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Natural antisense long non-coding RNAs (lncNATs) are involved in the regulation of gene expression in plants, modulating different relevant developmental processes and responses to various stimuli. We have identified and characterized two lncNATs (NAT1 UGT73C6 and NAT2 UGT73C6 , collectively NATs UGT73C6 ) from Arabidopsis thaliana that are transcribed from a gene fully overlapping UGT73C6, a member of the UGT73C subfamily of genes encoding UDP-glycosyltransferases (UGTs). Expression of both NATs UGT73C6 is developmentally controlled and occurs independently of the transcription of UGT73C6 in cis. Downregulation of NATs UGT73C6 levels through artificial microRNAs results in a reduction of the rosette area, while constitutive overexpression of NAT1 UGT73C6 or NAT2 UGT73C6 leads to the opposite phenotype, an increase in rosette size. This activity of NATs UGT73C6 relies on its RNA sequence and, although modulation of UGT73C6 in cis cannot be excluded, the observed phenotypes are not a consequence of the regulation of UGT73C6 in trans. The NATs UGT73C6 levels were shown to affect cell proliferation and thus individual leaf size. Consistent with this concept, our data suggest that the NATs UGT73C6 influence the expression levels of key transcription factors involved in regulating leaf growth by modulating cell proliferation. These findings thus reveal an additional regulatory layer on the process of leaf growth. In this work, we characterized at the molecular level two long non-coding RNAs (NATs UGT73C6 ) that are transcribed in the opposite direction to UGT73C6, a gene encoding a glucosyltransferase involved in brassinosteroid homeostasis in A. thaliana. Our results indicate that NATs UGT73C6 expression influences leaf growth by acting in trans and by modulating the levels of transcription factors that are involved in the regulation of cell proliferation., (© 2022 The Authors. The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.)

Objective: Epilepsy surgery can potentially cure drug-resistant epilepsy, but careful presurgical evaluation is vital to select patients who will profit from such an intervention. Many epilepsy surgery programs offer extensive presurgical evaluation including several days of video-EEG monitoring. Non-lesional epilepsy cases are rare among epilepsy surgery patients. We set up a lesion-orientated paediatric epilepsy surgery program for patients with clearly localized lesions with limited presurgical diagnostics, in particular, with a maximum of 48 hours of non-invasive EEG monitoring that did not necessarily include ictal EEGs., Methods: We retrospectively evaluated the outcome of patients who were operated on within our epilepsy surgery program with respect to seizure freedom., Results: Fifty-two children and adolescents with MRI lesions at a mean age of 8.27 ±4.83 years (range: 0.17-18.87) underwent a resective procedure. The most frequent surgery was a hemispherotomy. Overall seizure freedom was 81.8% after 12 months and 85.6% after a median observation period of 20.45 months. Seizure frequency was reduced >50% in all other patients. Preoperative recording of an ictal EEG on the side of surgery had no effect on postoperative seizure outcome (p= 0.697), nor did recording of epileptiform discharges on the ipsilateral (p= 0.538) and contralateral side (p= 0.147)., Significance: Our findings highlight the high success rate using a lesion-orientated epilepsy surgical approach with reduced presurgical video-EEG monitoring in the paediatric epilepsy population. Our data show that it is possible to reduce the complex pre-surgical work-up for epilepsy in children and adolescents by asking the basic question: "Is there any reason why the lesion should not be resected".

Cytokine signaling is transmitted by cell surface receptors which act as natural biological switches to control cellular functions such as immune reactions. Recently, we have designed synthetic cytokine receptors (SyCyRs) consisting of green fluorescent protein (GFP)- and mCherry-nanobodies fused to the transmembrane and intracellular domains of cytokine receptors. Following stimulation with homo- and heterodimeric GFP-mCherry fusion proteins, the resulting receptors phenocopied signaling induced by physiologically occurring cytokines. GFP and mCherry fusion proteins were produced in E. coli or CHO-K1 cells, but the overall yield and stability was low. Therefore, we applied two alternative multimerization strategies and achieved immunoglobulin Fc-mediated dimeric and coiled-coil GCN4pII-mediated trimeric assemblies. GFP- and/or mCherry-Fc homodimers activated synthetic gp130 cytokine receptors, which naturally respond to Interleukin 6 family cytokines. Activation of these synthetic gp130 receptors resulted in STAT3 and ERK phosphorylation and subsequent proliferation of Ba/F3-gp130 cells. Half-maximal effective concentrations (EC50) of 8.1 ng/ml and 0.64 ng/ml were determined for dimeric GFP-Fc and mCherry-Fc, respectively. This is well within the expected EC50 range of the native cytokines. Moreover, we generated tetrameric and hexameric GFP-mCherry-Fc fusion proteins, which were also biologically active. This highlighted the importance of close juxtaposition of two cytokine receptors for efficient receptor activation. Finally, we used a trimeric GCN4pII motif to generate homo-trimeric GFP and mCherry complexes. These synthetic cytokines showed improved EC50 values (GFP3: 0.58 ng/ml; mCherrry3: 0.37 ng/ml), over dimeric Fc fused variants. In conclusion, we successfully generated highly effective and stable multimeric synthetic cytokine receptor ligands for activation of synthetic cytokine receptors., Competing Interests: The authors have declared that no competing interests exist.