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Metals are ubiquitous on Earth and have become necessary for living organisms like bacteria. However, there is a subtle balance between the life required amount and toxicity resulting from an overload of metal ions. The intracellular level of these ions must be tightly regulated. A strategy often used by cells is to regulate uptake and efflux of metal ions by expressing speci c proteins involved in this traffic. In the bacterium Escherichia coli, nickel and cobalt toxicity is counterbalanced by action of the RcnA efflux pump that permits excess Ni2+ and Co2+ ions to be excluded out of cells. Its expression is under the control of the nickel and cobalt inducible RcnR metalloregulator that represses rcnA transcription in absence of metals. Recently, we identi ed a new gene that we called rcnB (formerly yohN ) involved in this system. This gene is a part of the rcnAB operon and as rcnA, is involved in nickel and cobalt homeostasis. But, unlike rcnA, a mutant strain lacking rcnB is more resistant to nickel and cobalt. This phenotype is associated to a lower accumulation of these ions in the cells. rcnB gene encodes a small monomeric protein of 10 kDa which is found in the periplasmic fraction. Until now, databases do not contain any described homologs. RcnB displayed any Ni2+ or Co2+ binding capacity. On the other hand, RcnB can bind both Cu+ and Cu2+ ions in vitro. It is equally true in vivo and a link with copper homeostasis was found based on the copper resistance systems of E. coli. This is of particular interest because copper confers particular redox properties to proteins which could be directly related to the biological function of the RcnB protein. Further studies are required to precise the role of RcnB but this work provides the identifi cation and characterization of the first member of a new family of accessory proteins tied to metal ions efflux pumps., Les métaux sont omniprésents sur la planète et sont devenus nécessaires à tous les organismes vivants au cours de l'évolution. Cependant, la quantité de métal présent dans les cellules doit être très finement régulée car une quantité trop importante de métaux peut générer une toxicité. Pour assurer cette homéostasie, les cellules doivent être capables de réguler l'import et l'efflux des métaux par l'expression dynamique de protéines spécifi ques impliquées dans ces flux ioniques. Chez la bactérie modèle Escherichia coli, la toxicité du nickel et du cobalt est neutralisée par l'action de la pompe RcnA, protéine membranaire capable d'exporter les ions Ni2+ et Co2+ hors de la cellule. La production de RcnA est contrôlée par le métallorégulateur RcnR capable de détecter ces ions en cas d'excès et d'induire, en réponse à ce signal, l'expression du gène rcnA. Récemment, notre équipe a identi fié un nouveau gène que nous avons rebaptisé rcnB (précédemment yohN ). Ce gène fait partie de l'opéron rcnAB et est, comme rcnA, impliqué dans l'homéostasie des ions Ni2+ et Co2+. A l'inverse de rcnA, une souche mutante délétée de rcnB est plus résistante au nickel et au cobalt et cela est directement corrélée à une plus faible accumulation de métal dans les cellules. Le gène rcnB code pour une petite protéine monomérique de 10 kDa localisée dans le périplasme et ne possédant pas d'homologues décrits à l'heure actuelle. Aucune fixation des ions Ni2+ et Co2+ n'a été mise en évidence pour la protéine RcnB. Elle peut en revanche interagir avec les ions Cu+ et Cu2+ aussi bien in vitro que in vivo et un lien avec les systèmes de résistance au cuivre de E. coli a été mis en évidence. Or, le cuivre confère des propriétés d'oxydo-réduction particulières aux protéines qui pourraient être directement liées à la fonction de RcnB. Des investigations complémentaires sont nécessaires pour préciser la fonction exacte de RcnB, mais ce travail a mis en évidence le premier membre d'une nouvelle famille de protéines accessoires associées aux pompes d'efflux des ions métalliques.

The purpose of this study was to measure the serum cobalt levels and their correlation with clinical and radiological findings in patients with metal on metal hip articulating surfaces.Forty-one patients with metal on metal hip arthroplasty were reviewed retrospectively at mean follow-up of 12.9 months. Serum cobalt levels were determined for each patient by atomic absorption spectrometry at the maximal follow-up and were compared to a control group (19 patients). Two patients and one control subjects also performed exercise on a treadmill in order to appreciate the influence of physical activity on serum cobalt levels.The metal on metal group presented higher serum cobalt levels than those of the control group (p0.0001). There was no correlation between serum cobalt and clinical and radiological findings at the exception of patient age (n = 40, r = 0.37). However, when the follow-up was greater than 18 months, mean serum cobalt was significantly higher compared to a follow-up less than 18 months. The physical exercise test led to a moderate elevation (around 10 p. 100) of cobalt in the two patients but not in the control subject.The interpretation of an elevated cobalt serum level is difficult. Cobalt-containing drugs, other implants, excess of activity and diseases (renal failure) may influence serum cobalt level. In this study, the high serum cobalt levels seem not linked to a failure of the implant, mainly because of the short follow-up. They could rather be attributed to an increase of the patient activity beginning 18 months after the surgery. Because potential long-term cobalt toxicity and carcinogenicity is not well known, careful medical follow-up should be emphasized specially in young patients.

The concentrations of cobalt and chromium have been measured in the urine of 22 patients who had received uncemented hip prostheses made from cobalt, chromium and molybdenum alloy, and in 21 patients who had not received prostheses, using absorption spectrophotometry. The results were statistically analysed using the Wilcoxon test. The levels of cobalt in the urine in the patients with uncemented prostheses did not show a significant increase, indicating that there is no greater risk of cobalt toxicity. The concentration of chromium was significantly raised (p less than 10(-2) in the urine of patients with uncemented prostheses, and in three of these patients significant corrosion had occurred.