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Abstract High quality label-free imaging of oocytes and early embryos is essential for accurate assessment of their developmental potential, a key element of assisted reproduction procedures. To achieve this goal, we propose full-field optical coherence microscopy (FF-OCM), constructed as a compact module fully integrated with a commercial wide-field fluorescence microscope. Our system achieves optical sectioning in wide-field, high in-plane resolution of 0.5 µm, and high sensitivity to backscattered light. To demonstrate its imaging capabilities, we study live mouse oocytes and embryos at all important stages of meiotic maturation and early embryogenesis. Our system enables visualization of intracellular structures, which are not visible in common bright-field microscopy, i.e., internal structure of nuclear apparatus, cytoskeletal filaments, cellular cortex, cytoplasmic protrusions, or zona pellucida features. Additionally, we visualize and quantify intracellular dynamics like cytoplasmic stirring motion, nuclear envelope fluctuations and nucleolus mobility. Altogether, we demonstrate that FF-OCM is a powerful tool for research in developmental biology that also holds great potential for non-invasive time-lapse monitoring of oocyte and embryo quality in assisted reproduction.

Abstract Cytoplasmic polyadenylation plays a vital role in gametogenesis; however, the participating enzymes and substrates in mammals remain unclear. Using knockout and knock-in mouse models, we describe the essential role of four TENT5 poly(A) polymerases in mouse fertility and gametogenesis. TENT5B and TENT5C play crucial yet redundant roles in oogenesis, with the double knockout of both genes leading to oocyte degeneration. Additionally, TENT5B-GFP knock-in females display a gain-of-function infertility effect, with multiple chromosomal aberrations in ovulated oocytes. TENT5C and TENT5D both regulate different stages of spermatogenesis, as shown by the sterility in males following the knockout of either gene. Finally, Tent5a knockout substantially lowers fertility, although the underlying mechanism is not directly related to gametogenesis. Through direct RNA sequencing, we discovered that TENT5s polyadenylate mRNAs encoding endoplasmic reticulum-targeted proteins essential for gametogenesis. Sequence motif analysis and reporter mRNA assays reveal that the presence of an endoplasmic reticulum-leader sequence represents the primary determinant of TENT5-mediated regulation.

Abstract Profound tuberculosis (TB)-induced metabolic changes reflected in the blood metabolomic profile may provide an opportunity to identify specific markers of Mycobacterium tuberculosis infection. Using targeted liquid chromatography tandem mass spectrometry, we compared the levels of 30 small metabolites, including amino acids and derivatives, and small organic compounds in serum and M.tb antigen-stimulated whole blood cultures of active TB children, latent TB (LTBI) children, nonmycobacterial pneumonia (NMP) children, and healthy controls (HCs) to assess their potential as biomarkers of childhood TB. We found elevated levels of leucine and kynurenine combined with reduced concentrations of citrulline and glutamine in serum and blood cultures of TB and LTBI groups. LTBI status was additionally associated with a decrease in valine levels in blood cultures. The NMP metabolite profile was characterized by an increase in citrulline and glutamine and a decrease in leucine, kynurenine and valine concentrations. The highest discriminatory potential for identifying M.tb infection was observed for leucine detected in serum and kynurenine in stimulated blood cultures. The use of targeted metabolomics may reveal metabolic changes in M.tb-infected children, and the obtained results are a proof of principle of the usefulness of metabolites in the auxiliary diagnosis of TB in children.

Today, endless discourses about Large Panel System (LPS) buildings take place to ask whether the residents should be concerned about their flats being demolished. Taking into account: turning from the "machine for housing" created by Le Corbusier to modern construction, sudden disasters like gas explosion, progressive collapse, planned demolitions, the trend can result in the creation of huge amounts of waste. LPS buildings are usually located in areas of dense urban development. Recycling of these wastes is very problematic. Because concrete load bearing panels seem to be in good condition, there is urgent need to evaluate the possibility of recycling them, e.g. as coarse aggregate in a new concrete mix. Self-Compacting Concrete (SCC) seems promising with its practical feature that it does not have to be vibrated during concreting, which in turn leads to a reduction in noise, making it a perfect material for investments in areas of dense urban development. Such solution would allow to eliminate the transport of waste, and at the same time to reduce it. The aim of the research is to preliminary evaluate the properties of Recycled Coarse Aggregate (RCA), acquired from the Construction and Demolition Waste (C&DW) of LPS residential building. The results of the sieve analysis in the products of 4-8 mm showed little irregular grains content, amounted to 5.9 % of the selected representative sample. Because no study before analysed RCA from LPS, there is a need of further research focusing on properties of acquired aggregate and concrete. The novelty of the manuscript lies in conducting preliminary research towards the recycling LPS for new SCC.

RNA polyadenosine tails are important for the export, translation and stability of mRNAs and play a role in non-coding RNA biogenesis. Here the authors measure yeast poly(A) tail lengths by direct RNA sequencing, revealing its dynamics in yeast exonuclease, deadenylase and poly(A) polymerase mutants.

This paper predicts the abrasion resistance of a cementitious composite containing granite powder and fly ash replacing up to 30% of the cement weight. For this purpose, intelligent artificial neural network (ANN) models were used and compared. A database was built based on mix composition, curing time, and curing method. The model developed to predict the abrasion resistance of the cementitious composites containing granite powder and fly ash was shown to be accurate. It was proved by the very high values of the accuracy parameters that were above 0.93 in the case of the coefficient of the determination R2 and very low values of the errors, which were about 10% in the case of mean average percentage error. This method can be used especially for designing cement mortars with granite powder and fly ash additives replacing cement in a range from 0 to 30% of its weight. These mortars can be used for floors in industrial buildings.

TUTase mediated uridylation of mRNA promotes degradation. Here, Scheer, de Almeida et al. show that Arabidopsis TUTase URT1 interacts directly with the translation inhibitor and decay factor DECAPPING5 and suppresses siRNA biogenesis by preventing accumulation of deadenylated mRNAs

Regulating polyadenylation is important for mRNA stability, which can in turn affect B cell maturation and humoral immune responses. Here the authors use Nanopore poly(A) sequencing to explore the importance of the cytoplasmic poly(A) polymerase TENT5C, particularly in the production of immunoglobulins.

Large panel system (LPS) buildings, during the 1950s–1990s, were the most widespread among precast systems and are still a huge part of some national housing stocks. Disadvantages in characteristics, poor quality of all construction stages, sudden disasters and poor retrofit and repair techniques can result in demolishing some LPS buildings and thus the creation of huge amounts of waste. There is urgent need of evaluating the possibility of reusing the elements of LPS buildings. The novelty of the article is based on the formulation of research gaps in terms of recycling these buildings and the justification of such need. The authors reviewed the current state of knowledge regarding characteristics of LPS buildings, directions of retrofitting, durability and their locations within cities. The possibility of recycling elements of LPS buildings was analysed. The findings are that concrete load bearing panels, proved to be in satisfactory conditions in terms of strength and durability, have the possibility to be recycled into recycled coarse aggregate (RCA), so such waste could be beneficial for concrete. However, there are research gaps of such an application highlighted in the text. The authors suggest that conducting comprehensive basic research in this area could have significant benefits for the next generation of researchers and engineers.

Artykuł ma formę pracy przeglądowej, której celem jest zaprezentowanie historii i dawnych technik uprawy roślin z rodzaju Citrus L. w Europie, w tym w Polsce. Metodyka badawcza polegała na analizie materiałów archiwalnych, obejmujących historyczne traktaty z zakresu ogrodnictwa i rolnictwa. Należą do nich rozprawy powstałe na przestrzeni XVI-XVIII w. m.in. autorstwa: G. B. Ferrariego, J. Commelina, J. van der Groena, J. Ch. Volkamera, J. S. Elsholtza, oraz W. Sierakowskiego, A. P. Jabłonowskiej czy J. K. Kluka. Opisano nowożytne dzieje hodowli roślin cytrusowych w Europie oraz przedstawione w traktatach metody, obejmujące uprawę w gruncie oraz w pojemnikach i sposoby zimowania. Scharakteryzowano także rodzaje wznoszonych w tym celu drewnianych i murowanych oranżerii oraz wykorzystywanych donic.

Summary: Osteoblasts orchestrate bone formation through the secretion of type I collagen and other constituents of the matrix on which hydroxyapatite crystals mineralize. Here, we show that TENT5A, whose mutations were found in congenital bone disease osteogenesis imperfecta patients, is a cytoplasmic poly(A) polymerase playing a crucial role in regulating bone mineralization. Direct RNA sequencing revealed that TENT5A is induced during osteoblast differentiation and polyadenylates mRNAs encoding Col1α1, Col1α2, and other secreted proteins involved in osteogenesis, increasing their expression. We postulate that TENT5A, possibly together with its paralog TENT5C, is responsible for the wave of cytoplasmic polyadenylation of mRNAs encoding secreted proteins occurring during bone mineralization. Importantly, the Tent5a knockout (KO) mouse line displays bone fragility and skeletal hypomineralization phenotype resulting from quantitative and qualitative collagen defects. Thus, we report a biologically relevant posttranscriptional regulator of collagen production and, more generally, bone formation.

Fibre-reinforced polymer materials (FRP) are increasingly used to reinforce structural elements. Due to this, it is possible to increase the load-bearing capacity of polymer, wooden, concrete, and metal structures. In this article, the authors collected all the crucial aspects that influence the behaviour of concrete elements reinforced with FRP. The main types of FRP, their characterization, and their impact on the load-carrying capacity of a composite structure are discussed. The most significant aspects, such as type, number of FRP layers including fibre orientation, type of matrix, reinforcement of concrete columns, preparation of a concrete surface, fire-resistance aspects, recommended conditions for the lamination process, FRP laying methods, and design aspects were considered. Attention and special emphasis were focused on the description of the current research results related to various types of concrete reinforced with FRP composites. To understand which aspects should be taken into account when designing concrete reinforcement with composite materials, the main guidelines are presented in tabular form.

Comparative transcriptional analyses require appropriate and precise normalization. Here we describe a modified transcription run-on (TRO) method, named quantitative TRO (qTRO), that allows quantification of nascent transcription activity. The most critical improvement it introduces is a new standardization method for RNA isolation and hybridization steps, enabling transcription activity quantification and comparative biological analysis. We used this technique with chromatin immunoprecipitation to investigate RNA polymerase I (RNAPI) transcription activity and its rDNA gene profiles in Saccharomyces cerevisiae. We designed a set of new oligonucleotide probes complementary to nascent ribosomal RNA (rRNA) transcripts and standardized their hybridization strength. The qTRO method could be successfully implemented to study RNAPI transcription in response to oxidative stress and in two mutant strains with impaired rRNA synthesis.

To avoid time-consuming, costly, and laborious experimental tests that require skilled personnel, an effort has been made to formulate the depth of wear of fly-ash concrete using a comparative study of machine learning techniques, namely random forest regression (RFR) and gene expression programming (GEP). A widespread database comprising 216 experimental records was constructed from available research. The database includes depth of wear as a response parameter and nine different explanatory variables, i.e., cement content, fly ash, water content, fine and coarse aggregate, plasticizer, air-entraining agent, age of concrete, and time of testing. The performance of the models was judged via statistical metrics. The GEP model gives better performance with R2 and ρ equals 0.9667 and 0.0501 respectively and meet with the external validation criterion suggested in the previous literature. The k-fold cross-validation also verifies the accurateness of the model by evaluating R2, RSE, MAE, and RMSE. The sensitivity analysis of GEP equation indicated that the time of testing is the influential parameter. The results of this research can help the designers, practitioners, and researchers to quickly estimate the depth of wear of fly-ash concrete thus shortening its ecological susceptibilities that push to sustainable and faster construction from the viewpoint of environmentally friendly waste management.

Biopolymeric Chitosan-Carrageenan nanocomposites 66.6–231.82 nm in size containing the chemical fungicide mancozeb (nano CSCRG-M) were synthesized following a green chemistry approach. The physicochemical study of nanoparticles (NPs) was accomplished using a particle size analyzer, SEM and FTIR. TEM exhibited clover leaf-shaped nanoparticles (248.23 nm) with mancozeb on the inside and entrapped outside. Differential scanning calorimetry and TGA thermogravimetry exhibited the thermal behaviour of the nanoform. Nano CSCRG-1.5 at 1.5 ppm exhibited 83.1% inhibition against Alternaria solani in an in vitro study and performed as well as mancozeb (84.6%). Complete inhibition was exhibited in Sclerotinia sclerotiorum at 1.0 and 1.5 ppm with the nanoformulation. The in vivo disease control efficacy of mancozeb-loaded nanoparticles against A. solani in pathogenized plants was found to be relatively higher (79.4 ± 1.7) than that of commercial fungicide (76 ± 1.1%) in pot conditions. Nanomancozeb showed superior efficacy for plant growth parameters, such as germination percentage, root–shoot ratio and dry biomass. The nanoformulation showed higher cell viability compared to mancozeb in Vero cell cultures at 0.25 and 0.50 mg/mL in the resazurin assay. CSCRG-0.5 showed slow-release behavior up to 10 h. Thus, these green nano-based approaches may help combat soil and water pollution caused by harmful chemical pesticides.

Artificial intelligence and machine learning are employed in creating functions for the prediction of self-compacting concrete (SCC) strength based on input variables proportion as cement replacement. SCC incorporating waste material has been used in learning approaches. Artificial neural network (ANN) support vector machine (SVM) and gene expression programming (GEP) consisting of 300 datasets have been utilized in the model to foresee the mechanical property of SCC. Data used in modeling consist of several input parameters such as cement, water–binder ratio, coarse aggregate, fine aggregate, and fly ash (FA) in combination with the superplasticizer. The best predictive models were selected based on the coefficient of determination (R2) results and model validation. Empirical relation with mathematical expression has been proposed using ANN, SVM, and GEP. The efficiency of the models is assessed by permutation features importance, statistical analysis, and comparison between regression models. The results reveal that the proposed machine learning models achieved adamant accuracy and has elucidated performance in the prediction aspect.

FAM46C is one of the most frequently mutated genes in multiple myeloma (MM), but its molecular function remains unknown. Here the authors show that FAM46C is a poly(A) polymerase and that loss of function of FAM46C drives multiple myeloma through the destabilisation of ER response transcripts.

Historical urban parks are particularly valuable in terms of their ecology, as they offer habitats to many valuable flora and fauna. A study was conducted in Felin Manor Park, Lublin, southeastern Poland, dating back to the end of the nineteenth and the first half of twentieth century. The aim of the study was to assess the suitability of this type of urban park for the proecological design of urban green areas that will comply with the legal requirements for historic gardens. The investigation demonstrate that long-living native tree species (e.g., Tilia cordata, Quercus robur, and Fraxinus excelsior) are the best preserved plants in this park. The dendroflora of the park is, however, characterized by a great variety of species flowering between early spring and fall. These reward species ensure continuity of the food base for insects throughout the vegetation season. A floristic inventory facilitated development of a concept for the restoration of the park combining cultural (historical, educational) and ecological functions. Tilia cordata was used for line planting arrangements along the park boundary. It is a component of the alley design along the driveway and is one of the most valuable nectar reward species. The survey allowed the definition of future design guidelines, i.e., the preservation of the historical park structures (formed by long-living native species) and complementation of the park dendroflora with species that are attractive for bees and other insect pollinators. To supplement their food base, it is recommended that a “bee garden” with herbaceous reward plants should be created.

Antimony is a toxic metalloid with poorly understood mechanisms of toxicity and uncertain carcinogenic properties. By using a combination of genetic, biochemical and DNA damage assays, we investigated the genotoxic potential of trivalent antimony in the model organism Saccharomyces cerevisiae. We found that low doses of Sb(III) generate various forms of DNA damage including replication and topoisomerase I-dependent DNA lesions as well as oxidative stress and replication-independent DNA breaks accompanied by activation of DNA damage checkpoints and formation of recombination repair centers. At higher concentrations of Sb(III), moderately increased oxidative DNA damage is also observed. Consistently, base excision, DNA damage tolerance and homologous recombination repair pathways contribute to Sb(III) tolerance. In addition, we provided evidence suggesting that Sb(III) causes telomere dysfunction. Finally, we showed that Sb(III) negatively effects repair of double-strand DNA breaks and distorts actin and microtubule cytoskeleton. In sum, our results indicate that Sb(III) exhibits a significant genotoxic activity in budding yeast.

Polyaniline (PANI) was synthesized chemically with the modified rapid mixing protocol in the presence of sulfuric acid of various concentrations. A two-step synthetic procedure was utilized maintaining low-temperature conditions. Application of the modified rapid mixing protocol allowed obtaining a material with local ordering. A higher concentration of acid allowed obtaining a higher yield of the reaction. Structural characterization performed with Fourier-transform infrared (FTIR) analysis showed the vibration bands characteristic of the formation of the emeraldine salt in both products. Ultraviolet–visible light (UV–Vis) spectroscopy was used for the polaronic band and the p–p* band determination. The absorption result served to estimate the average oxidation level of PANI by comparison of the ratio of the absorbance of the polaronic band to that of the π–π* transition. The absorbance ratio index was higher for PANI synthesized in a more acidic solution, which showed a higher doping level for this polymer. For final powder products, particle size distributions were also estimated, proving that PANI (5.0 M) is characterized by a larger number of small particles; however, these particles can more easily agglomerate and form larger structures. The X-ray diffraction (XRD) patterns revealed an equilibrium between the amorphous and semicrystalline phase in the doped PANI. A higher electrical conductivity value was measured for polymer synthesized in a higher acid concentration. The time-of-flight secondary ion mass spectrometry (TOF-SIMS) analysis showed that the molecular composition of the polymers was the same; hence, the difference in properties was a result of local ordering.