Running title: Maximal loading of MCM2/4 in late G1

Once-per-cell cycle replication is regulated through the assembly onto chromatin of multisubunit protein complexes that license DNA for a further round of replication. Licensing consists of the loading of the hexameric MCM2-7 complex onto chromatin during G1 phase and is dependent on the licensing factor Cdt1. In vitro experiments have suggested a two-step binding mode for minichromosome maintenance (MCM) proteins, with transient initial interactions converted to stable chromatin loading. Here, we assess MCM loading in live human cells using an in vivo licensing assay on the basis of fluorescence recovery after photobleaching of GFP-tagged MCM protein subunits through the cell cycle. We show that, in telophase, MCM2 and MCM4 maintain transient interactions with chromatin, exhibiting kinetics similar to Cdt1. These are converted to stable interactions from early G1 phase. The immobile fraction of MCM2 and MCM4 increases during G1 phase, suggestive of reiterative licensing. In late G1 phase, a large fraction of MCM proteins are loaded onto chromatin, with maximal licensing observed just prior to S phase onset. Fluorescence loss in photobleaching experiments show subnuclear concentrations of MCM-chromatin interactions that differ as G1 phase progresses and do not colocalize with sites of DNA synthesis in S phase.Fil: Symeonidou, Ioanna Eleni. University of Patras. School of Medicine. Laboratory of General Biology; Grecia;Fil: Kotsantis, Panagiotis. University of Patras. School of Medicine. Laboratory of General Biology; Grecia;Fil: Roukos, Vassilis. University of Patras. School of Medicine. Laboratory of General Biology; Grecia;Fil: Rapsomaniki, Maria Anna. University of Patras. School of Medicine. Laboratory of General Biology; Grecia;Fil: Grecco, Hernan Edgardo. Max Planck Institute of Molecular Physiology. Department of Systemic Cell Biology; Alemania; Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Física de Buenos Aires; Argentina;Fil: Bastiaens, Philippe. Max Planck Institute of Molecular Physiology. Department of Systemic Cell Biology; Alemania;Fil: Taraviras, Stavros. University of Patras. School of Medicine. Laboratory of Physiology; Grecia;Fil: Lygerou, Zoi. University of Patras. School of Medicine. Laboratory of General Biology; Grecia

CLAPNQ: Cohesive Long-form Answers from Passages in Natural Questions for RAG systems

Retrieval Augmented Generation (RAG) has become a popular application for large language models. It is preferable that successful RAG systems provide accurate answers that are supported by being grounded in a passage without any hallucinations. While considerable work is required for building a full RAG pipeline, being able to benchmark performance is also necessary. We present ClapNQ, a benchmark Long-form Question Answering dataset for the full RAG pipeline. ClapNQ includes long answers with grounded gold passages from Natural Questions (NQ) and a corpus to perform either retrieval, generation, or the full RAG pipeline. The ClapNQ answers are concise, 3x smaller than the full passage, and cohesive, with multiple pieces of the passage that are not contiguous. RAG models must adapt to these properties to be successful at ClapNQ. We present baseline experiments and analysis for ClapNQ that highlight areas where there is still significant room for improvement in grounded RAG. CLAPNQ is publicly available at https://github.com/primeqa/clapnqComment: 25 page

Exposure to the ROCK inhibitor fasudil promotes gliogenesis of neural stem cells in vitro

Fasudil is a clinically approved Rho-associated protein kinase (ROCK) inhibitor that has been used widely to treat cerebral consequences of subarachnoid hemorrhage. It is known to have a positive effect on animal models of neurological disorders including Parkinson's disease and stroke. However, its cellular effect on progenitor populations and differentiation is not clearly understood. While recent studies suggest that fasudil promotes the mobilization of neural stem cells (NSCs) from the subventricular zone in vivo and promotes the differentiation of the C17.2 cerebellar neuroprogenitor line in vitro, it is unclear whether fasudil is involved in the differentiation of primary NSCs. Here, we tested the effect of fasudil on mouse NSCs in vitro, and observed increased gliogenesis in NSCs derived from lateral ventricles. Upon treatment, fasudil promoted characteristics of neurogenesis including phenotypic changes in neural outgrowth and interkinetic nuclear-like movements as an immediate response, while Sox2 expression was maintained and GFAP expression increased. Moreover, the gliogenic response to fasudil medium was observed in both early postnatal and adult NSC cultures. Taken together, our results show that fasudil promotes the differentiation of NSCs into astroglial lineage, suggesting that it could be used to develop novel vitro gliogenesis models and regulate differentiation for neural repair

Beyond HER2 and trastuzumab: heterogeneity, systems biology, and cancer origin research may guide the future for personalized treatment of very early but aggressive breast cancer

J Clin Onco

Twenty-one-gene assay: challenges and promises in translating personal genomics and whole-genome scans into personalized treatment of breast cancer

J Clin Onco