Site-specific functionalization of proteins by organopalladium reactions

A new carbon-carbon bond has been regioselectively introduced into a target position (position 32 or 174) of the Ras protein by two types of organopalladium reactions (Mizoroki-Heck and Sonogashira reactions). Reaction conditions were screened by using a model peptide, and the stability of the Ras protein under the reaction conditions was examined by using the wild-type Ras protein. Finally, the iF-Ras proteins containing a 4-iodo-L-phenylalanine residue were subjected to organopalladium reactions with vinylated or propargylated biotin. Site-specific biotinylations of the Ras protein were confirmed by Western blot and LC-MS/MS.     

Impact psychologique de l'horaire de travail et de la participation aux taches familiales chez des couples à double carrière.

The present research examined the joint impact of characteristics of the work schedule and the participation in household work on psychological distress and parental role strain as well as stress crossover. A total of 93 dual-earner couples with at least one child living at home participated. Multiple regression analyses revealed that predictive variables of the psychological distress or the parental role strain differed for men and women and that the parental role strain was related to the psychological distress for both men and women. Results partially support the hypothesis of stress crossover since no characteristics of work schedule of the husband's and wife's were related to parental role strain of the spouses, but women's participation in the children's care was associated with men's higher parental role strain. These results underline the importance of studying strains associated with specific social roles of women and men with respect to effects of both work schedule and family strains related to psychological distress. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Mineral admixtures in mortars: Effect of inert materials on short-term hydration

This work is the first part of an overall project the aim of which is the development of general mix design rules for concrete containing different kinds of mineral admixtures. The separation of the different physical effects responsible for the modification of cement hydration, when chemically inert quartz powders are used in mortars, is presented. The phenomenological approach, based on semi-adiabatic calorimetry, is only associated with first-order phenomena, and the study excludes the complex physicochemical details involved in the chemistry of cement. The results, obtained for a wide range of fineness (between 180 and 2000 m²/kg) and replacement rates (up to 75%), show that short-term degrees of hydration in mortars containing a chemically inert mineral admixture (quartz) are always higher than for a reference mortar. This study confirms that cement hydration is enhanced by inert mineral admixtures. The two main physical effects responsible for the modification of the hydration of cement are identified as the dilution effect and heterogeneous nucleation. The dilution effect of the cement is highlighted with a coarse mineral admixture. The heterogeneous nucleation effect, although it increases with fineness of mineral admixtures, presents an optimum depending on the replacement rate. In the following part of this work, these results will be used for the development of an empirical model allowing us to quantify both physical effects.     

NMR-Guided Repositioning of Non-Steroidal Anti-Inflammatory Drugs into Tight Junction Modulators

Bioavailability is a major bottleneck in the clinical application of medium molecular weight therapeutics, including protein and peptide drugs. Paracellular transport of these molecules is hampered by intercellular tight junction (TJ) complexes. Therefore, safe chemical regulators for TJ loosening are desired. Here, we showed a potential application of select non-steroidal anti-inflammatory drugs (NSAIDs) as TJ modulators. Based on our previous observation that diclofenac and flufenamic acid directly bound various PDZ domains with a broad specificity, we applied solution nuclear magnetic resonance techniques to examine the interaction of other NSAIDs and the first PDZ domain (PDZ1) of zonula occludens (ZO)-1, ZO-1(PDZ1). Inhibition of ZO-1(PDZ1) is expected to provide loosening of the epithelial barrier function because the domain plays a crucial role in maintaining TJ integrity. Accordingly, diclofenac and indomethacin were found to decrease the subcellular localization of claudin (CLD)-2 but not occludin and ZO-1 at the apicolateral intercellular compartment of Madin–Darby canine kidney (MDCK) II cells. These NSAIDs exhibited 125–155% improved paracellular efflux of fluorescein isothiocyanate insulin for the Caco-2 cell monolayer. We propose that these NSAIDs can be repurposed as drug absorption enhancers for peptide drugs.     

Phenotypic abnormalities observed in aged cloned mice from embryonic stem cells after long-term maintenance

Somatic/embryonic stem cell cloning has made it possible to produce an individual genomically identical to another individual. However, the cloned animals have a variety of abnormalities caused by the aberrant gene modification, with insufficient reprogramming in cloning. We previously reported abnormalities in cloned mice at birth. In this study, we examined what abnormalities could be seen in cloned mice after long-term maintenance. The aged cloned mice showed multiple abnormalities: increase of body weight, some phenotypic abnormalities in the kidneys, testes and thymus, and lower urea nitrogen in their serum biochemical values. The kidneys of all cloned mice were hypertrophied, with a metamorphic or whitish appearance. The multiple lesions, including the enlarged renal pelvis and distension of the renal veins in histology, might be the result of urine accumulation by urinary tract obstruction. The testes of the cloned mice were atrophied, and showed no sperm formation in histology. In contrast, the thymus was rather hypertrophied, and a comparably increased number of lymphocytes were observed in the medulla, consisting mainly of T cells. By conducting a progeny test between the cloned mice, it was confirmed that these abnormalities in the aged cloned mice were not transmitted to their offspring, indicating that the incomplete reprogramming in clones might be in part responsible for the abnormalities detected in aged clones. These results indicate that the postnatal abnormalities observed in aged cloned mice are varied and can be restored through the germ line.     

Role of the nature of reaction products in the differing behaviours of fine glass powders and coarse glass aggregates used in concrete

This paper deals with the incorporation of glass cullet in cement-based materials. The aim is to help understand the differing behaviours observed depending on the particle size of the glass: the use of fine powders usually improves the concrete properties due to pozzolanic reaction, while coarse aggregates are generally detrimental for concrete due to alkali-silica reaction. It is shown that the lack of swelling of gels resulting from the reaction of glass fines can be partly due to the nature of these gels. The comparative investigation carried out on reaction products resulting from glass grains of various sizes, in the presence of both portlandite Ca(OH)₂ and C₃S (tricalcium silicate: 3CaO · SiO₂), has shown that glass aggregates produce detrimental gels while glass fines produce gels that help to improve concrete properties.     

Analysis of biphasic lubrication of articular cartilage loaded by cylindrical indenter

Combination of theoretical biphasic analyses and corresponding experimental measurements for articular cartilage has successfully revealed the fundamental material properties and time-depending mechanical behaviors of articular cartilage containing plenty of water. The insight of load partitioning between solid and fluid phases advanced the prediction of the frictional behavior of articular cartilage. One of the recent concerns about biphasic finite element (FE) analysis seems to be a dynamic and physiological condition in terms of mechanical functionality as a load-bearing for articular joint system beyond material testing, which has mainly focused on time-dependent reaction force and deformation in relatively small and low speed compression. Recently, the biphasic FE model for reciprocating sliding motion was applied to confirm the frictional effect on the migrating contact area. The results indicated that the model of a cylindrical indenter sliding over the cartilage surface remarkably sustained the higher proportion of fluid load support than a condition without migrating contact area, but the effectiveness of constitutive material properties has not been sufficiently evaluated for sliding motion. In our present study, at the first stage, the compressive response of the articular cartilage was examined by high precision testing machine. Material properties for the biphasic FE model, which included inhomogeneous apparent Young's modulus of solid phase along depth, strain-dependent permeability and collagen reinforcement in tensile strain, were estimated in cylindrical indentation tests by the curve fitting between the experimental time-dependent behavior and FE model simulation. Then, the biphasic lubrication mechanism of the articular cartilage including migrating contact area was simulated to elucidate functionality as a load-bearing material. The results showed that the compaction effect on permeability of solid phase was functional particularly in the condition without the migrating contact area, whereas in sliding condition the compaction effect did not clearly show its role in terms of the proportion of fluid load support. The reinforcement of solid phase, which represented the collagen network in the tissue, improved the proportion of fluid load support especially in the sliding condition. Thus, a functional integration of constitutive mechanical properties as a load-bearing was evaluated by FE model simulation in this study.     

Garlic extract and its selected organosulphur constituents promote ileal immune responses ex vivo

The effects of garlic extract and three organosulphur compounds of garlic on intestinal immune responses in mice were investigated. Peyer’s patch (PP) cells were isolated from mice orally administered with garlic extracts or one of three organosulphur compounds (alliin, allicin, diallyl disulphide (DADS)). PP cells isolated from mice that had been orally injected with ethanol extract significantly produced interferon (IFN)-γ and interleukin (IL)-4. IL-2 production in PP cells was significantly reduced by hot-water and ethanol extracts from garlic. PP cells from mice administered with two organosulphur compounds, alliin or DADS (5 mg/kg/day), could produce IL-2, IFN-γ and IL-4, whereas allicin showed moderate activity. The enhancement activity of IL-2 and IFN-γ productions in PP cells by DADS was higher than those obtained by administration of alliin or allicin. Comprehensive analyses of genetic profiles in PP tissue from mice administered with ethanolic extracts, allicin or alliin revealed that oral administration of samples increased 68–144 genes and decreased 50–52 genes by ?1.8-fold. Analyses of clustering profiles of microarrays indicated that ethanol extract and alliin upregulated the expression of IFN-γ. These data showed that garlic and its organosulphur compounds stimulate de novo IFN-γ biosynthesis in PP cells, thereby promoting ileal immune responses.