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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]

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.

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.

Background and Objectives : It is well established that patients with cognitive impairment are at a higher risk of falls. However, the impact of coexisting neuropsychiatric symptoms on the overall risk of falls in hospitalized geriatric individuals with and without dementia has not been extensively studied. This cross-sectional study will assess the association between neuropsychiatric symptoms and fall risk in geriatric individuals analyzed by sex. Materials and Methods : A total of 234 patients, both with and without dementia, admitted to the geriatric ward at Leszek Giec Upper-Silesian Medical Centre of the Silesian Medical University in Katowice, Poland, between January 2019 and January 2020 were included in this study. The Neuropsychiatric Inventory-Questionnaire was used to assess the presence of neuropsychiatric symptoms. Increased fall risk was defined by Berg scores of ≤40. Results : The mean age of the study group was 80.7 ± 6.6, and women accounted for 62.8% of the study population. Apathy was the most common neuropsychiatric symptom, affecting 58.1% of patients, and it was the most common symptom among people with dementia, affecting 67.80% of patients. The receiver operating characteristics curve analysis revealed that a high fall risk was significantly associated with the total number of neuropsychiatric symptoms (≥4) and the total intensity of these symptoms (≥6). For women, high fall risk was associated with three or more neuropsychiatric symptoms and a total neuropsychiatric symptom intensity score of at least 6. For men, the association of high fall risk with the total number of NPS was not significant; a total NPS intensity score of 10 or more was associated with high fall risk. Multivariate logistic regression analysis identified associations with fall risk for hallucinations. Conclusions : Our results suggest that the presence of neuropsychiatric symptoms, particularly hallucinations is associated with an increased risk of falls in geriatric inpatients. In addition, the cumulative total of NPS and their cumulative intensity are both independently associated with an increased risk of falls. These results suggest that fall prevention strategies should include the management of neuropsychiatric symptoms in hospitalized geriatric individuals.

Phenotype-gene association studies can uncover disease mechanisms for translational research. Association with multiple phenotypes or clinical variables in complex diseases has the advantage of increasing statistical power and offering a holistic view. Existing multi-variate association methods mostly focus on SNP-based genetic associations. In this paper, we extend and evaluate two adaptive Fisher's methods, namely AFp and AFz, from the p -value combination perspective for phenotype-mRNA association analysis. The proposed method effectively aggregates heterogeneous phenotype-gene effects, allows association with different data types of phenotypes, and performs the selection of the associated phenotypes. Variability indices of the phenotype-gene effect selection are calculated by bootstrap analysis, and the resulting co-membership matrix identifies gene modules clustered by phenotype-gene effect. Extensive simulations demonstrate the superior performance of AFp compared to existing methods in terms of type I error control, statistical power and biological interpretation. Finally, the method is separately applied to three sets of transcriptomic and clinical datasets from lung disease, breast cancer, and brain aging and generates intriguing biological findings.

Based on the current state of the BPPV field, there are no guidelines that specify an angular head movement's velocity (AHMV) during diagnostic maneuvers of BPPV. The aim of this study was to evaluate the impact of AHMV during diagnostic maneuvers on proper BPPV diagnosis and therapy. The analysis covered the results obtained in 91 patients with a positive result of the Dix-Hallpike (D-H) maneuver or the roll test. The patients were divided into four groups based on values of AHMV (high 100-200°/s and low 40-70°/s) and the BPPV type (posterior: PC-BPPV or horizontal: HC-BPPV). The parameters of the obtained nystagmuses were analyzed and compared to AHMV. There was a significant negative correlation between AHMV and latency of nystagmus in all study groups. Furthermore, there was a significant positive correlation between AHMV and both maximum slow phase velocity and average frequency of nystagmus in the PC-BPPV groups, whereas it was not observed in the HC-BPPV patients. Complete relief of symptoms was reported after 2 weeks and was better in patients diagnosed with maneuvers performed with high AHMV. High AHMV during the D-H maneuver allows the nystagmus to be more visible, increasing the sensitivity of diagnostic tests and is crucial for a proper diagnosis and therapy.

DiGeorge syndrome is associated with microdeletion of chromosome 22q11. Hypoplasia of the thymus, hypoparathyroidism, facial malformations and cardiac defects as well as learning difficulties are typical features of the disease. On the other hand hypocalcemia related to hypoparathyroidism is not present in every patient and can develop later and be persistent or transient and is often masked by the other signs or symptoms. We described a 13-year-old boy diagnosed with DiGeorge syndrome, after a few years of nonspecific signs and symptoms, and a microarray examination performed because myopathy was suspected on the basis of elevated creatine kinase activity. Only after molecular confirmation of DiGeorge syndrome the patient was referred to a pediatric endocrinologist and proper therapy started. Looking back to his medical history, low calcium levels were at least 2 times reported in the medical records, the child had learning difficulties, speech disturbances, and submucosal cleft palate suspicion. In conclusion it is important to educate general practitioners and pediatricians to check the serum calcium levels in patients presenting with nonspecific, muscular signs and symptoms.

Hadfield steel, under unit pressure conditions, strengthens itself by forming a high density dislocation structure, which results in increased resistance to dynamic impact wear. However, under abrasion conditions, the homogeneous microstructure of the cast steel is insufficient to achieve the expected service life. The aim of the research is to conduct a comparative analysis of the material in its as-delivered state and after two-stage heat treatment (isothermal annealing followed by re-austenitisation). It was found that after isothermal annealing of X120Mn12 grade steel at a temperature of 510 °C, a microstructure with a complex morphology consisting of colonies of fine-grained pearlite, (Fe,Mn) 3 C carbides distributed along the grain boundaries of the former austenite and needle-like (Fe,Mn) 3 C carbides was obtained in the austenite matrix. The subsequent thermal treatment of the steel with the use of supersaturating annealing at 900 °C resulted in a heterogeneous microstructure consisting of evenly distributed globular carbide precipitations in a matrix of considerably finer austenite grains in comparison with the as-delivered original state. As a result of the final microstructural changes achieved, a 16.4% increase in abrasion resistance was obtained compared to the delivered condition.

While overproduction of recombinant secretory proteins (rs-Prots) triggers multiple changes in the physiology of the producer cell, exposure to suboptimal growth conditions may further increase that biological response. The environmental conditions may modulate the efficiency of both the rs-Prot gene transcription and translation but also the polypeptide folding. Insights into responses elicited by different environmental stresses on the rs-Prots synthesis and host yeast physiology might contribute to a better understanding of fundamental biology processes, thus providing some clues to further optimise bioprocesses. Herein, a series of batch cultivations of Yarrowia lipolytica strains differentially metabolically burdened by the rs-Prots overproduction have been conducted. Combinations of different stress factors, namely pH (3/7) and oxygen availability (kLa 28/110 h -1 ), have been considered for their impact on cell growth and morphology, substrate consumption, metabolic activity, genes expression, and secretion of the rs-Prots. Amongst others, our data demonstrate that a highly metabolically burdened cell has a higher demand for the carbon source, although presenting a compromised cell growth. Moreover, the observed decrease in rs-Prot production under adverse environmental conditions rather results from the emergence of a less-producing cell subpopulation than from the decrease of the synthetic capacity of the whole cell population.

Background and Objectives : Anticholinergic drug use in the pharmacotherapy of elderly persons is common despite the increased risk of side effects. We examined the prevalence of anticholinergic drug use and total anticholinergic drug burden among patients admitted to an acute care geriatric ward in Poland. Materials and Methods : Cross-sectional study of 329 subjects hospitalized at the geriatric ward. Patient condition was assessed with a comprehensive geriatric assessment. The Anticholinergic Cognitive Burden (ACB) scale was used to estimate the total anticholinergic load. Results : Mean patient age was 79.61 ± 6.82 years. 40.73% of them were burdened with at least one anticholinergic drug. The clinically significant anticholinergic burden was observed in 13.98% of subjects. Patients with dementia, risk of falls, and severe disability had significantly higher total ACB scores compared to other groups. The receiver operating characteristics (ROC) curve revealed that the total ACB score ≥ 1 was significantly associated with dementia and the risk of falls. Total ACB score ≥ 2 was significantly associated with severe disability. Conclusions : Patients admitted to an acute care geriatric ward had an anticholinergic cognitive burden score comparable to other patient populations. We found associations at both low and elevated levels of anticholinergic burden with dementia and risk of falls. At elevated anticholinergic burden levels, we found associations with severe disability. Despite recommendations against the use of anticholinergics in older adults these medications are still commonly prescribed. Further study is necessary to define the characteristics of anticholinergic medication most closely associated with negative outcomes in elderly populations.

Purpose: In patients with metastatic castration-resistant prostate cancer (mCRPC), resistance to androgen receptor (AR)-targeted therapies, such as enzalutamide, remains an issue. Inactivation of inhibitory PTEN activates PI3K/AKT signaling and contributes to resistance to androgen deprivation therapy and poor outcomes. Therefore, dual targeting of AR and PI3K/AKT pathways may limit tumor growth and reverse resistance., Patients and Methods: In this phase I study (NCT02215096), patients with PTEN-deficient mCRPC who progressed on prior enzalutamide received once-daily enzalutamide 160 mg plus PI3Kβ inhibitor GSK2636771 at 300 mg initial dose, with escalation or de-escalation in 100-mg increments, followed by dose expansion. Primary objectives were to evaluate safety/tolerability, determine the recommended phase II dose, and assess the 12-week non-progressive disease (PD) rate., Results: Overall, 37 patients were enrolled; 36 received ≥1 dose of GSK2636771 (200 mg: n = 22; 300 mg: n = 12; 400 mg: n = 2) plus 160 mg enzalutamide. Dose-limiting toxicities occurred in 5 patients (200 mg: n = 1; 300 mg: n = 2, 400 mg: n = 2). No new or unexpected adverse events or evidence of drug-drug interaction were observed. At the recommended dose of GSK2636771 (200 mg) plus enzalutamide, the 12-week non-PD rate was 50% (95% confidence interval: 28.2-71.8, n = 22); 1 (3%) patient achieved a radiographic partial response lasting 36 weeks. Four of 34 (12%) patients had prostate-specific antigen reduction of ≥50%., Conclusions: Although there was acceptable safety and tolerability with GSK2636771 plus enzalutamide in patients with PTEN-deficient mCRPC after failing enzalutamide, limited antitumor activity was observed., (©2021 American Association for Cancer Research.)

Thanks to the continuous development of light-curing resin composites it is now possible to print permanent single-tooth restorations. The purpose of this study was to compare resin composites for milling -Gandio Blocks (GR), Brilliant Crios (CR) and Enamic (EN) with resin composite for 3D printing-Varseo Smile Crown plus (VSC). Three-point bending was used to measure flexural strength (σ f ) and flexural modulus (E f ). The microhardness was measured using a Vickers method, while fractographic, microstructural, texture and fractal dimension (FD) analyses were performed using SEM, optical microscope and picture analysis methods. The values of σ f ranged from 118.96 (±2.81) MPa for EN to 186.02 (±10.49) MPa for GR, and the values of E f ranged from 4.37 (±0.8) GPa for VSC to 28.55 (±0.34) GPa for EN. HV01 ranged from 25.8 (±0.7) for VSC to 273.42 (±27.11) for EN. The filler content ranged from 19-24 vol. % for VSC to 70-80 vol. % for GR and EN. The observed fractures are typical for brittle materials. The correlation between FD of materials microstructure and E f was observed. σ f of the printed resin depends on layers orientation and is significantly lower than σ f of GR and CR. E f of the printed material is significantly lower than E f of blocks for milling.

Electrically-Assisted Forming (EAF) techniques are interesting and promising for the automotive industry. Electrically-assisted tensile tests were carried out on specimens of AA7075 aluminum alloy in different states of hardening, namely T6 (the as-received state) and the supersaturated solid solution state. All the tests were carried out in quasi-static conditions under the application of direct electric current (DC) in the range of 90 to 540 A. The experimental results showed that with a DC density of 10 A/mm 2 the uniform strain and strain at fracture increased when the AA7075 was in the supersaturated solid solution state. A correlation between the mechanical results and microstructural features analysed through transmission electron microscopy was assessed. An explanation of the investigated phenomena based on the electron wind theory, heterogeneous microscale Joule heating and the Portevin-Le Chatelier (PLC) effect was finally proposed.

Animal behaviors are often composed of distinct alternating behavioral states. Neuromodulatory signals are thought to be critical for establishing stable behavioral states and for orchestrating transitions between them. However, we have only a limited understanding of how neuromodulatory systems act in vivo to alter circuit performance and shape behavior. To address these questions, we have investigated neuromodulatory signaling in the context of Caenorhabditis elegans egg-laying. Egg-laying activity cycles between discrete states-short bursts of egg deposition (active phases) that alternate with prolonged quiescent periods (inactive phases). Here using genetic, pharmacological and optogenetic approaches for cell-specific activation and inhibition, we show that a group of neurosecretory cells (uv1) located in close spatial proximity to the egg-laying neuromusculature direct the temporal organization of egg-laying by prolonging the duration of inactive phases. We demonstrate that the modulatory effects of the uv1 cells are mediated by peptides encoded by the nlp-7 and flp-11 genes that act locally to inhibit circuit activity, primarily by inhibiting vesicular release of serotonin from HSN motor neurons. This peptidergic inhibition is achieved, at least in part, by reducing synaptic vesicle abundance in the HSN motor neurons. By linking the in vivo actions of specific neuropeptide signaling systems with the generation of stable behavioral outcomes, our study reveals how cycles of neuromodulation emanating from non-neuronal cells can fundamentally shape the organization of a behavioral program.

Low-pressure cold spraying is a newly developed technology with high application potential. The aim of this study was to investigate potential application of this technique for producing a new type of transparent conductive oxide films target. Cold spraying technique allows the manufacture of target directly on the backing plate; therefore the proposed sputtering target has a form of Sn+In 2 O 3 coating sprayed onto copper substrate. The microstructure and properties of the feedstock powder prepared using three various methods as well as the deposited ones by low-pressure cold spraying coatings were evaluated, compared, and analysed. Produced cermet Sn+In 2 O 3 targets were employed in first magnetron sputtering process to deposit preliminary, thin, transparent conducting oxide films onto the glass substrates. The resistivity of obtained preliminary films was measured and allows believing that fabrication of TCO (transparent conducting oxide) films using targets produced by cold spraying is possible in the future, after optimization of the deposition conditions., Competing Interests: The authors declare no competing interests regarding the publication of this paper.

A study of the interaction between five gemini amphiphilic valine-based pseudopeptides (GAPs) differing by the length of the central aliphatic spacer linking two amino acid subunits, and a model bacterial membrane lipid, 1,2-dimyristoyl-sn-glycero-3-phospho-rac-(1-glycerol) (DMPG), is here presented. Pure DMPG, pure GAPs and mixed GAPs/DMPG monolayers were formed at the air-water interface using Langmuir technique. The properties of the Langmuir films were investigated using surface pressure measurements, polarization modulation-infrared reflection-absorption spectroscopy, and Brewster angle microscopy. The atomic level information concerning the orientation of molecules in the monolayer and hydration of the polar headgroups was obtained from molecular dynamics simulations. It was demonstrated that the length of the central spacer in the GAPs structure is important for the properties of the mixed films; the disorganization of the membrane increases with the length of the spacer. The latter point is important for developing possible antimicrobial agents based on GAPs., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Introduction and Aim. Exceptional properties of the NiTi archwires may be jeopardized by the oral cavity; thus its long-term effect on the mechanical and physiochemical properties of NiTi archwires was the aim of work. Material and Methods . Study group comprised sixty 0.016 × 0.022 NiTi archwires from the same manufacturer evaluated (group A) after the first 12 weeks of orthodontic treatment. 30 mm long pieces cut off from each wire prior to insertion formed the control group B. Obeying the strict rules of randomization, all samples were subjected to microscopic evaluation and nanoindentation test. Results. Both groups displayed substantial presence of nonmetallic inclusions. Heterogeneity of the structure and its alteration after usage were found in groups B and A, respectively. Conclusions. Long-term, reliable prediction of biomechanics of NiTi wires in vivo is impossible, especially new archwires from the same vendor display different physiochemical properties. Moreover, manufacturers have to decrease contamination in the production process in order to minimize risk of mutual negative influence of nickel-titanium archwires and oral environment., Competing Interests: The authors declared that they have no conflict of interests.

The interaction between five gemini amphiphilic pseudopeptides (GAPs) differing by the length of the central spacer and a model membrane lipid, 1,3-bis[1,2-dimyristoyl-sn-glycero-3-phospho]-sn-glycerol (cardiolipin) were studied with the aim to evaluate their possible antimicrobial properties. To this end, monomolecular films were formed at the air/water interface with pure cardiolipin or cardiolipin/GAPs mixtures; film properties were determined using surface pressure and surface potential measurements, as well as polarization-modulation infrared reflection-absorption spectroscopy. Moreover, to better understand the GAPs-phospholipid interaction at the molecular level, molecular dynamics simulations were performed. The results obtained indicate that the length of the central spacer has an effect on the interaction of GAPs with cardiolipin and on the properties of the lipid film. The GAPs with the longer linkers can be expected to be useful for biological membrane modification and for possible antimicrobial applications.

Two p-tert-butylcalix[4]arene derivatives bearing one or two nalidixic acid groups connected to the lower rim of p-tert-butylcalix[4]arene through the propylenic spacer were studied upon interaction with model bacterial membranes. Indeed, these derivatives were developed recently as new macrocyclic antibiotic carriers for antibacterial therapy. To obtain molecular level information about the interaction between the calixarene conjugates and a membrane lipid, atomistic molecular dynamics simulation, as well as surface pressure, surface potential, polarization modulation infrared reflection-absorption spectroscopy, and Brewster angle microscopy studies of 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine (DMPE)-calixarene derivative films were performed. The results obtained indicate that the interaction between the calixarene derivatives and DMPE occurs via the phosphate and carbonyl groups present in the lipid. Although both calixarene derivatives increase the chain tilt and conformational disordering of the DMPE molecules, these effects are more important in the case of the monosubstituted derivative. Importantly, the two derivatives have an opposite impact on hydration of the phosphoglyceride polar head.

Drosophila is increasingly used for understanding the neural basis of behavior through genetically targeted manipulation of specific neurons. The primary approach in this regard has relied on the suppression of neuronal activity. Here, we report the results of a novel approach to find and characterize neural circuits by expressing neuronal activators to stimulate subsets of neurons to induce behavior. Classical electrophysiological studies demonstrated that stimulation of command neurons could activate neural circuits to trigger fixed action patterns. Our method was designed to find such command neurons for diverse behaviors by screening flies in which random subsets of brain cells were activated. We took advantage of the large collection of Gal4 lines from the NP project and crossed 835 Gal4 strains with relatively limited Gal4 expression in the brain to flies carrying a UAS transgene encoding TRPM8, a cold-sensitive ion channel. Low temperatures opened the TRPM8 channel in Gal4-expressing cells, leading to their excitation, and in many cases induced overt behavioral changes in adult flies. Paralysis was reproducibly observed in the progeny of crosses with 84 lines, whereas more specific behaviors were induced with 24 other lines. Stimulation performed using the heat-activated channel, TrpA1, resulted in clearer and more robust behaviors, including flight, feeding, and egg-laying. Through follow-up studies starting from this screen, we expect to find key components of the neural circuits underlying specific behaviors, thus providing a new avenue for their functional analysis.

Many feeding behaviours are the result of stereotyped, organized sequences of motor patterns. These patterns have been the subject of neuroethological studies, such as electrophysiological characterization of neurons governing prey capture in toads. However, technical limitations have prevented detailed study of the functional role of these neurons, a common problem for vertebrate organisms. Complexities involved in studies of whole-animal behaviour can be resolved in Drosophila, in which remote activation of brain cells by genetic means enables us to examine the nervous system in freely moving animals to identify neurons that govern a specific behaviour, and then to repeatedly target and manipulate these neurons to characterize their function. Here we show neurons that generate the feeding motor program in Drosophila. We carried out an unbiased screen using remote neuronal activation and identified a critical pair of brain cells that induces the entire feeding sequence when activated. These 'feeding neurons' (here abbreviated to Fdg neurons for brevity) are also essential for normal feeding as their suppression or ablation eliminates sugar-induced feeding behaviour. Activation of a single Fdg neuron induces asymmetric feeding behaviour and ablation of a single Fdg neuron distorts the sugar-induced feeding behaviour to become asymmetric, indicating the direct role of these neurons in shaping motor-program execution. Furthermore, recording neuronal activity and calcium imaging simultaneously during feeding behaviour reveals that the Fdg neurons respond to food presentation, but only in starved flies. Our results demonstrate that Fdg neurons operate firmly within the sensorimotor watershed, downstream of sensory and metabolic cues and at the top of the feeding motor hierarchy, to execute the decision to feed.

Aim of the Study: Evaluation of the influence of the introduced structure modification in porous ceramic grafts on TiO2 base on overgrowing with bone tissue, in examinations with use of scanning microscopy and X-rays was the subject of the examinations., Material and Method: New ceramic materials based on TiO2 with high values of mechanical resistance, large sintering degree and biocompatibility in in vitro conditions were prepared. Those properties cause that they are worth interest as potential osteosubstitutive materials. Two kinds of grafts were created from ceramics based on TiO2: with compact and porous structure. The introduced structure modification - macroporosity - had the purpose to give osteoconductive properties to the grafts, to evoke processes favorable for bone tissue forming. In examinations of the local reaction of bone tissue after implantation of the formed porous grafts, degree of their osteointegration, the essential issue is the evaluation of the settling of the inner spaces with supporting tissues. Samples of the tested compact and porous materials in the form of cylinders were implanted in femoral bones of rabbits for a period till 6 months. The surfaces of grafts and the degree of their settling with supporting tissues were evaluated in cross-sections of the implants with light and scanning microscopic methods and they were confirmed in X-ray tests., Results: Analysis of the obtained data showed that, the surface of solid ceramic on TiO2 base was covered mainly with increasing with a time of observation quantities of extracellular substance and lamellas of bone. The macrporous structure of porous ceramic on TiO2 base enabled settling of the inner spaces of graft with supporting tissue cells, partly in the 1st month, more intensively in the 3rd and it was not completed in the 6th month after implantation., Conclusions: Porous grafts in the form of ceramic foam on TiO2 base showed osteoconductive properties, though process of colonization after 6th month observation was not completed and the condition of the cells inside of the implant was reduced.

In this research, the interaction between a membrane phospholipid, 1,2-dilauroyl-sn-glycero-3-phosphocholine (DLPC), and a p-tert-butylcalix[4]arene derivative bearing 6-aminopenicillanic acid (Calix), conceived as a possible drug carrier, was studied. The Langmuir film balance technique was used to measure surface pressure and electrical surface potential of pure and mixed Calix/DLPC monolayers spread on water at different temperatures. Phospholipase A2 (PLA2) activity was used as well to detect the impact of the calixarene derivative on the monolayer properties. Interaction between the molecules in mixed monolayers has been described quantitatively using thermodynamic functions. Interestingly, low amounts of Calix introduce ordering in the lipid film. This effect may be analogous to that of cholesterol interacting with phospholipids. A lower activity of PLA2 observed with the Calix/DLPC films compared to pure DLPC may be related to structural modifications of the mixed systems., (Copyright © 2012 Elsevier B.V. All rights reserved.)

We conducted an international phase 2 trial to evaluate 2 dose levels of ofatumumab, a human CD20 mAb, combined with fludarabine and cyclophosphamide (O-FC) as frontline therapy for chronic lymphocytic leukemia (CLL). Patients with active CLL were randomized to ofatumumab 500 mg (n = 31) or 1000 mg (n = 30) day 1, with fludarabine 25 mg/m(2) and cyclophosphamide 250 mg/m(2) days 2-4, course 1; days 1-3, courses 2-6; every 4 weeks for 6 courses. The first ofatumumab dose was 300 mg for both cohorts. The median age was 56 years; 13% of patients had a 17p deletion; 64% had β2-microglobulin > 3.5 mg/L. Based on the 1996 National Cancer Institute Working Group (NCI-WG) guidelines, the complete response (CR) rate as assessed by an independent review committee was 32% for the 500-mg and 50% for the 1000-mg cohort; the overall response (OR) rate was 77% and 73%, respectively. Based on univariable regression analyses, β2-microglobulin and the number of O-FC courses were significantly correlated (P < .05) with CR and OR rates and progression-free survival (PFS). The most frequent Common Terminology Criteria (CTC) grade 3-4 investigator-reported adverse events were neutropenia (48%), thrombocytopenia (15%), anemia (13%), and infection (8%). O-FC is active and safe in treatment-naive patients with CLL, including high-risk patients. This trial was registered at www.clinicaltrials.gov as NCT00410163.

Activity-dependent modifications in synapse structure play a key role in synaptic development and plasticity, but the signaling mechanisms involved are poorly understood. We demonstrate that glutamatergic Drosophila neuromuscular junctions undergo rapid changes in synaptic structure and function in response to patterned stimulation. These changes, which depend on transcription and translation, include formation of motile presynaptic filopodia, elaboration of undifferentiated varicosities, and potentiation of spontaneous release frequency. Experiments indicate that a bidirectional Wnt/Wg signaling pathway underlies these changes. Evoked activity induces Wnt1/Wg release from synaptic boutons, which stimulates both a postsynaptic DFz2 nuclear import pathway as well as a presynaptic pathway involving GSK-3beta/Shaggy. Our findings suggest that bidirectional Wg signaling operates downstream of synaptic activity to induce modifications in synaptic structure and function. We propose that activation of the postsynaptic Wg pathway is required for the assembly of the postsynaptic apparatus, while activation of the presynaptic Wg pathway regulates cytoskeletal dynamics.

The Wingless pathway plays an essential role during synapse development. Recent studies at Drosophila glutamatergic synapses suggest that Wingless is secreted by motor neuron terminals and binds to postsynaptic Drosophila Frizzled-2 (DFz2) receptors. DFz2 is, in turn, endocytosed and transported to the muscle perinuclear area, where it is cleaved, and the C-terminal fragment is imported into the nucleus, presumably to regulate transcription during synapse growth. Alterations in this pathway interfere with the formation of new synaptic boutons and lead to aberrant synaptic structures. Here, we show that the 7 PDZ protein dGRIP is necessary for the trafficking of DFz2 to the nucleus. dGRIP is localized to Golgi and trafficking vesicles, and dgrip mutants mimic the synaptic phenotypes observed in wg and dfz2 mutants. DFz2 and dGRIP colocalize in trafficking vesicles, and a severe decrease in dGRIP levels prevents the transport of endocytosed DFz2 receptors to the nucleus. Moreover, coimmunoprecipitation experiments in transfected cells and yeast two-hybrid assays suggest that the C terminus of DFz2 interacts directly with the PDZ domains 4 and 5. These results provide a mechanism by which DFz2 is transported from the postsynaptic membrane to the postsynaptic nucleus during synapse formation and implicate dGRIP as an essential molecule in the transport of this signal.

At vertebrate neuromuscular junctions (NMJs), Agrin plays pivotal roles in synapse development, but molecules that activate synapse formation at central synapses are largely unknown. Members of the Wnt family are well established as morphogens, yet recently they have also been implicated in synapse maturation. Here we demonstrate that the Drosophila Wnt, Wingless (Wg), is essential for synapse development. We show that Wg and its receptor are expressed at glutamatergic NMJs, and that Wg is secreted by synaptic boutons. Loss of Wg leads to dramatic reductions in target-dependent synapse formation, and new boutons either fail to develop active zones and postsynaptic specializations or these are strikingly aberrant. We suggest that Wg signals the coordinated development of pre- and postsynaptic compartments.

Background: Membrane-associated guanylate kinases (MAGUKs), such as Discs-Large (DLG), play critical roles in synapse maturation by regulating the assembly of synaptic multiprotein complexes. Previous studies have revealed a genetic interaction between DLG and another PDZ scaffolding protein, SCRIBBLE (SCRIB), during the establishment of cell polarity in developing epithelia. A possible interaction between DLG and SCRIB at synaptic junctions has not yet been addressed. Likewise, the biochemical nature of this interaction remains elusive, raising questions regarding the mechanisms by which the actions of both proteins are coordinated., Results: Here we report the isolation of a new DLG-interacting protein, GUK-holder, that interacts with the GUK domain of DLG and which is dynamically expressed during synaptic bouton budding. We also show that at Drosophila synapses DLG colocalizes with SCRIB and that this colocalization is likely to be mediated by direct interactions between GUKH and the PDZ2 domain of SCRIB. We show that DLG, GUKH, and SCRIB form a tripartite complex at synapses, in which DLG and GUKH are required for the proper synaptic localization of SCRIB., Conclusions: Our results provide a mechanism by which developmentally important PDZ-mediated complexes are associated at the synapse.

Ras proteins are small GTPases with well known functions in cell proliferation and differentiation. In these processes, they play key roles as molecular switches that can trigger distinct signal transduction pathways, such as the mitogen-activated protein kinase (MAPK) pathway, the phosphoinositide-3 kinase pathway, and the Ral-guanine nucleotide dissociation stimulator pathway. Several studies have implicated Ras proteins in the development and function of synapses, but the molecular mechanisms for this regulation are poorly understood. Here, we demonstrate that the Ras-MAPK pathway is involved in synaptic plasticity at the Drosophila larval neuromuscular junction. Both Ras1 and MAPK are expressed at the neuromuscular junction, and modification of their activity levels results in an altered number of synaptic boutons. Gain- or loss-of-function mutations in Ras1 and MAPK reveal that regulation of synapse structure by this signal transduction pathway is dependent on fasciclin II localization at synaptic boutons. These results provide evidence for a Ras-dependent signaling cascade that regulates fasciclin II-mediated cell adhesion at synaptic terminals during synapse growth.

Although abnormal processing of beta-amyloid precursor protein (APP) has been implicated in the pathogenic cascade leading to Alzheimer's disease, the normal function of this protein is poorly understood. To gain insight into APP function, we used a molecular-genetic approach to manipulate the structure and levels of the Drosophila APP homolog APPL. Wild-type and mutant forms of APPL were expressed in motoneurons to determine the effect of APPL at the neuromuscular junction (NMJ). We show that APPL was transported to motor axons and that its overexpression caused a dramatic increase in synaptic bouton number and changes in synapse structure. In an Appl null mutant, a decrease in the number of boutons was found. Examination of NMJs in larvae overexpressing APPL revealed that the extra boutons had normal synaptic components and thus were likely to form functional synaptic contacts. Deletion analysis demonstrated that APPL sequences responsible for synaptic alteration reside in the cytoplasmic domain, at the internalization sequence GYENPTY and a putative G(o)-protein binding site. To determine the likely mechanisms underlying APPL-dependent synapse formation, hyperexcitable mutants, which also alter synaptic growth at the NMJ, were examined. These mutants with elevated neuronal activity changed the distribution of APPL at synapses and partially suppressed APPL-dependent synapse formation. We propose a model by which APPL, in conjunction with activity-dependent mechanisms, regulates synaptic structure and number.