New direct assay of free protein S antigen applied to diagnosis of protein S deficiency

Congenital deficiencies of protein S (PS) are associated with thrombophilia. Their characterization and classification have been hampered by the complex physiology of the protein C-protein S system and the poor standardization and reliability of laboratory assays. The free active form of protein S is usually determined by immunoassay using polyclonal antibodies in the plasma supernate after polyethyleneglycol (PEG) precipitation. A new one step ELISA using two monoclonal antibodies specific for distinct epitopes of the free form of protein S has been developed for the direct measurement of free PS in untreated plasma. We have tested two ELISA assays for free PS. One assay was based on the PEG precipitation (Asserachrom PS, Stago, Asnières, France) whereas the other was a one step ELISA assay (Asserachrom free PS, Stago). Values were obtained in 35 PS deficient patients recruited among 500 consecutive patients evaluated by the laboratory for diagnosis of congenital disorders of coagulation. Values were compared to those obtained in 50 patients with no PS deficiency matched for age and sex with the PS deficient patients as well as in 33 normal subjects and in 12 pregnant women. Strong correlation was found between the two tests (r = 0.81, p < 10(-5)) in the entire population (n = 130), as well as in the separate groups. The new one step ELISA was more accurate than the PEG free PS determination. Determination of PS activity and antigens allowed us to separate quantitative and qualitative deficiencies. Among the qualitative deficiencies, isolated decrease in PS activity was the most frequent defect observed (66%). This fact questions the substitution of PS activity assays by the one step antigenic free PS ELISA assay.     

Incorporation and distribution of saturated and unsaturated fatty acids into nuclear lipids of hepatic cells

Liver nuclear incorporation of stearic (18∶0), linoleic (18∶2n−6), and arachidonic (20∶4n−6) acids was studied by incubation in vitro of the [1-¹⁴C] fatty acids with nuclei, with or without the cytosol fraction at different times. The [1-¹⁴C] fatty acids were incorporated into the nuclei as free fatty acids in the following order: 18∶0>20∶4n−6≫18∶2n−6, and esterified into nuclear lipids by an acyl-CoA pathway. All [1-¹⁴C] fatty acids were esterified mainly to phospholipids and triacylglycerols and in a minor proportion to diacylglycerols. Only [1-¹⁴C] 18∶2n−6-CoA was incorporated into cholesterol esters. The incorporation was not modified by cytosol addition. The incorporation of 20∶4n−6 into nuclear phosphatidylcholine (PC) pools was also studied by incubation of liver nuclei in vitro with [1-¹⁴C]20∶4n−6-CoA, and nuclear labeled PC molecular species were determined. From the 15 PC nuclear molecular species determined, five were labeled with [1-¹⁴C]20∶4n−6-CoA: 18∶0–20∶4, 16∶0–20∶4, 18∶1–20∶4, 18∶2–20∶4, and 20∶4–20∶4. The highest specific radioactivity was found in 20∶4–20∶4 PC, which is a minor species. In conclusion, liver cell nuclei possess the necessary enzymes to incorporate exogenous saturated and unsaturated fatty acids into lipids by an acyl-CoA pathway, showing specificity for each fatty acid. Liver cell nuclei also utilize exogenous 20∶4n−6-CoA to synthesize the major molecular species of PC with 20∶4n−6 at the sn-2 position. However, the most actively synthesized is 20∶4–20∶4 PC, which is a quantitatively minor component. The labeling pattern of 20∶4–20∶4 PC would indicate that this molecular species is synthesized mainly by the de novo pathway.     

A knowledge-based analysis of global function computation

Consider a distributed system N in which each agent has an input value and each communication link has a weight. Given a global function, that is, a function f whose value depends on the whole network, the goal is for every agent to eventually compute the value f (N). We call this problem global function computation. Various solutions for instances of this problem, such as Boolean function computation, leader election, (minimum) spanning tree construction, and network determination, have been proposed, each under particular assumptions about what processors know about the system and how this knowledge can be acquired. We give a necessary and sufficient condition for the problem to be solvable that generalizes a number of well-known results (Attyia et al. in J ACM 35(4):845–875, 1988; Yamashita and Kameda in IEEE Trans Parallel Distrib Syst 7(1):69–89, 1996; Yamashita and Kameda in IEEE Trans Parallel Distrib Syst 10(9):878–887, 1999). We then provide a knowledge-based (kb) program (like those of Fagin et al. (Reasoning about knowledge, MIT Press, Cambridge, 1995, Distrib Comput 10(4):199–225, 1997)) that solves global function computation whenever possible. Finally, we improve the message overhead inherent in our initial kb program by giving a counterfactual belief-based program (Halpern and Moses in Distrib Comput 17(2):91–106, 2004) that also solves the global function computation whenever possible, but where agents send messages only when they believe it is necessary to do so. The latter program is shown to be implemented by a number of well-known algorithms for solving leader election.     

High repetition rate collisional soft X-ray lasers based on grazing incidence pumping

We discuss the demonstration of gain-saturated high repetition rate table-top soft X-ray lasers producing microwatt average powers at wavelengths ranging from 13.9 to 33 nm. The results were obtained heating a precreated plasma with a picosecond optical laser pulse impinging at grazing incidence onto a precreated plasma. This pumping geometry increases the energy deposition efficiency of the pump beam into the gain region, making it possible to saturate soft X-ray lasers in this wavelength range with a short pulse pump energy of only 1 J at 800-nm wavelength. Results corresponding to 5-Hz repetition rate operation of gain-saturated 14.7-nm Ni-like Pd and 32.6-nm line Ne-like Ti lasers pumped by a table-top Ti:sapphire laser are reported. We also discuss results obtained using a 1 /spl omega/1054-nm prepulse and 2 /spl omega/527-nm short pulse from a Nd:glass pump laser. This work demonstrates the feasibility of producing compact high average power soft X-ray lasers for applications.     

VPI-FP: an integrative information system for factory planning

Nowadays, one of the main challenges in factory planning is the consistent and coherent information modelling along planning processes. Despite the current efforts in the fields of virtual production as well as digital and virtual factory, planning and simulation applications mostly support only the analysis and the optimisation of single planning aspects. However, to match nowadays challenges, planners require solutions that provide an integrated view to evaluate planning scenarios in advance and to achieve increasing production quality and efficiency. The concept of virtual production intelligence (VPI) provides a basic concept for such an integrative information system that enables planners to integrate, to aggregate and to analyse data gathered during one planning project as well as to compare different projects. In this study, we present such an information system for factory planning using the concept of the VPI. The focus lies in particular on the information modelling as well as the information integration and evaluation. Therefore, the study presents theoretical basics and implementations of the VPI platform within a precise application scenario in factory planning. This is to process and provide a consolidated information base along the whole planning process to support factory planning projects.     

A sorption rate hypothesis for the increase in H2 permeability of palladium-silver (Pd-Ag) membranes caused by air oxidation

Hydrogen permeation measurements were performed at 300 °C for 25-μm cold-rolled Pd-Ag 25 wt% membranes before and after air oxidation at the same temperature as permeation. The air oxidation resulted in enhanced H₂ permeation through the membrane, as well as a roughening of the surface with the formation of surface grains and defects. The protruding grains can be leveled off by exposure to H₂ but the surface defects cannot. These microstructure changes are only on the membrane surfaces and do not create transmembrane defects that would allow permeation for gas species other than H₂. The H₂ permeability of the oxidized membrane increased by 25-90% compared to that of the as-received film at the same permeation condition, and the membranes retained perfect H₂ selectivity over N₂. The percent improvement of H₂ permeability decreases with increasing H₂ feed pressure. A new sorption kinetics hypothesis is proposed to elucidate the increase in H₂ permeability of Pd-Ag membranes caused by oxidation. H₂ solubility and sorption rate results were presented to test the new hypothesis. It is found that air oxidation does not change the H₂ solubility in Pd-Ag membranes, but enhances the H₂ sorption kinetics significantly. The extent of kinetics enhancement also decreases with increasing H₂ pressures. The much faster sorption equilibrium implies higher effective H₂ diffusivity at the Pd-Ag membrane surface for the oxidized sample and a higher transfer rate of atomic hydrogen from surface/sub-surface to the membrane bulk that contributes to the increase of H₂ permeability observed in experiments.     

Hsp100 Molecular Chaperone ClpB and Its Role in Virulence of Bacterial Pathogens

This review focuses on the molecular chaperone ClpB that belongs to the Hsp100/Clp subfamily of the AAA+ ATPases and its biological function in selected bacterial pathogens, causing a variety of human infectious diseases, including zoonoses. It has been established that ClpB disaggregates and reactivates aggregated cellular proteins. It has been postulated that ClpB’s protein disaggregation activity supports the survival of pathogenic bacteria under host-induced stresses (e.g., high temperature and oxidative stress), which allows them to rapidly adapt to the human host and establish infection. Interestingly, ClpB may also perform other functions in pathogenic bacteria, which are required for their virulence. Since ClpB is not found in human cells, this chaperone emerges as an attractive target for novel antimicrobial therapies in combating bacterial infections.     

Prodynorphin and Proenkephalin in Cerebrospinal Fluid of Sporadic Creutzfeldt–Jakob Disease

Proenkephalin (PENK) and prodynorphin (PDYN) are endogenous opioid peptides mainly produced in the striatum and, to a lesser extent, in the cerebral cortex. Dysregulated metabolism and altered cerebrospinal fluid (CSF) levels of PENK and PDYN have been described in several neurodegenerative diseases. However, no study to date investigated these peptides in the CSF of sporadic Creutzfeldt–Jakob disease (sCJD). Using liquid chromatography-multiple reaction monitoring mass spectrometry, we evaluated the CSF PDYN- and PENK-derived peptide levels in 25 controls and 63 patients with sCJD belonging to the most prevalent molecular subtypes (MM(V)1, VV2 and MV2K). One of the PENK-derived peptides was significantly decreased in each sCJD subtype compared to the controls without a difference among subtypes. Conversely, PDYN-derived peptides were selectively decreased in the CSF of sCJD MV2K, a subtype with a more widespread overall pathology compared to the sCJD MM(V)1 and the VV2 subtypes, which we confirmed by semiquantitative analysis of cortical and striatal neuronal loss and astrocytosis. In sCJD CSF PENK and PDYN were associated with CSF biomarkers of neurodegeneration but not with clinical variables and showed a poor diagnostic performance. CSF PDYN and PENK-derived peptides had no significant diagnostic and prognostic values in sCJD; however, the distinct marker levels between molecular subtypes might help to better understand the basis of phenotypic heterogeneity determined by divergent neuronal targeting.