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Anuran saxiphilins (Sxphs) are "toxin sponge" proteins thought to prevent the lethal effects of small-molecule neurotoxins through sequestration. Here, we present a protocol for the expression, purification, and characterization of Sxphs. We describe steps for using thermofluor, fluorescence polarization, and isothermal titration calorimetry assays that probe Sxph:saxitoxin interactions using a range of sample quantities. These assays are generalizable and can be used for other paralytic shellfish poisoning toxin-binding proteins. For complete details on the use and execution of this protocol, please refer to Chen et al. (2022).1.

The purpose of this work was to determine the essence of the influence of the impact velocity (5, 7, and 9 m/s) on Hertzian stresses and the erosion mechanism of ferritic-pearlitic S355J2 steel. The investigations were carried out using a slurry pot tester. S355J2 steel showed a strong sensitivity to changes in impact velocity. A significant increase in erosion rate was observed at a velocity of 9 m/s. This increase was 5-fold and over 15-fold compared to velocities of 7 m/s and 5 m/s, respectively. The study of Hertzian stress is crucial in erosion research because it helps understand how impact energy is absorbed by the eroded material and the mechanisms that cause surface wear. A linear increase in mean contact pressure and maximum shear stress was observed with increasing impact velocity. The mean contact pressure increased from 4.3 GPa to 5.5 GPa and the maximum shear stress increased from 2.0 GPa to 2.5 GPa. The kinetic energy of the solid particles that hits the eroded steel is distributed in the contact area, which leads to various deformations and wear mechanisms. The primary type of deformation was fatigue degradation of the surface layers of the eroded steel. The high kinetic energy of solid particles contributed to the formation of plastic deformations and strongly deformed steel flakes. Higher impact velocities generally result in greater forces and contact stresses on the material surface. This led to the intensification of plastic deformation in the contact areas and increased the Hertzian stresses.

Abstract Cytoplasmic mRNA decay is effected by exonucleolytic degradation in either the 5’ to 3’ or 3’ to 5’ direction. Pervasive terminal uridylation is implicated in mRNA degradation, however, its functional relevance for bulk mRNA turnover remains poorly understood. In this study, we employ genome-wide 3’-RACE (gw3’-RACE) in the model system fission yeast to elucidate the role of uridylation in mRNA turnover. We observe widespread uridylation of shortened poly(A) tails, promoting efficient 5’ to 3’ mRNA decay and ensuring timely and controlled mRNA degradation. Inhibition of this uridylation process leads to excessive deadenylation and enhanced 3’ to 5’ mRNA decay accompanied by oligouridylation. Strikingly we found that uridylation of poly(A) tails and oligouridylation of non-polyadenylated substrates are catalysed by different terminal uridyltransferases Cid1 and Cid16 respectively. Our study sheds new light on the intricate regulatory mechanisms underlying bulk mRNA turnover, demonstrating the role of uridylation in modulating mRNA decay pathways.

Abstract Aging-related biochemical changes in nerve cells lead to dysfunctional synapses and disrupted neuronal circuits, ultimately affecting vital processes such as brain plasticity, learning, and memory. The imbalance between excitation and inhibition in synaptic function during aging contributes to cognitive impairment, emphasizing the importance of compensatory mechanisms. Fear conditioning-related plasticity of the somatosensory barrel cortex, relying on the proper functioning and extensive up regulation of the GABAergic system, in particular interneurons containing somatostatin, is compromised in aging (one-year-old) mice. The present research explores two potential interventions, taurine supplementation, and environmental enrichment, revealing their effectiveness in supporting learning-induced plasticity in the aging mouse brain. They do not act through a mechanism normalizing the Glutamate/GABA balance that is disrupted in aging. Still, they allow for increased somatostatin levels, an effect observed in young animals after learning. These findings highlight the potential of lifestyle interventions and diet supplementation to mitigate age-related cognitive decline by promoting experience-dependent plasticity.

Abstract Condensins play important roles in maintaining bacterial chromatin integrity. In mycobacteria, three types of condensins have been characterized: a homolog of SMC and two MksB-like proteins, the recently identified MksB and EptC. Previous studies suggest that EptC contributes to defending against foreign DNA, while SMC and MksB may play roles in chromosome organization. Here, we report for the first time that the condensins, SMC and MksB, are involved in various DNA transactions during the cell cycle of Mycobacterium smegmatis (currently named Mycolicibacterium smegmatis). SMC appears to be required during the last steps of the cell cycle, where it contributes to sister chromosome separation. Intriguingly, in contrast to other bacteria, mycobacterial MksB follows replication forks during chromosome replication and hence may be involved in organizing newly replicated DNA.

Abstract Alterations in miRNA levels have been observed in various types of cancer, impacting numerous cellular processes and increasing their potential usefulness in combination therapies also in brain tumors. Recent advances in understanding the genetics and epigenetics of brain tumours point to new aberrations and associations, making it essential to continually update knowledge and classification. Here we conducted molecular analysis of 123 samples of childhood brain tumors (pilocytic astrocytoma, medulloblastoma, ependymoma), focusing on identification of genes that could potentially be regulated by crucial representatives of OncomiR-1: miR-17-5p and miR-20a-5p. On the basis of microarray gene expression analysis and qRTPCR profiling, we selected six (WEE1, CCND1, VEGFA, PTPRO, TP53INP1, BCL2L11) the most promising target genes for further experiments. The WEE1, CCND1, PTPRO, TP53INP1 genes showed increased expression levels in all tested entities with the lowest increase in the pilocytic astrocytoma compared to the ependymoma and medulloblastoma. The obtained results indicate a correlation between gene expression and the WHO grade and subtype. Furthermore, our analysis showed that the integration between genomic and epigenetic pathways should now point the way to further molecular research.

The current study presents the results of the chemical and the radiochemical characterization of phosphogypsum obtained from the former Wizów Chemical Plant in Poland. Phosphogypsum is a residue obtained from phosphoric acid production. Phosphogypsum mainly contains calcium sulfate dihydrate and impurities originating from the source phosphate rock, including toxic trace elements and natural radionuclides. The phosphogypsum stacks occupy a large area and are an environmental issue today. The project “Phosphogypsum Processing to Critical Raw Materials”, currently realized at the Institute of Nuclear Chemistry and Technology, focuses on the use of phosphogypsum from the Polish stacks as material to recover rare earth elements (REEs). The remaining gypsum matrix can be used as an inexpensive material in construction. The chemical and radiological characterization of the radioactive isotopes present in phosphogypsum was performed by using inductively coupled plasma mass spectrometry (ICP-MS) and the gamma spectrometry technique. The radioactivity of the Polish phosphogypsum was then compared with the radioactivity of the phosphogypsum waste present worldwide.

BackgroundChemoresistance of cancer cells, resulting from various mechanisms, is a significant obstacle to the effectiveness of modern cancer therapies. Targeting fibroblast growth factors (FGFs) and their receptors (FGFRs) is becoming crucial, as their high activity significantly contributes to cancer development and progression by driving cell proliferation and activating signaling pathways that enhance drug resistance.MethodsWe investigated the potential of honokiol and FGF ligand trap in blocking the FGF1/FGFR1 axis to counteract drug resistance. Using PEAQ-ITC, we verified direct interaction of honokiol with the FGFR1 kinase domain. We then demonstrated the effect of FGF1/FGFR1 inhibition on taltobulin resistance in cells expressing FGFR1. Finally, we generated drug-resistant clones by prolonged exposure of cells with negligible FGFR levels to taltobulin alone, taltobulin and honokiol, or taltobulin and FGF ligand trap.ResultsWe demonstrated for the first time a direct interaction of honokiol with the FGFR1 kinase domain, resulting in inhibition of downstream signaling pathways. We revealed that both honokiol and FGF ligand trap prevent FGF1-dependent protection against taltobulin in cancer cells expressing FGFR1. In addition, we showed that cells obtained by long-term exposure to taltobulin are resistant to both taltobulin and other microtubule-targeting drugs, and exhibit elevated levels of FGFR1 and cyclin D. We also found that the presence of FGF-ligand trap prevents the development of long-term resistance to taltobulin.ConclusionOur results shed light on how blocking the FGF1/FGFR1 axis by honokiol and FGF ligand trap could help develop more effective cancer therapies, potentially preventing the emergence of drug-resistant relapses.

The aim of this work was to develop and validate of method for the determination of topiramate (TPM) by a high-performance liquid chromatographic method coupled to triple-quadrupole mass spectrometry (LC-MS/MS) in MRM mode in the human plasma, saliva, and hair, and implementation of this method for the determination of TPM in patients with epilepsy. Saliva may as an alternative matrix for monitoring drug level, and hair drug content may be a reliable biomarker of the history of drug exposure, allowing to assess patient long-term compliance. Chromatographic separation was achieved in 3 min on a Kinetex analytical column (5 μm C18 100 Å, 100×2.1 mm) using an isocratic elution of acetonitrile and 10 mM ammonium acetate at a ratio of 80:20 (v/v) and a flow rate of 0.2 mL/min. Detection of TPM and internal standard (IS) (TPM-d12) was performed in negative ion mode (ESI−). The following transitions were used m/z 338 → 77.90; 338 → 95.90 for TPM and 350.3 → 78.20 for IS. The method showed to be selective, accurate, precise, and linear for TPM over the concentration ranges of 0.20-30 μg/mL (plasma, saliva), and 5.0-500.0 ng/mL (hair). The simple and robust LC-MS/MS method was successfully applied for the determination of TPM in patients with epilepsy.

Abstract Merkel cell polyomavirus (MCPyV) is associated with Merkel cell carcinoma (MCC). In tumor cells the MCPyV large T antigen (LT-Ag) is frequently found truncated and this is considered a major tumor-specific signature. The role of MCPyV in other, non-MCC tumours, is little known. Viral DNA and/or tumour-specific mutations have been sometimes detected in different tumours, but such data are not unequivocal and the involvement of the virus in the tumorigenesis is not clear. In a previous study, we demonstrated a significantly higher prevalence of MCPyV DNA in formalin fixed paraffin embedded (FFPE) porocarcinoma tissues compared to the normal skin. In the present study, we investigated the presence of truncating mutations in MCPyV LT-Ag coding region in porocarcinoma specimens. Using several overlapped PCR primer pairs, the complete LT-Ag sequence from two biopsies were obtained. No truncating mutations were detected. The lack of truncating mutations in LT-Ag sequence does not seem to support the role of MCPyV in porocarcinoma oncogenesis. However, an oncogenetic mechanism, different from that proposed for MCC and not associated with the LT-Ag mutations/deletions, cannot be excluded. Further studies of more sequences coding for LT-Ag would be needed to verify this hypothesis.

Abstract Galectins constitute a class of lectins that specifically interact with β-galactoside sugars in glycoconjugates and are implicated in diverse cellular processes, including transport, autophagy or signaling. Since most of the activity of galectins depends on their ability to bind sugar chains, galectins exert their functions mainly in the extracellular space or at the cell surface, which are microenvironments highly enriched in glycoconjugates. Galectins are also abundant inside cells, but their specific intracellular functions are largely unknown. Here we report that galectin-1, -3, -7 and -8 directly interact with the proteinaceous core of fibroblast growth factor 12 (FGF12) in the cytosol and in nucleus. We demonstrate that binding of galectin-1 to FGF12 in the cytosol blocks FGF12 secretion. Furthermore, we show that intracellular galectin-1 affects the assembly of FGF12-containing nuclear/nucleolar ribosome biogenesis complexes consisting of NOLC1 and TCOF1. Our data provide a new link between galectins and FGF proteins, revealing an unexpected glycosylation-independent intracellular interplay between these groups of proteins.

IntroductionDisgust sensitivity to body odors plays a role in a set of psychological mechanisms supposedly evolved to avoid pathogens. To assess individual differences in body odor disgust, we previously developed the body odor disgust scale (BODS) and validated it in English. The BODS presents six scenarios where disgust could be evoked by smells coming from an internal source and an external source. The present study aimed to validate the BODS in the Italian population and to find further evidence for its structural, construct, and criterion validity.MethodsWe used two large samples (N = 1,050, F = 527; and N = 402, F = 203, respectively) that were representative of the Italian population for sex and age.ResultsAcross these two studies, we confirmed the hypothesized bifactor structure, with all the items loading onto a general body odor disgust sensitivity factor, and on two specific factors related to the internal structure. In terms of construct validity, we found that the BODS converged with pathogen disgust sensitivity of the three-domain disgust scale (TDDS) but was distinct from a general propensity to experience negative emotions. The BODS showed criterion validity in predicting the behavioral intentions toward COVID-19 avoidance behavior, although it did not seem to be incrementally valid when compared to the TDDS pathogen subscale. We also established scalar measurement invariance of the BODS regarding gender and found that women display higher levels of BODS.DiscussionResults from the Italian version of the BODS indicate its structural, construct, nomological and criterion validity. Furthermore, our result on sex differences in disgust sensitivity are consistent with previous literature, and we discuss them in the broader context of cross-cultural and primate findings that points toward a possible evolutionary explanation of this difference.

American bullfrog (Rana castesbeiana) saxiphilin (RcSxph) is a high-affinity "toxin sponge" protein thought to prevent intoxication by saxitoxin (STX), a lethal bis-guanidinium neurotoxin that causes paralytic shellfish poisoning (PSP) by blocking voltage-gated sodium channels (NaVs). How specific RcSxph interactions contribute to STX binding has not been defined and whether other organisms have similar proteins is unclear. Here, we use mutagenesis, ligand binding, and structural studies to define the energetic basis of Sxph:STX recognition. The resultant STX "recognition code" enabled engineering of RcSxph to improve its ability to rescue NaVs from STX and facilitated discovery of 10 new frog and toad Sxphs. Definition of the STX binding code and Sxph family expansion among diverse anurans separated by ∼140 My of evolution provides a molecular basis for understanding the roles of toxin sponge proteins in toxin resistance and for developing novel proteins to sense or neutralize STX and related PSP toxins.

Primary intracranial melanoma is a very rare brain tumor, especially when accompanied by benign intramedullary melanocytoma. Distinguishing between a primary central nervous system (CNS) lesion and metastatic melanoma is extremely difficult, especially when the primary cutaneous lesion is not visible. Here we report a 13-year-old girl admitted to the Neurosurgery Department of the Institute of Polish Mother’s Health Centre in Lodz due to upper limb paresis. An intramedullary tumor of the cervical C3–C4 and an accompanying syringomyelic cavity C1–C7 were revealed. The child underwent partial removal of the tumor due to the risk of damage to spinal cord motor centers. The removed part of the tumor was diagnosed as melanocytoma. Eight months later, a neurological examination revealed paresis of the right sixth cranial nerve, accompanied by bilateral optic disc edema. Diagnostic imaging revealed a brain tumor. The girl underwent resection of both detected the tumors and an additional satellite lesion revealed during the surgery. The removed tumors were diagnosed as malignant melanomas in pathomorphological examination. Molecular analysis revealed NRASQ61K mutation in both the intracranial and the intramedullary tumor. It should be noted that in cases where available evidence is inconclusive, an integrative diagnostic process is essential to reach a definitive diagnosis.

This study's aim is to determine the consistency of student opinions regarding the importance of individual skills in three areas--people, practice, and perspective--with expert assessment. The study group consisted of five-year students accredited by IPMA Poland. The team of experts was comprised of recruiters, project management lecturers, assessors, and information technology (IT) industry project managers. Our research shows divergences in the approach to key competence that a project manager should have. This is understandable, given the specifics of the comparison. However, the analysis that has been conducted and presented will allow the curricula to be revised. The subsequent evaluation of curricula should address the changes in the labor market. Education should provide key competences, especially as modern project management requires a full spectrum of competences and approaches. Besides experience, project management is the most frequently pursued competence that is required in terms of staff recruitment criteria. The research resulting figures may be useful for recruiters, certification institutions, and universities.

Aims: We examined the cardiovascular effects of celiac disease (CeD) in a humanized mouse model, with a focus on vascular inflammation, endothelial dysfunction, and oxidative stress. Methods and results: NOD.DQ8 mice genetically predisposed to CeD were subjected to a diet regime and oral gavage to induce the disease (gluten group vs. control). We tested vascular function, confirmed disease indicators, and evaluated inflammation and oxidative stress in various tissues. Plasma proteome profiling was also performed.CeD markers were confirmed in the gluten group, indicating increased blood pressure and impaired vascular relaxation. Pro-inflammatory genes were upregulated in this group, with increased CD11b+ myeloid cell infiltration and oxidative stress parameters observed in aortic and heart tissue. However, heart function remained unaffected. Plasma proteomics suggested the cytokine interleukin-17A (IL-17A) as a link between gut and vascular inflammation. Cardiovascular complications were reversed by adopting a gluten-free diet. Conclusion: Our study sheds light in the heightened cardiovascular risk associated with active CeD, revealing a gut-to-cardiovascular inflammatory axis potentially mediated by immune cell infiltration and IL-17A. These findings augment our understanding of the link between CeD and cardiovascular disease providing clinically relevant insight into the underlying mechanism. Furthermore, our discovery that cardiovascular complications can be reversed by a gluten-free diet underscores a critical role for dietary interventions in mitigating cardiovascular risks associated with CeD.

Objective: The main objective of the study was to assess the level of Agile implementation in organisations in Poland and to determine the most commonly used Agile methods. The intention of this paper is to present the results of empirical research on technological tools supporting agile organisations in Poland and consider which digital competencies may favour their use. Research Design & Methods: This paper presents a review of the literature and the results of empirical research on the use of information technology in enterprises which follow Agile practices and approaches. Empirical research is based on an interview questionnaire, which contained questions about the level of Agile implementation in the organisation, Agile implementation and support areas, support factors, barriers to implementation, and implementation results. Findings: The paper presents the main IT tools used in agile enterprises and the results of research conducted in this field. The study shows that in 2019 companies mainly used the Agile approach for the creation of innovative services and (or) products. Research also indicates that, when it comes to Agile Good Practices, respondents most often use Scrum and Kanban. Practices such as DevOps, TDD, or SAFe were much less frequently used by the companies surveyed. Implications / Recommendations: The results of the research provide valuable information on three issues: implementation of the Agile approach in Polish enterprises, assessment of the use of technological bricks in Polish enterprises, and an analysis of good practices in Polish enterprises. The research also indicates directions for further research. Among others, it is recommended to repeat the research on the level of agility of the organisation in Poland, primarily to check whether the level of implementation has changed and whether the implementation has taken place in other key areas of the company. Contribution: Literature review and economic practice indicate that there is increasing popularity of the use of information technologies, not only in agile organisations but in general, requiring employees and managers to improve their digital competencies. This paper addresses the need to evaluate the current state of Agile implementation in the organisation and examines the use of technological bricks in Agile in both theoretical and empirical ways.

Summary: Anuran saxiphilins (Sxphs) are “toxin sponge” proteins thought to prevent the lethal effects of small-molecule neurotoxins through sequestration. Here, we present a protocol for the expression, purification, and characterization of Sxphs. We describe steps for using thermofluor, fluorescence polarization, and isothermal titration calorimetry assays that probe Sxph:saxitoxin interactions using a range of sample quantities. These assays are generalizable and can be used for other paralytic shellfish poisoning toxin-binding proteins.For complete details on the use and execution of this protocol, please refer to Chen et al. (2022).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Abstract Background NOL12 5′-3′ exoribonucleases, conserved among eukaryotes, play important roles in pre-rRNA processing, ribosome assembly and export. The most well-described yeast counterpart, Rrp17, is required for maturation of 5.8 and 25S rRNAs, whereas human hNOL12 is crucial for the separation of the large (LSU) and small (SSU) ribosome subunit rRNA precursors. Results In this study we demonstrate that plant AtNOL12 is also involved in rRNA biogenesis, specifically in the processing of the LSU rRNA precursor, 27S pre-rRNA. Importantly, the absence of AtNOL12 alters the expression of many ribosomal protein and ribosome biogenesis genes. These changes could potentially exacerbate rRNA biogenesis defects, or, conversely, they might stem from the disturbed ribosome assembly caused by delayed pre-rRNA processing. Moreover, exposure of the nol12 mutant to stress factors, including heat and pathogen Pseudomonas syringae, enhances the observed molecular phenotypes, linking pre-rRNA processing to stress response pathways. The aberrant rRNA processing, dependent on AtNOL12, could impact ribosome function, as suggested by improved mutant resistance to ribosome-targeting antibiotics. Conclusion Despite extensive studies, the pre-rRNA processing pathway in plants remains insufficiently characterized. Our investigation reveals the involvement of AtNOL12 in the maturation of rRNA precursors, correlating this process to stress response in Arabidopsis. These findings contribute to a more comprehensive understanding of plant ribosome biogenesis.

Chitosan is a very promising material for tissue model printing. It is also known that the introduction of chemical modifications to the structure of the material in the form of methacrylate groups makes it very attractive for application in the bioprinting of tissue models. The aim of this work is to study the characteristics of biomaterials containing chitosan (BCH) and its methacrylated equivalent (BCM) in order to identify differences in their usefulness in 3D bioprinting technology. It has been shown that the BCM material containing methacrylic chitosan is three times more viscous than its non-methacrylated BCH counterpart. Additionally, the BCM material is characterized by stability in a larger range of stresses, as well as better printability, resolution, and fiber stability. The BCM material has higher mechanical parameters, both mechanical strength and Young’s modulus, than the BCH material. Both materials are ideal for bioprinting, but BCM has unique rheological properties and significant mechanical resistance. In addition, biological tests have shown that the addition of chitosan to biomaterials increases cell proliferation, particularly in 3D-printed models. Moreover, modification in the form of methacrylation encourages reduced toxicity of the biomaterial in 3D constructs. Our investigation demonstrates the suitability of a chitosan-enhanced biomaterial, specifically methacrylate-treated, for application in tissue engineering, and particularly for tissues requiring resistance to high stress, i.e., vascular or cartilage models.

Parvovirus B19 (B19V) is a human pathogen belonging to the Parvoviridae family. It is widely diffused in the population and responsible for a wide range of diseases, diverse in pathogenetic mechanisms, clinical course, and severity. B19V infects and replicates in erythroid progenitor cells (EPCs) in the bone marrow leading to their apoptosis. Moreover, it can also infect, in an abortive manner, a wide set of different cell types, normally non-permissive, and modify their normal physiology. Differences in the characteristics of virus–cell interaction may translate into different pathogenetic mechanisms and clinical outcomes. Joint involvement is a typical manifestation of B19V infection in adults. Moreover, several reports suggest, that B19V could be involved in the pathogenesis of some autoimmune rheumatologic diseases such as rheumatoid arthritis (RA), juvenile idiopathic arthritis (JIA), systemic sclerosis (SSc), systemic lupus erythematosus (SLE), or vasculitis. This review provides basic information on the B19 virus, highlights characteristics of viral infection in permissive and non-permissive systems, and focuses on recent findings concerning the pathogenic role of B19V in rheumatologic diseases.

In recent years, there has been considerable focus on ensuring that energy is used in the most efficient manner possible. This is due to the fact that globally, over 70% of energy is generated from fossil fuels. Consequently, the matter of designing and utilizing materials that will negate energy losses within the construction industry is of paramount importance. Simultaneously, the necessity for a sustainable approach to the design and production of materials is strongly emphasized. This paper presents an innovative approach to the use of a combination of mineral and plant-based fillers in polyurethane foam technology as a thermal insulation material with the potential to be used in construction to reduce energy consumption. Polyurethane composites containing fly ash from biomass combustion and the addition of rice, sunflower, and buckwheat husks as plant fillers were proposed. The structure of the obtained materials was studied, and the most important physical properties were analyzed. These included apparent density, dimensional stability, water absorption, and the effects of UV radiation and water influence on the carbon, hydrogen, nitrogen, and oxygen content. Moreover, the mechanical properties of the materials were investigated, including compressive strength and brittleness. Additionally, the foams were subjected to flammability tests using a cone calorimeter. Furthermore, additional parameters were determined, including the limiting oxygen index and the vertical and horizontal flammability tests. The results demonstrate the beneficial effects of combining mineral and vegetable fillers in polyurethane foam.