PGRP-LB: An Inside View into the Mechanism of the Amidase Reaction

Peptidoglycan recognition proteins (PGRPs) are ubiquitous among animals and play pivotal functions in insect immunity. Non-catalytic PGRPs are involved in the activation of immune pathways by binding to the peptidoglycan (PGN), whereas amidase PGRPs are capable of cleaving the PGN into non-immunogenic compounds. Drosophila PGRP-LB belongs to the amidase PGRPs and downregulates the immune deficiency (IMD) pathway by cleaving meso-2,6-diaminopimelic (meso-DAP or DAP)-type PGN. While the recognition process is well analyzed for the non-catalytic PGRPs, little is known about the enzymatic mechanism for the amidase PGRPs, despite their essential function in immune homeostasis. Here, we analyzed the specific activity of different isoforms of Drosophila PGRP-LB towards various PGN substrates to understand their specificity and role in Drosophila immunity. We show that these isoforms have similar activity towards the different compounds. To analyze the mechanism of the amidase activity, we performed site directed mutagenesis and solved the X-ray structures of wild-type Drosophila PGRP-LB and its mutants, with one of these structures presenting a protein complexed with the tracheal cytotoxin (TCT), a muropeptide derived from the PGN. Only the Y78F mutation abolished the PGN cleavage while other mutations reduced the activity solely. Together, our findings suggest the dynamic role of the residue Y78 in the amidase mechanism by nucleophilic attack through a water molecule to the carbonyl group of the amide function destabilized by Zn²⁺.     

Synthesis and characterization of maltose fatty acid monoesters as biosurfactants

Maltose long-chain fatty acid esters (MFAE), esterified at the 6 and 6′ position, were synthesized with stearic, palmitic, myristic, and oleic groups. Synthesis yields were 15–20% based on initial maltose present, and structural confirmation was obtained using plasma desorption mass spectrometry and nuclear magnetic resonance spectroscopy. These surfactants have surface tensions in the range of 34–36 dyn/cm at their critical micelle concentrations (CMC) of approximately 10⁻⁵–10⁻⁶ mol/L. The increased chain lengths have a marked effect, reducing CMC values for MFAE by approximately three orders of magnitude over similar carbohydrate-based dodecyl chain sources. Within chain lengths between 14 and 18 carbons, the rate of change in CMC is significant and decreases with increasing chain length for MFAE. The melting points of MFAE are approximately 40°C, and the heat capacities range from 1.6 to 1.9 J/g·K. These numbers are comparable to those of sucrose esters, indicating their applicability in similar uses. However, because MFAE, unlike sucrose, possess an anomeric carbohydrate carbon position, these surfactants maintain their reducing nature and are susceptible to further derivatization. They are also synthesized from renewable, economical carbohydrates and lipids and may provide an excellent alternative to pertrochemical-derived products.     

Fitting polynomial surfaces to triangular meshes with Voronoi squared distance minimization

This paper introduces Voronoi squared distance minimization (VSDM), an algorithm that fits a surface to an input mesh. VSDM minimizes an objective function that corresponds to a Voronoi-based approximation of the overall squared distance function between the surface and the input mesh (SDM). This objective function is a generalization of the one minimized by centroidal Voronoi tessellation, and can be minimized by a quasi-Newton solver. VSDM naturally adapts the orientation of the mesh elements to best approximate the input, without estimating any differential quantities. Therefore, it can be applied to triangle soups or surfaces with degenerate triangles, topological noise and sharp features. Applications of fitting quad meshes and polynomial surfaces to input triangular meshes are demonstrated.     

The effect of cavity-filling mutations on the thermostability of Bacillus stearothermophilus neutral protease

Cavities in the hydrophobic core of the neutral protease ofBacillus stearothermophilus were analyzed using a threedimensionalmodel that was inferred from the crystal structure of thermolysin,the highly homologous neutral protease of B.thermoproteolyticus(85% sequence identity). Site–directed mutagenesis wasused to fill some of these cavities, thereby improving hydrophobicpacking in the protein interior. The mutations had small effectson the thermostability, even after drastic changes, such asLeu284Trp and Met168Trp. The effects on T50, the temperatureat which 50% of the enzyme is irreversibly inactivated in 30min, ranged from 0.0 to +0.4°C. These results can be explainedby assuming that the mutations have positive and negative structuraleffects of approximately the same magnitude. Alternatively,it could be envisaged that the local unfolding steps, whichrender the enzyme susceptible towards autolysis and which arerate limiting in the process of thermal inactivation, are onlyslightly affected by alterations in the hydrophobic core.     

The prediction of transport parameters in filamentous fermentation broths based on results obtained in pseudoplastic polymer solutions

Transport phenomena studies on single phase “model” fluids are of limited value in biochemical engineering if they cannot be translated to the heterogeneous systems encountered in real fermentation processes. In this paper we discuss the utility of polymer solutions as models of filamentous fermentation broths for evaluation of: pipeline friction factors and impeller power numbers (turbine and helical ribbon). To a first approximation, polymer solutions can serve as suitable models for the prediction of laminar flow pressure drop in pipelines and turbulent power consumption in stirred tanks. However, results obtained on polymer solutions do not directly apply to filamentous fermentation broths for predictions of laminar flow impeller power consumption and the transition point for turbulent flow in stirred tanks. These discrepancies are believed to result from the existence of a time dependent yield stress in filamentous fermentation broths.     

Characterization of Ceramics by X-Ray and Neutron Small-Angle Scattering

This paper reviews some recent advances in small-angle X-ray and neutron scattering methods and their application to address complex issues in ceramic systems of technological importance. It is shown how small-angle scattering (SAS) can be applied to ceramic systems in order to extract statistically representative microstructure information (e.g., void volume fraction size distributions, internal surface areas, pore morphologies) that complements the information obtained from diffraction methods, X-ray microtomography, or electron microscopy. It is demonstrated how SAS studies provide insights, not obtainable by other means, on the processing–microstructure–property relationships that frequently govern technological performance.     

Ergonomic contribution of ABLE exoskeleton in automotive industry

In PSA Peugeot Citroen factories, high precision requirements of workstations make them being manual. One of the main goal of the car manufacturer is to minimize the pain of workers while maintaining high efficiency of production lines. Consequently, assisting operators with an exoskeleton is a potential solution for improving ergonomics of painful workstations while respecting industrial constraints. To determine ergonomic performances of an exoskeleton, human joint angles and torques, ground reaction forces, and duration of operations are analysed for eight subjects performing a representative screwing task. Experiments were performed using ABLE upper-limb exoskeleton, developed by the French Atomic Energy Commission (CEA), which has the functionality to compensate arm and tools loads. Results show a clear reduction of the sum of the joints torques, up to 38.9%, given by ABLE supply and invite to make concrete the use of exoskeletons in car assembly lines.     

Tissue variation in hydrocarbon composition in the rabbit

This study, which deals with the distribution of hydrocarbons in seven types of rabbit tissues, was done for the purpose of providing information that might help shed light on the biological relevance of the hydrocarbons in mammalian metabolism. Liver, kidneys, brain, spleen, skeletal muscle, perinephric adipose, and a sample of blood serum were collected from a single animal for analysis of their hydrocarbon composition. The analytical methodology consisted of solvent extraction, saponification (adipose), elution chromatography on hydrated alumina, and combined gas chromatography-mass spectrometry. Hydrocarbons were detected in all of the tissues examined at concentrations estimated to range from 0.1 to 0.01% of the total lipid extracted. Three quite distinct distribution modes were recognized. The bulk of the identified components consisted of normal, saturated, nonterpenoid hydrocarbons in the C₁₆ to C₃₃ range. Squalene, phytene, phytadiene, and pristane were the only terpenoids detected. Nonterepenoid branched (iso andanteiso) hydrocarbons were identified unequivocally and in significant amounts in the muscle only. The adipose was the only tissue which was relatively rich in monoalkenes, and its overall hydrocarbon composition closely resembled that of the feed. The results of the study are not consistent with metabolic inertness. The observed qualitative and quantitative differences might reflect function and metabolic activities of the individual organs in a way yet to be elucidated. Presented in part at the AOCS Spring Meeting, New Orleans, April, 1976.     

Thermomechanical heat of deformation studies on cis-polybutadiene

Thermomechanical heat of torsional deformation measurements have been made on crosslinked cis-polybutadiene by means of a Calvet microcalorimeter operated at 30°C. When corrected for volume changes utilizing the Gaussian statistical theory of elasticity, the data gave a value for the relative energy contribution to the torsional couple, $\text{M}{}_{\mathrm{e}}\text{M}$, of 0.14 ± 0.02. Measurements were also made on a sample subjected to simple tensile deformations. The relative energy contribution to the tensile force ($\text{f}{}_{\mathrm{e}}\text{f}$) was found to agree within experimental error with the value obtained for $\text{M}{}_{\mathrm{e}}\text{M}$, and the two results gave an average value for din $\text{〈r}{}^2{}_0\text{〉}\text{d}\text{T}$ of 4.1 × 10^?4 K^?1.     

Structures and morphologies of cast and plastically strained polyamide 6 films as evidenced by confocal Raman microspectroscopy and atomic force microscopy

Both confocal Raman microspectroscopy and atomic force microscopy (AFM) have been undertaken to study the crystalline and the morphological aspects of cast PA 6 films at a sub-microscopic scale. The percentages of the different crystalline structures present within PA 6 cast films, i.e. the monoclinic α, the pseudo-hexagonal β, and the monoclinic γ, have been measured by confocal Raman microspectroscopy. In cast films, the prevailing structure is the β one. AFM has been used to characterize the morphology of the PA 6 films. Simultaneously, the deformed state has been considered as well. Our main interest has been to follow the evolution of the percentage of each crystalline structure as a function of the plastic deformation mechanisms which are responsible of the yielding of PA 6 films: shear banding for temperatures T lower than 160 °C and formation of fibrils for