In this study, we demonstrate that conducting polymers could be ideal materials for continuously managing humidity in a wide range of enclosed spaces. We demonstrate a simple battery-driven humidity control unit to manage the humidity in a closed environment and studied humidity-responsive nanocapsules using Zn-coordinated lipid nanovesicles. This study not only promises new applications for conducting polymers but also provides an easy approach for fabricating chambers with a controlled environment, which are often used by physicists, chemists, and biologists. In this study, we demonstrate that conducting polymers could be ideal materials for continuously managing humidity in a wide range of enclosed spaces. One method is the gas flow method in which a dry gas that was passed through a desiccant is mixed with a gas that has been passed through water. By controlling the ratio between the two gases, the humidity can be controlled.5 One of the advantages of this method is that an equilibrium can be quickly established , and the changes in the humidity can be continuous. However, this system is relatively complicated and bulky. Aside from the gas supply, a number of different moving parts and valves are required for the construction of the equipment required for this method. Furthermore, the method does not create a closed system, which limits its applicability to laboratory-scale experiments such as gas analysis. The second method predominantly used to precisely control humidity involves the use of saturated salt solutions such as lithium chloride or magnesium chloride.6 This method is highly accurate and relatively simple to perform, because the concentration of a saturated solution is fixed and the vapor pressure can be precisely determined for different temperatures. Because of its high accuracy, this method is used for the certification of humidity sensors. However, this method can lead to salt contaminating a closed system, which is detrimental for laboratory-scale experiments. Furthermore, the humidity value that can be obtained using this method are discontinuous, as the salt solution has to be changed so that different humidity values can be obtained. This can result in it taking several hours for an equilibrium to be achieved.The control of relative humidity (RH) levels in enclosed spaces is not only important for laboratory-scale experiments in physics,In this paper, we report on a simple approach that can be used to precisely control RH levels (from 20% to 90%) in a closed system in a continuous manner; this is achieved through the use of conducting polymer films. This approach involves no moving parts or chemicals that could contaminate an experimental environment, and an equilibrium is achieved within several minutes. By combining a humidity switch module using a printed circuit board, we were able to demonstrate a compact battery-driven humidity control unit that was less than 7 cm × 7 cm in size. We found that this could be used to control the humidity in an environmental chamber with a capacity of over 1 L. We also demonstrated the morphology change of humidity-responsive nanocapsules using Zn-coordinated lipid nanovesicles, which could potentially contribute to the controlled release of chemicals from nanocapsules under different humidity conditions.7 It is well known that PEDOT/PSS films readily absorb water from the air.8,9 The water uptake of PEDOT/PSS films is reported to have a negative effect on the stability and reproducibility of organic electronic devices because many organic semiconductors and metal electrodes are sensitive to moisture.10 However, the extremely strong hygroscopic behavior of PEDOT/PSS films has not attracted much attention. If a freshly prepared free-standing PEDOT/PSS film is placed into a glovebox that has 20% humidity, the weight of the film will increase over time and eventually reach saturation. It has been suggested that this film can absorb water even in very low-humidity conditions and that the hygroscopicity of PEDOT/PSS films could be stronger than that of desiccant silica gels (ca. 0.7 cm3) can absorb roughly 80 mg of water at 20% RH. This value is equivalent to the amount of water contained in 3.4 L of air at 100% RH (the saturated water vapor pressure at 25 °C is roughly 3.2 × 103 Pa). Using the ideal gas law, we find that the volume . In other words, the humidity of a closed system can be controlled using a PEDOT/PSS to gas volume ratio of only 1/5000. This calculation suggests that if one can control the amount of water present in a PEDOT/PSS film, an ideal material for managing the humidity in a closed system could be created.Our proposal utilizes the conducting polymer poly/polystyrene sulfonate (PEDOT/PSS) to control the humidity in a closed system. PEDOT/PSS has attracted a lot of attention owing to its potential for use in applications such as energy storage, energy conversion, and flexible electronics.ica gels . As such11–16 As a benchmark conducting polymer, PEDOT/PSS films exhibit among the highest electrical conductivities  and the electrical conductivity value is less sensitive to the humidity (the amount of water in the film).8 This is because the PEDOT nanocrystals form a layered structure. The spacing between the layers could change before and after the water absorption, but the crystal ordering and crystal size don't significantly affect by the amount of the water in the film  was applied to the film, its weight started to decrease and became saturated at ca. 3 mg. This can be attributed to the desorption of water from the polymer film. A further ca. 10 mg was lost when the current density was increased to 50 A cm−2 (1.0 A), and 18 mg in total was lost for a current density of 75 A cm−2 (1.5 A). Interestingly, the weight lost as a constant current density does not keep decreasing but will saturate at a constant value. Once a larger current was applied to the sample, the weight began to further decrease. However, when the current density decreased, the weight began to increase; this suggests that the PEDOT/PSS film began to absorb water from the environment. The weight of the film did not increase continuously, but rather became saturated at a certain value depending on the current density. At a current flow of zero, the weight of the film reverted to the initial value shown in 17 Note that the carrier mobility in the PEDOT component of the film was partly limited by the columbic interaction of the PSS−. This interaction may have screened the water molecules owing to the high dielectric constant. Once the water was baked out, the carrier mobility decreased, and the resistivity increased. For the same current density passing through the film, the sample temperature was higher during the cooling process owing to a larger resistivity. As a result, the amount of the water uptaken from the air is smaller. At a current density of zero, the temperature of the film should be the same as that of the room temperature, which would result in the weight of the film reverting to its initial value.In order to confirm whether water absorption and desorption occur in PEDOT/PSS films during current flow, we prepared a free-standing film with a thickness of e sample . The exp−2 pulse current at a duration of 60 s to the films. The weight of the films quickly decreased before returning to their initial weight after ca. 10 min. The absolute amount of water desorbed during the pulse current and the recovery time remained almost constant, which suggests that the PEDOT/PSS films are very stable. The water desorption of the PEDOT/PSS films during the current flow was faster than the water absorption from the atmosphere. −2, 5 °C for a current density of 50 A cm−2, and 14 °C for a current density of 75 A cm−2. Although a higher current density can desorb a greater amount of water, it is not preferable for humidity control in a closed system because it could raise the temperature of the system. In order to avoid any unwanted temperature changes from occurring during humidity control, we set the current density to being smaller than 40 A cm−2 in our experiment.In order to study the stability and reproducibility of water desorption and absorption of the PEDOT/PSS films during current flow, we applied a 75 A cmca. 20% was maintained as a starting point. As shown in −2 was applied to the film, the humidity increased and became saturated at ca. 30%. The saturated humidity was ca. 42% at a current density of 20 A cm−2, 62% at a current density of 25 A cm−2, and over 86% at a current density of 40 A cm−2. When the current density decreased from 40 to 20 A cm−2, the saturated humidity dropped to 49% owing to the water absorption of the PEDOT/PSS film. When we stopped applying the voltage to the PEDOT/PSS film, the humidity in the box dropped below 30% after 30 min. The change in temperature in the sealed box over the entirety of the measurement process was less than 5 °C. This result is consistent with the observation of the change in weight of the film shown in A PEDOT/PSS film whose dimensions were 2.6 cm × 5.0 cm × 290 μm was fixed inside a sealed box that had a volume of 0.5 L . The humvia an on/off switch; the switch will turn on the power supply to the PEDOT/PSS film when the humidity falls below a previously defined point , and it will turn it off when the humidity rises above a different previously defined point . We continuously conducted the humidity control experiment using a single set-up ; no gas exchange took place during the process. −2. We were able to use a battery-driven on/off switch on a printed circuit board to fabricate the proof-of-concept humidity controller, as shown in †The compact and simple structure of our device is an advantage. The high conductivity of the PEDOT/PSS film makes the device have low power consumption, such that it could be driven using a battery. The two-probe resistivity of the PEDOT/PSS–Au wire in our set-up was less than 0.3 Ω. A 1.5 V battery can produce a current density as high as 35 A cm18 In this paper, we investigate humidity-responsive nanocapsules in a closed plastic chamber using Zn-coordinated lipid nanovesicles . The encapsulation of functional molecules for daily care products and agricultural chemicals and its release is a currently much researched topic.The humidity control approach proposed in this study could be used in a closed system, which has the advantage that it allows us to study a controlled release of chemicals from nanocapsules, because no chemicals or gases are exchanged when the humidity is varied. The control of the release of chemicals from nanocapsules, which are usually accompanied by a morphological change in the nanocapsules under various stimuli, is important because of their applications to medicine, food, and daily care products. However, research into humidity-responsive nanocapsules is far less reported upon than conventional pH-, light-, and temperature-responsive nanocapsules.vesicles .19,20Figles 19,20. This stles 19,20. This reIn conclusion, we have demonstrated a novel approach to controlling the humidity in a closed system with an RH between 20% and 90%. In comparison to the gas flow method and the approach that uses saturated salt solutions, there are no moving parts in our method nor are there any chemicals that will contaminate the experimental environment; furthermore, equilibrium is very quick to achieve. Owing to the high electrical conductivity of conducting polymers, our humidity control unit consumes very little power. We believe that this will allow for humidity control units that can be powered by batteries to be developed. Our proposed device could also contribute to the study of the controlled release of substances for potential future applications in daily care products or agricultural chemicals. This study not only presents new potential applications for conducting polymers but also provides a new way to make more compact, simple, and low-cost environmental chambers.There are no conflicts to declare.RA-008-C8RA01776J-s001RA-008-C8RA01776J-s002


Then, the graphene-modified coating for building was prepared by using graphene-modified styrofoam emulsion. All the performance indexes of the coating are in line with the industry standards, and it still showed benign EMI shielding effect even when the graphene content was low. It is demonstrated that in situ polymerization technology and the application of graphene in resin coatings modification will promote the development of green coatings.With the aim of developing green coatings, styrene–acrylic emulsion has been widely used in architectural coatings due to its excellent environmental protection and energy conservation. Nevertheless, the lack of water and oxygen resistance of water-based styrofoam coatings has promoted various nanomaterials being studied for modification. To improve the performance of waterborne styrofoam coating, we introduced the graphene nanopowder and expected to enable it with the function of electromagnetic interference (EMI) shielding to reduce the damage of electromagnetic radiation. In this paper, the problem of poor interface compatibility between graphene and polymer resin was successfully addressed by in situ polymerization. In the process of pre-polymerization of styrene–acrylic emulsion monomer, graphene-modified styrene–acrylic emulsion was obtained by introducing graphene aqueous dispersion. The results showed that the styrene–acrylic emulsion with 4 wt% aqueous graphene dispersions exhibited the best dispersion stability, improved water and oxygen resistance, and the conductivity reached 1.89 × 10 In the field of green coatings, waterborne coatings which contain no or only a small amount of organic solvents, have the great advantages of low pollution, being tasteless, and causing little or almost no harm to human health and the environment ,2,3. Thuformance ,15,16. Mformance ,18, so eformance .2 foaming technique [Compared with traditional metal-based EMI shielding materials, the excellent properties of conductive polymer composites include electrical conductivity, chemical and temperature resistance, light weight, high specific surface area and large aspect ratio, which are all conducive to the application of EMI shielding . Among technique . Bontas echnique  modifiedechnique . The graechnique ,26. Therechnique  synthesiechnique ,28, it iThis work uniquely introduced in situ polymerization which provides an ideal strategy to overcome the interfacial incompatibility between graphene and the polymer resins; surfactants were added to improve the dispersion of graphene in water; and it successfully realized an eco-friendly method of preparing graphene-modified styrene–acrylic emulsion in aqueous solution. The results revealed that graphene modification can effectively improve the rheological properties of the styrene–acrylic emulsion along with the excellent EMI shielding effect. Finally, this graphene-modified styrene–acrylic emulsion was successfully applied as a waterborne architectural coating which shows excellent water resistance and low water permeability. The successful modification of graphene also affords the coating with good electromagnetic shielding performance. Each performance index of the graphene-modified styrene–acrylic emulsion-based coating reaches the industry standard (T/CNCIA 01004-2017) for application as waterborne graphene electromagnetic shielding coating for architecture. We expect the strategy will open up a new vista in fabricating graphene-modified styrene–acrylic emulsion and the application of waterborne coatings.3) were all purchased from Chemical Reagent Co., Ltd.  with A.R. grade. Ethylene glycol, Thickening agent HS300, Dispersing agent P-19, Wetting agent X-405, Titanium dioxide, White clay (4000 mesh), CaCO3 (1250 mesh), Hydroxyethyl cellulose (HEC), Coalescing agents, Flatting agent 3020, Antifoaming agents 3016, Thickening agent ASE-60 and Corrosion remover 981 were all provided from Fuzhou Aidite New Material Technology Co., Ltd. . All the above chemicals were used as received without further purification.Graphene , sodium carboxymethyl cellulose , Polyvinyl alcohol and sodium ligninsulfonate (SLS) were purchased from Xiamen Kaina Co., Clariant Chemicals Co., Ltd.  and Aladdin Reagents Co., respectively. Styrene (St), butyl acrylate (BA), methyl methacrylate (MMA), propyl methacrylate (n-PMA), alkoxyphenol ethoxylate , sodium polymethacrylate , ammonium persulfate (APS) and sodium bicarbonate  and sodium carboxymethylcellulose (CMC) as stabilizer through the following process . First, Distilled water (150 g) and OP-10 (26 g) were added to a three-necked flask, and then stirred at high speed until foaming. Four kinds of monomers, St (200 g), MMA (50 g), BA (210 g) and n-PMA (9 g), were mixed and added to the foamed emulsifier solution within 30 min. Then the aqueous graphene dispersion was continuously added at the same speed. The obtained mixture was stirred at high speed for 20 min, and then stirred at low speed for 10 min to obtain a pre-emulsion.3 in 12 g deionized water) were added to above mixture at 70 °C. Then the remaining pre-emulsion and the initiator–buffer mixed solution (1.5 g APS and 1.5 g NaHCO3 in 80 g deionized water) were dripped in within 3 h. Finally, an extra dose of initiator solution (0.6 g APS in 10 g deionized water) was continuously added dropwise at the same rate to produce a sufficient polymerization. Cool down naturally after 2 h of reaction.Deionized water (220 g), OP-10 (4 g) and PMA (15 g) were added to a four-necked flask in sequence and stirred at high speed until foaming. One-third of the pre-emulsion and the initiator–buffer mixed solution  The preparation of graphene-modified styrene–acrylic emulsion architectural coatings requires the preparation of premixed slurry; according to the ratio of premixed slurry shown on the left of (2) According to the ratio on the right of Among the above, thickening agent HS300 and thickening agent ASE-60 are increasing the viscosity of the coating, so it can better adhere to the substrate material. Dispersing agent P-19 is used to disperse the particles which need to be dispersed in waterborne coatings on the surface, so that it has better stability. Wetting agent X-405 is to moisten the substrate. Titanium dioxide, White clay (4000 mesh) and CaCO3 (1250 mesh) are fillers. Hydroxyethyl cellulose (HEC) can act as a binder and thickener. Coalescing agents are to make the coating better film. The purpose of adding flatting agent 3020 is to make the fluidity of the coating better and smoother. In the process of high-speed stirring, samples will be bubbles, so it needs to be anti-foamed by agent 3016 to stir more fully and reduce the production of bubbles after coating film.The graphene-modified styrene–acrylic architectural coating is denoted as G-St/Ac-C. The coating without graphene (St/Ac-C) was also prepared for comparison.−1. Rheological properties of styrene–acrylic emulsion were measured using a rheometer . A 40 mm stainless steel parallel plate was used for the tests and the plate gap was 1 mm. All experiments were carried out at a constant temperature of 25 °C. The Zeta potential of aqueous graphene dispersion was measured using a Malvern Zetaszier Nano-ZS; the instrument measures the particle size by the principle of Dynamic Light Scattering (DLS). Before the test, the sample was diluted with deionized water to a certain multiple before measurement. Surface resistivity was measured by using a four-probe setup . The electromagnetic shielding efficiency for graphene-modified styrene–acrylic architectural coating was measured with a shielding effectiveness tester (DR-S02) using the flange coaxial test device for shielding effectiveness.The X-ray Diffraction (XRD) patterns were performed on a Bruker D8 diffractometer with Cu-Kα radiation (λ = 0.15406 nm) over a range of 10–80°. The field-emission scanning electron microscope  and transmission electron microscopy  were carried out to analyze the morphologies and microstructures of the graphene in the styrene–acrylic emulsion. Confocal Raman microspectrometer (DXR2xi) with an Arion laser at the excitation wavelength of 532 nm was used to collect Raman spectra of the samples. To confirm the amount of carbon in the nanocomposites, thermogravimetric analysis (TGA) was performed  in air atmosphere. Fourier-transform infrared (FTIR) spectra were obtained using a Thermo Fisher Scientific Instruments Nicolet iS10 FTIR spectrometer at room temperature in the wavenumber range of 400–4000 cm−1 are corresponding to the characteristic peaks of G-band and 2D-band of graphene, respectively [G/I2D value reveals the number of graphene layers. Generally, graphene with an IG/I2D about 0.5~1 contains 1–2 layers, while multiple layers when IG/I2D = 1.5~3. G/I2D values of the three sites which indicates the graphene stacks with multiple layers other than bulk graphite. Thus, in the following polymerization process, we chose the well-stabilized aqueous graphene dispersion with PVA/CMC (2.67 wt%:1.33 wt%) to modify the styrene–acrylic emulsion.Thus, with the aid of 3 wt% surfactants including PVA (3 wt%), CMC (3 wt%), SLS (3 wt%), PVA/CMC (1.5 wt%:1.5 wt%), PVA/SLS (1.5 wt%:1.5 wt%) and CMC/SLS (1.5 wt%:1.5 wt%), the stability of the dispersion with 5 wt% graphene is remarkably improved. In contrast, when 5 wt% graphene was dispersed in pure water, it almost completely stratifies within one week. Noticeably different from the other surfactant systems with distinct precipitation after two months, the dispersion with PVA/CMC shows little stratification, indicating excellent stability a. Transmectively ,31,32. Tθ = 26° increases. Compared with the film formed from the pure styrene–acrylic emulsion (Dynamic light scattering (DLS), X-ray diffraction (XRD) and transmission electron microscopy (TEM) were performed to identify the successful modification of graphene. Compared to the pure styrene–acrylic emulsion, DLS results show that with the addition of graphene, the size of the particles in the emulsion increases in line with the increasing of graphene addition a. Furtheemulsion c, the fiemulsion d–h show −1 is assigned to styrene–acrylic emulsions [1580/I1600 reveals the graphene content at the corresponding site. As 1580/I1600 gradually decrease from site 3 to site 1, and there is almost no graphene dispersion at site 1 in the blue area. In −1 which are assigned to the vibration of -CH3, -CH2, C=O, C-O, respectively, and the peaks at 759–1059 cm−1 are characteristic peaks of butyl acrylate, which are consistent with the literature results [Raman mapping was performed to characterize the dispersion state and the relative content of graphene in the emulsion. mulsions . Thus, t results . Thus, tFor coatings, the rheological properties are critical. Thus, the rheological properties of the obtained emulsions with different graphene content were investigated a–d. Figu−2 S/cm, when the addition reaches 4.0 wt%. Then, the conductivity sharply drops to 3.62 × 10−3 S/cm at the addition of 4.4 wt%. In order to figure out the mechanism behind the conductivity change with the graphene addition, the exact graphene content in the graphene-modified styrene–acrylic emulsion was investigated by thermogravimetric analysis (TGA), SEM topography and Raman mapping image. As shown in Compared to the traditional nonconductive styrene–acrylic emulsion, the loading of graphene endows the modified emulsion with good electrical conductivity. 3 is distinctly improved in the G-St/Ac-C, indicating that the addition of graphene may facilitate the crystallization of CaCO3 to form more ordered arrangement. CaCO3, as a main filler, has an important influence on the properties of the architectural coating. The enhanced crystallinity of CaCO3 may also be beneficial to improve water resistance and reduce water permeability. Furthermore, To verify the practical application as a waterborne coating, the graphene-modified styrene–acrylic architectural coating (G-St/Ac-C) was prepared according to the industrial formulation . AmazingIn this study, aqueous graphene dispersion with high stability was readily prepared and the graphene-modified styrene–acrylic emulsion was successfully synthesized by in situ polymerization in aqueous solution through a facile eco-friendly method. The graphene was first dispersed into aqueous solution with the aid of polymeric surfactant stabilizer PVA and CMC. Then the aqueous graphene dispersion was added to the pre-emulsion with monomers and initiator. Through in situ polymerization, the graphene was successfully loaded into the styrene–acrylic emulsion. The optimum graphene addition amount (4.0 wt%) was explored through systematically investigating the rheological properties, conductive properties and the exact graphene content with different graphene additions. The graphene modification improved the rheological properties of the styrene–acrylic emulsion and endowed it with excellent electrical conductivity. The emulsion was further applied as graphene electromagnetic shielding coating for architecture and every performance reaches the industry standard. We hope this in situ polymerization method for graphene-modified styrene–acrylic emulsions will be a potential application in the field of environmentally friendly coatings.


The SEM morphology revealed a sphere-like fluffy structure, particles of the 0.4%rGO-10%SnO2-Fe2O3 composite were smaller and more compact with a specific surface area of 223.19 m2/g, the first discharge capacity of 1423.75 mAh/g, and the specific capacity could be maintained at 687.60 mAh/g even after 100 cycles. It exhibited a good ratio performance and electrochemical reversibility, smaller charge transfer resistance, and contact resistance, which aided in lithium-ion transport. Its superior electrochemical performance was due to the addition of graphene, which made the spherical particle size distribution more uniform, effectively lowering the volume expansion during the process of charging and discharging and improving the electrochemical cycle stability of the anode materials.To enhance the conductivity and volume expansion during the charging and discharging of transition metal oxide anode materials, rGO-SnO These materials also have the advantages of low working potential, simple charging and discharging cycles, and abundant reserves, which makes them potential candidates [The acceleration of social progress and industrialization will create an increasingly high demand for energy resources. At present, the commercial lithium-ion battery anode material is mainly graphite, which has a small theoretical specific capacity 372 mAh/g) and hence cannot meet the power supply demand of large equipment ,2,3. The mAh/g anndidates . Howeverndidates ,13,14,15ndidates ,17,18.2@Fe2O3 double-shell hollow spheres. For a current density of 100 mA/g, the specific discharge capacity decreased to 464 mAh/g after 46 cycles while increased to 1043 mAh/g after 190 cycles, showcasing better electrochemical performance. Chai et al. [2O3 nanostructured anode materials by simple pyrolysis deposition of ferrocene on stainless steel substrates. After 300 cycles at a current density of 500 mA/g, it could still retain a high reversible specific capacity of 1138 mAh/g.Cui et al.  used a ti et al.  used glui et al.  prepared+ embedding and removal, alleviating volume expansion [2O3/graphene nano-sheets by a simple and effective spray drying method, which showed a specific capacity of 711 mAh/g after 50 cycles of charging and discharging, and the chemical properties were quite improved. Qin et al. [2O3/N-doped graphene (Fe2O3/N-rGO) anode materials by a one-step in situ hydrothermal growth method. The porous structure and the introduction of curly GO greatly improved the electrochemical performance of the composite electrode, and the discharge-specific capacity was maintained at 1072.5 mAh/g after 100 cycles. Yang et al. [3O4 nanoparticle coated with graphene (GE-Co3O4) exhibited a high reversible specific capacity of 1100 mAh/g during the first 10 cycles, and the reversible specific capacity could be maintained at 1000 mAh/g even after 130 cycles, showing excellent cycling performance. Therefore, it exhibited great potential as an anode material for lithium storage. Lian et al. [2/graphene composites. The SnO2 nanoparticles could be uniformly distributed on the graphene substrate and showed a reversible specific capacity of 1304 mAh/g at a current density of 100 mA/g. Zhou et al. [+ embedding. Hu et al. [2 nanospheres with GO. This compound exhibited excellent electrochemical performance with a high reversible specific capacity of 1107 mAh/g after 100 cycles at a current density of 0.1 A/g.Due to the unique structure of graphene, it can show good electrochemical performance when used as the anode material of lithium-ion batteries ,24,25,26xpansion ,33,34,35xpansion  preparedn et al.  preparedg et al.  introducn et al.  developeu et al.  synthesiu et al. ,42,43,44u et al.  combinedu et al.  synthesi2-Fe2O3 composite materials. The phase composition and microstructure were determined by XRD and SEM. The charge and discharge performance, cycle performance, rate performance, and AC impedance of the materials were tested. The physical and chemical properties and internal mechanisms of the composite materials were explored.To enhance the conductivity and volume expansion in the charging and discharging cycles of transition metal oxide anode materials, this paper employed the in situ hydrothermal synthesis method to prepare rGO-SnO2-Fe2O3 composite metal oxide. The Sn content was calculated as SnO2, the mass percentage was fixed at 10%, and the graphene content was varied as 0.2%, 0.4%, 0.6%, and 0.8%, respectively.A certain amount of graphene oxide was added to deionized water and ultrasonic dispersion for 1 h, then the percentage of tin chloride and ferric chloride was dissolved. Then, sodium hydroxide solution (1 mol/L) was added during the mixing process until the solution pH reached approximately 9. After stirring for 4 h, the mixture was shifted to the high-pressure reaction kettle  for 4 h under 100 °C. It was further cooled to room temperature and allowed to stand for 8 h, washed, dried at 60 °C for 24 h, and lastly, vacuum roasted at 400 °C for 4 h to obtain the rGO-SnOThe prepared anode material was used as the active material, acetylene black was used as the conductive agent, and PVDF was used as the binder. The mass ratio was fixed as 8:1:1. N-methyl pyrrolidone (NMP) was used as the solvent to prepare the slurry. After coating, vacuum drying at 80 °C for 12 h, and stamping, the electrode sheet was ready for use.6/ethylene carbonate (EC) + dimethyl carbonate (DMC) (volume ratio was 1:1) mixture solution was used as the electrolyte. The battery was assembled according to a certain sequence of a positive shell, electrode sheet, diaphragm, lithium sheet, gasket, spring sheet, and negative shell. During the assembly process, the electrolyte was added to both sides of the diaphragm.The electrochemical performance tests of electrode materials were executed by using the half-cell test system, the prepared electrode sheet as the anode, the lithium sheet as the counter electrode and auxiliary electrode, the polypropylene microporous membrane  as the diaphragm, and 1 mol/L LiPFThe surface morphology of the anode material was observed and analyzed by a SSX-550 tungsten filament analytical scanning electron microscope .A X ‘Pert Pro MPD diffractometer PW3040/60 manufactured by Koninklijke Philips N.V., Amsterdam, The Netherlands was used. The Kα of the Cu target was the radiation source (wavelength 1.540598 A), the working voltage was 40 kV, the sweep speed was set as 6°/min, and the sweep range was 10–90°.The surface area tester (Micromeritics ASAP 2020) produced by Micromeritics  was used to calculate the specific surface area of the sample by Brunauer–Emmett–Teller (BET) isotherm method.A constant current charge/discharge test was carried out following the steps of static-constant current discharge-static-constant current charge-constant voltage charge. The test voltage window was set as 0.01–3 V. The charge and discharge current values were calculated from the theoretical specific capacity and the mass of the active substance.The instrument used for the cyclic voltammetry test was the CHI660D electrochemical workstation of Shanghai Chenhua Company, Shanghai, China. The test conditions were as follows: scan rate of 0.1 mV/s, the voltage scan range of 0–3.0 V.In the Electrochemical impedance spectroscopy test, the frequency range was 0.01 Hz–1 MHz, and the voltage vibration amplitude was 5 mV.2-Fe2O3 composite oxide samples with varying graphene contents, and the corresponding specific surface area is listed in 2-Fe2O3 composite oxide exhibited an irregular structure. After adding graphene, the rGO-10%SnO2-Fe2O3 composite materials exhibited a globular fluffy structure, and the granular complex was distributed on the surface of the gossamer graphene, in between the lamellae  [4+ to Sn2+ disappeared. The oxidation peaks of the first cycle appeared at 1.7 V and 2.0 V, corresponding to the transformation of Fe0 to Fe3+ and Sn2+ to Sn4+. The cyclic voltammetry curves overlapped well, suggesting higher reversibility and better stability.Cyclic voltametric curves of 10%SnOnversion ,50,51. IEI film) ,53, and 2-Fe2O3 composites with different graphene contents are shown in 2-Fe2O3, 0.6%rGO-10%SnO2-Fe2O3, 0.8%rGO-10%SnO2-Fe2O3, 0.2%rGO-10%SnO2-Fe2O3 and 10%SnO2-Fe2O3. After graphene addition, the resistance of the material decreased, which was closely related to the effect of graphene on the microstructure. Among them, when the graphene content was 0.4%, the 0.4%rGO-10%SnO2-Fe2O3 anode material possessed the smallest semi-circuit radius, indicating that it had the smallest contact resistance and charge transfer resistance, which was conducive to the transport of lithium ions and was consistent with the cycling performance results. With the amount of graphene increasing, as in 0.8%rGO-10%SnO2-Fe2O3, the charge transfer resistance was increased due to the agglomeration phenomenon and lower specific surface area.The AC impedance spectra of rGO-SnO2-Fe2O3 anode materials were prepared by using SnO2-Fe2O3 as the matrix and adding graphene oxide in varying contents. The structural characterization and electrochemical performance of the above materials were investigated, and the following conclusions were drawn: (1) The phase structure of the rGO-SnO2-Fe2O3 composite was SnO2, and the morphology was a globular fluffy structure. The granular complex was distributed on the surface and between the lamellae of the bubble graphene. The morphology of 0.4%rGO-10%SnO2-Fe2O3 composite was the best, and the spherical particles were smaller and more in numbers. The spherical particles were more closely packed, and the specific surface area was 223.19 m2/g. (2) The cycle performance of the 0.4%rGO-10%SnO2-Fe2O3 anode material was the best, the first discharge-specific capacity was 1423.75 mAh/g, and after 100 cycles, the capacity was maintained at approximately 687.60 mAh/g. The results of different rate tests showed that the specific capacity of the 0.4%rGO-10%SnO2-Fe2O3 anode material decreased with an increase in the rate, but the overall rate of decline was relatively slow. (3) 0.4%rGO-10%SnO2-Fe2O3 exhibited better electrochemical reversibility, smaller contact resistance, and charge transfer resistance. All these characteristics were beneficial to the transport of lithium ions and were consistent with the cycling performance results. (4) The excellent electrochemical performance of 0.4%rGO-10%SnO2-Fe2O3 can be attributed to the addition of graphene, which made the size distribution of spherical particles more uniform, effectively alleviating the volume expansion phenomenon during the process of charging and discharging. This has further enhanced the electrochemical cycle stability of anode materials. It can be seen from the experimental results that the addition of graphene can improve the specific capacity and cycle stability. However, it has no significant effect on the capacity attenuation in the first lap, which will be paid more attention in the later research.In this paper, rGO-SnO


Recently, it was associated with higher rates of progression to chronic osteomyelitis in pediatric patients with acute MSSA osteomyelitis, independent of the treatment choice. Thus, its detection could be important for management of certain MSSA infections. Broth microdilution at high inoculum is the gold standard for the detection of the CzIE. Yet, this method is lengthy (3–5 days), cumbersome, and expensive. We developed a nitrocefin-based rapid test (∼3h) which can identify the CzIE among MSSA isolates and that can be incorporated in the laboratory workflow. Previously, this rapid test was validated in MSSA isolates from adult patients with bacteremia. Here, we aimed to validate it in isolates from pediatric patients with acute MSSA osteomyelitisThe cefazolin (Cz) inoculum effect (CzIE) defined as a Cz MIC ≥ 16 µg/mL at high inoculum . Performance metrics were calculated for the complete data set and for specific BlaZ types.We included 206 MSSA isolates from the same number pediatric patients from Houston, TX (2011–2018). All patients were diagnosed with acute MSSA osteomyelitis. The CzIE was determined using broth microdilution at high inoculum with a cutoff of Cz MIC ≥ 16 µg/mL. Whole genome sequence analysis was performed using Illumina Hi-Seq. BlaZ type was determined according to predicted BlaZ residues at position 119 and 207. The nitrocefin rapid test was performed following the published protocol . The false negative and false positive rate of 9.2% and 13.8%, respectively, with an overall accuracy of 89%. There were no false positive results among blaZ negative strains. Among type A BlaZ MSSA, the sensitivity was 96.7%, and the specificity was 80%, while in type C BlaZ, they were 70.5% and 92.8%, respectively.The prevalence of the CzIE was 33%. Compared to the gold standard, the nitrocefin rapid test had a Performance metrics of the Nitrocefin-based Rapid Test in 206 MSSA isolates from pediatric patients with acute MSSA osteomyelitis compared to the Gold Standard of Broth Microdilution at high Inoculum.The nitrocefin-based rapid test was able to correctly detect the CzIE in isolates from pediatric patients with MSSA acute osteomyelitis, with an overall accuracy of 89%. Implementation of this test may contribute to therapeutic decisions in deep-seated MSSA infections.Sheldon L. Kaplan, MD, MeMed: Grant/Research Support|Pfizer: Grant/Research Support Jonathon C. McNeil, MD, Agency for Healthcare Research and Quality: Grant/Research Support|Allergan: Provided reagents for unrelated research|Nabriva: Site investigator for multicenter clinical trial Cesar A. Arias, MD, PhD, Entasis: Grant/Research Support|MeMed Diagnostics Ltd: Grant/Research Support|Merck: Grant/Research Support.


The presence of trigonal GeO2 under graphene was identified by Raman and XRD measurements. The oxidation of Ge leads to the formation of blisters under the graphene layer. It is suggested that oxidation of Ge is connected with the dissociation of water molecules and penetration of OH molecules or O to the Ge surface. It has also been found that the formation of blisters of GeO2 leads to a dramatic increase in the intensity of the graphene Raman spectrum. The increase in the Raman signal intensity is most likely due to the screening of graphene by GeO2 from the Ge(001) surface.The problem of graphene protection of Ge surfaces against oxidation is investigated. Raman, X-Ray diffraction (XRD), atomic force microscopy (AFM) and scanning electron microscopy (SEM) measurements of graphene epitaxially grown on Ge(001)/Si(001) substrates are presented. It is shown that the penetration of water vapor through graphene defects on Gr/Ge(001)/Si(001) samples leads to the oxidation of germanium, forming GeO Due to the tight graphene lattice, it may be expected that it should protect surfaces against oxidation well. In this work protection of germanium surfaces by graphene was investigated. The CVD growth of graphene on Ge001)/Si(001) substrates has been used for this purpose. This method of growth was proposed as a promising method for obtaining graphene layers on large area substrates [/Si(001) 2 . TherefoThe graphene growth on Ge(001)/Si(001) substrates is complicated due to the formation of germanium nano-facets underneath the graphene ,14. The Graphene films were synthesized in a 6-inch Aixtron Black Magic system using the CVD method on Ge(001)/Si(001) wafers which contained 3 μm thick Ge film. During the graphene deposition a pressure of 850 mbar was sustained, and the temperature was 900 °C. Methane gas mixed with Ar in the ratio of 1:200 was used as a carbon precursor. The CVD growth was performed on 2 inch wafers preceded by annealing the substrate under pure hydrogen to reduce native oxides.A scanning electron microscope (SEM)  equipped with a secondary-electron detector was used to observe the influence of water vapor on the graphene morphology.Room temperature Raman measurements were done using a Renishaw Via Raman micro-scope system with a 532 nm Nd:YAG laser as an excitation source, with a laser spot diameter of approximately 0.5 μm. The laser was focused on the sample using a ×100 objective and a numerical aperture NA = 0.9 with the average power of the laser being about 5 mW. Spatial Raman maps were performed in the area of 10 μm × 10 μm (15) with a distance of 1 micron between collected spectra. The Raman peaks were fitted with a single Lorentzian/Gaussian shape using the Renishaw WiRE.4.0 program .The atomic force microscopy (AFM) was measured on a Veeco Dimension V SPM with a NanoScope V controller. The surface images were flattened and plain fitted using Nanoscope analysis software. OTESPA Al coated probes with a constant force of ~42 N/m with a typical nominal tip radius of 7 nm were used for the AFM imaging. All the AFM measurements were performed in a normal atmosphere at room temperature.The measurement of the XRD curve was performed using a SmartLab diffractometer equipped with a 9-kV rotating Cu-anode.8 cm−2. The AFM image of one group of blisters is shown in After growth the 1 cm × 1 cm samples were subjected to a humid atmosphere (about 90%) at 50 °C for about 3 h or about 150 h. The samples were placed in a heated container filled with water upon the water surface. The temperature and humidity were measured in the simplest way using a thermometer and a hygrometer. After 3 h of water vapor treatment it was possible to observe regular spots of the size of a few micrometers on the surface of the Gr/Ge/Si(001) sample on the SEM. The SEM image of this surface is shown in The longer water vapor treatment, up to 150 h, dramatically changed the morphology of the surface of the Gr/Ge/Si(001). The SEM image of the surface is shown in −1, 166 cm−1 and 123 cm−1 shown in 2 [2 Raman modes appears approximately proportional to the thickness of the blisters. The fact that GeO2 blisters already appear after 3 h of water vapor treatment indicates that the influence of water vapor strongly speeds up the oxidation process of Ge. The samples kept in the “drawer” (room temperature and low humidity atmosphere) have not shown any occurrence of GeO2. Additionally, there is no sign in the spectra of Raman Si mode (520 cm−1), which indicates that the Ge layer is not completely dissolved under the blisters. As can be seen from the lower part of We wondered what the origin of the blisters on the surface of the Gr/Ge/Si(001) was. The Raman measurements indicate that in the blister locations, for both samples after 3 and 150 h of water vapor treatment, in addition to the intense Ge mode, the spectrum characteristic for germanium oxide occurs. The observed lines at 444 cmown in 2 . This spThe intensities of the Raman spectra of graphene are quite different in the region of blisters versus further away from them. The corresponding spectra are shown in 12 cm−2 before, to about 1.5 × 1012 cm−2 after, the water vapor treatment. The ratio of the D/D’ intensity provides information about the nature of the defects created by the water vapor. The ratio of D/D’ modes is about eleven for the untreated sample (virgin) (measured only in a few points because of a very long collection time) and changes to about seven for the sample after 150 h of water vapor treatment, as shown in the histogram in 3 defects in the virgin sample changes to domination by point defects in the water vapor treated sample. Some experimental points around a value of three to four of the D/D’ ratio /Si(001) sample for 150 h in the humid atmosphere results in a shift of a group of points in ent with . This ch2 blisters on the surface of graphene/Ge(001)/Si(001) surface indicate the presence of graphene defects which are responsible for process water vapor penetration and oxidation of the germanium surface. However, it is puzzling why the oxidation of germanium under graphene takes place so effectively. Exposing pure germanium without graphene to the same oxidation conditions does not lead to formation of GeO2 on the surface. The question arises as to why presence of graphene on the surface of germanium speeds up the process of oxidation. The answer can be found in previous experiments connected with the investigation of the interaction of water with the graphene monolayer. Electron energy loss spectroscopy measurements on water adsorption on quasi-freestanding monolayer graphene on Pt(111) have shown that water molecules at room temperature dissociate, giving rise to C–H vibrational bands [2O --- C–H + OH, or 2C + H2O --- 2(C–H) + O. Most likely, O atoms or OH molecules may go through graphene defects and will interact with the germanium surface, leading to faster oxidation of Ge by O or by OH according to reaction:Formation of GeOal bands . This in2 underneath the graphene identified by Raman measurements, proves that oxidation of Ge in these places takes place. Most likely, extended defects connected with poor bonding between graphene sheets may be responsible for sites of dissociation of water molecules and places of penetration of OH molecules to the Ge surface.A similar process was reported in the case of oxidation of Cu surfaces ,26. It w2 compared to Ge(001), is strongly enhanced. This increase is similar to the increase in the Raman signal by surface enhanced Raman spectroscopy (SERS) by metallic nanoparticles, which is a well-known effect [2 blisters as being highly conductive. The Raman signal from the area of the blisters is about one or two orders of magnitude stronger than the one outside of this area. This increase in the graphene-related Raman bands has already been reported recently when water penetrates underneath the graphene layer and detaches it from the Ge substrate [2 seems to be connected with the isolation of the graphene by the GeO2 from the Ge(001) surface. The 2D vs. G diagram shown in 2 is predominantly under tensile strain, which is in qualitative agreement with the AFM results shown in The intensity of the Raman spectra obtained from the graphene that is on GeOn effect ,31,32,33ubstrate . The low3 defects. Apparently, the hill-and-valley nano-structures which are positioned at 90° to each other and run along the <100> directions on the Ge(001) surface create the conditions for the formation of sp3 bonds between some Ge surface atoms and carbon atoms from the graphene lattice. The presence of water vapor may break these sp3 bonds and change them into graphene point defects. The histogram presented in 12 cm−2 before, to about 1.5 × 1012 cm−2 after, the water vapor treatment. These point defects may be vacancies and may have the character of five-fold and seven-fold rings called Stole-Wales defects [8 cm−2 which is several orders of magnitude lower than the density of point defects established from the Raman D/G ratio. That clearly proves that the defects responsible for the oxidation of germanium are not point defects. This is in agreement with the histogram presented in The Raman spectrum of graphene outside and inside the blister is characterized by a large D peak. The ratio of the intensity of the D to G peak being 1.5 for the virgin sample, changes to 1.9 for the water vapor treated sample as shown in  defects . However2. It is proposed that graphene defects are sites of dissociation of water molecules and penetration of OH molecules or O to the Ge surface and become the origin of oxidation processes. The formation of GeO2 under graphene, dramatically increases the intensity of the graphene Raman spectrum. It is believed that the increase is due to the screening of graphene by GeO2 from the Ge(001) surface.To conclude it has been shown that water vapor penetrates through defects in the graphene/Ge(001)/Si(001) layer and oxidizes underneath the germanium forming blisters of GeO


Caenorhabditis elegans, ablation of germline stem cells (GSCs) and activity changes of the conserved signaling mediators unc-43/CaMKII  and egl-8/PLCβ (phospholipase Cβ) also increase lifespan. Like IIS, these pathways depend on the conserved transcription factor daf-16/FOXO for lifespan extension, but how they functionally interact is unknown. Here, we show that altered unc-43/egl-8 activity further increases the lifespan of long-lived GSC-deficient worms, but not of worms that are long-lived due to a strong reduction-of-function mutation in the insulin/IGF1-like receptor daf-2. Additionally, we provide evidence for unc-43 and, to a lesser extent, egl-8 modulating the expression of certain collagen genes, which were reported to be dispensable for longevity of these particular daf-2 mutant worms, but not for other forms of longevity. Together, these results provide new insights into the conditions and potential mechanisms by which CaMKII- and PLCβ-signals modulate C. elegans lifespan.Reduction of insulin/insulin-like growth factor 1 (IGF1) signaling (IIS) promotes longevity across species. In the nematode  Caenorhabditis elegans and the fruit fly Drosophila melanogaster, provided key insights into the biology of lifespan regulation that apparently are also applicable to human aging [C. elegans to cause a dramatic, ~two-fold increase in lifespan and, subsequently, was confirmed to promote longevity in mammals [C. elegans, lifespan extension by reduced IIS, e.g., by reduction-of-function mutations in the daf-2 gene , is dependent on daf-16  [FOXO3 have been repeatedly and robustly associated with lifespan in humans [Studies in invertebrate model organisms, such as the roundworm an aging . A proto mammals . In C. edaf-2 mutation, additional mechanisms for lifespan extension have been described in C. elegans, including two pathways that can even further extend the extraordinary long lifespan of daf-2 mutant worms: the absence of germline stem cells (GSCs) [unc-43 [daf-2 lifespan raises the possibility that the germline- and the unc-43 signaling pathways are at least in part mechanistically different from the IIS/daf-2 pathway. Interestingly, both the germline- and the unc-43 pathway, just as the daf-2 pathway, depend on daf-16 for lifespan extension [daf-2 deficient animals, such as hlh-30 (TFEB), hsf-1 (HSF1), and skn-1 (NRF2) [unc-43 pathway has not yet been investigated.Beyond s (GSCs) ,4, and a [unc-43 . This fuxtension . Whether1 (NRF2) ,8,9, areGSC(−) and daf-2(−) C. elegans. For example, only GSC(−) worms require the conserved adaptor protein KRI-1 and the translation elongation factor TCER-1 for lifespan extension [GSC(−) longevity, it acts in the intestine and neurons to ensure daf-2(−) longevity [2+) levels by binding to Ca2+-calmodulin complexes [2+-signaling are conserved across metazoans and present in C. elegans, including multiple types of sarcoplasmic/endoplasmic reticulum or plasma membrane channels, G-protein coupled receptors (GPCRs), phospholipase C (PLC) enzymes, and Ca2+-pumps [egl-30, the γ-aminobutyric acid type B (GABAB) receptor gbb-1, and the PLCβ-ortholog egl-8, through their function in neurons, also contribute to intestinal daf-16 activation and longevity [Several lines of evidence indicate that activation and function of the FOXO-transcription factor DAF-16 differ between xtension ,11. Moreongevity . DAF-16/ongevity , includiongevity . CaMKII,omplexes . Many ad2+-pumps . Among tongevity ,18.unc-43 gain-of-function mutation, activate DAF-16 and further extend daf-2(−) lifespan, we hypothesized that the two pathways are largely overlapping. We tested this hypothesis in the present study and provide evidence for a novel longevity-promoting mechanism triggered by unc-43(gf) and also, albeit more weakly, by unc-43 and egl-8 loss, which may explain their different effects on various lifespan-extending pathways in C. elegans.Given that both, germline-ablation and E. coli OP50 [glp-1(e2144ts) mutation served as a genetic model for germline ablation [glp-1(ts) strains (referred to as GSC(−) in Results/Discussion), and corresponding glp-1(+) ) control strains were incubated at 25 °C for the first 24 h of postembryonic development and subsequently shifted to 20 °C for the remainder of the experiment. daf-2(e1370) worms and corresponding daf-2(+) control worms were continuously cultured at 20 °C. For experiments, all strains were routinely cultured on plates containing 20 µM 5-Fluoro-2′-deoxyuridine  from L4 onward to avoid progeny development and internal hatching in fertile strains carrying the unc-43(gf) mutation .Strains used in this study are listed in oli OP50 . Worms c[glp-1e214ts mutatablation . To elimWorms were synchronized by hypochlorite treatment, cultured under the appropriate temperature regimen (cf. above), and scored for survival every other day starting on days 8–10 of adulthood. From the late L4 stage onward, animals were maintained on 6 cm plates at a density of 40 worms/plate and every 10 days, they were transferred to fresh OP50-seeded FUDR-containing NG agar plates to prevent progeny development and desiccation, respectively. Worms were considered dead if they did not respond to gentle touching with a worm pick. Animals that showed a protruding vulva or had ruptured, died from internal progeny hatching (bagging), or escaped from the plate were censored. tert-butyl hydroperoxide (TBHP) and were prepared 12 h before starting the experiment.Worms were synchronized by hypochlorite treatment, cultured under the appropriate temperature regimen (cf. above), and transferred to assay plates on day 2 of adulthood . Survival was scored every 1–2 h. Oxidative stress assay plates contained 15.4 mM E. coli OP50 (for RNA-seq: 1600 worms/2 plates) and cultured at the required temperatures (cf. above). At the L4-stage, 500 worms per strain were manually transferred to two E. coli OP50-seeded 6 cm NG agar plates supplemented with 20 µM FUDR to inhibit germ cell proliferation and progeny production (for RNA-seq: 1500 worms to two 10 cm plates). At day 1 of adulthood, worms were harvested by washing them off their plates with M9 buffer. After additional washing with M9 and RNAse-free water, worms were suspended in 1 mL Trizol, snap-frozen in liquid nitrogen, and stored at −80 °C until RNA extraction. To obtain synchronized populations, gravid adults were treated with hypochlorite, and eggs were allowed to hatch in M9 overnight. ~700 L1 larvae per strain were plated on 10 cm NG agar plates seeded with concentrated RNA was extracted using Trizol and cleaned up with the Monarch Total RNA Miniprep Kit or RNA Cleanup Kit  according to the manufacturer’s instructions. Equal amounts of RNA extracted from three biological replicates were pooled for RNA-sequencing to mitigate batch effects.6 reads  were obtained, resulting in at least 15.7× genome coverage . A minimum of 94.5% of reads  could be mapped to the C. elegans reference genome (cf. below).RNA quality control, library preparation, and 50-bp paired-end RNA-sequencing on the Illumina HiSeq4000 platform were performed at Eurofins Genomics, Ebersberg, Germany. For each sample, a minimum of 31.6 × 10https://usegalaxy.eu (accessed on 30 May 2020) were used for the initial analysis steps: FastQC (v0.72) and MultiQC (v1.7) for initial quality control; Trimmomatic  for gentle trimming of reads; STAR (v2.7.2b) for alignment to the C. elegans reference genome (WS276 release); and featureCounts (v1.6.4) for read summarization [NOIseq  were defined as having a probability of differential expression >95%. Only DEGs that were detected in 20 repetitions of replicate simulation and that displayed an expression-fold chance ≥1.5 with each of the two normalization methods in each of the three repetitions of the entire NOIseq-analysis were considered further. PCA-analysis of CPM-filtered, TMM/UQUA-normalized RNA-seq count data was performed using the NOIseq PCA function. Heatmaps of filtered, normalized, log-transformed count data or of sample-to-sample distances were generated using the R package pheatmap (v1.0.12). Complete NOIseq-data is provided in NOIseq M and probability values for all DEGs, along with gene ontology (GO) term information.The following tools provided by the European Galaxy Server , at rization ,22,23. Arization ) and cusv2.34.0; ,26). Lowgenome WS6 releasephyper function in R. Representation factors were calculated as the number of overlapping genes divided by the expected number of overlapping genes in the two lists . For all calculations, the number of genes in the genome was set to 10,602, i.e., the number of genes in Wormbase WS276 that passed CPM-filtering during NOIseq-analysis. DEG-lists from published studies were, if necessary, converted to WBGene-IDs using WormMine  and adjusted to genes in WS276 using custom R-scripts and manual curation. The statistical significance of overlaps between gene lists was calculated as the hypergeometric probability of detecting at least as many common genes as observed in the two lists using the https://wormbase.org/tools/enrichment/tea/tea.cgi; accessed on 27 May 2021) [http://revigo.irb.hr/; accessed on 27 May 2021). For WormCat analysis [www.wormcat.com (accessed on 9 January 2022). Collagen and matrisome classification [http://CeColDB.permalink.cc/ (accessed on 3 January 2022) and http://ce-matrisome-annotator.permalink.cc/ (accessed on 4 January 2022) and custom R scripts.GO term enrichment analysis was performed using the enrichment tool provided by Wormbase (ay 2021) . The q-vay 2021) ; http://analysis , the resfication  was perfDaf-2(−) strains, which all formed 100% dauers when incubated at 25 °C, were allowed to recover from dauer by incubation at 15 °C for 120 h before classification. Experiments in daf-2(+) strains were either conducted in parallel with experiments in daf-2(−) strains or included daf-2(−) control plates to provide a reference for the typical dauer morphology.Synchronized (hypochlorite treatment) L1 larvae were plated at a density of ~50 larvae/6 cm plate (2 plates per strain), incubated at 20 °C or 25 °C, and inspected for the presence of dauers every 12 h. At the 60 h (20 °C) or 48 h (25 °C) timepoint, when wildtype worms reached adulthood, worms were morphologically classified as “dauer” or “non-dauer”. cdc-42, tba-1, and Y45F10D.4 expression [A total of 1 µg total RNA was reverse transcribed using LunaScript RT SuperMix . qPCR-reactions were performed in duplicates or triplicates in a 20 µL reaction volume on an CFX Connect Real-Time PCR Detection System  with iTaq Universal SYBR Green Supermix (Bio-Rad Laboratories). The thermal cycling protocol comprised one activation step at 95 °C for 3 min, followed by 40 cycles of denaturation at 95 °C for 10 s and combined annealing/extension at 60 °C for 30 s. Melting curve analysis was performed from 65 °C to 95 °C with 0.5 °C increments at 5 s per step. Data were analyzed by the ΔΔCt method, and target gene expression levels were normalized to the geometric means of pression ,33. PrimStatistical analysis was performed using Prism 5 or 9 . Details on the particular tests used are specified in the figure legends.C. elegans lifespan at least in part by activating unc-43/CaMKII, we generated a set of relevant double mutants and measured their lifespans. Thereby, we took advantage of a widely used genetic model for germline stem cell deficiency, glp-1(e2144ts) (hereafter referred to as GSC(−); [unc-43 alleles n498 and n498n1186 (referred to as unc-43(gf) and unc-43(−), respectively). All experiments were conducted in the presence of FUDR , to avoid internal progeny hatching in fertile unc-43(gf) worms and to facilitate comparison with similar previous studies that also took advantage of FUDR to prevent progeny development in fertile strains [GSC(+), i.e., otherwise wildtype worms, unc-43(gf) robustly extended lifespan  also consistently produced a small to moderate lifespan increase in otherwise wildtype worms  worms, unc-43(gf) substantially extended lifespan (67–92%), while unc-43(−) had a more modest effect  and unc-43(−) both trigger longevity-promoting mechanisms. Moreover, these mechanisms apparently are not, or are not maximally, triggered by germline ablation.To test the hypothesis that germline ablation extends  GSC(−);  unc-43 a strains ,18,34. I 36–87%; , consists 6–21%; . Similarunc-43 alleles, we investigated how loss of egl-8/PLCβ influences the lifespan of GSC(−) worms. As unc-43(−), egl-8(md1971), a putative loss-of-function allele, extended the lifespan of otherwise wildtype worms  further extended lifespan in GSC(−) worms  (egl-8(−), similar to unc-43(−) and unc-43(gf) appears to induce lifespan-extending programs that are not, or not fully, active in GSC(−) worms.In parallel with the  13–38%; , in agre 30–43%, . Similar8(e2917) . Thus, eunc-43(gf), unc-43(−) and egl-8(−) in intact and in GSC-deficient animals, we examined the effect of these alleles on the lifespan of worms harboring a strong reduction-of-function mutation in the insulin/IGF-1 like receptor daf-2  [daf-2(+)) animals, unc-43(gf) substantially and unc-43(−) more modestly extended lifespan  worms, however, unexpectedly in the light of published work by others [unc-43(gf) did not further extend but rather slightly decrease lifespan  did not significantly decrease daf-2(−) lifespan. Consistent with our previous observations (egl-8(−) alleles extended lifespan in daf-2(+) worms , similar to unc-43(−), had essentially no effect on daf-2(−) lifespan (unc-43(gf), unc-43(−) and egl-8(−) all trigger longevity-promoting pathways that are already highly active, or that are dispensable for longevity, in daf-2(−) worms.In addition to our lifespan analyses on e e1370) . As mentn 9–13%; . Moreovervations A, both e 27–39%; . Finallylifespan . In summunc-43 and egl-8 affect the resistance of GSC(−), daf-2(−) and otherwise wildtype worms to selected forms of physical stress. Specifically, we examined heat and TBHP-induced oxidative stress. Only in daf-2(−) background, unc-43(gf) slightly increased heat stress resistance, while only in otherwise wildtype worms, unc-43(−) conferred heat sensitivity (unc-43(gf) and unc-43(−) both consistently caused at least trends towards sensitivity in most or all backgrounds, with only the GSC(−); unc-43(gf) strains being clearly not sensitive (egl-8(−) strains consistently tended to be or were more resistant than controls to oxidative stress but more sensitive to heat stress, with the exception of daf-2(−); egl-8(−) strains, which did not consistently show heat sensitivity (unc-43(gf), unc-43(−) and egl-8(−) did not confer broad stress resistance but rather sensitivity to at least one of the two physical stressors examined.Longevity frequently correlates with stress resistance . Therefositivity . On the ensitive . All eglsitivity . In summunc-43(−) and egl-8(−) extended C. elegans lifespan more modestly, but in the same genetic background-dependent manner as unc-43(gf). To identify novel factors and pathways that may mediate unc-43′s and/or egl-8′s lifespan-regulatory functions in addition to daf-16, we performed an exploratory RNA-sequencing (RNA-seq) study in daf-2(−) and otherwise wildtype (daf-2(+)) worms with normal or altered unc-43/egl-8 activity. GSC(−) worms were omitted from this study since unc-43/egl-8 mutations changed their lifespan in essentially the same way as the lifespan of otherwise wildtype worms. Out of the two egl-8 loss-of-function alleles, md1971 was chosen, and hereafter, egl-8(−) refers to md1971 unless otherwise stated. Initial comparison of gene expression between daf-2(−) and wildtype worms revealed highly significant overlaps of differentially expressed genes (DEGs) from our study with published work [daf-2(−) control strains from each other, and strikingly, egl-8(−) and especially unc-43(gf) single mutant worms appeared closer to daf-2(−) in these analyses (unc-43(gf) and egl-8(−) in otherwise wildtype worms significantly overlapped with genes induced by daf-2(−) in our and in published studies (unc-43(gf)/egl-8(−)- and most to all lists of daf-2(−) repressed genes significantly overlapped with each other. Interestingly, in otherwise wildtype worms, unc-43(−) induced and repressed genes also significantly overlapped with most to all lists of daf-2(−) induced and -repressed genes (daf-2(−) worms, unc-43(gf), unc-43(−) and egl-8(−) seemed to support some gene expression changes already induced by daf-2(−), while opposing others (unc-43(gf), unc-43(−) and egl-8(−)-regulated DEGs to genes regulated by daf-2(−) in a daf-16 dependent manner [unc-43(gf) and egl-8(−) globally shift gene expression towards daf-2(−), and they are consistent with previously reported roles of unc-43(gf) and egl-8(−) as positive regulators of daf-16 [In our hands, hed work ,37, thusanalyses A–C. In l studies . Similared genes . In daf-g others . Similart manner ,39,40,41f daf-16 ,16,17,18unc-43(gf), unc-43(−) and egl-8(−) DEGs in otherwise wildtype and in daf-2(−) worms, we performed gene ontology (GO) term enrichment analysis [unc-43 and egl-8 in neurons, muscle, and immunity, respectively [unc-43(gf), unc-43(−) or egl-8(−) strains were also enriched among DEGs between daf-2(−) and wildtype worms, consistent with the previously observed overlaps between the respective DEG-lists (daf-2(+) strains was “structural component of cuticle”, followed by additional GO terms containing many collagen genes , and by multiple GO terms related to pathogen defense. “Structural component of cuticle” and pathogen defense-related GO terms were also enriched among upregulated DEGs of most to all daf-2(−) strains. On the other hand, unc-43(gf), unc-43(−) and egl-8(−) downregulated DEGs were most significantly and commonly enriched for pathogen defense related GO terms in all daf-2(+)- and in the daf-2(−); egl-8(−) strain, and for organic acid-metabolism-related GO terms in daf-2(−); unc-43(gf) and daf-2(−); unc-43(−) worms. Enrichment of similar functional categories was detected by a different analysis tool, WormCat [unc-43(gf), unc-43(−) and egl-8(−)-induced genes, “extracellular material: collagen”  allele analyzed by us, daf-2(e1370), at least not at the standard culture temperature of 20 °C at which we conducted our study [Unc-43(gf), unc-43(−), and egl-8(−) do not sensitize C. elegans to dauer entry at 20 °C or 25 °C (daf-2(−) collagens” ([unc-43(−), egl-8(−) and especially unc-43(gf) upregulated DEGs from daf-2(+), as well as among unc-43(gf) upregulated DEGs from daf-2(−) worms  appeared to up- and downregulate significant numbers of genes that are up- or downregulated by skn-1 under normal conditions (unc-43(gf), unc-43(−) and egl-8(−), especially in daf-2(+) backgrounds, revealed strong overrepresentation of cuticular collagens, as also seen for skn-1 upregulated (non-dauer longevity-associated) collagens ([unc-43(gf), unc-43(−), egl-8(−) and skn-1(−) induced DEG-sets, all of which showed particular enrichment of nematode-specific core matrisome genes  relative to otherwise wildtype worms, while induction in unc-43(−) and egl-8(−) was less consistent and/or weaker (GSC(−) and daf-2(−) worms, col-120, col-141, and col-176 upregulation by unc-43(gf) was repeatedly observed, while unc-43(−) did not trigger strong alterations (in daf-2(−) worms, egl-8(−) appeared to at least mildly suppress all four collagen genes examined (GSC(−) and daf-2(−) worms displayed at least trends towards elevated mRNA levels for most of the collagen genes examined (unc-43(gf), and to a smaller extent, unc-43(−) and egl-8(−), promote the expression of non-dauer longevity-associated collagens. Moreover, the effects of the three alleles appear strongest in GSC(+)/daf-2(+), and weakest, or even lost, in daf-2(−) background.Certain cuticle-forming collagens have been shown to be required for longevity, specifically under conditions that do not induce processes related to dulthood . Expressdulthood . Non-dauur study . Unc-43  at 20 °C) worms. This result may be explained by at least two, not mutually exclusive, models. First, unc-43(gf), unc-43(−) and egl-8(−) all may trigger processes that are already maximally induced by daf-2(−), but not by GSC(−) and wildtype worms. This model is supported by the fact that all three mutations induce similar gene expression changes than daf-2(−). Alternatively, processes triggered by unc-43(gf), unc-43(−) and egl-8(−) may not be critical to daf-2(e1370) longevity. Consistent with this possibility, unc-43(gf) and, to a lesser extent, unc-43(−) and egl-8(−), induce the expression of certain collagen genes, which are required for longevity under conditions that do not induce dauer-like traits in adults. These conditions include germline deficiency but not the daf-2(e1370) allele under our culture conditions of 20 °C [daf-16, by which in particular unc-43(gf), but also unc-43(−) and egl-8(−) modulate lifespan. Of note, overexpression of a single key collagen, such as col-120, also examined by us by qPCR, is sufficient for lifespan extension [daf-2(e1370), mild induction of collagen expression, which is still observed, at least in response to unc-43(gf), may not be functionally relevant. Clearly, it will be extremely interesting to directly examine the role of non-dauer longevity-associated collagens in unc-43(gf), unc-43(−) and egl-8(−) dependent C. elegans lifespan regulation and, eventually, to define the mechanisms that link unc-43/egl-8 to expression of these collagens. Although we have not yet experimentally confirmed or excluded the possibility that the known mediator of unc-43(gf)/egl-8(−) longevity, daf-16, regulates non-dauer longevity-associated collagen expression in the specific context of unc-43/egl-8 mutation, even though it does not do so in other contexts, an alternative candidate is emerging from the literature: skn-1 [unc-43(gf), unc-43(−) and egl-8(−) modulate the expression of similar gene sets, including similar collagens, than skn-1. A key finding of our study consists in the observation that altered of 20 °C . Thus, ixtension . Converse: skn-1 . In suppunc-43(−) and unc-43(gf) differ from each other in several details despite the use of the same alleles. In the case of unc-43(−), these discrepancies may be explained by different experimental conditions, i.e., different temperatures, and by the use of FUDR, both of which have been shown to affect lifespan in certain genetic backgrounds [unc-43(−) and unc-43(gf) alleles within the same experiments, which required the addition of FUDR not just to unc-43(gf) and corresponding control [unc-43(−) strains. Importantly, FUDR does not affect wildtype lifespan nor daf-2(−)’s and GSC(−)’s basic ability to promote longevity [Yet, our study and a previous report  on the lkgrounds . Specifikgrounds  and anal control , but alsongevity ,34,43.daf-2(e1370), rather than daf-2 RNAi [daf-2 activity may account for unc-43(gf) in our hands not extending the lifespan of worms with reduced IIS. Differential interactions of “strong”  and “weak”  daf-2 inhibition with other lifespan-regulatory mechanisms are not unprecedented in the literature, e.g., with somatic gonad signaling and collagen expression [daf-2 RNAi does not efficiently reduce daf-2 activity in neurons [daf-2(−)- and unc-43(gf) lifespan-extending factors daf-16 in neurons accounts for a fraction of daf-2(−) longevity [unc-43(gf) may trigger daf-16 regulated processes in neurons that are already active in daf-2(e1370), but not in daf-2 RNAi worms, which may explain why it further extends daf-2 RNAi- but not daf-2(e1370) lifespan. Beyond different assay temperatures, our choice of f-2 RNAi  to inhibdaf-2e170, rathepression ,45. More neurons ,46,47. Yongevity . Thus, uunc-43(−) and unc-43(gf) extend lifespan, at least in certain genetic backgrounds under certain experimental conditions, appears paradoxical at the first glance. On the other hand, the moderate to strong longevity increase of unc-43(−) and unc-43(gf) worms can be expected, based on the nature of the—relatively moderate to relatively large—changes in gene expression triggered by these two mutations. Indeed, as discussed above, unc-43(−) and unc-43(gf) regulated gene sets are similar to each other and to daf-2- and daf-16-regulated gene sets. In addition, both mutations, although to different extents, induce non-dauer longevity-associated collagens. How unc-43(−) and unc-43(gf) affect collagen expression and/or its indirect regulator skn-1 is currently unclear [daf-16 targets upon unc-43(−) loss, as observed in our RNA-seq study, represents additional, indirect effects of unc-43 on daf-16. Moreover, UNC-43′s direct contribution to DAF-16 activity appears to be dispensable for the maintenance of a normal/normally long lifespan in all genetic backgrounds tested, as unc-43(−), GSC(−); unc-43(−) and daf-2(−); unc-43(−) worms are long-lived, in contrast to daf-16(-), GSC(−); daf-16(-) and daf-2(−); daf-16(-) worms, which are short- to normal-lived [unc-43 loss may “overpower” (direct) DAF-16 regulation by UNC-43.The observation that both,  unclear . Previou unclear  suggesteal-lived . As propal-lived , GSC losegl-8 further extends wildtype lifespan. However, in contrast to previous work [egl-8 loss. Again, this difference may be explained by different experimental conditions, such as different egl-8(−) alleles (md1971 vs. n488), different assay temperatures (20 °C vs. 25 °C) and treatment with different chemicals to impose oxidative stress (TBHP vs. arsenite). Indeed, TBHP and arsenite have already been shown to trigger different transcriptional responses [egl-8 modulates lifespan cell non-autonomously through its function in neurons: egl-8 loss reduces neuronal secretion of insulin-like peptides, which in turn increases daf-16 activity in the intestine; in addition, evidence suggests that this pathway is further mediated by the EGL-8-generated second messenger diacylglycerol (DAG) and the DAG-dependent kinase dkf-2 [egl-8(−) mutation, while GSC(−) worms may display further intestinal daf-16 activation and lifespan extension. Results from our lifespan analyses are in line with this prediction. Conceptually, egl-8 loss also decreases the levels of another second messenger, inositol-1, 4, 5-trisphosphate and subsequently, intracellular Ca2+ levels, and CaMKII/UNC-43 activity [egl-8(−) and unc-43(−) interact to modulate daf-16 activity and C. elegans lifespan.In agreement with published studies ,17,18, wous work , we did esponses . In a cuse dkf-2 . This moactivity . Yet, adunc-43 and egl-8 can/cannot modulate lifespan in C. elegans and suggests collagen expression as a second lifespan-regulatory mechanism functioning downstream of unc-43 and egl-8, in addition to the known regulator daf-16.In summary, our study identified new genetic backgrounds in which


Brain-derived neurotrophic factor (BDNF) induces activation of the TrkB receptor and several downstream pathways , leading to neuronal survival, growth, and plasticity. It has been well established that TrkB signaling regulation is required for neurite formation and dendritic arborization, but the specific mechanism is not fully understood. The non-receptor tyrosine kinase c-Abl is a possible candidate regulator of this process, as it has been implicated in tyrosine kinase receptors’ signaling and trafficking, as well as regulation of neuronal morphogenesis. To assess the role of c-Abl in BDNF-induced dendritic arborization, wild-type and c-Abl-KO neurons were stimulated with BDNF, and diverse strategies were employed to probe the function of c-Abl, including the use of pharmacological inhibitors, an allosteric c-Abl activator, and shRNA to downregulates c-Abl expression. Surprisingly, BDNF promoted c-Abl activation and interaction with TrkB receptors. Furthermore, pharmacological c-Abl inhibition and genetic ablation abolished BDNF-induced dendritic arborization and increased the availability of TrkB in the cell membrane. Interestingly, inhibition or genetic ablation of c-Abl had no effect on the classic TrkB downstream pathways. Together, our results suggest that BDNF/TrkB-dependent c-Abl activation is a novel and essential mechanism in TrkB signaling. Regulation of dendritic arborization is essential for brain function since it determines the receptive field of neurons . ExtrinsThe process of dendritic growth and branching requires active reorganization of the actin cytoskeleton. Neurite outgrowth has been proposed to be directly regulated by lamellipodia and filopodia formation, which in turn are dependent on actin cytoskeleton dynamics mediated by Rho GTPases such as Rac1, Cdc42, and RhoA ,14. InteHere, we show that BDNF-induced dendritic branching requires c-Abl expression and activity. Interestingly, BDNF increases c-Abl activity and c-Abl-TrkB interaction. Although this process requires TrkB activity, it does not require the activation of MAPK, PI3K, and PLC-γ signaling pathways. These results suggest that c-Abl activation is a key downstream mechanism required for BDNF-TrkB-induced dendritic growth, and its mechanism of action does not involve the regulation of the classical signaling pathways regulated by BDNF/TrkB. To obtain further insight into the mechanism of action of c-Abl activity in the context of BDNF/TrkB signaling and function, we investigated the role of c-Abl in the regulation of BDNF/TrkB internalization and endocytic trafficking. We found that c-Abl activity was not required for BDNF-induced endocytosis or degradation of the TrkB receptor. However, we found that c-Abl was required for basal endocytosis of TrkB and that its activation increased the speed of TrkB retrograde traffic.Altogether, our results show that c-Abl activation downstream of BDNF/TrkB signaling mediates a cellular process leading to dendritic growth that does not involve the activation of classical signaling pathways triggered by BDNF/TrkB but is required in BDNF/TrkB-dependent dendritic growth.BDNF induces activation of TrkB, promoting dendritic growth ,15,33,34As shown in previous studies, neurons treated with BDNF showed increased dendritic branching compared to control neurons A. We assTo confirm the requirement of c-Abl expression in the dendritic arborization increase induced by BDNF, we used hippocampal neurons derived from c-Abl KO embryos. Seven DIV c-Abl KO and WT hippocampal neurons were treated with BDNF for 48 h, and primary neurites and branching points were evaluated. Consistent with the results obtained with c-Abl inhibitors, the absence of c-Abl expression in neurons reduced BDNF-induced dendritic growth to basal levels F. The Shabl/arg double-null mice exhibit defective neural tube morphology and gross alterations in neuroepithelial actin cytoskeleton structures, suggesting that c-Abl knockout neurons could have alterations in dendritic tree development -2,5-diphenyltetrazolium bromide (MTT) assay. Then, the MTT reagent (5 mg/mL) was added and incubated for 4 hrs. The medium was then removed and replaced with lysis buffer (50% Dimethylformamide and 20% SDS) overnight. Subsequently, the absorbance of the sample was measured at 620 and 570 nm.t-test or one-way ANOVA followed by appropriate multiple comparisons test depending on the number of groups used in each experiment. Details of the specific tests used, level of significance, and number of replicates are indicated in each figure legend. Statistical analyses were performed using GraphPad Prism 8 (Scientific Software). The significance level was p < 0.05 for all treatments.All data are presented as the mean ± Standard Error of the Mean (SEM). Sholl’s analysis curves were compared with two-way repeated measures ANOVA, followed by Bonferroni’s multiple comparisons. Statistical analyses were performed using a Student’s


This paper presents a rapid assessment of coronavirus disease 2019 (COVID-19) pandemic plans and explores the representation of culturally and linguistically diverse (CALD) communities in such plans. Four levels of pandemic plans were reviewed: regional, state, national and international.Discussions with representatives from four CALD communities informed the development of search and selection criteria for the COVID-19 plans, which were gathered and assessed using a CALD lens. Six COVID-19 pandemic plans that met the inclusion criteria were critically assessed.The reviewed plans did not report any CALD community voices, views or consultations with community groups in the development phase, nor did they acknowledge the diversity of CALD populations. A few plans noted the vulnerability of CALD communities, but none discussed the challenges CALD communities face in accessing health information or health services during the pandemic, or other structural barriers .Our analysis revealed major gaps in all pandemic plans in terms of engaging with immigrant or CALD communities. Policies and plans that address and consider the complex needs and challenges of CALD communities are essential. Collaboration between public health services, multicultural services and policy-makers is vital for the inclusion of this higher-risk population. In Australia, as in other developed nations, the coronavirus disease 2019 (COVID-19) pandemic has disproportionately affected culturally and linguistically diverse (CALD) communities. In many countries, immigrant communities have experienced higher rates of COVID-19 infection, hospitalization, severity of disease and death.  conducting the study. It explores whether the needs, expectations and challenges of CALD communities are represented in these plans. The terms “immigrant” and “CALD communities” are used interchangeably here. The study did not include a review of all relevant literature or research papers; rather, the focus was on pandemic plans and policies at multiple settings and levels .Pandemic plans at regional, state, national and international levels were selected, accessed and then critically assessed through a “CALD lens.”The selection criteria included plans that were:available online at the time of assessment;updated or published within the past 5 years; andapplicable to the region or state of the PHU undertaking the review, to a neighbouring state or to international plans published by the World Health Organization (WHO).As part of using a CALD lens, discussions with representatives from four CALD communities informed the development of the search and selection criteria. These key informants were e-mailed a series of questions. Four consumer representatives of local multicultural health services then discussed the e-mailed questions with the informants to finalize the assessment questions. The final questions were as follows:Does the plan describe a governance structure that includes CALD community representatives?Does the plan describe any consultation with CALD communities before or during the development of the plan?Does the plan outline how it reflects and embraces the diversity of CALD communities?Does the plan reference the challenges CALD communities encounter in accessing health systems?Does the plan describe how CALD communities would be involved in the oversight, implementation or review of the plan when it is operationalized?Six plans met the inclusion criteria and were critically reviewed . However, none of the documents discussed CALD community challenges in accessing health information or health services during the pandemic or other structural barriers such as social determinants of health. Also, the documents did not mention factors such as unemployment, crowded housing, visa status, low health literacy, racism and cultural beliefs. A few references were made to challenges in communication between health organizations and immigrant communities, but the plans tended to fall short of addressing solutions for overcoming barriers to reduce risk. Only two plans  talked about the necessity of providing translated information.In general, Australian policies and plans do not engage with CALD communities, and there is little data regarding the needs of these communities and the challenges they face in accessing health-care systems. This may be an indication of structural racism in the system. (Despite the health inequities faced by people from CALD communities, their voices and needs were not reflected in the pandemic plans assessed in this study. The plans failed to address embedded inequities, which are particularly important in health emergencies. It is recommended that CALD communities be included in the development and implementation of pandemic plans. Further research should be undertaken with diverse communities to enable effective public health actions for COVID-19 and future pandemics.


Adult adrenocortical carcinoma (ACC) is a rare and aggressive tumor in adults, usually associated with excessive steroid secretion. It is highly metastatic and has few therapeutic options and a poor prognosis. Here, we explore the hallmarks of ACC influenced by transcription factors and their target genes (regulons) to provide a prognostic overview of ACC biology. Using an in silico clinical data analysis approach, we assessed human transcriptomic data from publicly available datasets. We found four distinct clusters of regulons associated with good and worse prognoses associated with cell proliferation and/or immunologic activity. Some findings require further bench analyses, primarily focusing on worse prognostic regulons and their targets.NR5A1  and CENPA (Centromeric Protein A), that were consistently associated with overall survival for further downstream analyses. The CENPA regulon was the primary regulator of MKI-67 (a marker of proliferation KI-67), while the NR5A1 regulon is a well-described transcription factor (TF) in ACC tumorigenesis. We also found that the ZBTB4 (Zinc finger and BTB domain-containing protein 4) regulon, which is negatively associated with CENPA in our transcriptional regulatory network, is also a druggable anti-tumorigenic TF. We anticipate that the ACC regulons may be used as a reference for further investigations concerning the complex molecular interactions in ACC tumors.We reconstructed a transcriptional regulatory network for adrenocortical carcinoma (ACC) using transcriptomic and clinical data from The Cancer Genome Atlas (TCGA)-ACC cohort. We investigated the association of transcriptional regulatory units (regulons) with overall survival, molecular phenotypes, and immune signatures. We annotated the ACC regulons with cancer hallmarks and assessed single sample regulon activities in the European Network for the Study of Adrenal Tumors (ENSAT) cohort. We found 369 regulons associated with overall survival and subdivided them into four clusters: RC1 and RC2, associated with good prognosis, and RC3 and RC4, associated with worse outcomes. The RC1 and RC3 regulons were highly correlated with the ‘Steroid Phenotype,’ while the RC2 and RC4 regulons were highly correlated with a molecular proliferation signature. We selected two regulons,  TP53 mutation, and an upregulation of the WNT/β-Catenin pathway , were the regulons with the most associations   and Wound Healing signatures , and a significant negative correlation with leukocyte fraction , Macrophage Regulation , and Lymphocyte Infiltration Signature Score   . Interes × 10−5) .CENPA was the main regulator of the MKI-67 gene (MI = 0.77), the most common histopathologic marker of proliferation, followed by FOXM1 (MI = 0.59), MXD3 (MI = 0.48), and DNMT1 (MI = 0.34)  . MXD3 anPA Duals , and FOXCENPA was significantly associated with both the Steroid IP and the Proliferation IS  and clustered together with RC3  and in OS , in addition to the ENSAT cohort for OS   . In the CENPA’s activity were THRB (Thyroid Hormone Receptor Beta), STAT5B , and ZBTB4 (Zinc Finger And BTB Domain Containing 4) . All of them from RC1 related to good prognosis in the TCGA-ACC and ENSAT cohorts .In the TCGA-ACC cohort, the three regulons with higher negative Spearman correlation with  cohorts  and presThe TCGA-ACC cohort is the largest ACC cohort, thus being the most suitable for regulatory network inference. The inferred network had a good balance between positive and negative targets , with 61Here we present 369 regulons associated with OS, where 188 are related to a good prognosis and 181 to a worse ACC prognosis. Within these regulons, we identified subgroups with specific relationships for each set of molecular characteristics and functional annotations, resulting in four RCs. Regulon cluster 1 and RC3 are more related to the Steroid Phenotype, while RC2 and RC4 are more related to the molecular Proliferation Score as defined by Zheng et al. (2016) . As the RC1 and RC3 are related to Steroid IP, with RC1 associated with LSP and RC3 with HSP. These two clusters presented opposite profiles, both in activity and functional annotations. These clusters were at high activity in the immune and proliferation pathways, as previously shown for the Steroid Phenotype . The strWe also observed that RC2 and RC4 follow opposite patterns in terms of activity and characteristics. RC2, with a good prognosis, is related to low scores in the Proliferation IS, while RC4, with a poor prognosis, is related to high scores in this phenotype. We identified from the Hallmarks enrichment  that theTP53 gene, may alter the P53 pathway and its DNA repair activity [Our functional analysis using the MSigDb Hallmarks presented an overall panorama, providing insights into the biology and function of each group of inferred regulons. However, the MSigDb Hallmarks represent themes of gene sets rather than pathways , which mactivity ,64. The FLI1 (Friend Leukemia Virus Integration 1), STAT5A , and IKZF1 (Ikaros Family Zinc Finger Protein 1). For example, in our results, IKZF1 appeared related to good prognosis , and with the strongest association with the LSP  and the highest positive correlation with leukocyte fraction , Macrophage Regulation , and Lymphocyte Infiltration Signature Score  among the 369 regulons  , shows tregulons . FindingNR5A1 TF, an important player in adrenal development and ACC tumorigenesis [NR5A1 regulon correlated with low OS in the multivariate Cox and Kaplan-Meier analyses in the TCGA-ACC cohort, which was also confirmed in the ENSAT cohort , a common prognostic and proliferation marker widely used in cancer histopathology [Remarkably, we identified ormation . In thisathology .CENPA was described as relating to proliferation and prognosis in ovarian cancer [CENPA overexpression was also associated with proliferation and metastasis in kidney carcinoma by activating the WNT/β-Catenin signaling pathway [CENPA appeared positively enriched in this pathway , but moe cancer  showed tFOXM1 is assigned as a prognostic marker in ACC by Yuan et al. (2018) [It should be noted that our study had some limitations. Our selection strategy may have removed some interesting regulons, which are not listed in the 369 prognostic regulons, thereby excluding their possible role in the ACC regulatory scenario. For example, . (2018) . This TFIn conclusion, we have generated a regulatory network for ACC, evaluated the regulons inferred in concern to OS, clustered them in four RCs, and investigated how they relate to characteristics associated with worse outcomes like the steroid phenotype and the immune and proliferation pathways. The list of prognostic regulons, and their characterization, may open new research avenues and relevant questions to be answered in this hard-to-treat and aggressive malignancy.