Liquid Chromatography Orbitrap Mass Spectrometry Method for Analysis of Homocysteine and Related Aminothiols in Cereal Products

Liquid chromatography Orbitrap mass spectrometry method for quantification of biological aminothiols (cysteine, homocysteine, and glutathione) in cereal products has been developed. The assay is based on preliminary derivatization with N-(2-acridonyl)maleimide and high resolution accurate mass spectrometry with utilization of dl-Homocystine-3,3,3′,3′,4,4,4′,4′-d₈ (homocystine-d₈) as internal standard. The limits of quantification for homocysteine, cysteine, and glutathione are 19.44, 40.78, and 338.93 pg, respectively, per 10 μl injection. Intra- and inter-day precision expressed as relative standard deviations are in the range of 1.76 to 2.94 % and 1.06 to 4.13 %, respectively. The average recoveries were 98 % for Hcy, 87 % for Cys, and 92 % for GSH. Wheat, maize, and bakery products with different origin were analyzed. The content of Hcy in the investigated samples was found to be in range of 9–436 μg/100 g, Cys in range of 29–6,895 μg/100 g and GSH in range of 259–14,795 μg/100 g.     

Technical note: Evaluation of a triaxial accelerometer for monitoring selected behaviors in dairy calves

The objectives of this study were (1) to develop an algorithm for the acceleration sensor of the Smartbow Eartag (Smartbow GmbH, Weibern, Austria) to distinguish between postures (lying and standing or locomotion) and to detect 6 kinds of activities (milk intake, water intake, solid feed intake, ruminating, licking or sucking without milk intake, and other activities) in dairy calves and (2) to evaluate this sensor for identifying these behaviors in dairy calves compared with observations from video. Accelerometers were applied to the left ears of 15 preweaned Holstein dairy calves. Calves were kept in a group pen and received milk replacer from an automatic calf feeder. Based on 38 h of acceleration data and video observation, an algorithm was established to detect the predefined behaviors. Using cross-validation, video recordings were used to analyze whether a behavior was detected correctly by the developed algorithm. For posture, sensitivity (94.4%), specificity (94.3%), precision (95.8%), and accuracy (94.3%) were high. Cohen's kappa was calculated as 0.88. For the 6 defined activities, overall (i.e., aggregated for all activities) accuracy was 70.8% and kappa was calculated as 0.58. Some activities (e.g., ruminating, feed intake, other activities) were identified better than others. In conclusion, the developed algorithm based on the acceleration data of the Smartbow Eartag was successful in detecting lying behavior, rumination, feed intake, and other activities in calves, but further development of the underlying algorithm will be necessary to produce reliable results for milk and water intake.     

A signature protein-based method to distinguish Mediterranean water buffalo and foreign breed milk

A novel genetic variant at the α_s1-casein locus of water buffalo (WB), 8-residue shorter than its wild-type has been found and sequenced. The internal deletion of the peptide E³⁵KVNELsT⁴² was confirmed by the isolation of the junction peptide. The 8-residue deletion mutant has a molecular weight that is 919 Da less than that of the wild-type. The novel isoform with a unique f35-42 deletion could be the result of the skipping of exon 6, generating an exon 6-deleted variant of α_s1-casein. The wild-type and its shortened α_s1-casein forms were found to co-exist in many individual milk samples. In contrast, the 8-residue, internally deleted α_s1-casein variant did not occur in water buffaloes of the Mediterranean breed reared in Italy. Wild-type α_s1-casein has 6 to 8 phosphate groups (P) while the internally deleted form 6 and 7P per molecule.     

Fate of ivermectin residues in ewes' milk and derived products

The fate of ivermectin (IVM) residues was studied throughout the processing of daily bulk milk from 30 ewes (taken up to 33 d following subcutaneous administration of 0.2 mg IVM/kg b.w.) in the following milk products: yoghurt made from raw and pasteurized milk; cheese after pressing; 30- and 60-day ripened cheese; and whey, secondary whey and whey proteins obtained after cheese-making (albumin cheese). The concentration of the H2B1a component of IVM was analysed in these dairy products using an HPLC method with fluorescence detection. The mean recovery of the method was, depending on the matrix, between 87 and 100%. Limits of detection in the order of only 0.1 microg H2B1a/kg of product were achieved. Maximum concentrations of IVM were detected mostly at 2 d after drug administration to the ewes. The highest concentration of IVM was found on day 2 in 60-day ripened cheese (96 microg H2B1a/kg cheese). Secondary whey was the matrix with the lowest concentration of IVM (<0.6 microg H2B1a/ kg). Residue levels fell below the limits of detection between day 5 (for secondary whey) and day 25 (for all cheese samples). In the matrices investigated, linear correlations between daily concentrations of IVM, milk fat and solid content were evident. During yoghurt production, fermentation and thermal stability of IVM was observed. During cheese production, approximately 35% of the IVM, present in the raw (bulk) milk samples, was lost. From the results it was concluded that the processing of ewes' milk did not eliminate the drug residues under investigation. The consequences of IVM in the human diet were discussed. Milk from treated animals should be excluded from production of fat products like cheese for longer after treatment with IVM than for lower fat products.     

The proton iron-sulfur cluster environment of the [FeFe]-hydrogenase maturation protein HydF from Thermotoga neapolitana

[FeFe]-hydrogenases contain a complex [4Fe–4S]-2Fe cluster (H-cluster) and are able to efficiently reduce protons to H₂. Due to their potential exploitation for renewable energy production biotechnologies, significant efforts have been put into understanding the mechanisms driving the H-cluster assembly, which involves three conserved proteins. Among them, HydF works as scaffold upon which the H-cluster precursor is synthesized and carrier to deliver it to the hydrogenase, resulting in its activation. A FeS cluster binding sequence (CxHx₄₆-₅₃HCxxC) is conserved in all HydF proteins and should in principle provide four ligands to coordinate the Fe atom. However, we found that alternative metal coordination may exist in different HydF proteins and that only the three cysteines are strictly required, whereas the fourth ligand may vary and is, in any case, readily exchangeable. In this work we analyzed by EPR/HYSCORE the FeS cluster proton environment of HydF from Thermotoga neapolitana to determine the possible role of surrounding residues in the non-cysteinyl iron ligation of the protein.     

Experimental study of a mechanically ventilated double-skin façade with venetian sun-shading device: A full-scale investigation in controlled environment

The aim of this article is to present results of an experimental campaign performed on a full-scale facility provided with a double-skin façade. The behaviour of this architectural concept is tested under controlled climatic conditions. A summer case is scrutinised under different configurations: variation of the airflow through the double-skin façade and different angle of the solar shading device. This paper describes the experimental conditions, as well the test facility and the tested façade element. The results show the temperatures of the test cell and the façade and how they depend on the climatic conditions and the sun-shading device blade angles. One objective of this research was to measure and provide extensive data set detailing air and surface temperatures on the double-skin façade, together with airflow rates and air velocities. The experiments are fully described so that the results can be used for the validation of numerical models dealing with ventilated double-skin façades with venetian sun-shading device.     

Kennzeichnende Teilabschnitte der Entkohlungsreaktion beim O2-Aufblasen auf Fe-C-Schmelzen

Reaktionen während des Anlaufs. Brennfleckentkohlung während des linearen Abfalls. Schematische Temperaturprofile durch den Reaktionsraum. Modellvorstellungen zu den Vorgängen im Blasstrahl. Badentkohlung während des Auslaufs.     

Application of annealing for extension of WWER vessel lives

The safe operation of nuclear power plants (NPP) is dependent upon the assurance that the reactor pressure vessel will not fail in a brittle manner when the effects of radiation embrittlement are taken into account. The recovery of the properties of the irradiated materials is an important way of extending the operating life of a reactor vessel. The intent of this paper is to demonstrate the efficiency of thermal annealing for the recovery of reactor vessel material properties and to present the implications for extended service life.In order to substantiate the application of annealing to the extensior of the service life of vessels, detailed investigations were conducted which involved thermal annealing temperature and time, fast neutron fluence, and metallurgical factors (i.e. impurity contents) on the recovery of properties after the annealing of irradiated materials. Similar studies were continued to determine predictive methods for radiation embrittlement after repeated annealings.In May 19876 the first pilot annealing of a commercial reactor vessel (Novo-Voronezhskaya, III, NPP) was performed. The development of the annealing equipment and investigations performed to test the annealing process proved successful, and an improved safe operation period for the reactor vessel was thus attained providing for an extended service life.     

Blowing against the wind—An exploratory application of actor network theory to the analysis of local controversies and participation processes in wind energy

This paper analyses the deployment of wind power and the related local controversies using actor-network theory (ANT). ANT provides conceptual instruments for a fine-tuned analysis of the contingencies that condition a project’s success or failure by focusing on the micro-decisions that intertwine the material aspects of the technology, the site where it is implemented, the participation process, and the social relations in which they are embedded. By considering controversies as alternative efforts of competing networks of actors to ‘frame’ the reality and enroll others, ANT sheds light on the complex and political nature of planning a wind farm project, insofar as it consist in aligning material and human behaviours into a predictable scenario. ‘Overflows’ occur when actors do not conform to expectations, adopt conflicting positions and develop their own interpretations of the project, thus obliging designers to adapt their frames and change their plans. To demonstrate this framework, we apply it to the case of a wind farm project in the South of France, near Albi. Our analysis suggests a new approach to examining wind power projects in terms of the interaction between globally circulating technologies, unique characteristics of the site, the participation process and the social dynamics that emerge when these are combined.