Validation of the performance of a GMO multiplex screening assay based on microarray detection

A new screening method for the detection and identification of GMO, based on the use of multiplex PCR followed by microarray, has been developed and is presented. The technology is based on the identification of quite ubiquitous GMO genetic target elements first amplified by PCR, followed by direct hybridisation of the amplicons on a predefined microarray (DualChip^® GMO, Eppendorf, Germany). The validation was performed within the framework of a European project (Co-Extra, contract no 007158) and in collaboration with 12 laboratories specialised in GMO detection. The present study reports the strategy and the results of an ISO complying validation of the method carried out through an inter-laboratory study. Sets of blind samples were provided consisting of DNA reference materials covering all the elements detectable by specific probes present on the array. The GMO concentrations varied from 1% down to 0.045%. In addition, a mixture of two GMO events (0.1% RRS diluted in 100% TOPAS19/2) was incorporated in the study to test the robustness of the assay in extreme conditions. Data were processed according to ISO 5725 standard. The method was evaluated with predefined performance criteria with respect to the EC CRL method acceptance criteria. The overall method performance met the acceptance criteria; in particular, the results showed that the method is suitable for the detection of the different target elements at 0.1% concentration of GMO with a 95% accuracy rate. This collaborative trial showed that the method can be considered as fit for the purpose of screening with respect to its intra- and inter-laboratory accuracy. The results demonstrated the validity of combining multiplex PCR with array detection as provided by the DualChip^® GMO (Eppendorf, Germany) for the screening of GMO. The results showed that the technology is robust, practical and suitable as a screening tool.     

Deoxynivalenol in wheat,maize, wheat flour and pasta: surveys in Hungary in 2008–2015

Among Fusarium mycotoxins, deoxynivalenol (DON) is the most common contaminant in case of cereals and cereal-based foods in Hungary. In this study, Hungarian wheat (n = 305), maize (n = 108), wheat flour (n = 179) and pasta (n = 226) samples were analysed (N = 818). The samples were collected during 2008–2015 in Hungary. Applied methods of analysis were enzyme-linked immunosorbent assay and liquid-chromatography coupled with a mass spectrometer. Results were compared and evaluated with Hungarian weather data. Among cereal samples, in 2011, wheat was contaminated with DON (overall average ± standard deviation was 2159 ± 2818 µg kg^?1), which was above the maximum limit (ML). In case of wheat flour and pasta, no average values above ML were found during 2008–2015, but higher DON contamination could be observed in 2011 as well (wheat flour: 537 ± 573 µg kg^?1; pasta: 511 ± 175 µg kg^?1).     

Evaluierung und Optimierung einer doppelschaligen Fassade mit Hilfe von Simulationen

Bei großflächig verglasten, mehrgeschossigen Bürogebäuden liegen doppelschalige, hinterlüftete Glasfassaden nach wie vor im gestalterischen Trend. Die Konstruktionsdetails der Fassade haben einen erheblichen Einfluss auf die sommerliche Überhitzung der Luft im Fassadenzwischenraum (FZR), in dem auch der Sonnenschutz platziert ist. Es empfiehlt sich daher, die Details einer doppelschaligen Fassade im Hinblick auf die sommerlichen Temperaturverhältnisse zu optimieren. Die üblichen Methoden einer standardmäßigen bauphysikalischen Planung sind für eine derartige Optimierung nur sehr bedingt verwendbar. Im vorliegenden Beitrag wird anhand des Projektes Neues K & H Bank‐Gebäude in Budapest exemplarisch aufgezeigt, wie mit Hilfe von moderner Simulationstechnik die Ausbildung einer doppelschaligen Fassade evaluiert und anschließend optimiert werden kann. Evaluation and optimization of a double skin façade with the help of computational simulations. Natural ventilated double skin façades still prove to be trendy design choices for multi‐story office buildings with large glazed surfaces. The construction design details of these façades have a significant impact on the summer overheating of the air in the façade cavity, in which the shading is positioned also. Therefore it is recommended to investigate and optimize the parameters of the double skin façade concerning the summer temperature conditions. The usual methods of standard building physical planning have limited application possibilities for optimizing these kinds of systems. Therefore in the following article through the example of the K°&°H Bank, Budapest, we are going to explain how the modern simulation techniques can support the constructional design of double skin façades first by evaluation, then subsequently, by optimization.     

Breed-Specific Detection of Mangalica Meat in Food Products

A fast and reliable diagnostic system has been developed for the detection of Mangalica meat in foods. This qualitative test is based on a recombinase polymerase amplification which can be performed on the field, in situ, where it may be necessary to determine Mangalica content in food products at once. The required equipments for the procedure are pipettes, a portable homogenizer and a portable thermostat. DNA amplification is carried out at a constant temperature, and the detection is based on antibody reaction. The detection limit is one copy of the target sequence in 1 μl reaction volume. The test can be used for uncovering falsification of local brands on the spot within a very short (25–45 min) period of time. The present approach can be adopted for the detection of other food ingredients, if the species-specific target DNA sequence is known, e.g. in case of chicken, turkey, horse, and cattle.     

Stress Adaptation and the Brainstem with Focus on Corticotropin-Releasing Hormone

Stress adaptation is of utmost importance for the maintenance of homeostasis and, therefore, of life itself. The prevalence of stress-related disorders is increasing, emphasizing the importance of exploratory research on stress adaptation. Two major regulatory pathways exist: the hypothalamic–pituitary–adrenocortical axis and the sympathetic adrenomedullary axis. They act in unison, ensured by the enormous bidirectional connection between their centers, the paraventricular nucleus of the hypothalamus (PVN), and the brainstem monoaminergic cell groups, respectively. PVN and especially their corticotropin-releasing hormone (CRH) producing neurons are considered to be the centrum of stress regulation. However, the brainstem seems to be equally important. Therefore, we aimed to summarize the present knowledge on the role of classical neurotransmitters of the brainstem (GABA, glutamate as well as serotonin, noradrenaline, adrenaline, and dopamine) in stress adaptation. Neuropeptides, including CRH, might be co-localized in the brainstem nuclei. Here we focused on CRH as its role in stress regulation is well-known and widely accepted and other CRH neurons scattered along the brain may also complement the function of the PVN. Although CRH-positive cells are present on some parts of the brainstem, sometimes even in comparable amounts as in the PVN, not much is known about their contribution to stress adaptation. Based on the role of the Barrington’s nucleus in micturition and the inferior olivary complex in the regulation of fine motoric—as the main CRH-containing brainstem areas—we might assume that these areas regulate stress-induced urination and locomotion, respectively. Further studies are necessary for the field.     

Detection of Antimicrobial Peptides in Stratum Corneum by Mass Spectrometry

Antimicrobial and immunomodulatory peptides (AMPs) are considered as the key players in the maintenance of skin barrier functions. Here, we developed a novel approach for the examination of AMPs in the outermost layer of the epidermis, namely stratum corneum (SC). The SC sample collection by tape stripping was coupled with detection by highly specific and sensitive parallel reaction monitoring (PRM)-based mass spectrometry. We found that hexane-free processing of SC samples produced higher protein yield compared to hexane-based extraction. Of the 18 investigated peptides, 9 could be detected either in healthy or in inflamed skin specimens. Regarding the amount of S100A8, LCN2, LACRT and LYZ significant topographical differences were described among gland poor (GP), sebaceous gland rich (SGR) and apocrine gland rich (AGR) healthy skin regions. We applied a minimally invasive, reproducible approach for sampling, which can be assessed for research and diagnostic purposes and for monitoring the effectiveness of therapies in skin diseases.     

Ensemble Boosting and Bagging Based Machine Learning Models for Groundwater Potential Prediction

Water Resources Management - Due to the rapidly increasing demand for groundwater, as one of the principal freshwater resources, there is an urge to advance novel prediction systems to more...     

A Simple Environmentally-friendly Method for the Selective Synthesis of 1,5-benzodiazepine Derivatives using Zeolite Catalyst

The modified small pore size zeolite E4a has been found to be an efficient catalyst for the regioselective synthesis of 2,3-dihydro-1H-1,5-benzodiazepines from o-phenylenediamine and ketones. This method is simple, cheap, environmentally-friendly and give the benzodiazepines in high yield.     

A New Player in the Hippocampus: A Review on VGLUT3+ Neurons and Their Role in the Regulation of Hippocampal Activity and Behaviour

Glutamate is the most abundant excitatory amino acid in the central nervous system. Neurons using glutamate as a neurotransmitter can be characterised by vesicular glutamate transporters (VGLUTs). Among the three subtypes, VGLUT3 is unique, co-localising with other “classical” neurotransmitters, such as the inhibitory GABA. Glutamate, manipulated by VGLUT3, can modulate the packaging as well as the release of other neurotransmitters and serve as a retrograde signal through its release from the somata and dendrites. Its contribution to sensory processes (including seeing, hearing, and mechanosensation) is well characterised. However, its involvement in learning and memory can only be assumed based on its prominent hippocampal presence. Although VGLUT3-expressing neurons are detectable in the hippocampus, most of the hippocampal VGLUT3 positivity can be found on nerve terminals, presumably coming from the median raphe. This hippocampal glutamatergic network plays a pivotal role in several important processes (e.g., learning and memory, emotions, epilepsy, cardiovascular regulation). Indirect information from anatomical studies and KO mice strains suggests the contribution of local VGLUT3-positive hippocampal neurons as well as afferentations in these events. However, further studies making use of more specific tools (e.g., Cre-mice, opto- and chemogenetics) are needed to confirm these assumptions.