Fully automated chip‐based negative mode nanoelectrospray mass spectrometry of fructooligosaccharides produced by heterologously expressed levansucrase from Pseudomonassyringae pv. tomato DC3000

Pseudomonas syringae pathovars possess multiple levansucrases with still unclear specific roles for bacteria. We have cloned and expressed three levansucrase genes, lsc1, lsc2 and lsc3, from P. syringae DC3000 in Escherichia coli. Levansucrases synthesize a high molecular weight fructan polymer, levan, from sucrose and in the case of some levansucrases, fructooligosaccharides (FOS) with potential prebiotic effects are also produced. The ability of purified Lsc3 protein of DC3000 to synthesize FOS was tested using prolonged incubation time and varied concentrations of sugar substrates. Thin‐layer chromatography (TLC) analysis of reaction products disclosed formation of FOS from both sucrose and raffinose, revealing a new catalytic property for P. syringae levansucrases. In order to analyze Lsc3‐produced FOS in underivatized form, we optimized a novel method recently introduced in carbohydrate research, based on fully automated chip‐based nanoelectrospray ionization (nanoESI) high‐capacity ion trap mass spectrometry (HCT‐MS). Uding chip‐based nanoESI MS in negative ion mode, FOS, with degrees of polymerization up to five, were detected in reaction mixtures of Lsc3 with sucrose and raffinose. For confirmation, further structural analysis by tandem mass spectrometry (MS/MS) employing collision‐induced dissociation at low energies was performed. To validate the method, commercial inulin‐derived FOS preparations Orafti®P95 and Orafti®Synergy1, which are currently used as prebiotics, were used as controls. By chip‐based nanoESI HCT‐MS, similar FOS distribution was observed in these reference mixtures. Thereby, the obtained data allowed us to postulate that FOS produced by the Lsc3 protein of P. syringae DC3000 may be prebiotic as well. Copyright © 2009 John Wiley & Sons, Ltd.     

Yang–Mills effective action at high temperature

The European Physical Journal C - Yang–Mills theory undergoes a transition from a confined to a deconfined phase in the intermediate temperature regime, where perturbation is not applicable....     

Identification of Bremia lactucae and Oidium neolycopersici proteins extracted for intact spore MALDI mass spectrometric biotyping

Several proteomic approaches were applied to identify protein markers providing typical signals during intact cell/spore (IC/IS) MALDI‐TOF MS of two plant pathogens, namely Bremia lactucae (a downy mildew) and Oidium neolycopersici (a powdery mildew). First, proteins were extracted from intact spores of the microorganisms under conditions simulating their treatment prior to the mass spectrometric analysis. After a separation by electrophoresis and tryptic digestion, 198 and 140 proteins were identified in the B. lactucae and O. neolycopersici extracts, respectively. A large portion of them were found to be involved in the process of protein biosynthesis. For the first time, some proteins were assigned to characteristic signals in MS profiles of the investigated pathogens based on an agreement in the molecular mass. There were 9 and 10 proteins recognized, respectively, which could contribute significantly to the spectral patterns. These proteins were assigned tentatively to the following peaks in the MS profiles: (i) m/z 7828; 8593; 10 456; 11 312; 12 450; 12 763; 14 756 and 16 854 for B. lactucae; (ii) m/z 7709; 8895; 9504; 9952; 11 317; 14 082 and 14 839 for O. neolycopersici. We demonstrated the presence of ribosomal proteins and histones, which could be employed as markers in biotyping analyses for pathogen identification.     

An efficient mixed integer programming model for pairing containers in inland transportation based on the assignment of orders

The inland transportation takes a significant portion of the total cost that arises from intermodal transportation. In addition, there are many parties (shipping lines, haulage companies, customers) who share this operation as well as many restrictions that increase the complexity of this problem and make it NP-hard. Therefore, it is important to create an efficient strategy to manage this process in a way to ensure all parties are satisfied. This paper investigates the pairing of containers/orders in drayage transportation from the perspective of delivering paired containers on 40-ft truck and/or individual containers on 20-ft truck, between a single port and a list of customer locations. An assignment mixed integer linear programming model is formulated, which solves the problem of how to combine orders in delivery to save the total transportation cost when orders with both single and multiple destinations exist. In opposition to the traditional models relying on the vehicle routing problem with simultaneous pickups and deliveries and time windows formulation, this model falls into the assignment problem category which is more efficient to solve on large size instances. Another merit for the proposed model is that it can be implemented on different variants of the container drayage problem: import only, import–inland and import–inland–export. Results show that in all cases the pairing of containers yields less cost compared to the individual delivery and decreases empty tours. The proposed model can be solved to optimality efficiently (within half hour) for over 300 orders.     

Asymptotic Palm likelihood theory for stationary point processes

In the present paper, we propose a Palm likelihood approach as a general estimating principle for stationary point processes in $\mathbf{R}^d$ for which the density of the second-order factorial moment measure is available in closed form or in an integral representation. Examples of such point processes include the Neyman–Scott processes and the log Gaussian Cox processes. The computations involved in determining the Palm likelihood estimator are simple. Conditions are provided under which the Palm likelihood estimator is strongly consistent and asymptotically normally distributed.