The morphology of the articular surfaces of biological knee joints provides essential guidance for the construction of functional knee endoprostheses.

PURPOSE: In comparative examinations of kinematics of the knees of humans and pigs in flexional/extensional motion under compressive loads, the significant differential geometric essentials of articular guidance are elaborated to criticise the shaping of the articular surfaces of conventional knee-endoprostheses and to suggest constructional outlines that allow the endoprosthesis to adopt natural knee kinematics. Implantation is discussed with regard to the remaining ligamentous apparatus. METHODS: Twelve fresh pig knee joints and 19 preserved human knee joints were moved into several flexional/extensional positions. In each joint, the tibia and femur were repeatably caught by metal plates. After removing all ligaments, the tibia and femur were again caught in these positions, and their points of contact were marked on both articular surfaces. Along the marker points, a thin lead wire was glued onto each surface. The positions and shapes of the four contact lines were mapped by teleradiography. RESULTS: All contact lines were found to be plane curves. The medial and lateral planes were parallel, thus defining the joint's sagittal plane. In the human knee, as compared to the lateral, the medial femoral contact line was always shifted anteriorly by several millimetres. The tibial contact curve was laterally convex and medially concave. In the pig knees, the lateral and medial contact lines were asymmetrically placed. Both tibial curves were convex. CONCLUSIONS: Both knees represent cam mechanisms (with one degree of freedom) that produce rolling of the articular surfaces during the stance phase. Implantation requires preservation of the anterior cruciate ligament, and ligamentous balancing is disadvantageous.

Versican overexpression in human breast cancer lesions: known and new isoforms for stromal tumor targeting.

Proteoglycans play a key role in cancer development and progression by participating in the constitution of a specific fertile tumor microenvironment. As they are largely overexpressed in the malignant stroma, proteoglycans provide a reservoir of potential new targets for anticancer therapies, because they can serve to convey toxic payloads in the close proximity of cancer cells and subsequently destroy them. In this context, versican, a proteoglycan largely overexpressed in several solid cancers, bears the potential to be such an ideal target. As 4 main versican isoforms have been characterized, we sought to determine which isoform could represent the best target in human breast cancer. We used a series of 10 primary breast cancer lesions that were characterized as overexpressing the versican protein, when compared with matched normal breast tissues, using shotgun mass spectrometry and immunohistochemistry experiments. Quantitative polymerase chain reaction and western-blotting experiments were used to evaluate versican isoform expression in breast cancer/normal tissue pairs for which ARN quality was excellent. All known isoforms were significantly overexpressed in the malignant lesions, both at the mRNA and at the protein levels. In the course of this study, we also identified and cloned a new alternatively spliced versican isoform, referred to as V4, which was also found to be upregulated in human breast cancer. This study provides for the first time a comprehensive mRNA and protein analysis of versican isoforms expression in human breast tissues, and offers insights into which therapeutic strategy would be best suited to target versican in human breast cancer lesions.

Quantitative methods for food allergens: a review.

The quantitative detection of allergens in the food chain is a strategic health objective as the prevalence of allergy continues to rise. Food allergenicity is caused by proteins either in their native form or in forms resulting from food processing. Progress in mass spectrometry greatly opened up the field of proteomics. These advances are now available for the detection and the quantification of traces of allergenic proteins in complex mixtures, and complete the set of biological tests used until now, such as ELISA or PCR. We review methods classified according to their ability to simultaneously quantify and identify allergenic proteins and underline major advances in the mass-spectrometric methods.

Development of an absolute quantification method targeting growth hormone biomarkers using liquid chromatography coupled to isotope dilution mass spectrometry.

A method to perform absolute quantification of two biomarkers (IGF-1 and IGFBP-3) of growth hormone abuse has been developed. Isotope dilution is used with synthetically labelled peptides as internal standards. Peptide selection and multiple reaction monitoring design are discussed. A simple sample preparation based on the reduction and alkylation of cysteine residues followed by tryptic digestion provides a sufficient digestion of proteins. Serum samples fortified with increasing amounts of target proteins are analysed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) on a triple quadrupole mass spectrometer. Specificity is ensured by the selection of sequences with no homology in BLAST, as well as retention time deviation check, and ion ratio monitoring. Linearity is studied in terms of calibration curves. These curves for IGFBP-3 and IGF-1 are generated with mean slopes of 0.055 and 0.065, intercepts of 0.107 and -0.011, and with coefficients of correlation of 0.95 and 0.98, respectively. These curves result from the addition of proteins to the serum. Risks of variations related to potential matrix effects are therefore reduced, as well as probable variations related to the digestion steps. The working concentration ranges are 4-10 ng/microl for IGFBP-3 and 2-8 ng/microl for IGF-1. Preliminary data regarding repeatability show that relative standard deviations (RSDs) range between 13 and 32% for IGFBP-3 and between 7 and 29% for IGF-1.