Effect of P-glycoprotein on flavopiridol sensitivity

Flavopiridol is the first potent inhibitor of cyclin-dependent kinases (CDKs) to enter clinical trials. Little is known about mechanisms of resistance to this agent. In order to determine whether P-glycoprotein (Pgp) might play a role in flavopiridol resistance, we examined flavopiridol sensitivity in a pair of Chinese hamster ovary cell lines differing with respect to level of Pgp expression. The IC 50 s of flavopiridol in parental AuxB1 (lower Pgp) and colchicine-selected CHRC5 (higher Pgp) cells were 90.2 ± 6.6 nM and 117 ± 2.3 nM, respectively (P< 0.01), suggesting that Pgp might have a modest effect on flavopiridol action. Consistent with this hypothesis, pretreatment with either quinidine or verapamil (inhibitors of Pgp-mediated transport) sensitized CHRC5 cells to the antiproliferative effects of flavopiridol. Because of concern that colony forming assays might not accurately reflect cytotoxicity, we also examined flavopiridol-treated cells by trypan blue staining and flow cytometry. These assays confirmed that flavopiridol was less toxic to cells expressing higher levels of Pgp. Further experiments revealed that flavopiridol inhibited the binding of [3H]-azidopine to Pgp in isolated membrane vesicles, but only at high concentrations. Collectively, these results identify flavopiridol as a weak substrate for Pgp. © 2001 Cancer Research Campaign www.bjcancer.co

Quels enjeux pour la gestion des eaux urbaines ? Contribution à la formulation des services attendus par le système de gestion des eaux urbaines

International audienceLe système d'assainissement dont nous héritons s'est développé en couches successives, répondant chacune à des enjeux différents, et qui présentent une valeur patrimoniale très importante. Ces systèmes doivent aujourd'hui répondre à des enjeux de plus en plus diversifiés. Les techniques mises en ½uvre deviennent également de plus en plus diversifiées. Cette multiplication des fonctions et des dispositifs concourt à accroître le nombre d'organisations concernées et actives dans la gestion du système. Se pose alors la question de la coordination d'une pluralité d'organisations, gérant une pluralité d'ouvrages dans une grande diversité d'objectifs et d'intérêts. L'enjeu principal devient donc le développement d'un système durable de gestion des eaux urbaines qui se compose de dispositifs techniques et spatiaux, d'organisations en charge de ces dispositifs, et du bassin versant naturel correspondant au domaine d'influence. Cependant, pour gérer ce système, il faut dans un premier temps identifier les enjeux liés aux eaux urbaines. La multiplication des dispositifs et organisations, et le changement d'échelle pour considérer le système (ville + bassin versant naturel) nécessite en effet de repenser les services qu'il doit rendre. Cette communication propose une réponse sous forme de " marguerite des fonctions ". Les fonctions de service liées à la gestion des eaux urbaines et le mode d'obtention de la marguerite sont détaillés

Mutations on the FG surface loop of human papillomavirus type 16 major capsid protein affect recognition by both type-specific neutralizing antibodies and cross-reactive antibodies.

The aim of this study was to further characterize the conformational neutralizing epitopes present on the surface-exposed FG loop of human papillomavirus (HPV) type 16 L1 major capsid protein. We have generated previously two chimeric L1 proteins by insertion of a foreign peptide encoding an epitope of the hepatitis B core (HBc) antigen within the FG loop. In addition, three other chimeric L1 proteins were obtained by replacing three different FG loop sequences by the HBc motif and three others by point mutations. All these chimeric L1 proteins retained the ability to self-assemble into virus-like particles (VLPs), with the exception of the mutant with substitution of the L1 sequence 274-279 by the HBc motif. The eight chimeric VLPs were then analyzed for differential reactivity with a set of six HPV-16 and HPV-31 monoclonal antibodies that bound to conformational and linear epitopes. The binding patterns of these monoclonal antibodies confirmed that the FG loop contained or contributed to neutralizing conformational epitopes. The results obtained suggested that the H31.F7 antibody, an anti-HPV-31 cross-reacting and neutralizing antibody, recognized a conformational epitope situated before the 266-271 sequence. In addition, H16.E70 neutralizing antibody reactivity was reduced with L1 VLPs with an Asn to Ala point mutation at position 270, suggesting that Asn is a part of the epitope recognized by this antibody. This study contributes to the understanding of the antigenic structure of HPV-16 and -31 L1 proteins by confirming that the FG loop contributes to neutralizing epitopes and suggesting the existence of both type-specific and cross-reactive conformational epitopes within the FG loop

Agriculture and forest: A sustainable strategy in the Brazilian Amazon

Abstract Large-scale agriculture is increasing in anthropogenically modified areas in the Amazon Basin. Crops such as soybean, maize, oil palm, and others are being introduced to supply the world demand for food and energy. However, the current challenge is to enhance the sustainability of these areas by increasing efficiency of production chains and to improve environmental services. The Amazon Basin has experienced a paradigm shift away from the traditional slash-and-burn agricultural practices, which offers decision makers the opportunity to make innovative interventions to enhance the productivity in previously degraded areas by using trees to ecological advantage. This study describes a successful experiment integrating the production of soybean and paricá (Glycine max L. and Schizolobium amazonicum) based on previous research that indicated potential topoclimatic zones for planting paricá in the Brazilian state of Pará. This paper shows that a no-tillage system reduces the effects of drought compared to conventional tillage still used by many farmers in the region. The integrated system was implemented during the 2014/2015 season in 234.6 ha in the high-potential zone in the municipality of Ulianópolis, Pará. Both soybean and paricá were planted simultaneously. Paricá was planted in 5 m x 2 m inter-tree spacing totaling 228x10 3 trees per hectare and soybean, in 4 m x 100 m spacing, distributed in nine rows with a 0.45 m inter-row distance, occupying 80% of the area. The harvested soybean production was 3.4 t ha -1 , higher than other soybean monocultures in eastern Pará. Paricá benefited from soybean fertilization in the first year: It exhibited rapid development in height (3.26 m) and average diameter (3.85 cm). Trees and crop rotation over the following years is six years for forest species and one year for each crop. Our results confirm there are alternatives to the current production systems able to diminish negative impacts resulting from monoculture. In addition, the system provided environmental services such as reduced soil erosion and increased carbon stock by soil cover with no-tillage soybean cultivation. The soybean cover contributes to increased paricá thermal regulation and lower forestry costs. We concluded that innovative interventions are important to show local farmers that it is possible to adapt an agroforest system to large-scale production, thus changing the Amazon

Combined rotation scarf and Akin osteotomies for hallux valgus: a patient focussed 9 year follow up of 50 patients

<p>Abstract</p> <p>Background</p> <p>The Cochrane review of hallux valgus surgery has disputed the scientific validity of hallux valgus research. Scoring systems and surrogate measures such as x-ray angles are commonly reported at just one year post operatively but these are of dubious relevance to the patient. In this study we extended the follow up to a minimum of 8 years and sought to address patient specific concerns with hallux valgus surgery. The long term follow up also allowed a comprehensive review of the complications associated with the combined rotation scarf and Akin osteotomies.</p> <p>Methods</p> <p>Between 1996 and 1999, 101 patients underwent rotation scarf and Akin osteotomies for the treatment of hallux valgus. All patients were contacted and asked to participate in this study. 50 female participants were available allowing review of 73 procedures. The average follow up was over 9 years and the average age at the time of surgery was 57. The participants were physically examined and interviewed.</p> <p>Results</p> <p>Post-operatively, in 86% of the participants there were no footwear restrictions. Stiffness of the first metatarsophalangeal joint was reported in 8% (6 feet); 10% were unhappy with the cosmetic appearance of their feet, 3 feet had hallux varus, and 2 feet had recurrent hallux valgus. There were no foot-related activity restrictions in 92% of the group. Metatarsalgia occurred in 4% (3 feet). 96% were better than before surgery and 88% were completely satisfied with their post-operative result. Hallux varus was the greatest single cause of dissatisfaction. The most common adverse event in the study was internal fixation irritation. Hallux valgus surgery is not without risk and these findings could be useful in the informed consent process.</p> <p>Conclusions</p> <p>When combined the rotation scarf and Akin osteotomies are an effective treatment for hallux valgus that achieves good long-term correction with a low incidence of recurrence, footwear restriction or metatarsalgia. The nature of the osteotomies allows early return to normal shoes and activity without the need for postoperative immobilisation in a plaster cast.</p

Birbeck granule-like "organized smooth endoplasmic reticulum" resulting from the expression of a cytoplasmic YFP-tagged langerin

Langerin is required for the biogenesis of Birbeck granules (BGs), the characteristic organelles of Langerhans cells. We previously used a Langerin-YFP fusion protein having a C-terminal luminal YFP tag to dynamically decipher the molecular and cellular processes which accompany the traffic of Langerin. In order to elucidate the interactions of Langerin with its trafficking effectors and their structural impact on the biogenesis of BGs, we generated a YFP-Langerin chimera with an N-terminal, cytosolic YFP tag. This latter fusion protein induced the formation of YFP-positive large puncta. Live cell imaging coupled to a fluorescence recovery after photobleaching approach showed that this coalescence of proteins in newly formed compartments was static. In contrast, the YFP-positive structures present in the pericentriolar region of cells expressing Langerin-YFP chimera, displayed fluorescent recovery characteristics compatible with active membrane exchanges. Using correlative light-electron microscopy we showed that the coalescent structures represented highly organized stacks of membranes with a pentalaminar architecture typical of BGs. Continuities between these organelles and the rough endoplasmic reticulum allowed us to identify the stacks of membranes as a form of "Organized Smooth Endoplasmic Reticulum" (OSER), with distinct molecular and physiological properties. The involvement of homotypic interactions between cytoplasmic YFP molecules was demonstrated using an A206K variant of YFP, which restored most of the Langerin traffic and BG characteristics observed in Langerhans cells. Mutation of the carbohydrate recognition domain also blocked the formation of OSER. Hence, a "double-lock" mechanism governs the behavior of YFP-Langerin, where asymmetric homodimerization of the YFP tag and homotypic interactions between the lectin domains of Langerin molecules participate in its retention and the subsequent formation of BG-like OSER. These observations confirm that BG-like structures appear wherever Langerin accumulates and confirm that membrane trafficking effectors dictate their physiology and, illustrate the importance of molecular interactions in the architecture of intracellular membranes

Barcoding T Cell Calcium Response Diversity with Methods for Automated and Accurate Analysis of Cell Signals (MAAACS)

International audienceWe introduce a series of experimental procedures enabling sensitive calcium monitoring in T cell populations by confocal video-microscopy. Tracking and post-acquisition analysis was performed using Methods for Automated and Accurate Analysis of Cell Signals (MAAACS), a fully customized program that associates a high throughput tracking algorithm, an intuitive reconnection routine and a statistical platform to provide, at a glance, the calcium barcode of a population of individual T-cells. Combined with a sensitive calcium probe, this method allowed us to unravel the heterogeneity in shape and intensity of the calcium response in T cell populations and especially in naive T cells, which display intracellular calcium oscillations upon stimulation by antigen presenting cells

Facial and dental alterations according to the breathing pattern

There is controversy in the literature about possible interaction of the respiratory mode with the facial and dental structures. OBJECTIVES: The aim of this study was to perform a longitudinal assessment of the changes in facial and dental structures in Angle's Class II, division 1 malocclusion individuals, divided according to the respiratory pattern (predominantly nasal or mouth), at two distinct moments of craniofacial development. MATERIAL AND METHODS: Pogonium and nose measurements were made on the lateral cephalometric tracings (LS'-Pog', LS'-B', B'-Pog', Pog'-PogTeg', Line NB, Pog-NB, N'-Prn, Prn-NPog, N-Prn-Sn, Prn-Sn-LS). Dental measurements were made on the plaster models (distances between the tips of the canine cusps and the tips of mesial cusps of the first molars) of 40 individuals aged 10 to 14 years (moment 1) and 13 to 16 years (moment 2), 23 being nose breathers (NB) and 17 being predominantly mouth breathers (MB). RESULTS: The Student's-t test and two-way ANOVA with repeated measures were applied to indicate differences between the mean values of these variables according to the moments and/or respiratory mode. CONCLUSIONS: There were alterations in the facial measurements, without interference of the breathing pattern. However, the breathing pattern infuenced dental alterations