[Nephrolithiasis and primary hyperarathyroidism in pregnancy]. / Lithiase rénale et hyperparathyroïdie primaire au cours de la grossesse.

Diagnosis of hyperparathyroidism is unusual during pregnancy. The presence of a renal stone is a very exceptional finding. Hypercalcemia is often revealed by standard blood tests. We present here a clinical case of renal nephretic colic occurring during the third trimester of a fourth pregnancy. Our investigations led to the diagnosis of primary hyperparathyroidism which was successfully treated. Hyperparathyroidism during pregnancy is associated with a high incidence of fetal and maternal complications, essentially neonatal hypocalcemia. In utero death is rare but has been reported. Calcium metabolism during pregnancy is dependent on PTHrp. The surgical option is usually taken during the second trimester but can only be proposed during the third trimester in the event of medically resistant hypercalcemia.

Effect of HUVEC on human osteoprogenitor cell differentiation needs heterotypic gap junction communication.

Bone development and remodeling depend on complex interactions between bone-forming osteoblasts and other cells present within the bone microenvironment, particularly vascular endothelial cells that may be pivotal members of a complex interactive communication network in bone. Our aim was to investigate the interaction between human umbilical vein endothelial cells (HUVEC) and human bone marrow stromal cells (HBMSC). Cell differentiation analysis performed with different cell culture models revealed that alkaline phosphatase activity and type I collagen synthesis were increased only by the direct contact of HUVEC with HBMSC. This "juxtacrine signaling" could involve a number of different heterotypic connexions that require adhesion molecules or gap junctions. A dye coupling assay with Lucifer yellow demonstrated a functional coupling between HUVEC and HBMSC. Immunocytochemistry revealed that connexin43 (Cx43), a specific gap junction protein, is expressed not only in HBMSC but also in the endothelial cell network and that these two cell types can communicate via a gap junctional channel constituted at least by Cx43. Moreover, functional inhibition of the gap junction by 18alpha-glycyrrhetinic acid treatment or inhibition of Cx43 synthesis with oligodeoxyribonucleotide antisense decreased the effect of HUVEC cocultures on HBMSC differentiation. This stimulation could be mediated by the intercellular diffusion of signaling molecules that permeate the junctional channel.

A novel C-terminal kinesin is essential for maintaining functional acidocalcisomes in Trypanosoma brucei.

Kinesins are cytoskeletal motor proteins that play roles in a variety of fundamental cellular processes including cell division and the anterograde transport of vesicles and organelles. We purified, cloned, and functionally characterized in Trypanosoma brucei a new member of the C-terminal kinesin family, TbKIFC1. Kinetic constants of the recombinant motor domain of TbKIFC1 were estimated at 0.56 microm for the microtubule dissociation constant (K(d)) with a k(cat) of 0.2 s(-1). Immunolocalization analysis showed an association of TbKIFC1 with punctate structures. Because they were rapidly transported to the negative pole of the microtubule after NH(4)Cl treatment, these structures were considered to be associated with acidic vesicles. To determine the role of the kinesin in vivo, we produced an inducible kinesin-deficient strain by double-stranded RNA interference methodology. Mutant cells were loaded with the fluorescent reagent fura2/acetoxymethylester to measure intracellular free calcium ([Ca(2+)](i)). The resting [Ca(2+)](i) was unchanged in mutant cells; however, alkalinization of acidic vesicles induced by NH(4)Cl or nigericin was not followed by release of Ca(2+). These data and the relative importance of the ionomycin-releasable and the ionomycin-plus-NH(4)Cl-releasable Ca(2+) pools suggest a lower Ca(2+) content in acidocalcisomes and dysfunctional Ca(2+) release.

Elastin-derived protein coating onto poly(ethylene terephthalate). Technical, microstructural and biological studies.

Recently, it has been shown in our laboratory that certain proteins solubilized from elastin (ESP) formed a tight association with certain polymers such as elastin or polyethylene glycol terephthalate (PET) ... . The present paper deals with the description of the optimal chemical conditions of this unexpected association, its microstructure and its biological properties. A microstructural study of the composite ESP-PET material was performed using scanning and transmission electron microscopy. The thickness of the yield composite was evaluated (0.4-2 microm) but its imperviousness was unsatisfactory using ESP alone. So, we tentatively coated PET with the elastin-ESP complex. The microscopic views confirmed that the polymer was better filled by the organic matrix, the thickness of the layer being markedly improved (3 microm). Simultaneously, we attempted to verify whether the yielded composite retains the biological properties previously demonstrated with the 'Biopatches' and probably due to ESP. Thus, the culture of endothelial cells on an ESP-coating (with elastin or not) showed that a 100 microg/cm2 ESP concentration was able to promote endothelial cell growth in perfect conditions, maintaining their phenotypic character. While several physico-chemical determinations are in progress in our laboratory to identify and characterize the protein involved, a prototype of small-calibre vascular prosthesis was elaborated with elastin-ESP-PET composite and will be placed in a dog at the abdominal femoral junction to evaluate the in vivo performance of such an attractive material in artery restoration.

Development of RGD peptides grafted onto silica surfaces: XPS characterization and human endothelial cell interactions.

The attachment of human umbilical vein endothelial cells (HUVECs) on substrates that had been covalently grafted with the cell adhesion peptides Arg-Gly-Asp (RGD) was investigated. This approach was used to provide substrates that are adhesive to cells even in the absence of serum proteins and to cells that have had no prior treatment of the surface with proteins that promote cell adhesion. We wanted to improve control of cellular interactions with cell-adhesive materials by providing fixedly bound adhesion ligands. Silica was examined as a model surface. The peptides were grafted using three different steps: grafting of aminosilane molecules; reaction with a maleimide molecule; and immobilization of cell-binding peptides containing the RGD sequence. The RGD-grafted surface was characterized by X-ray photoelectron spectroscopy (XPS) and contact-angle measurements.

Unexpected original property of elastin derived proteins: spontaneous tight coupling with natural and synthetic polymers.

Elastin fibres can be decomposed into their constituting proteins using several processes; in particular by saponification of some bonds with KOH in aqueous tertiobutyl alcohol, elastin solubilized proteins - ESP- of 10 to 200 KDa were produced with a good yield (70-80%). It is demonstrated that some of these proteins were capable of tightly re-associating with the native elastin fibres and remained bound on the fibres, in spite of several drastic washes using 1 M Guanidinium, HCl at 37 degrees C for 1 h. At pH 4-5, approximately 30-40 microg ESP were retained per mg elastin. The same association is also shown to occur, under similar conditions, with Poly-ethylene-terephthalate, Poly-hexamethylene diamine-adipic acid but not with polyurethanes. The optimal conditions of the coupling were described as depending on ESP concentration, time, pH, ionic strength and Ca++. It was also shown that opposite pH conditions, i.e. pH 14, 0.5 M NaOH, could allow the retained proteins to desorb from polyesters. Hence, it will be possible to determine the sort of proteins which could be involved. This property of ESP allows us (1) to better understand the exceptional capacity of tissue repair certainly due to adhesive properties of the artificial connective matrices containing elastin or ESP, developed in our laboratory, (2) to provide a new approach for elucidating elastin microstructure and function, (3) especially to provide a new mode for coating certain fibres, yielding materials with bioactive and biofunctional qualities.