Expression patterns of Notch receptors and their ligands in human osteoarthritic and healthy articular cartilage

Notch pathway plays a pivotal role in cell fate determination. There is much interest surrounding its therapeutic potential, in osteoarthritis, but the expression profile of Notch-related molecules, as well as their relation with cartilage pathological parameters, remains unclear. The purpose of our study is to analyze the expression pattern of Notch family members, type II and type I collagen, in normal (healthy) and osteoarthritic human knee cartilage. Osteoarthritic cartilages were obtained from 3 patients undergoing a total knee replacement. Macroscopically normal cartilage was dissected from 3 human knees at the time of autopsy or surgery. Immunohistochemical staining was performed using Notch1,2,3 and 4, Delta, Jagged, type II collagen and type I collagen antibodies. In healthy cartilage, type II collagen was abundantly expressed while type I was absent. This latter increased proportionally to the osteoarthritic grade. Type II collagen expression remained intense in osteoarthritic cartilage. In healthy cartilage as well as in cartilage with minor lesions, Notch family member's proteins were not or just weakly expressed at the surface and in the cells. However, Notch molecules were over-expressed in osteoarthritic cartilage compared to healthy one. This expression pattern was different according to the cartilage zone and the severity of OA. Our data suggest that Notch signaling is activated in osteoarthritic cartilage, compared to healthy cartilage, with a much more abundant expression in the most damaged areas.     

First identification of small-molecule inhibitors of Pontin by combining virtual screening and enzymatic assay

The human protein Pontin, which belongs to the AAA+ (ATPases associated with various cellular activities) family, is overexpressed in several cancers and its silencing in vitro leads to tumour cell growth arrest and apoptosis, making it a good target for cancer therapy. In particular, high levels of expression were found in hepatic tumours for which the therapeutic arsenal is rather limited. The three-dimensional structure of Pontin has been resolved previously, revealing a hexameric assembly with one ADP molecule co-crystallized in each subunit. Using Vina, DrugScore and Xscore, structure-based virtual screening of 2200 commercial molecules was conducted into the ATP-binding site formed by a dimer of Pontin in order to prioritize the best candidates. Complementary to the in silico screening, a versatile and sensitive colorimetric assay was set up to measure the disruption of the ATPase activity of Pontin. This assay allowed the determination of inhibition curves for more than 20 top-scoring compounds, resulting in the identification of four ligands presenting an inhibition constant in the micromolar concentration range. Three of them inhibited tumour cell proliferation. The association of virtual screening and experimental assay thus proved successful for the discovery of the first small-molecule inhibitors of Pontin.     

Alkaline xylanases from Bacillus mojavensis A21: Production and generation of xylooligosaccharides

Medium composition and culture conditions for the xylanases production by Bacillus mojavensis A21 were optimized using two statistical methods: Plackett-Burman design applied to find the key ingredients and conditions for the best yield of enzyme production and Box-Behnken design used to optimize the value of the four significant variables: barley bran, NaCl, agitation, and cultivation time. The optimal conditions for higher production of xylanases were barley bran 18.66g/l, NaCl 1.04g/l, speed of agitation 176rpm and cultivation time 34.08h. Under these conditions, the xylanase experimental yield (7.45U/ml) closely matched the yield predicted by the statistical model (7.23U/ml) with R(2)=0.98. The medium optimization resulted in a 6.83-fold increase in xylanase production compared to that of the initial medium. Best xylanase activity was observed at the temperature of 50°C and at pH 8.0. The enzyme retained more 96% of its activity after 24h at pH ranges from 7.0 to 90.0. The enzyme preserved more 80% of its initial activity after 60min of pre-incubation from 30°C to 60°C. The main hydrolysis products yielded from corncob extracted xylan were xylobiose and xylotriose, suggesting the good potential of strain A21 in xylooligosaccharides production.     

Large gallanes and the PSEPT theory: a theoretical study of Ga n H n+2 clusters (n?=?7–9)

Do alanes Al(n)H(n+2) and gallanes Ga(n)H(n+2) satisfy the polyhedral skeletal electron pair theory (PSEPT)? Taking into account previous work on this question, this paper provides a convincing answer and proposes the reformulation of the (n + 1) electron pairs rule of Wade and Mingos (W-M) for such systems. Following recent studies of tetra-, penta-, hexa-, hepta-, octa-, and nonaalanes as well as valence-isoelectronic/related gallanes, in this paper we present an analysis of the hydrides of aluminum and gallium A(n)H(n+2) (A = Al, Ga and n = 7-9). The aim is still to examine the applicability of PSEPT, especially the W-M rule, to these clusters. Exploration of the total potential energy surfaces (PESs) of hepta-, octa-, and nonagallanes shows that the absolute minima have a nido-like polyhedron arrangement. Unlike the smaller Ga(n)H(n+2) (n = 4, 5, 6), it seems that the size of the cluster largely dictates its preferred geometry. Although none of them have closed (totally triangular) cages, these clusters exhibit significant compactness, comparable to borane double anions, B(n)H(n) (2-), which are the archetypes for the PSEPT theory.     

Muscle activation of the elbow flexor and extensor muscles during self-resistance exercises: comparison of unilateral maximal cocontraction and bilateral self-resistance

Muscle activation during self-resistance exercises was studied in 18 subjects performing (a) maximal unilateral isometric cocontractions of flexor and extensor muscles of the right elbow (UNI); (b) bilateral exercises consisting of maximal isometric extensions of the right elbow against the left elbow flexors (BiExt) and maximal isometric flexion of the right elbow against the left elbow extensors (BiFlex). Force production by the biceps brachii (BB), brachioradialis (BR), and triceps brachii (TB) during UNI, BiFlex, and BiExt were estimated by comparing the integrated surface electromyograms (iEMG) of BB, BR, and TB during UNI, BiExt, and BiFlex with the individual iEMG-force relationship determined from isometric contractions at 30, 60, and 100% maximal voluntary contraction during elbow flexion (MVCflex) or extension (MVCext) against a force transducer. During BiFlex for BB or BR and BiExt for TB, the values (mean ± SE) of BB-iEMG, BR-iEMG, and TB-iEMG were 74.0 ± 4.5, 76.6 ± 5.7, and 84.4 ± 4.5% iEMG at MVC (% iEMGmax). The forces were 86.0 ± 3.7% TB-Forcemax during BiExt, 74.1 ± 3.6% BB-Forcemax and 71.8 ± 4.0% BR-Forcemax during BiFlex. During UNI, BB-iEMG, BR-iEMG, and TB-iEMG were 59.9 ± 4.6, 53.4 ± 4.0, and 66.3 ± 4.7% iEMGmax, respectively. The forces during UNI (70.4 ± 4.0% TB-Forcemax, 60.4 ± 4.3% BB-Forcemax, and 49.2 ± 3.1% BR-Forcemax) were significantly lower than those during bilateral exercises. A 2-way analysis of variance (Muscle × Exercise) indicated that the effects of Muscle and Exercise upon % iEMGmax were significant (p < 0.05; p < 0.001, respectively). In conclusion, bilateral opposition exercises should be more effective in developing strength than cocontraction exercises, which correspond to a moderate activation level even for weak agonist muscle groups.     

Contact rearrangements form coupled networks from local motions in allosteric proteins

Allosteric proteins bind an effector molecule at one site resulting in a functional change at a second site. We hypothesize that networks of contacts altered, formed, or broken are a significant contributor to allosteric communication in proteins. In this work, we identify which interactions change significantly between the residue-residue contact networks of two allosteric structures, and then organize these changes into graphs. We perform the analysis on 15 pairs of allosteric structures with effector and substrate each present in at least one of the two structures. Most proteins exhibit large, dense regions of contact rearrangement, and the graphs form connected paths between allosteric effector and substrate sites in five of these proteins. In the remaining 10 proteins, large-scale conformational changes such as rigid-body motions are likely required in addition to contact rearrangement networks to account for substrate-effector communication. On average, clusters which contain at least one substrate or effector molecule comprise 20% of the protein. These allosteric graphs are small worlds; that is, they typically have mean shortest path lengths comparable to those of corresponding random graphs and average clustering coefficients enhanced relative to those of random graphs. The networks capture 60-80% of known allostery-perturbing mutants in three proteins, and the metrics degree and closeness are statistically good discriminators of mutant residues from nonmutant residues within the networks in two of these three proteins. For two proteins, coevolving clusters of residues which have been hypothesized to be allosterically important differ from the regions with the most contact rearrangement. Residues and contacts which modulate normal mode fluctuations also often participate in the contact rearrangement networks. In summary, residue-residue contact rearrangement networks provide useful representations of the portions of allosteric pathways resulting from coupled local motions.     

Direct and Indirect Relationships Between Specific Leaf Area, Leaf Nitrogen and Leaf Gas Exchange. Effects of Irradiance and Nutrient Supply

We present a series of competing path models relating interspecificpatterns between specific leaf area, leaf nitrogen content,net photosynthesis and stomatal conductance and test these againstdata from 22 species of herbaceous plants grown under controlledconditions with contrasting irradiance and nutrient supply rates.We then compare these results with two previous data sets, onebased on field measures and one based on glasshouse measures,to determine the robustness of the results. Only one model wasable to account for the patterns of direct and indirect effectsbetween the four variables to all data sets. In this model specificleaf area is the forcing variable that directly affects bothleaf nitrogen levels and net photosynthetic rates. Leaf nitrogenthen directly affects net photosynthetic rates which in turnthen affect stomatal conductance to water. Copyright 2001 Annalsof Botany Company Comparative ecology, modelling, path analysis, photosynthesis, plant strategies, SLA, specific leaf area, stomatal conductance     

Involvement of Na(+)/Ca(2+) exchanger in endothelial NO production and endothelium-dependent relaxation

Endothelial nitric oxide (NO) synthase (eNOS) is controlled by Ca(2+)/calmodulin and caveolin-1 in caveolae. It has been recently suggested that Na(+)/Ca(2+) exchanger (NCX), also expressed in endothelial caveolae, is involved in eNOS activation. To investigate the role played by NCX in NO synthesis, we assessed the effects of Na(+) loading (induced by monensin) on rat aortic rings and cultured porcine aortic endothelial cells. Effect of monensin was evaluated by endothelium-dependent relaxation of rat aortic rings in response to acetylcholine and by real-time measurement of NO release from cultured endothelial cells stimulated by A-23187 and bradykinin. Na(+) loading shifted the acetylcholine concentration-response curve to the left. These effects were prevented by pretreatment with the NCX inhibitors benzamil and KB-R7943. Monensin potentiated Ca(2+)-dependent NO release in cultured cells, whereas benzamil and KB-R7943 totally blocked Na(+) loading-induced NO release. These findings confirm the key role of NCX in reverse mode on Ca(2+)-dependent NO production and endothelium-dependent relaxation.     

Listening to Music during Warming-Up Counteracts the Negative Effects of Ramadan Observance on Short-Term Maximal Performance

Aim

The aim of the present study was to examine whether listening to music during warming-up might influence short-term maximal performance (STMP), cognitive anxiety, self-confidence, and enjoyment during Ramadan, and whether these affects might predict STMP.

Methods

Nine male physical education students (age: 21 ± 1.1 years; height: 1.8 ± 0.04 m; body mass: 83 ± 5 kg) volunteered to participate in the present study. A within-subjects design consisted of four experimental sessions: Two sessions occurred one week before Ramadan and two others took place during Ramadan. They were scheduled at 5 p.m. and were conducted as follows: After a 10-minute warm-up either with or without listening to music, each participant performed a 5-m multiple shuttle run test, after which he was asked to answer items intended to assess his affective state during the experimental task.

Results

Our findings revealed that STMP was lower during Ramadan than before Ramadan in the no-music condition. Additionally, it was found that STMP was higher in the music condition than in the no-music condition during Ramadan, and that STMP measured before Ramadan did not differ from that measured during Ramadan in the music condition. Regarding affects, the findings revealed that enjoyment was lower during Ramadan than before Ramadan in the music condition, and that cognitive anxiety was lower in the music condition than in the no-music condition before Ramadan. Self-confidence was not influenced by the experimental conditions.

Conclusion

This study showed that listening to music during warming-up not only would be beneficial for STMP in Ramadan fasters, but also would counteract the negative effects of Ramadan observance on STMP.     

Dendritic Cells Cause Bone Lesions in a New Mouse Model of Histiocytosis

Langerhans cell histiocytosis (LCH) is a rare disease caused by the clonal accumulation of dendritic Langerhans cells, which is often accompanied by osteolytic lesions. It has been reported that osteoclast-like cells play a major role in the pathogenic bone destruction seen in patients with LCH and these cells are postulated to originate from the fusion of DCs. However, due to the lack of reliable animal models the pathogenesis of LCH is still poorly understood. In this study, we have established a mouse model of histiocytosis- recapitulating human disease for osteolytic lesions seen in LCH patients. At 12 weeks after birth, severe bone lesions were observed in our multisystem histiocytosis (Mushi) model, when CD8α conventional dendritic cells (DCs) are transformed (MuTuDC) and accumulate. Most importantly, our study demonstrates that bone loss in LCH can be accounted for the transdifferentiation of MuTuDCs into functional osteoclasts both in vivo and in vitro. Moreover, we have shown that injected MuTuDCs reverse the osteopetrotic phenotype of oc/oc mice in vivo. In conclusion, our results support a crucial role of DCs in bone lesions in histiocytosis patients. Furthermore, our new model of LCH based on adoptive transfer of MuTuDC lines, leading to bone lesions within 1–2 weeks, will be an important tool for investigating the pathophysiology of this disease and ultimately for evaluating the potential of anti-resorptive drugs for the treatment of bone lesions.