Lean Product Development Performance Measurement Tool

The need of applying lean thinking to product development is becoming a must for the organisations to success in the current industry. This paper presents a tool that helps to define the actual status of the organisations in relation to the lean principles. Extensive literature highlighted the need of developing a tool focused on assessing the implementation of lean principles themselves, rather than quantitate metrics. Based on the Balanced Scorecard, four perspectives, with corresponding set of questions, were defined reflecting the enablers of the product development model proposed by the LeanPPD European project. A five-level scale was customised to score the different readiness levels that define the transformation into a full lean implementation. The tool was used to assess the current and desired lean situation of an aerospace company within the research environment and resulted to be accurate to define the starting condition of the company to adopt leaner practices

Lean Product Development Performance Measurement Tool

The need of applying lean thinking to product development is becoming a must for the organisations to success in the current industry. This paper presents a tool that helps to define the actual status of the organisations in relation to the lean principles. Extensive literature highlighted the need of developing a tool focused on assessing the implementation of lean principles themselves, rather than quantitate metrics. Based on the Balanced Scorecard, four perspectives, with corresponding set of questions, were defined reflecting the enablers of the product development model proposed by the LeanPPD European project. A five-level scale was customised to score the different readiness levels that define the transformation into a full lean implementation. The tool was used to assess the current and desired lean situation of an aerospace company within the research environment and resulted to be accurate to define the starting condition of the company to adopt leaner practices

In vitro effects of the endocrine disruptor p,p' DDT on human choriogonadotropin/luteinizing hormone receptor signalling

International audienceDichlorodiphenyltrichloroethane (p,p ' DDT) is an endocrine-disrupting chemical (EDC). Several studies showed an association between p,p ' DDT exposure and reprotoxic effects. We showed that p,p ' DDT was a positive allosteric modulator of human follitropin receptor (FSHR). In contrast, we demonstrated that p,p ' DDT decreased the cyclic AMP (cAMP) production induced by human choriogonadotropin (hCG). This study evaluated further the effects of p,p ' DDT on Gs-, beta-arrestin 2- and steroidogenesis pathways induced by hCG or luteinizing hormone (LH). We used Chinese hamster ovary cells line stably expressing hCG/LHR. The effects of 10-100 mu M p,p ' DDT on cAMP production and on beta-arrestin 2 recruitment were measured using bioluminescence and time-resolved resonance energy transfer technology. The impact of 100 mu M of p,p ' DDT on steroid secretion was analysed in murine Leydig tumor cell line (mLTC-1). In cAMP assays, 100 mu M p,p ' DDT increased the EC50 by more than 300% and reduced the maximum response of the hCG/LHR to hCG and hLH by 30%. This inhibitory effect was also found in human granulosa cells line and in mLTC-1 cells. Likewise, 100 mu M p,p ' DDT decreased the hCG- and hLH-promoted beta-arrestin 2 recruitment down to 14.2 and 26.6%, respectively. Moreover, 100 mu M p,p ' DDT decreased by 30 and 47% the progesterone secretion induced by hCG or hLH, respectively, without affecting testosterone secretion. This negative effect of p,p'DDT was independent of cytotoxicity. p,p ' DDT acted as a negative allosteric modulator of the hCG/LHR signalling. This emphasizes the importance of analyzing all receptor-downstream pathways to fully understand the deleterious effects of EDC on human health

Human amniotic fluid-based exposure levels of phthalates and bisphenol A mixture reduce INSL3/RXFP2 signaling

International audienc

Molecular Insights into the Transmembrane Domain of the Thyrotropin Receptor.

The thyrotropin receptor (TSHR) is a G protein-coupled receptor (GPCR) that is member of the leucine-rich repeat subfamily (LGR). In the absence of crystal structure, the success of rational design of ligands targeting the receptor internal cavity depends on the quality of the TSHR models built. In this subfamily, transmembrane helices (TM) 2 and 5 are characterized by the absence of proline compared to most receptors, raising the question of the structural conformation of these helices. To gain insight into the structural properties of these helices, we carried out bioinformatics and experimental studies. Evolutionary analysis of the LGR family revealed a deletion in TM5 but provided no information on TM2. Wild type residues at positions 2.58, 2.59 or 2.60 in TM2 and/or at position 5.50 in TM5 were substituted to proline. Depending on the position of the proline substitution, different effects were observed on membrane expression, glycosylation, constitutive cAMP activity and responses to thyrotropin. Only proline substitution at position 2.59 maintained complex glycosylation and high membrane expression, supporting occurrence of a bulged TM2. The TSHR transmembrane domain was modeled by homology with the orexin 2 receptor, using a protocol that forced the deletion of one residue in the TM5 bulge of the template. The stability of the model was assessed by molecular dynamics simulations. TM5 straightened during the equilibration phase and was stable for the remainder of the simulations. Our data support a structural model of the TSHR transmembrane domain with a bulged TM2 and a straight TM5 that is specific of glycoprotein hormone receptors

Dynamic modelling of the TSHR transmembrane domain.

<p><b>(a)</b> Cartoon representation of TSHR transmembrane domain, obtained by homology modelling with the OX2 template, before and after simulations. The initial model is green. The snapshots obtained after 10, 20, 40 and 60 ns MD simulations are white, slate, grey and purple, respectively. The ovals indicate the positions of the distortions in TM2 and TM5. (b) TM2 (left) and TM5 (right) of the OX2 template (magenta) superimposed with the initial TSHR model (green) and with the snapshots obtained after 10 and 40 ns MD simulations (white and grey, respectively). (c) Close-up view of the relative orientation of the E3.37 and N5.47 side chains in the snapshots obtained after 10, 20, 40 and 60 ns MD simulations. (d) Sequence alignment between OX2 and TSHR used for homology modelling. The / symbol in the OX2 sequence indicates interruption of the sequence due to either missing residues in the crystal structure or residues not used for structural constraints in the modelling procedure. Helices are indicated by grey shading. The residues modeled with helical constraints in ECL1 are in italic. The positions of the anchors are indicated by stars. The position of the deletion in TM5 of OX2 is red. The positions of the proline substitutions in TSHR are red and underlined.</p

Structure of selected GPCRs.

<p>Ribbon representation of the crystal structure of (a) S1P1R (PDB # 3V2Y) and (b) OX2 (PDB # 4S0V). TM2 and TM5 are green. Positions 2.59 and 5.50 are blue. The proline residues at positions 2.59 and 5.50 in OX2 are shown as sticks. Ovals indicate the positions of the helical distortions, if any.</p

Analysis of the MD simulations of the TSHR transmembrane domain.

<p>(a) Time evolution of the root mean squared deviations (RMSD) of the Cα atoms of the TSHR transmembrane domain (grey) or transmembrane helices only (blue). The insert shows the evolution of the distance between the Cδ atom of E3.37 and the Nδ1 atom of N4.47; (b) Time evolution of the bend angle of TM2 (grey) and TM5 (blue); (c) Backbone <i>i</i> to <i>i</i>-4 (closed symbols) and <i>i</i> to <i>i</i>-5 (open symbols) H-bonds in TM2. The percent of H-bonds was measured during the equilibration phase (triangles) and the production run (circles); (d) Backbone <i>i</i> to <i>i</i>-4 H-bonds in TM5 during the equilibration phase (triangles), the 35 first ns (squares) and the 25 last ns (circles) of the production run.</p

Glycosylation status of the TSHR mutants.

<p>Typical western blots of WT and mutated TSHRs, representative of three independent experiments. The identity of the bands was verified by treatment with endoH and PNGase. The band at 120 kDa corresponds to complex glycosylated receptors that are endoH resistant and PNGaseF-sensitive. The band at 95 kDa corresponds to high mannose-glycosylated receptors that are endoH and PNGaseF-sensitive.</p