The NF45/NF90 Heterodimer Contributes to the Biogenesis of 60S Ribosomal Subunits and Influences Nucleolar Morphology

The interleukin enhancer binding factors ILF2 (NF45) and ILF3 (NF90/NF110) have been implicated in various cellular pathways, such as transcription, microRNA (miRNA) processing, DNA repair, and translation, in mammalian cells. Using tandem affinity purification, we identified human NF45 and NF90 as components of precursors to 60S (pre-60S) ribosomal subunits. NF45 and NF90 are enriched in nucleoli and cosediment with pre-60S ribosomal particles in density gradient analysis. We show that association of the NF45/NF90 heterodimer with pre-60S ribosomal particles requires the double-stranded RNA binding domains of NF90, while depletion of NF45 and NF90 by RNA interference leads to a defect in 60S biogenesis. Nucleoli of cells depleted of NF45 and NF90 have altered morphology and display a characteristic spherical shape. These effects are not due to impaired rRNA transcription or processing of the precursors to 28S rRNA. Consistent with a role of the NF45/NF90 heterodimer in nucleolar steps of 60S subunit biogenesis, downregulation of NF45 and NF90 leads to a p53 response, accompanied by induction of the cyclin-dependent kinase inhibitor p21/CIP1, which can be counteracted by depletion of RPL11. Together, these data indicate that NF45 and NF90 are novel higher-eukaryote-specific factors required for the maturation of 60S ribosomal subunits

Cell Type-Specific Arousal-Dependent Modulation of Thalamic Activity in the Lateral Geniculate Nucleus

State-dependent thalamocortical activity is important for sensory coding, oscillations, and cognition. The lateral geniculate nucleus (LGN) relays visual information to the cortex, but the state-dependent spontaneous activity of LGN neurons in awake behaving animals remains controversial. Using a combination of pupillometry, extracellular, and intracellular recordings from identified LGN neurons in behaving mice, we show that thalamocortical (TC) neurons and interneurons are distinctly correlated to arousal forming two complementary coalitions. Intracellular recordings indicated that the membrane potential of LGN TC neurons was tightly correlated to fluctuations in pupil size. Inactivating the corticothalamic feedback to the LGN suppressed the arousal dependency of LGN neurons. Taken together, our results show that LGN neuronal membrane potential and action potential output are dynamically linked to arousal-dependent brain states in awake mice, and this might have important functional implications.Peer reviewe

Metamorphic graphite from Szendrőlád (Szendrő Mts., NE-Hungary) detected by simultaneous DTA-TG

Graphite, one of the polymorphic forms of carbon, has become a versatile industrial material of nowadays due to its particular attributes. It is used mainly in the automobile industry, metal extractive industry and in the high-tech industry. Moreover, it is also included in the list of critical raw materials for the EU. Our aim was to prove the presence of graphite by thermal analysis beyond X-ray powder diffraction (XRD) and Raman spectroscopy. Thermogravimetry yields comparable results with quantitative XRD. The formation conditions are described by Raman spectrometry and microscopy examinations of drill core samples from Szendrőlád (Szendrő Mts, NE-Hungary; (Szendrőlád Limestone Formation, middle-late Devonian, shelf-basin facies). Polished rock slabs were made for optical microscopy, scanning electron microscopy with energy dispersive spectrometry (SEM–EDS) and Raman spectroscopy. X-ray powder diffraction (XRD) and thermal analysis (DTA-TG) measurements were made on powders. Based on our results, the graphite is epigenetic; its quantity varies between 1.5–3 mass% in the samples. It was developed in 20–50 μm sized flakes, which are often arranged in > 300 μm sized aggregates. Graphite was formed during regional metamorphism from the organic matter-rich shales. The average formation temperature, calculated from the results of Raman spectroscopy, is around 410 °C (± 30 °C). The Raman measurements also indicated the presence of a partially graphitized (disordered graphite) material beside graphite

Fókuszált ionsugaras és elektronsugaras nano-megmunkálás fizikai-kémiai alapjainak vizsgálata különös tekintettel a potenciális nanoelektronikai alkalmazásokra = Investigation of physical and chemical basics of focused electron and ion beam nano-machining with special emphasis on possible applications in nanoelectronics

Jelen OTKA pályázat tette lehetővé, hogy unikális LEO 1540XB keresztsugaras rendszerünk installációs és betanulási periódusa után kutatva „vegyük birtokba” a pásztázó nanosugaras vizsgálati és megmunkálási technikákat (litográfia / kisenergiás SEM és mikroanalízis / mikroszkópia és nanoméretekben folytatott marás és mintázatok kialakítása FIB segítségével / valamint ion vagy elektronsugaras anyagleválasztás nanoméretekben gázok injektálásával (GIS). Elsőnek a napelemtechnológiában használt Mo és ZnO FIB porlasztási sajátosságait és effektiv hozamát vizsgáltuk. A Mo erős anizotrópiájával szemben a ZnO homogén porlódást mutatott. Következő lépésként a FIB segítségével pórusos Si multirétegben fotonikus kristály szerkezeteket alakítottunk ki, majd W nanovezeték leválasztását tanulmányoztuk FIB- és SEM technikával. Az ionokkal leválasztott W réteg egyenletes leválási sebességet mutatott, míg az elektronsugár energiája és beesési szöge erősen hatott az így leválasztott réteg morfológiájára, bizonyítva a szekunder elektron emisszió hatását. In-situ és ex-situ ellenállásméréseket folytattunk, szobahőmérsékleten és hőkezelés közben. Az ionokkal leválasztott réteg ohmos jellegű volt, és hőkezelés után irreverzibilis ellenállás változást mutatott, ami W kristályosodásra utal. Az elektronsugárral leválasztott rétegen nemlineáris I-V karakterisztikát mértünk. A fenti kutatások tapasztalatai számos további kutatást segítettek, melyekről a „hasznosítás” pontban lesz szó. | The focused beam methods of shaping (lithography, milling, deposition), observation, and characterization of nanoscale objects had to be established in the MTA-MFA. After the installation and the basic training period of the new FEG-SEM/FIB cross beam instrument - being unique in Hungary – a combination of systematic training and research had to be carried out. As an example, basic FIB cutting and shaping properties and parameters of materials used in solar cell research (Mo and ZnO) were investigated. In the case of Mo layers strong sputtering anisotropy could be observed, while the ZnO showed uniform sputtering. As a next step FIB feature milling of photonic crystal structures followed. Optimization of FIB and SEM deposition of W nanowires had been carried out. The FIB deposited nanowires showed constant sputtering rate, while thickness of W layers deposited with e-beam of different energy and angle of incidence showed strong influence of secondary electrons. In-situ and ex-situ electrical resistivity measurement of the nanowires had been carried out at RT and elevated temperatures. The FIB deposited layers showed ohmic character,and a irreversible change of resistivity after heat treatment,- due to crystallization of W. The SEM deposited layers showed quasi-linear and nonlinear I-V characteristics. The methodology based on the results of the research contributed to several other research, mentioned in the section „usability”

A quantitative metric for the comparative evaluation of optical clearing protocols for 3D multicellular spheroids

3D multicellular spheroids quickly emerged as in vitro models because they represent the in vivo tumor environment better than standard 2D cell cultures. However, with current microscopy technologies, it is difficult to visualize individual cells in the deeper layers of 3D samples mainly because of limited light penetration and scattering. To overcome this problem several optical clearing methods have been proposed but defining the most appropriate clearing approach is an open issue due to the lack of a gold standard metric. Here, we propose a guideline for 3D light microscopy imaging to achieve single-cell resolution. The guideline includes a validation experiment focusing on five optical clearing protocols. We review and compare seven quality metrics which quantitatively characterize the imaging quality of spheroids. As a test environment, we have created and shared a large 3D dataset including approximately hundred fluorescently stained and optically cleared spheroids. Based on the results we introduce the use of a novel quality metric as a promising method to serve as a gold standard, applicable to compare optical clearing protocols, and decide on the most suitable one for a particular experiment. © 2021 The Author

Stable carbon and nitrogen isotopes identify nuanced dietary changes from the Bronze and Iron Ages on the Great Hungarian Plain

The Great Hungarian Plain (GHP) served as a geographic funnel for population mobility throughout prehistory. Genomic and isotopic research demonstrates non-linear genetic turnover and technological shifts between the Copper and Iron Ages of the GHP, which influenced the dietary strategies of numerous cultures that intermixed and overlapped through time. Given the complexities of these prehistoric cultural and demographic processes, this study aims to identify and elucidate diachronic and culture-specific dietary signatures. We report on stable carbon and nitrogen isotope ratios from 74 individuals from nineteen sites in the GHP dating to a ~ 3000-year time span between the Early Bronze and Early Iron Ages. The samples broadly indicate a terrestrial C 3 diet with nuanced differences amongst populations and through time, suggesting exogenous influences that manifested in subsistence strategies. Slightly elevated δ 15 N values for Bronze Age samples imply higher reliance on protein than in the Iron Age. Interestingly, the Füzesabony have carbon values typical of C 4 vegetation indicating millet consumption, or that of a grain with comparable δ 13 C ratios, which corroborates evidence from outside the GHP for its early cultivation during the Middle Bronze Age. Finally, our results also suggest locally diverse subsistence economies for GHP Scythians