Uridine Metabolism in HIV-1-Infected Patients: Effect of Infection, of Antiretroviral Therapy and of HIV-1/ART-Associated Lipodystrophy Syndrome

Background Uridine has been advocated for the treatment of HIV-1/HAART-associated lipodystrophy (HALS), although its metabolism in HIV-1-infected patients is poorly understood. Methods Plasma uridine concentrations were measured in 35 controls and 221 HIV-1-infected patients and fat uridine in 15 controls and 19 patients. The diagnosis of HALS was performed following the criteria of the Lipodystrophy Severity Grading Scale. Uridine was measured by a binary gradient-elution HPLC method. Analysis of genes encoding uridine metabolizing enzymes in fat was performed with TaqMan RT-PCR. Results Median plasma uridine concentrations for HIV-1-infected patients were 3.80 µmol/l (interquartile range: 1.60), and for controls 4.60 µmol/l (IQR: 1.8) (P = 0.0009). In fat, they were of 6.0 (3.67), and 2.8 (4.65) nmol/mg of protein, respectively (P = 0.0118). Patients with a mixed HALS form had a median plasma uridine level of 4.0 (IC95%: 3.40-4.80) whereas in those with isolated lipoatrophy it was 3.25 (2.55-4.15) µmol/l/l (P = 0.0066). The expression of uridine cytidine kinase and uridine phosphorylase genes was significantly decreased in all groups of patients with respect to controls. A higher expression of the mRNAs for concentrative nucleoside transporters was found in HIV-1-infected patients with respect to healthy controls. Conclusions HIV-1 infection is associated with a decrease in plasma uridine and a shift of uridine to the adipose tissue compartment. Antiretroviral therapy was not associated with plasma uridine concentrations, but pure lipoatrophic HALS was associated with significantly lower plasma uridine concentrations

Staged induction of HIV-1 glycan–dependent broadly neutralizing antibodies

A preventive HIV-1 vaccine should induce HIV-1–specific broadly neutralizing antibodies (bnAbs). However, bnAbs generally require high levels of somatic hypermutation (SHM) to acquire breadth, and current vaccine strategies have not been successful in inducing bnAbs. Because bnAbs directed against a glycosylated site adjacent to the third variable loop (V3) of the HIV-1 envelope protein require limited SHM, the V3-glycan epitope is an attractive vaccine target. By studying the cooperation among multiple V3-glycan B cell lineages and their coevolution with autologous virus throughout 5 years of infection, we identify key events in the ontogeny of a V3-glycan bnAb. Two autologous neutralizing antibody lineages selected for virus escape mutations and consequently allowed initiation and affinity maturation of a V3-glycan bnAb lineage. The nucleotide substitution required to initiate the bnAb lineage occurred at a low-probability site for activation-induced cytidine deaminase activity. Cooperation of B cell lineages and an improbable mutation critical for bnAb activity defined the necessary events leading to breadth in this V3-glycan bnAb lineage. These findings may, in part, explain why initiation of V3-glycan bnAbs is rare, and suggest an immunization strategy for inducing similar V3-glycan bnAbs

The role of resveratrol on skeletal muscle cell differentiation and myotube hypertrophy during glucose restriction

Glucose restriction (GR) impairs muscle cell differentiation and evokes myotube atrophy. Resveratrol treatment in skeletal muscle cells improves inflammatory-induced reductions in skeletal muscle cell differentiation. We therefore hypothesised that resveratrol treatment would improve muscle cell differentiation and myotube hypertrophy in differentiating C2C12 myoblasts and mature myotubes during GR. Glucose restriction at 0.6 g/L (3.3 mM) blocked differentiation and myotube hypertrophy versus high-glucose (4.5 g/L or 25 mM) differentiation media (DM) conditions universally used for myoblast culture. Resveratrol (10 μM) treatment increased SIRT1 phosphorylation in DM conditions, yet did not improve differentiation when administered to differentiating myoblasts in GR conditions. Resveratrol did evoke increases in hypertrophy of mature myotubes under DM conditions with corresponding elevated Igf-I and Myhc7 gene expression, coding for the ‘slow’ type I MYHC protein isoform. Inhibition of SIRT1 via EX-527 administration (100 nM) also reduced myotube diameter and area in DM conditions and resulted in lower gene expression of Myhc 1, 2 and 4 coding for ‘intermediate’ and ‘faster’ IIx, IIa and IIb protein isoforms, respectively. Resveratrol treatment did not appear to modulate phosphorylation of energy-sensing protein AMPK or protein translation initiator P70S6K. Importantly, in mature myotubes, resveratrol treatment was able to ameliorate reduced myotube growth in GR conditions over an acute 24-h period, but not over 48–72 h. Overall, resveratrol evoked myotube hypertrophy in DM conditions while favouring ‘slower’ Myhc gene expression and acutely ameliorated impaired myotube growth observed during glucose restriction

Modeling of Damage in Composite Materials Submitted to Off Axis Tests: Application to a Ceramic Woven Fabric SiC-SiC Composite

Woven fabric composites show a damage behavior which can be compared to that of cross-ply laminates concerning transverse cracking. The particularity of woven fabric composites is to have two networks of cracks oriented by the weft and the warp threads. The originality of this paper is to propose a vectorial approach with two damage variables instead of a tensorial one to model the behavior of woven fabric composites. A meso-macro approach is applied. A comparison of numerical simulations and off axis tests leads to an assessment of the accuracy of the proposed modeling methodology

Organ preservation at low temperature: a physical and biological problem

Before reporting the preliminary results obtained by our group, we first review the main problems to be solved in the preservation of organs at very low temperature, before being transplanted. This cryopreservation is being presently explored in order to increase the preservation tiine of transplants and to contribute to a better control of the donor recipient compatibility. We recall that, for the isolated cells to be preserved at nitrogen liquid temperatures, as now successfully performed at industrial scale, it is necessary to immerse the cells in a solution containing more or less t,oxical additives (so-called cryopro tect ants). Furthermore cooling and warming rates must be specific of each type of cells. We then show that cryo preservation could be extrapolated to whole organs by means of vitrification, the only way to avoid any ice crystallization. This vitrification will be the result of two directions of research, the one on the elaboration of cryoprotective solutions, the least toxic possible, the other on the obtention of high enough and homogeneous cooling and warming rates. After having briefly summarized the state of research on the heart and kidneys of small mammals, we present the first results that we have obtained on perfusion at 4 $^{\circ}$C and the auto-transplantation of rabbit kidneys, on the toxicity of a new cryoprotectant, 2,3-butanediol, on the heart rate, and on the cooling of experimental models of organs.Avant de présenter les résultats préliminaires obtenus par notre groupe, nous passons d'abord en revue les principaux problèmes à résoudre pour conserver à très basse température des organes en vue de leur transplantation. Cette cryopréservation est une voie de recherche actuellement explorée pour augmenter la durée de conservation des greffons et permettre ainsi de mieux contrôler la compatibilité donneur-receveur. Nous rappelons que la conservation des cellules isolées à la température de l'azote liquide, actuellement réalisée avec succès à l'échelle industrielle, ne peut se faire qu'en présence de substances plus ou moins toxiques dites cryoprotectrices, et à condition de respecter des vitesses de refroidissement et de réchauffement adaptées à chaque type de cellule. Nous montrons ensuite que l'extension de la cryopréservation au cas des organes entiers ne pourra se faire qu'au moyen de la vitrification, seule solution pour éviter toute formation de glace. Cette vitrification sera l'aboutissement de 2 axes de recherche, l'un sur l'élaboration de solutions cryoprotectrices les moins toxiques possibles, l'autre sur l'obtention de vitesses de refroidissement et de réchauffement suffisamment élevées et homogènes. Après avoir brièvement résumé l'état des recherches sur le coeur et le rein de petits mammifères, nous présentons les premiers résultats que nous avons obtenus sur la perfusion à 4 $^{\circ}$C et l'autotransplantation de reins de lapin, sur la toxicité sur le coeur de rat d'un nouveau cryoprotecteur le 2,3-butanediol, et sur le refroidissement de systèmes modèles expérimentaux d'organes