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Summary: Adar null mutant mouse embryos die with aberrant double-stranded RNA (dsRNA)-driven interferon induction, and Adar Mavs double mutants, in which interferon induction is prevented, die soon after birth. Protein kinase R (Pkr) is aberrantly activated in Adar Mavs mouse pup intestines before death, intestinal crypt cells die, and intestinal villi are lost. Adar Mavs Eifak2 (Pkr) triple mutant mice rescue all defects and have long-term survival. Adenosine deaminase acting on RNA 1 (ADAR1) and PKR co-immunoprecipitate from cells, suggesting PKR inhibition by direct interaction. AlphaFold studies on an inhibitory PKR dsRNA binding domain (dsRBD)-kinase domain interaction before dsRNA binding and on an inhibitory ADAR1 dsRBD3-PKR kinase domain interaction on dsRNA provide a testable model of the inhibition. Wild-type or editing-inactive human ADAR1 expressed in A549 cells inhibits activation of endogenous PKR. ADAR1 dsRNA binding is required for, but is not sufficient for, PKR inhibition. Mutating the ADAR1 dsRBD3-PKR contact prevents co-immunoprecipitation, ADAR1 inhibition of PKR activity, and co-localization of ADAR1 and PKR in cells.

Abstract Background Genes, principal units of genetic information, vary in complexity and evolutionary history. Less-complex genes (e.g., long non-coding RNA (lncRNA) expressing genes) readily emerge de novo from non-genic sequences and have high evolutionary turnover. Genesis of a gene may be facilitated by adoption of functional genic sequences from retrotransposon insertions. However, protein-coding sequences in extant genomes rarely lack any connection to an ancestral protein-coding sequence. Results We describe remarkable evolution of the murine gene D6Ertd527e and its orthologs in the rodent Muroidea superfamily. The D6Ertd527e emerged in a common ancestor of mice and hamsters most likely as a lncRNA-expressing gene. A major contributing factor was a long terminal repeat (LTR) retrotransposon insertion carrying an oocyte-specific promoter and a 5′ terminal exon of the gene. The gene survived as an oocyte-specific lncRNA in several extant rodents while in some others the gene or its expression were lost. In the ancestral lineage of Mus musculus, the gene acquired protein-coding capacity where the bulk of the coding sequence formed through CAG (AGC) trinucleotide repeat expansion and duplications. These events generated a cytoplasmic serine-rich maternal protein. Knock-out of D6Ertd527e in mice has a small but detectable effect on fertility and the maternal transcriptome. Conclusions While this evolving gene is not showing a clear function in laboratory mice, its documented evolutionary history in Muroidea during the last ~ 40 million years provides a textbook example of how a several common mutation events can support de novo gene formation, evolution of protein-coding capacity, as well as gene’s demise.

Background The master cast is the gold standard for the control and eventual adjustment of restorations produced by conventional procedures. Some digital workflow bypasses the master cast and relies completely on the precision of the CAD/CAM restoration. Aim To examine the reproducibility of the margins of CAD/CAM restorations generated from a single digital scan. Also, to check the readiness of these restorations for delivery directly after fabrication without adjustment on a master cast and thereby eliminate the need for the master cast. Methods A total of 18 metal substructures made from cobalt chrome alloy were fabricated utilizing a single STL file. The circumference was divided into eight zones. The vertical marginal discrepancy (VMD) was measured at each zone of each metal substructure, with optical microscopy at ×200 magnification. Results Measurements of vertical marginal discrepancy were in a range of (−94: 300) with a mean of 62 ± 60 μm. A one-way ANOVA test revealed that the mean VMD is significantly different among the 18 substructures (F17, 1,134 = 63.948, p < 0.001). Conclusion Although all the received substructures were fabricated from the same scan file, they were not identical and varied widely, and they were going outside the acceptable range in some zones. Within the limitations of this study, the marginal fit can be improved by extraoral adjustments on the master cast. Thus, skipping the master cast deprives the dentist of delivering a restoration of higher quality.

Statement of problem It has been shown that selective etching improves the bond strength of some self-adhesive resin cements to enamel. The same has yet to be determined with dentin pre-treatment. Purpose To evaluate the tensile bond strength of two self-adhesive resin cements after two dentin surface pre-treatments, and also to analyze the cement/dentin interface. Material and Methods One hundred and twelve human third molars were extracted. The teeth were distributed into seven groups (n = 16). Maxcem Elite Chroma (MAX) (Kerr, Scafati, Italy) and Relyx U200 (RLX) (3M ESPE, Neuss, Germany) were used without pre-treatment or with two dentin pre-treatments (polyacrylic acid or phosphoric acid). A conventional etch-and-rinse (EAR) luting cement, NX3 Nexus (NX3) (Kerr, Scafati, Italy), was used as an external control group. Before testing, all specimens were stored in distilled water for 24 hours. Three specimens from each group were prepared for scanning electron microscopy observation (SEM). A tensile bond strength test (TBS) was performed for the remaining samples. The data were statistically analyzed using the Kruskal–Wallis test and Pairwise comparisons using the Wilcoxon rank sum test. Results MAX without pre-treatment and with phosphoric acid etching attained statistically similar bond strengths to NX3 (P > 0.05). There was a statistical difference (P = 0.00488) between RLX without pre-treatment (5.62 MPa) and NX3 (10.88 MPa). Phosphoric acid pre-treatment increases the bond strength values of RLX to a strength that is comparable to NX3 (P > 0.05). The lowest tensile bond strength (TBS) was attained after the application of polyacrylic acid with MAX (1.98 MPa). No statistical differences were found between the RLX bond strength values after polyacrylic acid treatment and RLX without pre-treatment or NX3 (P > 0.05). SEM observations disclosed an enhanced potential of the self-adhesive cements to infiltrate into dentin tubules and form resin tags when applied after phosphoric acid pre-treatment. The failure mode was dominantly adhesive. Conclusions On dentin, the self-adhesive resin cement MAX might be an effective alternative to conventional resin cement. Etching the dentin with phosphoric acid does not have a negative effect on the bond strength of MAX to dentin. On the other hand, phosphoric acid improved the bond strength of RLX when compared to EAR cement.

Italian, annual and perennial ryegrasses are the most important grass species cultivated for seed production in the Czech Republic. The straw remaining after seeds threshing can be used as low quality forage, bedding material or as soil amendment. However, there are no comprehensive information concerning grass straw yields and quality in central Europe. The aim of this study was to investigate the Italian (IR) and perennial (PR) ryegrass straw yield and quality. These parameters were measured for two harvest years at two locations. IR did not provide statistically higher straw yield than PR (7.50 and 7.17 t.ha–1 respectively, p = 0.314), but dry matter (DM) content immediatelly after threshing was higher than for PR (0.42 and 0.38 respectively, p = 0.03). IR straw had the same content of ash and hemicellulose (mean 69.8 and 216 g.kg–1 DM, p = 0.22 and 0.57), but less crude protein (64.1 and 81.4 g.kg–1 DM, p = 0.003), more fibre (399 and 382 g.kg–1 DM, p = 0.02), NDF (656 and 629 g.kg–1 DM, p = 0.03) and ADF (442 and 412 g. kg–1 DM, p = 0.000) when compared with PR straw. Most of the parameters were affected by the year (with exception of crude protein) and by location (with exception of yield, DM content, ash and crude protein). There were no differences in mineral nutrient contents between species. The highest values were found in potassium (19.8 g.kg–1 DM) what is comparable with average quality hay. Phosphorus, calcium and magnesium contents (1.42, 2.96 and 1.02 g.kg–1 DM) were substantially lower than in hay. Data concerning ryegrasses straw yield and quality from the USA are comparable with our results and can be used for European conditions.

This paper deals with use of machine vision for the needs of automated inspections based on virtual instrumentation using a visualization tool named Vision Builder for Automated Inspection (VBAI) – National Instruments (NI). An application for camera tests of dimensions, shapes and presence is designed in experimental parts. This work is focused on the control, detection and distribution properties of internal holes.

Skeletochronology is a widely used method for age determination in amphibians. This method is based mainly on the histological examination of the finger bones. However, the lengths of utilized severed fingers have not been specified in previous studies. The objective of this study was to analyse the structure of line arrested growth (LAGs) involving taking only the last two phalanges of a finger, and using the entire finger of Alpine newts (Mesotriton alpestris). Altogether 432 fingers were taken from four localities in the Czech Republic during the newt breeding period. The first group (group A) contained fingers that consisted of the last two phalanges (330 samples), and the second (group B) contained complete fingers with all phalanges (102 samples). All fingers were processed using standard histological methods and stained with hematoxylin-eosin. Phalange cross-section slides were made, and the ages of the individuals were determined by the number of LAGs. From two phalanges it is determine the age of 17.87% of newts; however, age determination was successful in 49.01% of newts when using whole fingers. Age determination success rate differences between groups were significant (P < 0.001). This is a histological study and it is recommended utilizing whole fingers in future Caudata amphibian screening age studies.Errata to this article were published in: Acta Universitatis Agriculturae et Silviculturae Mendelianae Brunensis, 2015, 63(3): 1079–1079. http://dx.doi.org/10.11118/actaun201563031079

Fluorescence Lifetime Correlation Spectroscopy (FLCS) is a variant of fluorescence correlation spectroscopy (FCS), which uses differences in fluorescence intensity decays to separate contributions of different fluorophore populations to FCS signal. Besides which, FLCS is a powerful tool to improve quality of FCS data by removing noise and distortion caused by scattered excitation light, detector thermal noise and detector afterpulsing. We are providing an overview of, to our knowledge, all published applications of FLCS. Although these are not numerous so far, they illustrate possibilities for the technique and the research topics in which FLCS has the potential to become widespread. Furthermore, we are addressing some questions which may be asked by a beginner user of FLCS. The last part of the text reviews other techniques closely related to FLCS. The generalization of the idea of FLCS paves the way for further promising application of the principle of statistical filtering of signals. Specifically, the idea of fluorescence spectral correlation spectroscopy is here outlined.

Fluorescence correlation spectroscopy (FCS) is a single molecule technique used mainly for determination of mobility and local concentration of molecules. This review describes the specific problems of FCS in planar systems and reviews the state of the art experimental approaches such as 2-focus, Z-scan or scanning FCS, which overcome most of the artefacts and limitations of standard FCS. We focus on diffusion measurements of lipids and proteins in planar lipid membranes and review the contributions of FCS to elucidating membrane dynamics and the factors influencing it, such as membrane composition, ionic strength, presence of membrane proteins or frictional coupling with solid support.