Preliminary results of the recombinase polymerase amplification technique for the detection of Haemonchus contortus from Hungarian field samples

Haemonchus contortus is a parasitic nematode of small ruminants responsible for significant economic losses and animal health concerns globally. Detection of gastrointestinal nematode (GIN) infection in veterinary practice typically relies on microscopy-based methods such as the faecal egg count and morphological identification of larval culture. However, mixed co-infections are common and species-specific identification is typically time-consuming and expertise-intensive. Compounded by increasing anthelmintic resistance, there is an urgent need to implement the molecular diagnosis of GIN in the livestock industry, preferably in field settings. Advances in isothermal amplification techniques including recombinase polymerase amplification (RPA) assays could improve this. Yet, constraints in RPA kit availability and amplicon detection systems limit the use of this technology in point of care settings. In this study, we present an early-stage, proof-of-concept demonstration of RPA targeting the internal transcribed spacer (ITS2) region of H. contortus. Having tested against eight closely related nematodes and also against five farm isolates in Eastern Hungary, preliminary results derived from a comparative analysis of 3 primer sets showed the assay detects H. contortus DNA and has a limit of detection of 10 ng/μl. We also tested an end-result naked eye detection system using various DNA binding dyes, of which EvaGreen® dye was successful for a qualitative RPA detection that could be adaptable at farm sites. [Abstract copyright: Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.

Membrane-grafted asymmetric organocatalyst for an integrated synthesis-separation platform

In this work we introduce a sustainable membrane-based synthesis–separation platform for enantioselective organocatalysis. An azido derivatized cinchona-squaramide bifunctional catalyst was synthesized and subsequently grafted to the surface of a polybenzimidazole-based nanofiltration membrane. The favorable effect of the covalent graftingdue to the change in geometry and increased secondary interactionson the catalytic activity due to conformational changes was confirmed by quantum chemical calculations. Asymmetric <i>Michael</i> and aza-<i>Michael</i> reactions of 1,3-dicarbonyl and indole, pyrazole, and triazole derivatives to β-nitrostyrene were performed with as high as 99% enantiomeric excess. This report on the enantioselective aza-<i>Michael</i> reaction of pyrazoles and triazoles opens new frontiers in the application of squaramide-based cinchona catalysts. A catalytic membrane cascade reactor was developed for an integrated synthesis–purification process allowing at least 98% product and substrate recovery, and quantitative in situ solvent recycling. The sustainability of the synthetic methodology was assessed through E-factor and carbon footprint

Differential gene expression and physiological changes during acute or persistent plant virus interactions may contribute to viral symptom differences

Viruses have different strategies for infecting their hosts. Fast and acute infections result in the development of severe symptoms and may cause the death of the plant. By contrast, in a persistent interaction, the virus can survive within its host for a long time, inducing only mild symptoms. In this study, we investigated the gene expression changes induced in CymRSV-, crTMV-, and TCV-infected Nicotiana benthamiana and in PVX- and TMV-U1-infected Solanum lycopersicum plants after the systemic spread of the virus by two different high-throughput methods: microarray hybridization or RNA sequencing. Using these techniques, we were able to clearly differentiate between acute and persistent infections. We validated the gene expression changes of selected genes by Northern blot hybridization or by qRT-PCR. We show that, in contrast to persistent infections, the drastic shut-off of housekeeping genes, downregulation of photosynthesis-related transcripts and induction of stress genes are specific outcomes with acute infections. We also show that these changes are not a consequence of host necrosis or the presence of a viral silencing suppressor. Thermal imaging data and chlorophyll fluorescence measurements correlated very well with the molecular changes. We believe that the molecular and physiological changes detected during acute infections mostly contribute to virus symptom development. The observed characteristic physiological changes associated with economically more dangerous acute infections could serve as a basis for the elaboration of remote monitoring systems suitable for detecting developing virus infections in crops. Moreover, as molecular and physiological changes are characteristics of different types of virus lifestyles, this knowledge can support risk assessments of recently described novel viruses

Sport és Integráció, avagy az SNI-s és hátrányos helyzetű diákok integrálása és rekreálódása egy iskolai sportnap lehetőségein belül

INST: L_135Szakdolgozatomban a kalocsai iskolák negyedik évfolyamos diákjainak szerveztem egy rendezvényt integrációs jellegel, ahol taglaltam a sajátos nevelési igény, az integráció, az inklúzió, a fogyatékosság és a rekreáció fogalmát. Ezek alapján hoztam létre egy olyan inklúzív rekreációs sportnapot, egy olyan olvasztó tégelyt, ahol minden diák egyenlőnek számít. Ahol sportolhatnak két napon keresztül közösen