Acrylic Hydrogels Research Articles

Effective removal of malachite green dye by eco-friendly pectin-grafted poly(acrylamide-co-sodium acrylate) hydrogel from aqueous solutions

Effective removal of malachite green dye by eco-friendly pectin-grafted poly(acrylamide-co-sodium acrylate) hydrogel from aqueous solutions

Utilizing acrylic acid polymer hydrogel for 3-D quality assurance in CyberKnife radiotherapy

Dosimetry tools play a crucial role in radiotherapy as they are essential for recording and validating intricate 3-D dose distributions. This research introduces and examines a novel acrylic acid polymer hydrogel (ACAPHG) dosimeter composition designed for 3-D dose verification and quality assurance in the context of radiotherapy treatment. A phantom made of an 80 mm diameter cylindrical glass container was utilized. The phantom contained the hydrogel, which served as the medium for radiation exposure. The water equivalent hydrogel within the phantom was subjected to irradiation by a CyberKnife robotic radiotherapy system. An optical computed tomography (OCT) scanner with sub-millimeter resolution was used to obtain imaging data. The dose distribution of a CyberKnife robotic SRS/SBRT treatment plan for a brain cancer patient was compared to that of the hydrogel's OCT scan using 2-D and 3-D gamma analysis with a criterion of 3% dose difference and 3 mm distance-to-agreement. A gamma pass rate of 94.1% for the 2-D gamma analysis and a pass rate of 99% for the 3-D gamma analysis were calculated within the region at which the treatment planning system data drops to 20% of the maximum dose. The use of the ACAPHG dosimeter in conjunction with the described setup suggests that it has the potential to offer an accurate 3-D verification of complex dose distributions in SRS/SBRT radiotherapy treatments. By employing the ACAPHG dosimeter and utilizing OCT scanning, this dosimeter enables the assessment and validation of intricate dose distributions in these advanced radiotherapy treatments.

New Ibuprofen Cystamine Salts With Improved Solubility and Anti-Inflammatory Effect.

Two novel ibuprofen cystamine salts (IBU-CYS 1 and IBU-CYS 2) are synthesized by coupling the anion of ibuprofen with cystamine dihydrochloride in 1 : 1 and 2 : 1 ratio to improve the solubility and bioavailability of ibuprofen. The salts are characterized by 1HNMR, FT-IR and UV-Vis spectroscopy, differential scanning calorimetry (DSC), thermogravimetry (TGA, DTA) and X-ray diffraction measurements. IBU-CYS 1 and IBU-CYS 2 show higher solubility (6.11 and 7.81 mg/mL) compared to ibuprofen (0.04 mg/mL) in water. IBU-CYS2 was encapsulated into 2-hydroxyethyl methacrylate: poly (ethylene glycol) acrylate hydrogels for enhanced delivery. The in vitro studies in PBS (pH 7.4) indicate that the salts are effective in relieving inflammatory responses induced by lipopolysaccharide in RAW264.7 macrophage cells (nitrite inhibition percentages of IBU-CYS 1, IBU-CYS 2 and ibuprofen: approximately 34.29, 27.03 and 31.50 respectively) while indicating no cytotoxicity. Therefore, these salts may be promising candidates for the development of effective formulations of this drug.

Synthesis and application of novel sodium carboxy methyl cellulose-g-poly acrylic acid carbon dots hydrogel nanocomposite (NaCMC-g-PAAc/ CDs) for adsorptive removal of malachite green dye

This study aimed on synthesizing novel sodium carboxy methyl cellulose-g-poly acrylic acid carbon dots hydrogel nanocomposite (NaCMC-g-PAAc/ CDs) to remove malachite green (MG) dye from aqueous media. The characterisation of functionalities, morphology, particle size and crystallography of NaCMC-g-PAAc/ CDs nanocomposite both before and after adsorption. The results of the characterization analysis revealed the presence of numerous ionic functionalities such as carboxyl, amino and hydroxyl groups in nanocomposite. The morphological study revealed the presence of porosity, heterogenous and amorphous nature of adsorbent favoring the adsorption of MG. Furthermore, elemental analysis also confirmed the presence of carbon and oxygen as main elemental composition of nanocomposite. The temperature of 30 °C and the nanocomposite (having pHPZC of 3) dose of 0.06g at pH 10 results in achieving 96.92% of MG dye removal. The experimental data fits closely with pseudo-second and Freundlich models. The enhanced MG dye adsorption by NaCMC-g-PAAc/ CDs was attributed to different mechanisms such as electrostatic or H-bonding as well as pi-pi interactions. Thermodynamically, the studied process was spontaneous and is of endothermic kind. Overall, the study reveals adsorptive property of novel NaCMC-g-PAAc/ CDs nanocomposite as a promising adsorbent for MG dye adsorption from water.

Enhanced antifouling and anti-swarming properties poly (sulfobetaine methacrylate-co-2-hydroxy-3-phenoxypropyl acrylate) hydrogel coatings for urinary catheters

Catheter-associated urinary tract infection (CAUTI) remains an unsolved challenge to date, particularly with the emergence and rapid spread of antimicrobial-resistant bacterial pathogens. Despite extensive research, a catheter coating that can offer intrinsic resistance to host protein deposition, bacterial biofilm formation, and swarming is still urgently required. Zwitterionic hydrogel coatings due to their superior lubricity and antifouling properties represent a promising candidate, but their weak mechanical stability in water and poor resistance to bacterial swarming migration limit their application in urinary catheters for infection control. In this research, we describe the fabrication of a multifunctional catheter coating by copolymerizing zwitterionic sulfobetaine methacrylate (SBMA) polymers and a swarming inhibitor material, 2-hydroxy-3-phenoxypropyl acrylate (HPA). The introduction of polyHPA (PHPA) effectively impeded the uncontrolled swelling behavior of the zwitterionic PSBMA hydrogel, resulting in enhanced mechanical stability. Moreover, the copolymer coating retains the antifouling and anti-swarming properties of the homopolymers when challenged with fibrinogen, Escherichia coli, and Proteus mirabilis. The HPA content significantly correlated with its anti-adhesion activity against fibrinogen and biofilm, and the coating with an SBMA: HPA monomer feed molar ratio of 4:1 showed the best antifouling activity, reducing fibrinogen deposition by about 40 % and biofilm coverage by around fourfold compared to the uncoated polydimethylsiloxane (PDMS) surface. Furthermore, the copolymer coating also exhibited no cytotoxicity, suggesting it as a promising catheter coating for preventing CAUTI.

Osmotic energy harvesting using acrylic acid hydrogel PET membrane

In this study, we investigated the osmotic energy harvesting using a cation-selective membrane. The cation-selective membrane was synthesized by the incorporation of acrylic acid hydrogel in a porous support membrane. FTIR analysis and SEM images confirmed the presence of the acrylic acid hydrogel rods inside the porous support material. The exposure of the acrylic acid hydrogel PET (AP) membrane to a concentration gradient culminated in the generation of electric power. The AP membrane was evaluated in regard to the following parameters: EDiff, Io, Pmax and t+. The maximum power obtained with the membrane was 1.10 μW at the 40-fold concentration gradient (test area∽ 100 mm2). Increasing the concentration gradient increased the power output. However, a decrease in power output was observed after a certain value of the concentration gradient. An enhancement in power output was noted as the fraction of acrylic acid within the membrane was augmented. Additionally, the cation transference number also increased owing to a high charge density and a reduction in the mesh size. The AP membrane was also investigated in acidic and basic media. The time-dependent study revealed the superior chemical stability of the AP membrane.

Preparation and Stability Study of an Injectable Hydrogel for Artificial Intraocular Lenses.

Currently available intraocular lenses (IOLs) on the market often differ significantly in elastic modulus compared to the natural human lens, which impairs their ability to respond effectively to the tension of the ciliary muscles for focal adjustment after implantation. In this study, we synthesized a polyacrylamide-sodium acrylate hydrogel (PAH) through the cross-linking polymerization of acrylamide and sodium acrylate. This hydrogel possesses excellent biocompatibility and exhibits several favorable properties. Notably, the hydrogel demonstrates high transparency (94%) and a refractive index (1.41 ± 0.07) that closely matches that of the human lens (1.42). Additionally, it shows strong compressive strength (14.00 kPa), good extensibility (1400%), and an appropriate swelling ratio (50 ± 2.5%). Crucially, the tensile modulus of the hydrogel is 2.07 kPa, which closely aligns with the elastic modulus of the human lens (1.70-2.10 kPa), enabling continuous focal adjustment under the tension exerted by the ciliary muscles.

Functional-hydrogel-based electronic-skin patch for accelerated healing and monitoring of skin wounds

Conductive hydrogels feature reasonable electrical performance as well as tissue-like mechanical softness, thus positioning them as promising material candidates for soft bio-integrated electronics. Despite recent advances in materials and their processing technologies, however, facile patterning and monolithic integration of functional hydrogels (e.g., conductive, low-impedance, adhesive, and insulative hydrogels) for all-hydrogel-based soft bioelectronics still poses significant challenges. Here, we report material design, fabrication, and integration strategies for an electronic-skin (e-skin) patch based on functional hydrogels. The e-skin patch was fabricated by using photolithography-compatible functional hydrogels, such as poly(2-hydroxyethyl acrylate) (PHEA) hydrogel (substrate), Ag flake hydrogel (interconnection; conductivity: ∼571.43 S/cm), poly(3,4-ethylenedioxythiophene:polystyrene) (PEDOT:PSS) hydrogel (working electrode; impedance: ∼69.84 Ω @ 1 Hz), polydopamine (PDA) hydrogel (tissue adhesive; shear strength: ∼725.1 kPa), and poly(vinyl alcohol) (PVA) hydrogel (encapsulation). The properties of these functional hydrogels closely resemble those of human tissues in terms of water content and Young's modulus, enabling stable tissue-device interfacing in dynamically changing physiological environments. We demonstrated the efficacy of the e-skin patch through its application to accelerated healing and monitoring of skin wounds in mouse models − efficient fibroblast migration, proliferation, and differentiation promoted by electric field (EF) stimulation and iontophoretic drug delivery, and monitoring of the accelerated healing process through impedance mapping. The all-hydrogel-based e-skin patch is expected to create new opportunities for various clinically-relevant tissue interfacing applications.

Optical characterization of a new composition of acrylic acid hydrogel dosimeter for quality assurance in radiotherapy treatment

Optical characterization of a new composition of acrylic acid hydrogel dosimeter for quality assurance in radiotherapy treatment

Strong Acid Enabled Comprehensive Training of Poly (Sodium Acrylate) Hydrogel Networks

AbstractThe design of admirable hydrogel networks is of both practical and fundamental importance for diverse applications of hydrogels. Herein a general strategy of acid‐assisted training is designed to enable multiple improvements of conventional poly (sodium acrylate) networks for hydrogels. Hydrophobic homogeneous crosslinked poly (sodium acrylate) hydrogels are prepared to verify the strategy. The multiple improvements of poly (sodium acrylate) networks are simply achieved by immersing the hydrogel networks into 4 M H2SO4 solutions. The introduced acids would induce transformation of poly (sodium acrylate) into poly (acrylic acid) at hydrogel surface, which constructs dynamic hydrogen bonding interactions to tighten the network. The acid‐containing poly (sodium acrylate) hydrogels newly generate anti‐swelling and self‐healing performance, and show mechanical improvement. The internal poly (sodium acrylate) of the pristine acid‐containing hydrogels is further fully transformed via acid‐infiltration after following cyclic stretch/release training to significantly improve the mechanical performance. The Young's modulus, stress, and toughness of the fully‐trained hydrogels are 187.6 times, 35.6 times, and 5.4 times enhanced, respectively. The polymeric networks retain isotropic in fully‐trained hydrogels to ensure superior stretchability of 8.6. The acid‐assisted training performance of the hydrogels can be reversibly recovered by NaOH neutralization. The acid‐assisted training strategy here is general for poly (sodium acrylate) hydrogels.

Strong Acid Enabled Comprehensive Training of Poly (Sodium Acrylate) Hydrogel Networks.

The design of admirable hydrogel networks is of both practical and fundamental importance for diverse applications of hydrogels. Herein a general strategy of acid-assisted training is designed to enable multiple improvements of conventional poly (sodium acrylate) networks for hydrogels. Hydrophobic homogeneous crosslinked poly (sodium acrylate) hydrogels are prepared to verify the strategy. The multiple improvements of poly (sodium acrylate) networks are simply achieved by immersing the hydrogel networks into 4 M H2SO4 solutions. The introduced acids would induce transformation of poly (sodium acrylate) into poly (acrylic acid) at hydrogel surface, which constructs dynamic hydrogen bonding interactions to tighten the network. The acid-containing poly (sodium acrylate) hydrogels newly generate anti-swelling and self-healing performance, and show mechanical improvement. The internal poly (sodium acrylate) of the pristine acid-containing hydrogels is further fully transformed via acid-infiltration after following cyclic stretch/release training to significantly improve the mechanical performance. The Young's modulus, stress, and toughness of the fully-trained hydrogels are 187.6 times, 35.6 times, and 5.4 times enhanced, respectively. The polymeric networks retain isotropic in fully-trained hydrogels to ensure superior stretchability of 8.6. The acid-assisted training performance of the hydrogels can be reversibly recovered by NaOH neutralization. The acid-assisted training strategy here is general for poly (sodium acrylate) hydrogels.

Immobilization of Laccase in β-Cyclodextrin Composite Hydrogel for Efficient Degradation of Dye Pollutants

A stable and efficient biocatalyst was prepared by encapsulating Trametes versicolor laccase using an acrylic acid-grafted β-cyclodextrin hydrogel (Lac-CD-PAA). Scanning electron microscopy and nitrogen adsorption-desorption experiments showed that there were regularly distributed channels in the spongy Lac-CD-PAA. In addition, a large number of mesopores and macropores existed in the wall of the hydrogel lamellae. This network structure reduced the diffusion resistance of the hydrogel to the target substrate. The relative activity of the resulting Lac-CD-PAA could be maintained at 35.8% after six cycles of use. Lac-CD-PAA exhibited higher thermal and chemical stability compared to free laccase. The negative charge on the surface of Lac-CD-PAA gives it the ability to pretreat cationic dyes. In six consecutive methylene blue decolorization tests, Lac-CD-PAA decolorized better than free laccase. The results showed that the prepared β-cyclodextrin-based composite hydrogel was a good carrier for laccase.

Gellan Gum grafted poly acrylic acid hydrogels: Synthesis and adsorption studies

Gellan Gum (GG) grafted poly-acrylic acid (GG-g-pAAc) hydrogels of different composition were synthesized by changing the concentration of GG through free radical polymerization. The hydrogels were characterized through SEM, TGA, and FTIR. The FTIR spectra confirmed the grafting of GG into the network of polyacrylic acid and evidenced the GG-g-pAAc formation. The rough surface nature of the hydrogels was elaborated through SEM analysis provided the information about the materials to be utilized for removal purpose. The thermal stability was confirmed by TGA analysis and the hydrogels were found stable upto a high temperature range. With an efficiency of 96%, Basic Orange-2 dye (BO-2) was successfully removed from wastewater using synthesized hydrogels as an adsorbent. The adsorption parameters were calculated using a number of different kinetic models. According to the findings, the pseudo-second order kinetics model best suited to the kinetics data, while the Langmuir isotherm provided the most comprehensive explanation of the process of adsorption. BO-2 was removed to the greatest extent (257.5 mg/g). Additionally, the results showed that the adsorption process on synthesized hydrogels was exothermic with values of ΔH = −13.2, −219.4 and −12.9 kJ/mol for pAAc, 10%GG-g-pAAc, and 20%GG-g-pAAc respectively. The adsorption process is spontaneous, with negative values of ΔG = −8.2, −214.4 and −7.947 kJ/mol for pAAc, 10%GG-g-pAAc and 20%GG-g-pAAc at 293K, respectively. The adsorption process on synthesized hydrogels showed no dis-orderness with negative values of ΔS = −17.2, −461.16 and −16.42 kJ/mol for pAAc, 10%GG-g-pAAc and 20%GG-g-pAAc, respectively.

3D printing assisted surface patterning process on acrylated hydrogels for contact guidance of fibroblasts

Generating stable and customizable topography on hydrogel surfaces with contact guidance potential is critical as it can direct/influence cell growth. This necessitates the development of new techniques for surface patterning of the hydrogels. We report on the design of a square grid template for surface patterning hydrogels. The template was 3-D printed and has the diameter of a well in a 24-well plate. Hyaluronic acid methacrylate (HA) hydrogel precursor solutions were cast on the 3D printed template's surface, which generated 3D square shape topographies on the HA hydrogel surface upon demolding. The 3D Laser Microscopy has shown the formation of a periodic array of 3D topographies on hydrogel surfaces. 3D Laser and Electron Microscopy Imaging have revealed that this new method has increased the surface area and exposed the underlying pore structure of the HA hydrogels. To demonstrate the method's versatility, we have successfully applied this technique to generate 3D topography on two more acrylate hydrogel formulations, gelatin Methacrylate and polyethylene glycol dimethacrylate. Human neonatal dermal fibroblast cells were used as a model cell line to evaluate the cell guidance potential of patterned HA hydrogel. Confocal fluorescence microscopy imaging has revealed that the 3D surface topographies on HA hydrogels can guide and align the actin filaments of the fibroblasts presumably due to the contact guidance mechanism. The newly developed methodology of 3D topography generation in acrylate hydrogels may influence the cell responses on hydrogel surfaces which can impact biomedical applications such as tissue engineering, wound healing, and disease modeling.

Cellulose fiber derived from sugarcane bagasse and polyethylene glycol/acrylic acid/ branched polyethylenimine-based hydrogel composite prepared by gamma irradiation: A platform for mercury (II) ions adsorption

In this study, cellulose was successfully purified from sugarcane bagasse by alkali treatment and bleaching process. Then, 2-10 wt% of purified cellulose was added into polyethylene glycol and acrylic acid-based hydrogel composite. 25 kGy of gamma irradiation dose was employed to form the gel. With the presence of various cellulose contents, no significant change of functional group was therefore observed. The hydrogel composite was thermally resisted up to 250°C. The microstructure of composite exhibited as an interconnected porosity throughout hydrogel network. Cellulose was located inside hydrogel composite. The gel fraction and porosity for all of composites were reported to be 92-97% and 73-81%, respectively. The swelling behavior was very rapid within initial stage (5 hours) and then, it presented as a plateau region. The presence of purified cellulose significantly offered high dimensional stability. After that, in order to enhance the ability for Hg2+ adsorption, bPEI was coated onto the surface of hydrogel composite. FTIR was employed to evaluate the presence of N-H stretching. At the neutral pH, it defined as an optimal region for adsorption. The efficiency and capacity for Hg2+ adsorption were reported to be 55.17% and 82.94 mg/g in 24 hours, respectively. This was related to the pseudo-second-order kinetic model due to R2 of 0.9941 indicating the chemical adsorption process. Furthermore, EDX analysis confirmed that the existence of Hg was presented onto hydrogel composite surface. It was remarkably noted that purified cellulose from sugarcane bagasse was successfully inserted into hydrogel composite by gamma irradiation technique. It provided a great promise as a platform for mercury ion adsorption.

Fructose-Based Non-Isocyanate Polyurethane/Poly (Sodium Acrylate) Hydrogels: Design, Synthesis and Environmental Applications

Fructose-Based Non-Isocyanate Polyurethane/Poly (Sodium Acrylate) Hydrogels: Design, Synthesis and Environmental Applications

Thermochromic Hydrogels with Adjustable Transition Behavior for Smart Windows.

With the fast economic development and accelerating urbanization, more and more skyscrapers made entirely of concrete and glass are being constructed. To keep a comfortable indoor environment, massive energy for air conditioning or heating appliances is consumed. A huge amount of heat (>30%) is gained or released through glass windows. Using smart windows with the capability to modulate light is an effective way to reduce building energy consumption. Thermochromic hydrogel is one of the potential smart window materials due to its excellent thermal response, high radiation-blocking efficiency, cost-effectiveness, biocompatibility, and good uniformity. In this work, polyhydroxypropyl acrylate (PHPA) hydrogels with controllable lower critical solution temperature (LCST) were prepared by photopolymerization. The transition temperature and transition rate under "static transition" conditions were investigated. Unlike "static" conditions in which the transition temperature was not affected by the initial and final temperature and heating/cooling ramp, the transition temperature varied with the rate of temperature change under dynamic conditions. The "dynamic" transition temperature of the PHPA hydrogel gradually increased with the increase of the heating rate. It was the result of the movement of the molecular chains lagging behind the temperature change when the temperature change was too fast. The results of the solar irradiation experiment by filling PHPA hydrogels into double glazing windows showed that the indoor temperature was about 15 °C lower than that of ordinary glass windows, indicating that it can significantly reduce the energy consumption of air conditioning. In addition, a wide range of adjustable transition temperatures and fast optical response make PHPA hydrogels potentially applicable to smart windows.

Hygroscopy as an Indicator of Specific Surface Area in Polymer Materials.

Specific surface area (SSA) is an integral characteristic of the interfacial surface in poly-disperse systems, widely used for the assessment of technological properties in polymer materials and composites. Hygroscopic water content (Wh) is an obligate indicator of dispersed materials prior to any analysis of their chemical composition. This study links both indicators for the purpose of the express assessment of SSA using widely available Wh data, on the example of natural (starch, cellulose) and synthetic (acrylic hydrogels) polymer materials. The standard BET analysis of SSA using water vapor desorption was chosen as a reference method. In contrast to the known empirical correlations, this study is based on the fundamental thermodynamic theory of the disjoining water pressure for the connection of the analyzed quantities. The statistical processing of the results for the new methodology and the standard BET method showed their good compliance in a wide range of SSA from 200 to 900 m2/g. The most important methodological conclusion is the possibility of an accurate physically based calculation of hydrophilic SSA in polymer materials using their Wh data at a known relative humidity in the laboratory.

A stimuli-responsive hydrogel for reversible information storage, encryption and decryption

Smart hydrogel materials, known for their sensitivity to external stimuli, exhibit a reversible dynamic response and find applications in diverse fields, particularly in information storage. Despite significant efforts in this domain, developing a hydrogel with high-resolution, repeatable recording, and robust information encryption/decryption capabilities still remains a challenge. In this study, we synthesized a polymer hydrogel, namely polyvinyl alcohol-n-isopropylacrylamide-octadecyl polyoxyethylene ether acrylate hydrogel (PPNS), which features multiple hydrogen bonds through copolymerization, by using N-isopropylacrylamide, polyvinyl alcohol, and octadecyl polyoxyethylene ether acrylate (SGA15) as raw materials. The PPNS hydrogel demonstrated outstanding high-resolution, repeatable recording capabilities, enabling reversible recording, encryption, and decryption of information using anhydrous ethanol as the inducer. Varying the SGA15 monomer concentration revealed that the PPNS-2% hydrogel, prepared with 2% SGA15, outperformed the other hydrogels in terms of information recording and encryption/decryption when immersed in anhydrous ethanol and deionized water. Furthermore, the PPNS-2% hydrogel exhibited the ability to undergo multiple information cycles while maintaining excellent mechanical properties even after 25 cycles. Notably, ethanol served as a specialized ink for inscribing different patterns on the hydrogel surface for information recording. The recorded information could be erased through water wiping or ethanol volatilization, enabling reversible information recording, encryption, and decryption. Due to their responsive and dynamic nature of PPNS hydrogels are positions them as promising candidates for use as innovative information storage platforms.

Preparation and Characterization of Acrylic and Methacrylic Phospholipid-Mimetic Polymer Hydrogels and Their Applications in Optical Tissue Clearing.

The 2-methacryloyloxyethyl phosphorylcholine (MPC) polymers are mimetic to phospholipids, being widely used as biocompatible polymers. In our previous study, MPC polymer hydrogels proved more effective for optical tissue clearing compared to acrylamide (AAm) polymer hydrogels. In the present study, 2-acryloyloxyethyl phosphorylcholine (APC) was synthesized and employed to create hydrogels for a comparative analysis with methacrylic MPC-based hydrogels. APC, an acrylic monomer, was copolymerized with AAm in a similar reactivity. In contrast, MPC, as a methacrylic monomer, demonstrated higher copolymerization reactivity than AAm, leading to a spontaneously delayed two-step polymerization behavior. This suggests that the polymer sequences and network structures became heterogeneous when both methacrylic and acrylic monomers, as well as crosslinkers, were present in the copolymerization system. The molecular weight of the APC polymers was considerably smaller than that of the MPC polymers due to the formation of mid-chain radicals and subsequent β-scission during polymerization. The swelling ratios in water and strain sweep profiles of hydrogels prepared using acrylic and methacrylic compounds differed from those of hydrogels prepared using only acrylic compounds. This implies that copolymerization reactivity influences the polymer network structures and crosslinking density in addition to the copolymer composition. APC-based hydrogels are effective for the optical clearing of tumor tissues and are applicable to both passive and electrophoretic methods.