Search Results (15)

Dictyostelium is a unique model used to study the complex and interactive cyclic nucleotide signaling pathways that regulate multicellular development. Dictyostelium grow as individual single cells, but in the absence of nutrients, they initiate a multicellular developmental program. Central to this is secreted cAMP, a primary GPCR-response signal. Activated cAMP receptors at the cell surface direct a number of downstream signaling pathways, including synthesis of the intracellular second messengers cAMP and cGMP. These, in turn, activate a series of downstream targets that direct chemotaxis within extracellular cAMP gradients, multicellular aggregation, and, ultimately, cell-specific gene expression, morphogenesis, and cytodifferentiation. Extracellular cAMP and intracellular cAMP and cGMP exhibit rapid fluctuations in concentrations and are, thus, subject to exquisite regulation by both synthesis and degradation. The Dictyostelium genome encodes seven phosphodiesterases (PDEs) that degrade cyclic nucleotides to nucleotide 5’-monophosphates. Each PDE has a distinct structure, substrate specificity, regulatory input, cellular localization, and developmentally regulated expression pattern. The intra- or extra-cellular localizations and enzymatic specificities for cAMP or cGMP are essential for degradative precision at different developmental stages. We discuss the diverse PDEs, the nucleotide cyclases, and the target proteins for cAMP and cGMP in Dictyostelium. We further outline the major molecular, cellular, and developmental events regulated by cyclic nucleotide signaling, with emphasis on the input of each PDE and consequence of loss-of-function mutations. Finally, we relate the structures and functions of the Dictyostelium PDEs with those of humans and in the context of potential therapeutic understandings. Full article

The increase in exploitation durability of two-layer cast rolls with the working layer made of high chromium cast iron allows one to significantly improve the quality of rolled metal as well as to increase the economic efficiency of the manufacturing process. However, it is severely hindered due to the massiveness of castings, the impossibility of both evaluating mechanical properties along the depth of the working layer, and providing the structural uniformity of the working surface and the decrease in stresses. In our research, aiming to enhance the exploitation durability of sheet rolls, it is recommended to achieve structural uniformity by CuMg alloying, which increases the concentration of copper up to 2.78 wt.% in certain zones and, owing to the accelerated austenite decomposition at a high temperature during the cool-down of the castings, led to the reduction in excessive strength and the level of heat stresses in the castings. We propose the regimes of cyclic heat treatments which, due to the decomposition of retained austenite and the fragmentation of structure, control the level of hardness to reduce and uniformize the level of stresses along the length of a barrel. A further improvement in the predictions of exploitation durability using XGboost method, which was performed based on the chemical composition of the working layer of high-chromium cast iron and heat treatment parameters, requires taking into account the factors characterizing exploitation conditions of specific rolling mills and the transformations of structural-phase state of the surface obtained by a non-destructive control method. As the controlled parameter, the hardness measured on the roll’s surface is recommended, while the gradient change in mechanical properties along the working layer depth can be feasibly analyzed by a magnetic method of coercive force measuring. Full article

Cu₂Se is a promising thermoelectric (TE) material due to its low cost, Earth abundance, and high thermoelectric properties. However, the biggest problem of Cu₂Se is its unstable chemical properties. In particular, under the action of an electric field or gradient temperature field, the chemical potential of copper ions inside the material increases. When the external field is strong enough, the chemical potential of copper ions at the negative end of the material reaches the chemical potential of elemental copper. Under these conditions, copper ions must precipitate out, causing Cu₂Se to be unstable, and making it unsuitable for use in applications. In this study, we prepared Cu_2−xMn_xSe (x = 0, 0.02, 0.04 and 0.06) series bulk materials by vacuum melting–annealing and sintered by spark plasma sintering (SPS). We investigated the effects of Mn doping on the composition, microstructure, band structure, scattering mechanism, thermoelectric properties, and stability of Cu₂Se. The results show that Mn doping can adjust the carrier concentration, promote the stabilization of the β-phase structure and improve the electrical properties of Cu₂Se. When x = 0.06, the highest power factor (PF) value of Cu_1.94Mn_0.06Se at 873 K was 1.62 mW m⁻¹ K⁻². The results of carrier scattering mechanism analysis based on the conductivity ratio method show that the sample doped with Mn and pure Cu₂Se had the characteristics of ionization impurity scattering, and the scattering factor was 3/2. However, the deterioration in thermal conductivity was large, and a superior zT value needs to be obtained. The cyclic test results of high-temperature thermoelectric properties show that Mn doping can hinder Cu⁺ migration and improve its thermoelectric stability, which preliminarily verifies the feasibility of using the stable zirconia mechanism to improve the thermoelectric stability of Cu₂Se. Full article

Oxygen plays important roles in regulating various biological activities under physiological and pathological conditions. However, the response of cells facing temporal variation in oxygen microenvironments has seldom been studied due to technical limitations. In this paper, an integrated approach to studying hypoxic response under cyclic oxygen gradients is developed. In the experiments, a cell culture system based on a microfluidic device is constructed to generate cyclic oxygen gradients with desired periods by alternately introducing gases with specific compositions into the microfluidic channels next to the cell culture channel separated by thin channel walls. Observation of the hypoxic responses is performed using real-time fluorescence imaging of dyes sensitive to extra- and intracellular oxygen tensions as well as intracellular calcium concentrations. Cellular hypoxic responses of human aortic smooth muscle cells (AoSMCs) and lung carcinoma epithelium (A549) cells, including intracellular oxygen and calcium levels, are measured. The results show that the two types of cells have different hypoxic responses to the applied cyclic oxygen gradients. With the capability of real-time cellular response monitoring under cyclic oxygen gradients, the developed approach provides a useful scheme to investigate hypoxic responses in vitro under microenvironments mimicking various in vivo physiological and pathological conditions. Full article

The spherical diffusion that occurs when using ultramicroelectrodes (i.e., electrodes with a characteristic size of 1–10 µm) contributes to a higher mass transfer rate. This leads to equalization of the depletion rates of the near-electrode layer due to the electrochemical reaction and to the supply of the product from the solution depth. This is the reason why, for ultramicroelectrodes, a limiting size of the spherical layer exists in which the concentration gradient is localized (diffusion layer). Thus, a stationary mass transfer mode is achieved, which is expressed in the sigmoidal CV curve’s shape. In ultramicroelectrode arrays, when the diffusion hemispheres are separated, a steady-state diffusion is realized. However, with a decrease in the interelectrode distance, which leads to the diffusion spheres intersection, a mixed regime arises, which is not fully time-independent. The resulting voltammogram’s shape change can serve as an analytical signal in the study of substances with differing diffusion coefficients, since the diffusion layer growth rate and, consequently, the area of intersection of neighboring spheres, depends on it. This work shows the applicability of voltammetry using ensembles of ultramicroelectrodes operating in the transient mode for the analysis of mixtures of electrochemically active compounds with close electrode reaction parameters, such as exchange currents and electrode potential. Ferrocenemethanol esters are used as an example. The applicability of cyclic voltammetry on the UME array for analysis of mixtures was illustrated by means of finite element modelling. The reliability of the modelling results was experimentally proved for ferrocenemethanol esters with glycine and triglycine. Full article

A strain gradient was created by punching a hole in the center of a stretched elastic polydimethylsiloxane membrane to determine the effect of different strains on cultured human keratocytes (HK). In this study, two stretching methods were used: continuous stretching and cyclic stretching. Continuous stretching is relatively static, while acyclic stretching is relatively dynamic. These methods, respectively, represented the effects of high intraocular pressure and rubbing of the eyes on corneal cells. Image processing codes were developed to observe the effects of stress concentration, shear stress, continuous stretching, and cyclic stretching on HKs. The results demonstrate that stretching and shear stress are not conducive to the proliferation of corneal cells and instead cause cell death. A 10% strain had greater inhibitory effects than a 3% strain on cell proliferation. Cell survival rates for continuous stretching (static) were higher than those for cyclic stretching (dynamic). The stretching experiment revealed that cyclic stretching has a greater inhibitory effect on the growth and proliferation of corneal cells than continuous stretching. Accordingly, it shows that cyclic loading is more harmful than high intraocular pressure (static loading) to corneal cells. Full article

Distributed fibre optical sensing (DFOS) is increasingly used in civil engineering research. For reinforced concrete structures, almost continuous information concerning the deformations of embedded reinforcing bars can be obtained. This information enables the validation of basic and conventional assumptions in the design and modelling of reinforced concrete, particularly regarding the interaction of concrete and reinforcing bars. However, this relatively new technology conceals some difficulties, which may lead to erroneous interpretations. This paper (i) discusses the selection of sensing fibres for reinforced concrete instrumentation, accounting for strain gradients and local anomalies caused by stress concentrations due to the reinforcing bar ribs; (ii) describes suitable methods for sensor installation, strain acquisition and post-processing of the data, as well as determining and validating structurally relevant entities; and (iii) presents the results obtained by applying DFOS with these methods in a variety of experiments. The analysed experiments comprise a reinforced concrete tie, a pull-out test under cyclic load, and a flexural member in which the following mechanical relevant quantities are assessed: the initial strain state in reinforcing bars, normal and bond shear stresses, deflections as well as forces. These applications confirm the benefit of DFOS to better understand the bond behaviour, but also demonstrate that its application is intricate and the results may lead to erroneous conclusions unless evaluated meticulously. Full article

The accumulation characteristics of surface chloride in concrete in different zones are different in the marine environment. A series of laboratory experiments were conducted to investigate the surface chloride and permeation characteristics of concrete in a simulated marine environment. The experimental results indicated that the surface chloride and chloride profiles of concrete in different zones of marine environment decreased in the following order: tidal zone > splash zone > submerged zone > atmospheric zone. The width of the ascent zone of Cl⁻ concentration at tidal and splash zones was far less than that of the influential depth of moisture transport (IDMT), and the range of convection zone was dependent on the IDMT. Cl⁻ at splash and tidal zones penetrated into concrete as a bulk liquid by non-saturated permeation driven by a humidity gradient. The change of chloride profiles in concrete along the altitudinal gradient was consistent with that of the cyclic water absorption amount (CWAA). The transport rate of chloride was the highest at the highest point of the tide. Full article

Cyclic methylsiloxanes D4, D5, D6 (also called cyclic silicones) are widely used in various dermatological products and cosmetics, both for children and adults. As a result of their unique physicochemical properties, the production of cyclic methylsiloxanes has greatly increased over the last few years, which has resulted in increased exposure to mankind. The validated quantitative for gas chromatography-flame ionization detector (GC-FID) analysis with using the transdermal diffusion system with vertical Franz cells demonstrated that ex vivo human skin is not a barrier to cyclic siloxanes. D4, D5, and D6 have a specific affinity to stratum corneum (SC) (especially D6), and can even diffuse into the deeper layers of the skin (epidermis (E) and dermis (D)), or into the receptor fluid as well. An important achievement of this work was the observation of the characteristic ratio partitioning D4, D5, and D6 in skin layers and receptor fluid (RF). The studies have shown that, in order to thoroughly understand the mechanism, it is important to determine not only the differences in the amounts of cumulated doses in total in all skin layers and receptor fluid, but also the mutual ratios of analyte concentrations existing between matrices. For example, in the case of the stratum corneum, the cumulative doses of D4, D5, and D6 were 27.5, 63.9, and 67.2 µg/cm²/24 h, respectively, and in the epidermis, they were 6.9, 29.9, and 10.7 µg/cm²/24 h, respectively, which confirmed the highest affinity of D6 to stratum corneum as the amount diffused into the epidermis was 2.8 times smaller compared to D5. The calculated epidermis-to-stratum corneum ratios of analyte concentrations also confirm this. The largest ratio was identified for D5 (E/SC = 47), followed by D4 (E/SC = 25), and finally by D6 (E/SC = 16). The analysis of the next stage of diffusion from epidermis to dermis revealed that in dermis the highest cumulative dose was observed for D5 (13.9 µg/cm²/24 h), while the doses of D4 and D6 were similar (5.1 and 5.3 µg/cm²/24 h). Considering the concentration gradient, it can be concluded that the diffusion of D5 and D6 occurs at a similar level, while D4 diffuses at a much higher level. These observations were also confirmed by the dermis-to-epidermis concentration ratios. The final stage of diffusion from dermis to the receptor fluid indicated that D4 was able to permeate easily, while D5 exhibited a difficult diffusion and the diffusion of D6 was limited. The receptor fluid-to-dermis concentration ratios (RF/D) were calculated for D4, D5, and D6: 80, 53, and 17, respectively. Our results also revealed the increased risk of D4 and D5 absorption into the blood and lymphatic systems, whereas D6 demonstrated the lowest risk. Therefore, we can argue that, among the three tested compounds, D6 is the safest one that can be used in dermatological, cosmetic, and personal care products. This study demonstrates that the stratum corneum, epidermis, and dermis can be also considered reservoirs of cyclic methylsiloxanes. Therefore, these compounds can demonstrate potential long-term bioaccumulation, and can be absorbed to the bloodstream in a long-term and uncontrolled process. Full article

Knowledge of how the biochemical composition of the bovine oviduct is altered due to the oviduct anatomy or the presence of an embryo is lacking. Thus, the aim of this study was to assess the effect of (І) oviduct anatomy and (ІІ) embryo presence on oviductal fluid (OF) protein, amino acid, and carbohydrate composition. Cross-bred beef heifers (n = 19) were synchronized and those in standing estrus were randomly allocated to a cyclic (non-bred) or pregnant (artificially inseminated) group. All heifers were slaughtered on Day 3 after estrus. The oviducts ipsilateral to the corpus luteum from each animal were isolated, straightened and cut, separating ampulla and isthmus. Each portion was flushed with 500 µl of PBS enabling recovery of the oocyte/embryo. Recovered unfertilized oocytes (cyclic group) and embryos (8-cell embryos; pregnant group) were located in the isthmus of the oviduct. Samples of flushing medium from the isthmus and ampulla were used for proteomic (n = 2 per group), amino acid (n = 5), and carbohydrate (n = 5) analysis. For proteomic analysis, total protein from cyclic and pregnant samples were labelled with different cyanine fluorescent probes and separated according to the isoelectric point using immobilized pH gradient strips (pH 3–10, 17 cm, Protean^® IEF cell system, Bio Rad). Second dimension was performed in a polyacrylamide gel (12%) in the presence of SDS using a Protean II XL system (Bio Rad). Images were obtained with a Typhoon 9410 scanner and analyzed with Progenesis SameSpots software v 4.0. Amino acid content in the OF was determined by high performance liquid chromatography (HPLC). Glucose, lactate, and pyruvate were quantified using microfluorometric enzyme-linked assays. For the proteomic assessment, the results of the image analysis were compared by ANOVA. For both amino acid and carbohydrate analyses, statistical analysis was carried out by 2-way ANOVA with the Holm-Sidak nonparametric post hoc analysis. On Day 3 post-estrus, OF composition varied based on (І) anatomical region, where isthmic metabolites were present in lower (i.e., lactate, glycine, and alanine) or higher (i.e., arginine) concentrations compared to the ampulla; and (ІІ) embryo presence, which was correlated with greater, arginine, phosphoglycerate kinase 1, serum albumin, α-1-antiproteinase and IGL@ protein concentrations. In conclusion, data indicate that the composition of bovine OF is anatomically dynamic and influenced by the presence of an early embryo. Full article

Ta–Al multilayer coatings were fabricated through cyclical gradient concentration deposition by direct current magnetron co-sputtering. The as-deposited coatings presented a multilayer structure in the growth direction. The oxidation behavior of the Ta–Al multilayer coatings was explored. The results specified that Ta-rich Ta–Al multilayer coatings demonstrated a restricted oxidation depth after annealing at 600 °C in 1% O₂–99% Ar for up to 100 h. This was attributed to the preferential oxidation of Al, the formation of amorphous Al-oxide sublayers, and the maintenance of a multilayer structure. By contrast, Ta₂O₅ formed after exhausting Al in the oxidation process in an ambient atmosphere at 600 °C which exhibited a crystalline Ta₂O₅-amorphous Al-oxide multilayer structure. Full article

Myeloid phagocytes have evolved to rapidly recognize invading pathogens and clear them through opsonophagocytic killing. The adenylate cyclase toxin (CyaA) of Bordetella pertussis and the edema toxin (ET) of Bacillus anthracis are both calmodulin-activated toxins with adenylyl cyclase activity that invade host cells and massively increase the cellular concentrations of a key second messenger molecule, 3’,5’-cyclic adenosine monophosphate (cAMP). However, the two toxins differ in the kinetics and mode of cell entry and generate different cAMP concentration gradients within the cell. While CyaA rapidly penetrates cells directly across their plasma membrane, the cellular entry of ET depends on receptor-mediated endocytosis and translocation of the enzymatic subunit across the endosomal membrane. We show that CyaA-generated membrane-proximal cAMP gradient strongly inhibits the activation and phosphorylation of Syk, Vav, and Pyk2, thus inhibiting opsonophagocytosis. By contrast, at similar overall cellular cAMP levels, the ET-generated perinuclear cAMP gradient poorly inhibits the activation and phosphorylation of these signaling proteins. Hence, differences in spatiotemporal distribution of cAMP produced by the two adenylyl cyclase toxins differentially affect the opsonophagocytic signaling in myeloid phagocytes. Full article

Monolithic Hf–Si–N coatings and multilayered Hf–Si–N coatings with cyclical gradient concentration were fabricated using reactive direct current magnetron cosputtering. The structure of the Hf–Si–N coatings varied from a crystalline HfN phase, to a mixture of HfN and amorphous phases and to an amorphous phase with continuously increasing the Si content. The multilayered Hf₄₈Si₃N₄₉ coatings exhibited a mixture of face-centered cubic and near-amorphous phases with a maximal hardness of 22.5 GPa, a Young’s modulus of 244 GPa and a residual stress of −1.5 GPa. The crystalline phase-dominant coatings exhibited a linear relationship between the hardness and compressive residual stress, whereas the amorphous phase-dominant coatings exhibited a low hardness level of 15–16 GPa; this hardness is close to that of Si₃N₄. Various oxides were formed after annealing of the Hf–Si–N coatings at 600 °C in a 1% O₂–99% Ar atmosphere. Monoclinic HfO₂ formed after Hf₅₄N₄₆ annealing and amorphous oxide formed for the oxidation-resistant Hf₃₂Si₁₉N₄₉ coatings. The oxidation behavior with respect to the Si content was investigated by using transmission electron microscopy and X-ray photoelectron spectroscopy. Full article

Ru-riched and equiatomic Ru–Al multilayered thin films were fabricated on Si and Inconel 617 substrates. These thin films exhibited a multilayered structure that is caused by stacking cyclical gradient concentration through cosputtering. X-ray diffraction analysis indicated that the as-deposited Ru–Al multilayers comprised Ru and RuAl phases. Oxidation that is caused by annealing atmospheres and elements diffused from substrates was investigated. The results indicated that the inward diffusion of O at 600 °C in a 1% O₂–99% Ar atmosphere was restricted by the formation of an amorphous Al-oxide sublayer, and inward diffusion of O at 800 °C in air was limited by the formation of a crystalline Al₂O₃ scale. Additionally, the outward diffusion of elements from Inconel 617 penetrated the unoxidized parts of the 800 °C–annealed Ru–Al multilayers. Full article

In this study, equiatomic Ru–Zr coatings were deposited on Si wafers at 400 °C by using direct current magnetron cosputtering. The plasma focused on the circular track of the substrate holder and the substrate holder rotated at speeds within 1–30 rpm, resulting in cyclical gradient concentration in the growth direction. The nanoindentation hardness levels of the as-deposited Ru–Zr coatings increased as the stacking periods of the cyclical gradient concentration decreased. After the coatings were annealed in a 1% O₂–99% Ar atmosphere at 600 °C for 30 min, the internally oxidized coatings shifted their respective structures to a laminated structure, misaligned laminated structure, and nanocomposite, depending on their stacking periods. The effects of the stacking period of the cyclical gradient concentration on the mechanical properties and structural evolution of the annealed Ru–Zr coatings were investigated in this study. Full article