פרסומים

Latent 5' splice sites, highly abundant in human introns, are not normally used. This led to the proposal of a quality control mechanism, Suppression of Splicing (SOS), which protects cells from splicing at the numerous intronic latent sites, and whose activation can generate nonsense mRNAs. SOS was shown to be independent of Nonsense-Mediated mRNA Decay (NMD). Efforts to decipher the SOS mechanism revealed a pivotal role for initiator-tRNA, independent of protein translation. Recently, nucleolin (a multifunctional protein) was found to directly and specifically bind the initiator-tRNA in the nucleus and was shown to be a protein component of SOS, enabling an updated model of the SOS mechanism. Importantly, SOS is abrogated under stress and in cancer (e.g., in breast cancer cells and gliomas), generating thousands of nonsense mRNAs due to activation of latent splicing. The resulting affected human genes cover a variety of functional groups, including genes involved in cell proliferation and differentiation. Furthermore, in oligodendroglioma, the extent of activation of latent splicing increases with the severity of the cancer. Interesting examples are genes expressing aberrant nonsense mRNAs in both breast cancer and glioma, due to latent splicing activation. These findings highlight the unexplored potential of such aberrant isoforms as novel targets for cancer diagnosis and therapies.

Heavy precipitation events (HPEs) can lead to deadly and costly natural disasters and are critical to the hydrological budget in regions where rainfall variability is high and water resources depend on individual storms. Thus, reliable projections of such events in the future are needed. To provide high-resolution projections under the RCP8.5 scenario for HPEs at the end of the 21 st century, and to understand the changes in sub-hourly to daily rainfall patterns, weather research and forecasting (WRF) model simulations of 41 historic HPEs in the eastern Mediterranean are compared with "pseudo global warming" simulations of the same events. This paper presents the changes in rainfall patterns in future storms, decomposed into storms’ mean conditional rain rate, duration, and area. A major decrease in rainfall accumulation (-30% averaged across events) is found throughout future HPEs. This decrease results from a substantial reduction of the rain area of storms (-40%) and occurs despite an increase in the mean conditional rain intensity (+15%). The duration of the HPEs decreases (-9%) in future simulations. Regionally maximal 10-min rain rates increase (+22%), whereas over most of the region, long-duration rain rates decrease. The consistency of results across events, driven by varying synoptic conditions, suggests that these changes have low sensitivity to the specific synoptic evolution during the events. Future HPEs in the eastern Mediterranean will therefore likely be drier and more spatiotemporally concentrated, with substantial implications on hydrological outcomes of storms. Plain Language Summary Heavy precipitation events are large storms that can recharge freshwater reservoirs, but can also lead to hazardous outcomes such as flash floods. Therefore, understanding the impacts of climate change on such storms is critical. Here, a weather model similar to those used in weather forecasts is used to simulate heavy precipitation events in the eastern Mediterranean. A large collection of storms is simulated in pairs: (1) historic storms, selected for their high impact, and (2) the same storms placed in a global warming scenario projected for the end of the 21 st century. Using these simulations we ask how present-day storms would look like were they to occur at the warmer end of the 21 st century. The future storms are found to produce much less rainfall compared to their historic counterparts. This decrease in rainfall is attributed mainly to the reduction in the area covered by storms’ rainfall, and happens despite increasing rainfall intensities. These results suggest that the region will be drier in the future with larger dry areas during storms; however, over short durations, it would rain more intensely over contracted areas-increasing local hazards associated with heavy precipitation events.

Previously, we reported that RelA protein facilitates Hfq-mediated mRNA-sRNA regulation by binding sRNAs carrying a Shine-Dalgarno-like GGAG sequence. In turn, sRNA-Hfq monomers are stabilized, enabling the attachment of more Hfq subunits to form a functional hexamer. Here, using CLIP-seq, we present a global analysis of RelA-bound RNAs showing that RelA interacts with sRNAs as well as with mRNAs carrying a GGAG motif. RelA binding of mRNAs carrying GGAG at position -7 relative to the initiation codon (AUG) inhibits translation by interfering with the binding of 30S ribosomes. The extent of inhibition depends on the distance of GGAG relative to the AUG, as shortening the spacing between GGAG and AUG abrogates RelA-mediated inhibition. Interestingly, RelA binding of target mRNAs carrying GGAG in the coding sequence or close to AUG facilitates target gene regulation by sRNA partners that lack GGAG. However, translation inhibition caused by RelA binding of mRNAs carrying GGAG at position -7 relative to the AUG renders sRNA-mRNA basepairing regulation ineffective. Our study indicates that by binding RNAs carrying GGAG the ribosome-associated RelA protein inhibits translation of specific newly synthesized incoming mRNAs or enables basepairing regulation by their respective sRNA partners, thereby introducing a new regulatory concept for the bacterial response.

Cytochrome c oxidase (COX), a multimeric protein complex, is the final electron acceptor in the mitochondrial electron transfer chain. Primary COX deficiency, caused by mutations in either mitochondrial DNA or nuclear-encoded genes, is a heterogenous group of mitochondrial diseases with a wide range of presentations, ranging from fatal infantile to subtler. We previously reported a patient with primary COX deficiency due to a pathogenic variant in COX4I1 (encoding the common isoform of COX subunit 4, COX4-1), who presented with bone marrow failure, genomic instability, and short stature, mimicking Fanconi anemia (FA). In the present study, we demonstrated that accumulative DNA damage coincided primarily with proliferative cells in the patient's fibroblasts and in COX4i1 knockdown cells. Expression analysis implicated a reduction in DNA damage response pathways, which was verified by demonstrating impaired recovery from genotoxic insult and decreased DNA repair. The premature senescence of the COX4-1-deficient cells prevented us from undertaking additional studies; nevertheless, taken together, our results indicate replicative stress and impaired nuclear DNA damage response in COX4-1 deficiency. Interestingly, our in vitro findings recapitulated the patient's presentation and present status.

Insulin-like growth factor-1 (IGF-1) and its binding proteins and receptors are widely expressed in the central nervous system (CNS), proposing IGF-1-induced neurotrophic actions in normal growth, development, and maintenance. However, while there is convincing evidence that the IGF-1 system has specific endocrine roles in the CNS, the concept is emerging that IGF-I might be also important in disorders such as ischemic stroke, brain trauma, Alzheimer's disease, epilepsy, etc., by inducing neuroprotective effects towards glutamate-mediated excitotoxic signaling pathways. Research in rodent models has demonstrated rescue of pathophysiological and behavioral abnormalities when IGF-1 was administered by different routes, and several clinical studies have shown safety and promise of efficacy in neurological disorders of the CNS. Focusing on the relationship between IGF-1-induced neuroprotection and glutamate-induced excitatory neurotoxicity, this review addresses the research progress in the field, intending to provide a rationale for using IGF-I clinically to confer neuroprotective therapy towards neurological diseases with glutamate excitotoxicity as a common pathological pathway.

OBJECTIVE: Cellular senescence limits tumourigenesis by blocking the proliferation of premalignant cells. Additionally, however, senescent cells can exert paracrine effects influencing tumour growth. Senescent cells are present in premalignant pancreatic intraepithelial neoplasia (PanIN) lesions, yet their effects on the disease are poorly characterised. It is currently unknown whether senolytic drugs, aimed at eliminating senescent cells from lesions, could be beneficial in blocking tumour development. DESIGN: To uncover the functions of senescent cells and their potential contribution to early pancreatic tumourigenesis, we isolated and characterised senescent cells from PanINs formed in a Kras-driven mouse model, and tested the consequences of their targeted elimination through senolytic treatment. RESULTS: We found that senescent PanIN cells exert a tumour-promoting effect through expression of a proinflammatory signature that includes high Cox2 levels. Senolytic treatment with the Bcl2-family inhibitor ABT-737 eliminated Cox2-expressing senescent cells, and an intermittent short-duration treatment course dramatically reduced PanIN development and progression to pancreatic ductal adenocarcinoma. CONCLUSIONS: These findings reveal that senescent PanIN cells support tumour growth and progression, and provide a first indication that elimination of senescent cells may be effective as preventive therapy for the progression of precancerous lesions.

Irradiation-induced alopecia and dermatitis (IRIAD) are two of the most visually recognized complications of radiotherapy, of which the molecular and cellular basis remains largely unclear. By combining scRNA-seq analysis of whole skin-derived irradiated cells with genetic ablation and molecular inhibition studies, we show that senescence-associated IL-6 and IL-1 signaling, together with IL-17 upregulation and CCR6(+) -mediated immune cell migration, are crucial drivers of IRIAD. Bioinformatics analysis colocalized irradiation-induced IL-6 signaling with senescence pathway upregulation largely within epidermal hair follicles, basal keratinocytes, and dermal fibroblasts. Loss of cytokine signaling by genetic ablation in IL-6(-/-) or IL-1R(-/-) mice, or by molecular blockade, strongly ameliorated IRIAD, as did deficiency of CCL20/CCR6-mediated immune cell migration in CCR6(-/-) mice. Moreover, IL-6 deficiency strongly reduced IL-17, IL-22, CCL20, and CCR6 upregulation, whereas CCR6 deficiency reciprocally diminished IL-6, IL-17, CCL3, and MHC upregulation, suggesting that proximity-dependent cellular cross talk promotes IRIAD. Therapeutically, topical application of Janus kinase blockers or inhibition of T-cell activation by cyclosporine effectively reduced IRIAD, suggesting the potential of targeted approaches for the treatment of dermal side effects in radiotherapy patients.

Oogenesis produces functional eggs and is essential for fertility, embryonic development, and reproduction. The zebrafish ovary is an excellent model to study oogenesis in vertebrates, and recent studies have identified multiple regulators in oocyte development through forward genetic screens, as well as reverse genetics by CRISPR mutagenesis. However, many developmental steps in oogenesis, in zebrafish and other species, remain poorly understood, and their underlying mechanisms are unknown. Here, we take a genomic approach to systematically uncover biological activities throughout oogenesis. We performed transcriptomic analysis on five stages of oogenesis, from the onset of oocyte differentiation through Stage III, which precedes oocyte maturation. These transcriptomes revealed thousands of differentially expressed genes across stages of oogenesis. We analyzed trends of gene expression dynamics along oogenesis, as well as their expression in pair-wise comparisons between stages. We determined their functionally enriched terms, identifying uniquely characteristic biological activities in each stage. These data identified two prominent developmental phases in oocyte differentiation and traced the accumulation of maternally deposited embryonic regulator transcripts in the developing oocyte. Our analysis provides the first molecular description for oogenesis in zebrafish, which we deposit online as a resource for the community. Further, the presence of multiple gene paralogs in zebrafish, and the exclusive curation by many bioinformatic tools of the single paralogs present in humans, challenge zebrafish genomic analyses. We offer an approach for converting zebrafish gene name nomenclature to the human nomenclature for supporting genomic analyses generally in zebrafish. Altogether, our work provides a valuable resource as a first step to uncover oogenesis mechanisms and candidate regulators and track accumulating transcripts of maternal regulators of embryonic development.

OBJECTIVES: In this study, we aimed to develop a novel, sustained release varnish (SRV) for voice prostheses (VP) releasing chlorhexidine (CHX), for the prevention of biofilm formation caused by the common oral bacteria Streptococcus mutans on VP surfaces. METHODS: This study was performed in an in vitro model as a step towards future in vivo trials. VPs were coated with a SRV containing CHX (SRV-CHX) or SRV alone (placebo-SRV) that were daily exposed to S. mutans. The polymeric materials of SRV were composed of ethylcellulose and PEG-400. Biofilm formation was assessed by DNA quantification (qPCR), crystal violet staining, confocal laser scanning microscopy (CLSM), scanning electron microscopy (SEM), and kinetics experiments. RESULTS: The amount of DNA in the biofilms formed by S. mutans on VP surfaces coated once with SRV-CHX (1.024 ± 0.218 ng DNA/piece) was 58.5 ± 8.8% lower than that of placebo-SRV-coated VPs (2.465 ± 0.198 ng DNA/piece) after a 48-h exposure to S. mutans (p = 0.038). Reduced biofilm mass on SRV-CHX-coated VPs was visually confirmed by CLSM and SEM. CV staining of SRV-CHX single-coated VPs that have been exposed to S. mutans nine times showed a 98.1 ± 0.2% reduction in biofilm mass compared to placebo-SRV-coated VPs (p = 0.003). Kinetic experiments revealed that SRV-CHX triple-coated VPs could delay bacterial growth for 23 days. CONCLUSIONS: Coating VPs with SRV-CHX has an inhibitory effect on biofilm formation and prevents bacterial growth in their vicinities. This study is a proof-of-principle that paves the way for developing new clinical means for reducing both VPs' bacterial biofilm formation and device failure.

This paper proposes a formal definition of reanalysis, while emphasizing the importance of the distinction between two different kinds of reanalysis: those in which the change is confined to the grammatical level, and those in which it is confined to the semantic level. After tracing the history of a negative counterfactual conditional marker in Hebrew and Aramaic which underwent both syntactic and semantic reanalyses, the paper assesses the concept of reanalysis with focus on the following questions: Is reanalysis a single, clearly-defined phenomenon, and if so, what is its nature? Is it merely a descriptive label for a certain observable state of affairs, or does it explain diachronic changes? Alternatively, perhaps it is a theoretical constraint, a theoretical requirement that linguistic change must be associated with specific environments where reanalysis can take place? A detailed analysis of the marker and its evolution yields the following broad hypothesis: Reanalysis of a linguistic form does not change the truth conditions of the proposition that contains it, regardless of whether the reanalysis is on the grammatical level or on the semantic level.

Metacognition is an invaluable part of instruction of higher order thinking (HOT). The goal of this article is to review previous studies about teachers’ knowledge and professional development (PD) in the area of metacognitive instruction in the context of teaching HOT. Part A of the article reviews 25 empirical studies assembled through a scoping review. Although each individual study consists of significant findings, our analysis indicates that, as a field, the study of teachers’ and pre-service teachers’ knowledge in this area is still rather preliminary and exploratory. The review draws several conclusions regarding the nature of the research in this area. However, lack of a common conceptual framework and research instruments precludes the possibility of drawing meaningful general conclusions from the findings of the 25 studies. Part B centers on 8 empirical and theoretical studies addressing the same conceptual framework centering on meta-strategic knowledge (MSK). The findings demonstrate the significance of metacognition in general, and MSK in particular for teachers’ ability to teach HOT, showing that it can be developed in both pre-service and in-service teachers’ education and PD. The findings highlight several specific characteristics of teachers’ knowledge and learning processes in this area. Yet, the findings also show that metacognition is rarely addressed in a satisfactory manner in large-scale efforts to teach HOT and that MSK is mostly neglected in PD programs for teaching inquiry learning. The implications for research and practice of HOT and metacognition are discussed.

Metacognition is an invaluable part of instruction of higher order thinking (HOT). The goal of this article is to review previous studies about teachers’ knowledge and professional development (PD) in the area of metacognitive instruction in the context of teaching HOT. Part A of the article reviews 25 empirical studies assembled through a scoping review. Although each individual study consists of significant findings, our analysis indicates that, as a field, the study of teachers’ and pre-service teachers’ knowledge in this area is still rather preliminary and exploratory. The review draws several conclusions regarding the nature of the research in this area. However, lack of a common conceptual framework and research instruments precludes the possibility of drawing meaningful general conclusions from the findings of the 25 studies. Part B centers on 8 empirical and theoretical studies addressing the same conceptual framework centering on meta-strategic knowledge (MSK). The findings demonstrate the significance of metacognition in general, and MSK in particular for teachers’ ability to teach HOT, showing that it can be developed in both pre-service and in-service teachers’ education and PD. The findings highlight several specific characteristics of teachers’ knowledge and learning processes in this area. Yet, the findings also show that metacognition is rarely addressed in a satisfactory manner in large-scale efforts to teach HOT and that MSK is mostly neglected in PD programs for teaching inquiry learning. The implications for research and practice of HOT and metacognition are discussed.

Presented herein are recommendations for use of nirmatrelvir/ritonavir in patients with epilepsy, as issued by the Steering Committee of the Israeli chapter of the International League Against Epilepsy. The recommendations suggest that patients on moderate-to-strong enzyme-inducing antiseizure medications (ASMs) and everolimus should not be treated with nirmatrelvir/ritonavir; rectal diazepam may be used as an alternative to buccal midazolam; doses of ASMs that are cytochrome P450 (CYP3A4) substrates might be adjusted; and patients treated with combinations of nirmatrelvir/ritonavir and ASMs that are CYP3A4 substrates or lamotrigine should be monitored for drug efficacy and adverse drug reactions.