Gamma irradiation alters bluetongue virus protein antigen.

Bluetongue virus (BTV) antigen, prepared for a monoclonal antibody (MAb)-based competitive enzyme-linked immunosorbent assay (C-ELISA), was exposed to 1, 2, 3, 4, 5 and 6 Mrad of gamma irradiation. The major group-specific BTV protein (VP7) reactive with the Mab was altered at higher doses of radiation, as revealed by immunoblotting studies. As well, a reduction in immunoreactivity was noted when irradiated antigen was used in the ELISA.     

Computed tomographic study of the posterior condylar angle in arthritic knees: its use in the rotational positioning of the femoral implant of total knee prostheses

The epicondylar axis is a reliable reference to check the rotation of the femoral implant in total knee prostheses (TKPs). However, during the operation it seems easier to use the posterior condylar axis as a landmark. The angle between these two axes is called the posterior condylar angle (PCA). The aim of this study was to measure the PCA in arthritic knees to assess the reliability of the posterior condylar axis as a reference for the control of the rotation of the femoral implant and to look for correlation with other radiological measurements. This prospective study consisted of 103 arthritic knees (81 varus, 22 valgus) before a TKP had been done in 103 patients (75 women, 28 men). The assessment of the PCA was made by computed tomographic scanning (CT). The HKA, HKS and HKT angles were measured on the pangonogram. The posterior condylar axis was internally rotated with respect to the epicondylar axis. The average value for all the patients was 2.65° degrees with a range from 0° to 7°. The PCA was significantly increased in the valgus knees. There was no correlation between the angles on the pangonogram and the posterior condylar axis. While the preoperative assessment of the PCA by CT scanning is reliable, the results obtained indicate the marked variability in its value. If one wishes to use the posterior condylar axis as a guide for rotation, it is therefore necessary to assess the PCA for each patient using adjustable jigs according to the value obtained. No measurement on standard radiographs allowed an extrapolation of the value of the PCA, and CT scanning seems to be the preferable radiological examination.

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Electronic Supplementary Material The french version of this article is available in the form of electronic supplementary material and can be obtained by using the Springer Link server located at

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Etude tomodensitométrique de l'angle condylien postérieur dans les genoux arthrosiques. Intérêt dans le positionnement en rotation de l'implant fémoral dans les prothèses totales de genou

Résumé L'axe épicondylien est une référence fiable pour le contrôle de la rotation de l'implant fémoral dans les prothèses totales de genou (PTG). Mais, lors de l'intervention, il semble plus facile d'utiliser l'axe condylien postérieur comme repère. L'angle entre ses deux axes est appelé angle condylien postérieur (ACP). Le but de cette étude était de mesurer l'ACP dans les genoux arthrosiques, d'évaluer la fiabilité de l'axe condylien postérieur comme référence pour le réglage de la rotation de l'implant fémoral, de rechercher une corrélation avec d'autres mesures radiologiques. Une étude prospective comportant 103 genoux arthrosiques (81 varus et 22 valgus), avant PTG a été effectuée, chez 103 patients (75 femmes et 28 hommes). L'évaluation de l'ACP a été faite par examen tomodensitométrique (TDM). Les angles HKA, HKS et HKT ont été mesurés sur le pangonogramme. L'axe condylien postérieur était en rotation interne par rapport à l'axe épicondylien. La valeur moyenne pour tous les patients était de 2.65°, avec des valeurs de 0 à 7°. La valeur de l'angle CP augmentait avec une différence significative dans le groupe des genu valgum. Il n'y avait pas de corrélation entre les angles du pangonogramme et l'ACP. Si l'évaluation pré-opératoire de l'ACP par TDM est fiable, les résultats obtenus mettent en évidence une variabilité importante de sa valeur. Il faut donc, si l'on veut utiliser l'axe condylien postérieur comme repère de rotation, évaluer pour chaque patient l'ACP, et utiliser un ancillaire réglable reportant la valeur obtenue. Aucune mesure sur des radiographies standard ne permettant d'extrapoler la valeur de l'ACP, la TDM semble l'examen radiologique de choix.

     

Detection of a biliary cell membrane glycoprotein in the serum of cholangiocarcinoma-bearing rats. Possible relevance to the membrane proteins used as serum tumor markers in humans

Certain markers used in the diagnosis and monitoring of human adenocarcinomas, such as carcinoembryonic antigen are membrane glycoproteins normally absent from the serum. How those proteins may reach the blood after the neoplastic transformation remains debated. In this work, we show that cholangiocarcinoma induced by 3'-methyl-4-dimethylaminoazobenzene in the rat provides some insight into the mechanisms implicated in this process. We observed that the extracellular matrix of all cholangiocarcinomas tested contained large amounts of a glycoprotein identified by a monoclonal antibody termed B10, and previously characterized as an integral membrane protein normally restricted to the apical domain of epithelial cell plasma membrane. The extracellular deposition of the B10-binding glycoprotein in cholangiocarcinomas was associated with the appearance of detectable levels of the protein in the serum, and an abnormal membrane expression of the protein, which was detected on both apical and basal plasma membrane domains of neoplastic biliary cells. We postulate that neoplastic transformation of biliary cells leads to an inappropriate membrane expression of the B10-binding protein, which in turn, results in the extracellular release of the protein and in its diffusion into the blood. The characteristic desmoplastic stroma of cholangiocarcinoma offers the opportunity to easily visualize the release process. A comparable mechanism is likely to explain how certain membrane glycoproteins used as serum markers in human malignancy may reach the blood.     

Detection of Antiendothelial Cell Antibodies by an Enzyme-Linked Immunosorbent Assay Using Antigens from Cell Lysate: Minimal Interference with Antinuclear Antibodies and Rheumatoid Factors

The objective of the present work was to set up a routine test adapted to screening for antiendothelial cell antibodies (AECAs) in serum samples with minimal interference from antinuclear antibodies (ANAs) or rheumatoid factors (RFs). We compared the titers of AECAs titrated following two enzyme-linked immunosorbent assays (ELISAs): (i) an ELISA with ethanol-fixed EA.hy926 monolayers as the antigenic substrate and (ii) an ELISA with nucleus-depleted lysates prepared from EA.hy926 cells and normalized for protein (1.0 to 1.7 mg/ml) and DNA (≤0.1 μg/ml) contents as a surrogate substrate (postnuclear supernatant ELISA [PNS-ELISA]). The AECA titers in 51 serum samples, including 28 samples containing ANAs, were compared. A significantly positive correlation (r = 0.77; P < 0.001) between the two series was shown only for the ANA-negative serum samples. Conversely, ANAs or RFs in samples were shown not to interfere in tests for AECAs by the PNS-ELISA. AECAs recognize their antigenic targets in postnuclear supernatants, which is representative of the endothelial antigenic content, with improvement of the reliability of the assay, a prerequisite to application of the assay for their evaluation in clinical practice.     

Radical-initiated copolymers of styrene and p-formylstyrene, 1. Solution copolymerization and characterization

Kinetics of solution homopolymerization of p-formylstyrene

1 System name: 4-vinylbenzaldehyde.

(A) and copolymerization with styrene (S) have been examined at 60°C in N,N-dimethylformamide with 2,2′-azoisobutyronitrile as initiator. For p-formylstyrene, k_p/k = 0,135 L^1/2 · mol^?1/2 · s^?1/2 was estimated from the plot of monomer consumption versus time, i.e. a value greater than that for styrene (K_p and K_t are the rate constants of propagation and termination and termination, respectively). Reactivity ratios have been determined to be r_A = 3,0 and r_S = 0.2, indicating the reactivity of p-formylstyrene to be higher than that of styrene. Moreover, from initial copolymerization rates, the cross termination factor was calculated and found to depend upon the copolymer composition. Some solution properties of the homo- and copolymers have been analysed: solubility of (co)polymers, their molecular weight and its distribution (by gel permeation chromatography), and their composition and monomer sequence distribution (using ¹H and ¹³C NMR spectroscopy).     

Distinctive alterations of invasiveness, drug resistance and cell-cell organization in 3D-cultures of MCF-7, a human breast cancer cell line, and its multidrug resistant variant

Growth of human tumor cells as three-dimensional (3D) multicellular spheroids modifies their invasive properties. Here we study the differences in the biological features of MCF-7, a human breast cancer cell line, and its multidrug resistant variant (MDR-MCF-7) cultured as spheroids or as monolayers. Three-dimensional culture decreased the proliferative rate of both cell lines, reduced the drug sensitivity of MCF-7 cells and did not affect the resistance of MDR-MCF-7 cells. Transmission electron microscopic studies and intercellular junctions labeling showed that MCF-7 spheroids had a junctional system involving E-cadherin, tight-junctions and desmosomes. In MDR-MCF-7 cell spheroids, cell cohesion was mostly due to membrane interdigitations. MDR-MCF-7 cells, but not their parental counterpart, displayed a higher invasive potential when cultured as spheroids, as shown in the Boyden chamber assay. 3D-induced invasiveness was correlated with serine protease and plasminogen activator (PA) secretion. MCF-7 cells did not show any tendency to invade, whatever the mode of culture. These results show that 3D-cultures as spheroids distinctively altered structural features of parental and MDR-MCF-7 cells. In MCF-7 cells, 3D-culture increased cell-cell contacts and drug resistance; in MDR-MCF-7 cells, it induced invasive properties.     

In vitro degradation of silk fibroin

A significant need exists for long-term degradable biomaterials which can slowly and predictably transfer a load-bearing burden to developing biological tissue. In this study Bombyx mori silk fibroin yarns were incubated in 1mg/ml Protease XIV at 37 degrees C to create an in vitro model system of proteolytic degradation. Samples were harvested at designated time points up to 12 weeks and (1) prepared for scanning electron microscopy (SEM), (2) lyophilized and weighed, (3) mechanical properties determined using a servohydraulic Instron 8511, (4) dissolved and run on a SDS-PAGE gel, and (5) characterized with Fourier transform infrared spectroscopy. Control samples were incubated in phosphate-buffered saline. Fibroin was shown to proteolytically degrade with predictable rates of change in fibroin diameter, failure strength, cycles to failure, and mass. SEM indicated increasing fragmentation of individual fibroin filaments from protease-digested samples with time of exposure to the enzyme; particulate debris was present within 7 days of incubation. Gel electrophoresis indicated a decreasing amount of the silk 25 kDa light chain and a shift in the molecular weight of the heavy chain with increasing incubation time in protease. Results support that silk is a mechanically robust biomaterial with predictable long-term degradation characteristics.     

Functional interactions between the VRP1–LAS17 and RHO3–RHO4 genes involved in actin cytoskeleton organization in Saccharomyces cerevisiae

The RGD1 gene from Saccharomyces cerevisiae, which encodes a GTPase-activating protein for the Rho3 and Rho4 small G proteins, exhibits synthetic lethality with the VRP1 and LAS17 genes. Their products are proline-rich proteins that interact with both actin and myosins to ensure polarized growth. By testing functional links, we found that the VRP1 and LAS17 genes are potent suppressors of the rho3Delta mutation. In particular, they restore the polarization of actin patches in rho3Delta cells. Moreover, the vrp1Delta and las17Delta mutations were found to display a similar pattern of genetic interactions with specific actin-linked genes. These mutations also increase the sensitivity to activated forms of both Rho3p and Rho4p. These data support our working model, in which the VRP1 and LAS17 genes define a cellular complex that works in concert with the RHO3-RHO4 signaling pathway in yeast polarized growth. In addition, other observations lead us to propose that Rvs167p may act as a linking protein between the two cellular elements.