradiation on non-isothermally crystallized low density polyethylene (LDPE) has been studied by thermal-mechanical analysis (TMA) and transmission and scanning electron microscopy (TEM and SEM, respectively). Analysis of the TMA curves of the irradiated samples shows that the radiation-induced crosslinking of the macromolecular chains in the dose range 0–10 Mrad causes an increase, decrease and partial freezing of the mechanical microstresses located at the phase surfaces in the bulk of the polymer. These effects are explained in terms of the physical kinetics and macromolecular mobility. At temperatures above 120 °C., gamma-irradiated LDPE with doses above 5 Mrad develops a physical state corresponding to a high elastic state with thermotropic mesophases. Below 5 Mrad, controllable changes of the melting entropy and enthalpy of the crystalline regions and the free energy of the crystalline lamellae can be achieved. The TEM and SEM study shows an increase in the degree of co-operation of the sublevels of the superstructural molecular organization with the irradiation dose, even in areas that undergo plastic microdestruction.