Physicochemical properties of calcific deposits isolated from porcine bioprosthetic heart valves removed from patients following 2-13 years function

The purpose of this study was to characterize the physicochemical properties of calcific deposits that cause the failure of tissue-derived heart valve bioprostheses. This was done in an effort to understand the mechanism of pathologic biomineralization in the cardiovascular system and potentially prevent deterioration of bioprostheses. Calcific deposits taken from 10 failed bioprosthetic valves that had been implanted in patients for 2-13 years were characterized by chemical analysis, x-ray diffraction, FTIR spectroscopy, scanning electron microscopy, polarized light microscopy, and solubility measurements. The combined results identified the biomineral as an apatitic calcium phosphate salt with substantial incorporation of sodium, magnesium and carbonate. The average Ca/PO4 ratio for this "young" pathologic biomineral was approximately 1.3, considerably lower than approximately 1.7 found in mature atherosclerotic plaque biomineral and mature skeletal biomineral, both of which approximate hydroxyapatite in composition. Deproteinated calcific deposits from bioprostheses had thermodynamic solubilities comparable to those of both atherosclerotic plaque, typical pathologic biomineral and hydrolyzed octacalcium phosphate (OCP, Ca4H(PO4)3 x 2.5 H2O), a proposed precursor phase to biomineral apatite. This later finding, together with chemical composition and structural details of the bioprosthetic deposits themselves, supports a mechanism of cardiovascular calcification in which OCP plays a crucial role in the formation of the final apatitic phase. This suggests an approach toward prevention of bioprosthetic tissue calcification through control of the formation of the kinetically favored OCP precursor and/or its transformation into bioapatite.     

The Effect of High and Variable Glucose on the Viability of Endothelial Cells Co-Cultured with Smooth Muscle Cells

Diabetes mellitus causes endothelial dysfunction. The aim of this study was to investigate the effect of normal (5 mmol/L), high (20 mmol/L), and fluctuating (5 and 20 mmol/L changed every day) glucose concentration in the culture medium on the viability of human umbilical vein endothelial cells (HUVECs) co-cultured with human umbilical artery smooth muscle cells (HUASMCs). The cultures were conducted on semi-permeable flat polysulfone (PSU) fibronectin-coated membranes immobilized in self-made inserts. The insert contained either HUVECs on a single membrane or HUASMCs and HUVECs on two membranes close to each other. Cultures were conducted for 7 or 14 days. Apoptosis, mitochondrial potential, and the production of reactive oxygen species and lactate by HUVECs were investigated. The results indicate that fluctuations in glucose concentration have a stronger negative effect on HUVECs viability than constant high glucose concentration. High and fluctuating glucose concentrations slow down cell proliferation compared to the culture carried out in the medium with normal glucose concentration. In conclusion, HUASMCs affect the viability of HUVECs when both types of cells are co-cultured in medium with normal or variable glucose concentration.     

HIV-2 Neutralization Sensitivity in Relation to Co-Receptor Entry Pathways and Env Motifs

HIV-2, compared to HIV-1, elicits potent and broadly neutralizing antibodies, and uses a broad range of co-receptors. However, both sensitivity to neutralization and breadth of co-receptor use varies between HIV-2 isolates, and the molecular background is still not fully understood. Thus, in the current study, we have deciphered relationships between HIV-2 neutralization sensitivity, co-receptor use and viral envelope glycoprotein (Env) molecular motifs. A panel of primary HIV-2 isolates, with predefined use of co-receptors, was assessed for neutralization sensitivity using a set of HIV-2 Env-directed monoclonal antibodies and co-receptor indicator cell lines. Neutralization sensitivity of the isolates was analysed in relation target cell co-receptor expression, in addition to amino acid motifs and predicted structures of Env regions. Results showed that HIV-2 isolates were more resistant to neutralizing antibodies when entering target cells via the alternative co-receptor GPR15, as compared to CCR5. A similar pattern was noted for isolates using the alternative co-receptor CXCR6. Sensitivity to neutralizing antibodies appeared also to be linked to specific Env motifs in V1/V2 and C3 regions. Our findings suggest that HIV-2 sensitivity to neutralization depends both on which co-receptor is used for cell entry and on specific Env motifs. This study highlights the multifactorial mechanisms behind HIV-2 neutralization sensitivity.     

A method for sensorless on-line vibration monitoring of inductionmachines

This paper proposes a method for the sensorless online vibration monitoring of induction machines based on the relationship between the current harmonics in the machine and their related vibration harmonics. Initially, the vibration monitoring system records two baseline measurements of current and vibration with the machine operating under normal conditions. The baseline data is then evaluated to determine the critical frequencies to monitor online. Once these frequencies are determined, the baseline vibration measurement is simply used to scale the current harmonic signal to an estimated vibration level. Based on theoretical analysis, simulation results and the experimental results shown here, a linear relationship between the current harmonics and vibration level can be assumed. The results of two experiments on a three-phase 230 V 10 hp induction motor operating under no load are discussed and show the feasibility of this method for sensorless online vibration monitoring     

Estimation of the intracavity power levels for a hybrid free-electron laser device

A hybrid free-electron laser device (called a GYROFEL), employing axial bunching in a solenoidal field configuration, has been analyzed to determine potential operational power levels. A small signal gain model was utilized to estimate the intracavity electric field strengths attained at equilibrium in such a device. The results indicate that out-coupled power levels approaching 1 MW might be achievable for electron beams near the 1 MeV energy range.     

Learning from polar ice core research

An understanding of long-term climate changes, with some surprises, is emerging from analysis of ice core samples.     

Multi‐Site Functionalization of Protein Scaffolds for Bimetallic Nanoparticle Templating

The use of biological scaffolds to template inorganic material offers a strategy to synthesize precise composite nanostructures of different sizes and shapes. Proteins are unique biological scaffolds that consist of multiple binding regions or epitope sites that site‐specifically associate with conserved amino acid sequences within protein‐binding partners. These binding regions can be exploited as synthesis sites for multiple inorganic species within the same protein scaffold, resulting in bimetallic inorganic nanostructures. This strategy is demonstrated with the scaffold protein clathrin, which self‐assembles into spherical cages. Specifically, tether peptides that noncovalently associate with distinct clathrin epitope sites, while initiating simultaneous synthesis of two inorganic species within the assembled clathrin protein cage, are designed. The flexibility and diversity of this unique biotemplating strategy is demonstrated by synthesizing two types of composite structures (silver–gold mixed bimetallic and silver–gold core–shell nanostructures) from a single clathrin template. This noncovalent, Template Engineering Through Epitope Recognition, or TEThER, strategy can be readily applied to any protein system with known epitope sites to template a variety of bimetallic structures without the need for chemical or genetic mutations.     

Nanoparticle probes and mid-infrared chemical imaging for DNA microarray detection

To date most mid-infrared spectroscopic studies have been limited, due to lack of sensitivity, to the structural characterization of a single oligonucleotide probe immobilized over the entire surface of a gold-coated slide or other infrared substrate. By contrast, widely used and commercially available glass slides and a microarray spotter that prints approximately 120-μm-diameter DNA spots were employed in the present work. To our knowledge, mid-infrared chemical imaging (IRCI) in the external reflection mode has been applied in the present study for the first time to the detection of nanostructure-based DNA microarrays spotted on glass slides. Alkyl amine-modified oligonucleotide probes were immobilized on glass slides that had been prefunctionalized with succinimidyl ester groups. This molecular fluorophore-free method entailed the binding of gold-nanoparticle-streptavidin conjugates to biotinylated DNA targets. Hybridization was visualized by the silver enhancement of gold nanoparticles. The adlayer of silver, selectively bound only to hybridized spots in a microarray, formed the external reflective infrared substrate that was necessary for the detection of DNA hybridization by IRCI in the present proof-of-concept study. IRCI made it possible to discriminate between diffuse and specular external reflection modes. The promising qualitative results are presented herein, and the implications for quantitative determination of DNA microarrays are discussed.     

Crystallinity and accessibility of fibrous carboxymethylcellulose by pad-roll technology

Statistically designed experiments have been made on partially carboxymethylated cotton fabric. The effect of the reaction parameters (concentration of sodium hydroxide and mono-chioroacetic acid and period of time) on the crystallinity (measured by x-ray diffraction), on the iodine sorption and on the disorder of amorphous fraction (calculated from iodine sorption capacity) are presented by polynomial models. © 1996 John Wiley & Sons, Inc.     

The Effect of processing on the content and composition of free sterols and sterol esters in soybean oil

The content and composition of free sterols and sterol esters in crude soybean oil and in oils from different stages of two continuous refining systems were determined. The sterols were isolated by preparative thin layer chromatography and analyzed by gas chromatography with cholesterol as an internal standard. The free sterols in one of the degummed oils amounted to 3.1 mg/g and were diminished to 1.8 mg/g oil by the De Laval Short-Mix refining process. The content of free sterols of the other degummed oil was reduced from 3.4 to 1.6 mg/g oil by the Zenith process. The greatest reduction of sterol content was caused by the treatment with bleaching earth. The sterol esters accounted for 0.6 mg/g of the degummed oil, and only very small changes were observed during the processes. However, changes in the composition of fatty acids of the sterol esters were found. These changes might indicate a selective deacylation of sterol esters or an interesterification during the refining processes. The composition of sterols in free and esterified form were different. Campesterol, stigmasterol and sitosterol were obtained in both free and esterified form, but Δ7 stigmasterol was only found in esterified form. Only small changes in the percentage distribution of the sterols occurred during the processes. Present adress Food Technology Division, ALFA-LAVAL,S-14700 Tumba, Sweden