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The paper presents the study of microstructure and corrosion resistance of composite coatings (Al+Al2O3 and Cu+Al2O3) deposited by Low Pressure Cold Spraying method (LPCS). The atmospheric corrosion resistance was examined by subjecting the samples to cyclic salt spray and Kesternich test chambers, with NaCl and SO2 atmospheres, respectively. The selected tests allowed reflecting the actual working conditions of the coatings. The analysis showed very satisfactory results for copper coatings. After eighteen cycles, with a total time of 432 hours, the samples show little signs of corrosion. Due to their greater susceptibility to chloride ions, aluminium coatings have significant corrosion losses.

Pearlitic steels containing from some 0,8 to 0,95% C belong to the group of unalloyed steels intended for cold drawing or rolling. One of the problems discussed in literature is cracking of pearlitic steel subjected to plastic working, caused by high brittleness of the lamellar precipitations of hard cementite. This issue is extremely important because it affects significantly reduce fatigue strength. The paper presents proposals to modify the process of heat treatment, results in getting a steel with spheroidal structure characterized by better plastic properties, in order to eliminate this problem.

Thermal spraying methods are commonly used to regenerate damaged surface or change materials surface properties. One of the newest methods is cold spraying, where coating is deposited of material in the solid state. Therefore shape and size of the powder particles are very important parameters. The article presents the influence of copper powder morphology on mechanical properties of the coatings (adhesion, hardness, Young's modulus) deposited with the Low Pressure Cold Spraying method on the AA1350 aluminium alloy substrate. The coatings were deposited using two commercially available copper powders with spherical and dendritic morphology and granulation of -40+10 µm. The bond strength of coatings was determined with the pull off method, the hardness with the Vickers method at load of 2.94 N, while the Young's modulus through measurement of nanoindentation. Microstructure of the coatings was analysed using the light and scanning electron microscopy (SEM). Shape of the powder influences mechanical properties of the coating significantly. The coatings deposited with dendritic powder had low mechanical properties, hardness of the 81 HV0.3 order and adhesion of about 4 MPa. However changing powder morphology to spherical increased hardness of the coating to 180 HV0.3 and adhesion to 38.5 MPa. [ABSTRACT FROM AUTHOR]

Four types of nonvoltage-activated potassium channels in the body-wall muscles of Drosophila third instar larvae have been identified by the patch-clamp technique. Using the inside-out configuration, tetraethylammonium (TEA), Ba2+, and quinidine were applied to the cytoplasmic face of muscle membranes during steady-state channel activation. The four channels could be readily distinguished on the basis of their pharmacological sensitivities and physiological properties. The KST channel was the only type that was activated by stretch. It had a high unitary conductance (100 pS in symmetrical 130/130 mM KCl solution), was blocked by TEA (Kd approximately 35 mM), and was the most sensitive to Ba2+ (complete block at 10(-4) M). A Ca(2+)-activated potassium channel, KCF.72 pS (130/130) mM KCl), was gated open at greater than 10(-8) m Ca2+, was the least sensitive to Ba2+ Kd of approximately 3 mM) and TEA (Kd of approximately 100 mM), and was not affected by quinidine. K2 was a small conductance channel of 11 pS (130/2 KCl, pipette/bath), and was very sensitive to quinidine, being substantially blocked at 0.1 mM. It also exhibited a half block at approximately 0.3 mM Ba2+ and approximately 25 mM TEA. A fourth channel type, K3, was the most sensitive to TEA (half block less than 1 mM). It displayed a partial block to Ba2+ at 10 mM, but no block by 0.1 mM quinidine. The blocking effects of TEA, Ba2+ and quinidine were reversible in all channels studied. The actions of TEA and Ba2+ appeared qualitatively different: in all four channels, TEA reduced the apparent unitary conductance, whereas Ba2+ decreased channel open probability. [ABSTRACT FROM AUTHOR]

Heme oxygenase-1 (HO-1, HMOX1) degrades heme protecting cells from heme-induced oxidative damage. Beyond its well-established cellular functions, heme has emerged as a stabilizer of G-quadruplexes. These secondary DNA structures interfere with DNA replication. We recently revealed that nuclear HO-1 colocalizes with DNA G-quadruplexes and promotes their removal. Here, we investigate whether HO-1 safeguards cells against replication stress. Experiments were conducted in control and HMOX1-deficient HEK293T cell lines. Immunostaining unveiled that DNA G-quadruplexes accumulated in the absence of HO-1, the effect that was further enhanced in response to δ-aminolevulinic acid (ALA), a substrate in heme synthesis. This was associated with replication stress, as evidenced by an elevated proportion of stalled forks analyzed by fiber assay. We observed the same effects in hematopoietic stem cells isolated from Hmox1 knockout mice and in a lymphoblastoid cell line from an HMOX1-deficient patient. Interestingly, in the absence of HO-1, the speed of fork progression was higher, and the response to DNA conformational hindrance less stringent, indicating dysfunction of the PARP1-p53-p21 axis. PARP1 activity was not decreased in the absence of HO-1. Instead, we observed that HO-1 deficiency impairs the nuclear import and accumulation of p53, an effect dependent on the removal of excess heme. We also demonstrated that administering ALA is a more specific method for increasing intracellular free heme compared to treatment with hemin, which in turn induces strong lipid peroxidation. Our results indicate that protection against replication stress is a universal feature of HO-1, presumably contributing to its widely recognized cytoprotective activity., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Transcriptional regulatory networks (TRNs) associated with recombinant protein (rProt) synthesis in Yarrowia lipolytica are still under-described. Yet, it is foreseen that skillful manipulation with TRNs would enable global fine-tuning of the host strain's metabolism towards a high-level-producing phenotype. Our previous studies investigated the transcriptomes of Y. lipolytica strains overproducing biochemically different rProts and the functional impact of transcription factors (TFs) overexpression (OE) on rProt synthesis capacity in this species. Hence, much knowledge has been accumulated and deposited in public repositories. In this study, we combined both biological datasets and enriched them with further experimental data to investigate an interplay between TFs and rProts synthesis in Y. lipolytica at transcriptional and functional levels. Technically, the RNAseq datasets were extracted and re-analyzed for the TFs' expression profiles. Of the 140 TFs in Y. lipolytica , 87 TF-encoding genes were significantly deregulated in at least one of the strains. The expression profiles were juxtaposed against the rProt amounts from 125 strains co-overexpressing TF and rProt. In addition, several strains bearing knock-outs (KOs) in the TF loci were analyzed to get more insight into their actual involvement in rProt synthesis. Different profiles of the TFs' transcriptional deregulation and the impact of their OE or KO on rProts synthesis were observed, and new engineering targets were pointed.

Objectives : Even though BPPV is one of the most common causes of vertigo, it is often underdiagnosed and omitted in the diagnosis of patients reporting vertigo. The aim of the study was to establish a diagnostic pattern useful in patients admitted due to vertigo, based on the most common clinical characteristics of patients suffered from posterior canal BPPV (PC-BPPV), horizontal canal BPPV with geotropic (HCG-BPPV) and apogeotropic nystagmus (HCA-BPPV). Methods : The analysis covered the results obtained in 105 patients with a positive result of the Dix-Hallpike maneuver or the supine roll test. The patients were divided into 3 groups based on the BPPV type: gr.1:PC-BPPV (60%); gr.2: HCG-BPPV (27%); gr.3: HCA-BPPV (13%). Patients before the diagnostic maneuvers filled the questionnaire concerning their symptoms and previous diseases. Results : Almost all patients had vertigo during turning over in bed and the character of the symptoms was paroxysmal. The answers to questions about the type of head movement evoked vertigo and how long vertigo lasted were differentiating. The percentages of correct diagnosis speculated by the combined answers were 69.6% in PC-BPPV, 61.8% in HCG-BPPV, and 80% in HCA-BPPV. Conclusions : Basing on those observations there is presented the diagnostic schedule which could be useful in dizziness examination. The above results indicate that a properly collected interview with the patient allows for a high percentage of accurate diagnosis.

With the current progress in the 'design' and 'build' stages of the 'design-build-test-learn' cycle, many synthetic biology projects become 'test-limited'. Advances in the parallelization of microbes cultivations are of great aid, however, for many species down-scaling leaves a metabolic footprint. Yarrowia lipolytica is one such demanding yeast species, for which scaling-down inevitably leads to perturbations in phenotype development. Strictly aerobic metabolism, propensity for filamentation and adhesion to hydrophobic surfaces, spontaneous flocculation, and high acidification of media are just several characteristics that make the transfer of the micro-scale protocols developed for the other microbial species very challenging in this case. It is well recognized that without additional 'personalized' optimization, either MTP-based or single-cell-based protocols are useless for accurate studies of Y. lipolytica phenotypes. This review summarizes the progress in the scaling-down and parallelization of Y. lipolytica cultures, highlighting the challenges that occur most frequently and strategies for their overcoming. The problem of Y. lipolytica cultures down-scaling is illustrated by calculating the costs of micro-cultivations, and determining the unintentionally introduced, thus uncontrolled, variables. The key research into culturing Y. lipolytica in various MTP formats and micro- and pico-bioreactors is discussed. Own recently developed and carefully pre-optimized high-throughput cultivation protocol is presented, alongside the details from the optimization stage. We hope that this work will serve as a practical guide for those working with Y. lipolytica high-throughput screens., (© 2024. The Author(s).)

Background: In the era of rationally designed synthetic biology, heterologous metabolites production, and other counter-nature engineering of cellular metabolism, we took a step back and recalled that 'Mother(-Nature) knows best'. While still aiming at synthetic, non-natural outcomes of generating an 'over-production phenotype' we dug into the pre-designed transcriptional programs evolved in our host organism-Yarrowia lipolytica, hoping that some of these fine-tuned orchestrated programs could be hijacked and used. Having an interest in the practical outcomes of the research, we targeted industrially-relevant functionalities-stress resistance and enhanced synthesis of proteins, and gauged them over extensive experimental design's completion., Results: Technically, the problem was addressed by screening a broad library of over 120 Y. lipolytica strains under 72 combinations of variables through a carefully pre-optimized high-throughput cultivation protocol, which enabled actual phenotype development. The abundance of the transcription program elicitors-transcription factors (TFs), was secured by their overexpression, while challenging the strains with the multitude of conditions was inflicted to impact their activation stratus. The data were subjected to mathematical modeling to increase their informativeness. The amount of the gathered data prompted us to present them in the form of a searchable catalog - the YaliFunTome database ( https://sparrow.up.poznan.pl/tsdatabase/ )-to facilitate the withdrawal of biological sense from numerical data. We succeeded in the identification of TFs that act as omni-boosters of protein synthesis, enhance resistance to limited oxygen availability, and improve protein synthesis capacity under inorganic nitrogen provision., Conclusions: All potential users are invited to browse YaliFunTome in the search for homologous TFs and the TF-driven phenotypes of interest., (© 2024. The Author(s).)

Controllable regulatory elements, like inducible, titratable promoters, are highly desired in synthetic biology toolboxes. A set of previously developed erythritol-inducible promoters along with an engineered Yarrowia lipolytica host strain were shown to be a very potent expression platform. In this study, we push the previously encountered limits of the synthetic promoters' titratability (by the number of upstream motifs) by using a compatible transcription factor, Euf1, as the promoter titrator. Overexpression of spliced EUF1 turned out to be very efficient in promoting expression from the compatible promoter, however, the erythritol-inducible character of the promoter was then lost. Analysis of the EUF1's splicing pattern suggests that the intron removal is promoted in the presence of erythritol, but is not dependent on it. The 3D structures of spliced versus unspliced Euf1 were modeled, and ligand-binding strength was calculated and compared. Furthermore, the EUF1-dependent expression profile under different chemical stimulants was investigated. Depletion of carbon source was identified as the significant factor upregulating the expression from the Euf1-dependent promoter (2-10-fold). Considering these findings and transcriptomics data, a new mechanism of the Euf1-regulated promoter action is proposed, involving a 'catabolite repression' transcription factor-Adr1, both acting on the same ERY-inducible promoter., (© The Author(s) 2024. Published by Oxford University Press on behalf of FEMS.)

Hearing loss is one of the most common causes of disability worldwide. The aim of the study was to compare the demographic structure and the results of hearing tests in people qualified for hearing aids over the last 25 years. The material covered 1246 patients qualified for hearing aids in the years 1996-2001 and 2016-2021. Patients were divided into two groups according to the time of qualifying for hearing aids. Group 1 (G1) consisted of 759 people qualified in the years 1996-2001, and Group 2 (G2) comprised 487 people qualified in the years 2016-2021. Statistical analysis was performed on the results of pure tone threshold audiometry and the demographic structure in both groups. Patients in G1 had statistically significantly elevated hearing thresholds (HT) in the air conduction range at frequencies from 2000 to 8000 Hz in relation to G2 patients. The opposite situation was observed for the bone conduction threshold. G2 patients had significantly elevated bone conduction HT at frequencies from 250 to 1000 Hz compared to G1 patients. The age structure in both groups was similar; however, the gender distribution was statistically significantly different. In G1 women accounted for 40%, and in G2, they became the dominant gender (53%). Over the last twenty years, there has been a change in the structure of patients qualified for hearing aids. Although the age has remained similar, today, patients decide to use hearing aids at an earlier stage of hearing loss than 25 years ago. Modern women began to use hearing aids much more often.

Resistance to environmental stress and synthesis of recombinant proteins (r-Prots) are both complex, strongly interconnected biological traits relying on orchestrated contribution of multiple genes. This, in turn, makes their engineering a challenging task. One of the possible strategies is to modify the operation of transcription factors (TFs) associated with these complex traits. The aim of this study was to examine the potential implications of selected five TFs (HSF1-YALI0E13948g, GZF1-YALI0D20482g, CRF1-YALI0B08206g, SKN7-YALI0D14520g, and YAP-like-YALI0D07744g) in stress resistance and/or r-Prot synthesis in Yarrowia lipolytica. The selected TFs were over-expressed or deleted (OE/KO) in a host strain synthesizing a reporter r-Prot. The strains were subjected to phenotype screening under different environmental conditions (pH, oxygen availability, temperature, and osmolality), and the obtained data processing was assisted by mathematical modeling. The results demonstrated that growth and the r-Prot yields under specific conditions can be significantly increased or decreased due to the TFs' engineering. Environmental factors "awakening" individual TFs were indicated, and their contribution was mathematically described. For example, OE of Yap-like TF was proven to alleviate growth retardation under high pH, while Gzf1 and Hsf1 were shown to serve as universal enhancers of r-Prot production in Y. lipolytica. On the other hand, KO of SKN7 and HSF1 disabled growth under hyperosmotic stress. This research demonstrates the usefulness of the TFs engineering approach in the manipulation of complex traits and evidences newly identified functions of the studied TFs. KEY POINTS: • Function and implication in complex traits of 5 TFs in Y. lipolytica were studied. • Gzf1 and Hsf1 are the universal r-Prots synthesis enhancers in Y. lipolytica. • Yap-like TF's activity is pH-dependent; Skn7 and Hsf1 act in osmostress response., (© 2023. The Author(s).)

Reconstructive surgery after surgical treatment of neoplasms in the head and neck region is always a challenge. Many factors are responsible for the success of reconstruction. The anatomy of the facial region is complex, which significantly influences the aesthetic effect of the reconstruction. Moreover, many patients undergo postoperative radiotherapy after surgical treatment, which affects the range of reconstructive techniques. The aim of this study is to review current reconstructive methods in the craniofacial region, using bone-anchored implants to attach nasal prostheses. The article also comprises the authors' own experience with successful single-stage, Vistafix 3 osseointegrated implants for the attachment of an external nasal prosthesis in a 51-year-old man after surgical removal of squamous cell carcinoma of the nose and paranasal sinuses. The literature search for articles regarding implants in craniofacial reconstructions was performed using the three following databases: Scopus, Web of Science and MEDLINE (through PubMed), and follows the Preferred Reporting Items for Systematic Reviews and Meta-Analyses Statement (PRISMA). A systematic literature search was set for 2018-2023 and retrieved 92 studies. From them, 18 articles were included in the review.