Structure of the interleukin-5 receptor complex exemplifies the organizing principle of common beta cytokine signaling.

Cytokines regulate immune responses by binding to cell surface receptors, including the common subunit beta (ßc), which mediates signaling for GM-CSF, IL-3, and IL-5. Despite known roles in inflammation, the structural basis of IL-5 receptor activation remains unclear. We present the cryo-EM structure of the human IL-5 ternary receptor complex, revealing architectural principles for IL-5, GM-CSF, and IL-3. In mammalian cell culture, single-molecule imaging confirms hexameric IL-5 complex formation on cell surfaces. Engineered chimeric receptors show that IL-5 signaling, as well as IL-3 and GM-CSF, can occur through receptor heterodimerization, obviating the need for higher-order assemblies of ßc dimers. These findings provide insights into IL-5 and ßc receptor family signaling mechanisms, aiding in the development of therapies for diseases involving deranged ßc signaling.

Influence of the Type of Biocompatible Polymer in the Shell of Magnetite Nanoparticles on Their Interaction with DPPC in Two-Component Langmuir Monolayers.

Magnetite nanoparticles (MNPs) are attractive nanomaterials for applications in magnetic resonance imaging, targeted drug delivery, and anticancer therapy due to their unique properties such as nontoxicity, wide chemical affinity, and intrinsic superparamagnetism. Their functionalization with polymers such as chitosan or poly(vinyl alcohol) (PVA) can not only improve their biocompatibility and biodegradability but it also plays an important role in their interactions with biological cells. In this work, the effect of the functionalization of MNPs with chitosan, PVA, and their blend on model cell membranes formed from 1,2-dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC) using a Langmuir technique was studied. The studies performed showed that the type of biocompatible polymer in the MNP shell plays a crucial role in the effectiveness of its adsorption process into the model cell membrane. Modification of MNPs with chitosan facilitates significantly more effective adsorption than coating them with PVA or with a chitosan and PVA blend. The presence of all the investigated MNPs in the DPPC monolayer at low concentrations does not affect its thermodynamic state, fluidity, or morphology, which is promising in terms of their biocompatibility. On the other hand, their high concentration (molar fraction above ≈0.05) exerts a disruptive effect on the model cell membrane and results in their aggregation, leading probably to the loss of their superparamagnetic properties essential for nanomedicine.

Photochemical stability of chitosan films doped with cannabis oil.

The effect of UV radiation from three different sources on chitosan (CS) films containing the addition of 10% by weight of cannabis oil was investigated. Cannabis oil (CBD) alone exposed to UV is unstable, but its photostability significantly increases in the chitosan matrix. The course of photochemical reactions, studied by FTIR spectroscopy, is slow and inefficient in chitosan with CBD, even under high-energy UV sources. The research also included chitosan films with CBD cross-linked with dialdehyde starch (DAS). Using AFM microscopy and contact angle measurements, the morphology and surface properties of prepared chitosan films with CBD were investigated, respectively. It was found that CBD embedded in CS is characterized by the best photostability under the influence of an LED emitting long-wave radiation. Using a monochromatic and polychromatic UV lamp (HPK and UV-C) emitting high-energy radiation, gradual degradation accompanied by oxidation was observed, both in the CS chains and in the CBD additive. Additionally, changes in surface properties are observed during UV irradiation. It was concluded that CS protects CBD against photodegradation, and a further improvement in photochemical stability is achieved after system cross-linking with DAS.

Three-Dimensional Segmentation Assisted with Clustering Analysis for Surface and Volume Measurements of Equine Incisor in Multidetector Computed Tomography Data Sets.

Dental diagnostic imaging has progressed towards the use of advanced technologies such as 3D image processing. Since multidetector computed tomography (CT) is widely available in equine clinics, CT-based anatomical 3D models, segmentations, and measurements have become clinically applicable. This study aimed to use a 3D segmentation of CT images and volumetric measurements to investigate differences in the surface area and volume of equine incisors. The 3D Slicer was used to segment single incisors of 50 horses' heads and to extract volumetric features. Axial vertical symmetry, but not horizontal, of the incisors was evidenced. The surface area and volume differed significantly between temporary and permanent incisors, allowing for easy eruption-related clustering of the CT-based 3D images with an accuracy of >0.75. The volumetric features differed partially between center, intermediate, and corner incisors, allowing for moderate location-related clustering with an accuracy of >0.69. The volumetric features of mandibular incisors' equine odontoclastic tooth resorption and hypercementosis (EOTRH) degrees were more than those for maxillary incisors; thus, the accuracy of EOTRH degree-related clustering was >0.72 for the mandibula and >0.33 for the maxilla. The CT-based 3D images of equine incisors can be successfully segmented using the routinely achieved multidetector CT data sets and the proposed data-processing approaches.

Biochemical and biophysical characterization of natural polyreactivity in antibodies.

To become specialized binders, antibodies undergo a process called affinity maturation to maximize their binding affinity. Despite this process, some antibodies retain low-affinity binding to diverse epitopes in a phenomenon called polyreactivity. Here we seek to understand the molecular basis of this polyreactivity in antibodies. Our results highlight that polyreactive antigen-binding fragments (Fabs) bind their targets with low affinities, comparable to T cell receptor recognition of autologous classical major histocompatibility complex. Extensive mutagenic studies find no singular amino acid residue or biochemical property responsible for polyreactive interaction, suggesting that polyreactive antibodies use multiple strategies for engagement. Finally, our crystal structures and all-atom molecular dynamics simulations of polyreactive Fabs show increased rigidity compared to their monoreactive relatives, forming a neutral and accessible platform for diverse antigens to bind. Together, these data support a cooperative strategy of rigid neutrality in establishing the polyreactive status of an antibody molecule.

Polymer-Based Nanoparticles as Drug Delivery Systems for Purines of Established Importance in Medicine.

Many purine derivatives are active pharmaceutical ingredients of significant importance in the therapy of autoimmune diseases, cancers, and viral infections. In many cases, their medical use is limited due to unfavorable physicochemical and pharmacokinetic properties. These problems can be overcome by the preparation of the prodrugs of purines or by combining these compounds with nanoparticles. Herein, we aim to review the scientific progress and perspectives for polymer-based nanoparticles as drug delivery systems for purines. Polymeric nanoparticles turned out to have the potential to augment antiviral and antiproliferative effects of purine derivatives by specific binding to receptors (ASGR1-liver, macrophage mannose receptor), increase in drug retention (in eye, intestines, and vagina), and permeation (intranasal to brain delivery, PEPT1 transport of acyclovir). The most significant achievements of polymer-based nanoparticles as drug delivery systems for purines were found for tenofovir disoproxil in protection against HIV, for acyclovir against HSV, for 6-mercaptopurine in prolongation of mice ALL model life, as well as for 6-thioguanine for increased efficacy of adoptively transferred T cells. Moreover, nanocarriers were able to diminish the toxic effects of acyclovir, didanosine, cladribine, tenofovir, 6-mercaptopurine, and 6-thioguanine.

Fabrication and characterization of new levan@CBD biocomposite sponges as potential materials in natural, non-toxic wound dressing applications.

Wound healing is a complex process; therefore, new dressings are frequently required to facilitate it. In this study, porous bacterial levan-based sponges containing cannabis oil (Lev@CBDs) were prepared and fully characterized. The sponges exhibited a suitable swelling ratio, proper water vapor transmission rate, sufficient thermal stability, desired mechanical properties, and good antioxidant and anti-inflammatory properties. The obtained Lev@CBD materials were evaluated in terms of their interaction with proteins, human serum albumin and fibrinogen, of which fibrinogen revealed the highest binding effect. Moreover, the obtained biomaterials exhibited antibacterial activity against Staphylococcus aureus and Pseudomonas aeruginosa, as well as being non-hemolytic material as indicated by hemolysis tests. Furthermore, the sponges were non-toxic and compatible with L929 mouse fibroblasts and HDF cells. Most significantly, the levan sponge with the highest content of cannabis oil, in comparison to others, retained its non-hemolytic, anti-inflammatory, and antimicrobial properties after prolonged storage in a climate chamber at a constant temperature and relative humidity. The designed sponges have conclusively proven their beneficial physicochemical properties and, at the preliminary stage, biocompatibility as well, and therefore can be considered a promising material for wound dressings in future in vivo applications.

An application of the density standard and scaled-pixel-counting protocol to assess the radiodensity of equine incisor teeth affected by resorption and hypercementosis: preliminary advancement in dental radiography.

BACKGROUND: Equine Odontoclastic Tooth Resorption and Hypercementosis (EOTRH) syndrome is a dental disease where the radiographic signs may be quantified using radiographic texture features. This study aimed to implement the scaled-pixel-counting protocol to quantify and compare the image structure of teeth and the density standard in order to improve the identification of the radiographic signs of tooth resorption and hypercementosis using the EOTRH syndrome model. METHODS AND RESULTS: A detailed examination of the oral cavity was performed in 80 horses and maxillary incisor teeth were evaluated radiographically, including an assessment of the density standard. On each of the radiographs, pixel brightness (PB) was extracted for each of the ten steps of the density standard (S1-S10). Then, each evaluated incisor tooth was assigned to one of 0-3 EOTRH grade-related groups and annotated using region of interest (ROI). For each ROI, the number of pixels (NP) from each range was calculated. The linear relation between an original X-ray beam attenuation and PB was confirmed for the density standard. The NP values increased with the number of steps of the density standard as well as with EOTRH degrees. Similar accuracy of the EOTRH grade differentiation was noted for data pairs EOTRH 0-3 and EOTRH 0-1, allowing for the differentiation of both late and early radiographic signs of EOTRH. CONCLUSION: The scaled-pixel-counting protocol based on the use of density standard has been successfully implemented for the differentiation of radiographic signs of EOTRH degrees.

Correction: Górski et al. Application of Two-Dimensional Entropy Measures to Detect the Radiographic Signs of Tooth Resorption and Hypercementosis in an Equine Model. Biomedicines 2022, 10, 2914.

In the original article [...].

Bio-Approach for Obtaining Enantiomerically Pure Clopidogrel with the Use of Ionic Liquids.

Clopidogrel is a chiral compound widely used as an antiplatelet medication that lowers the risk of blood clots, strokes, and heart attacks. The main aim of the study presented herein was to obtain (S)-clopidogrel, which is commercially available in treatments, via the kinetic resolution of racemic clopidogrel carboxylic acid with the use of lipase from Candida rugosa and a two-phase reaction medium containing an ionic liquid. For this purpose, the enantioselective biotransformation of clopidogrel carboxylic acid and chiral chromatographic separation with the use of a UPLC-MS/MS system were optimized. The best kinetic resolution parameters were obtained by using a catalytic system containing lipase from Candida rugosa OF as a biocatalyst, cyclohexane and [EMIM][BF4] as a two-phase reaction medium, and methanol as an acyl acceptor. The enantiomeric excess of the product was eep = 94.21% ± 1.07 and the conversion was c = 49.60% ± 0.57%, whereas the enantioselectivity was E = 113.40 ± 1.29. The performed study proved the possibility of obtaining (S)-clopidogrel with the use of lipase as a biocatalyst and a two-phase reaction medium containing an ionic liquid, which is in parallel with green chemistry methodology and does not require environmentally harmful conditions.

Chitosan Composites Containing Boron-Dipyrromethene Derivatives for Biomedical Applications.

The work is devoted to preparing and characterizing the properties of photosensitive composites, based on chitosan proposed for photodynamic therapy. Chitosan films with a 5% addition of two BODIPY dyes were prepared by solution casting. These dyes are dipyrromethene boron derivatives with N-alkyl phthalimide substituent, differing in the presence of iodine atoms in positions 2 and 6 of the BODIPY core. The spectral properties of the obtained materials have been studied by infrared and UV-vis absorption spectroscopy and fluorescence, both in solutions and in a solid state. Surface properties were investigated using the contact angle measurement. The morphology of the sample has been characterized by Scanning Electron and Atomic Force Microscopy. Particular attention was paid to studying the protein absorption and kinetics of the dye release from the chitosan. Adding BODIPY to the chitosan matrix leads to a slight increase in hydrophilicity, higher structure heterogeneity, and roughness, than pure chitosan. The presence of iodine atoms in the BODIPY structure caused the bathochromic effect, but the emission quantum yield decreased in the composites. It has been found that BODIPY-doped chitosan interacts better with human serum albumin and acidic α-glycoprotein than unmodified chitosan. The release rate of dyes from films immersed in methanol depends on the iodine present in the structure.

Ensemble of Heterogeneous Base Classifiers for Human Gait Recognition.

Human gait recognition is one of the most interesting issues within the subject of behavioral biometrics. The most significant problems connected with the practical application of biometric systems include their accuracy as well as the speed at which they operate, understood both as the time needed to recognize a particular person as well as the time necessary to create and train a biometric system. The present study made use of an ensemble of heterogeneous base classifiers to address these issues. A Heterogeneous ensemble is a group of classification models trained using various algorithms and combined to output an effective recognition A group of parameters identified on the basis of ground reaction forces was accepted as input signals. The proposed solution was tested on a sample of 322 people (5980 gait cycles). Results concerning the accuracy of recognition (meaning the Correct Classification Rate quality at 99.65%), as well as operation time (meaning the time of model construction at <12.5 min and the time needed to recognize a person at <0.1 s), should be considered as very good and exceed in quality other methods so far described in the literature.

Facile repurposing of peptide-MHC-restricted antibodies for cancer immunotherapy.

Monoclonal antibodies (Abs) that recognize major histocompatability complex (MHC)-presented tumor antigens in a manner similar to T cell receptors (TCRs) have great potential as cancer immunotherapeutics. However, isolation of 'TCR-mimic' (TCRm) Abs is laborious because Abs have not evolved the structurally nuanced peptide-MHC restriction of αß-TCRs. Here, we present a strategy for rapid isolation of highly peptide-specific and 'MHC-restricted' Abs by re-engineering preselected Abs that engage peptide-MHC in a manner structurally similar to that of conventional αß-TCRs. We created structure-based libraries focused on the peptide-interacting residues of TCRm Ab complementarity-determining region (CDR) loops, and rapidly generated MHC-restricted Abs to both mouse and human tumor antigens that specifically killed target cells when formatted as IgG, bispecific T cell engager (BiTE) and chimeric antigen receptor-T (CAR-T). Crystallographic analysis of one selected pMHC-restricted Ab revealed highly peptide-specific recognition, validating the engineering strategy. This approach can yield tumor antigen-specific antibodies in several weeks, potentially enabling rapid clinical translation.

The Postural and Body Surface Temperature Response of Leisure Horses to Lunging with Selected Lunging Aids.

Incorporating lunging into a horse's daily routine aims to enhance fitness, physical condition, and specific skills or exercises when using lunging aids (LAs). To assess the effectiveness of lunging, non-contact technologies like geometric morphometrics and infrared thermography can be employed. This study seeks to evaluate lunging efficiency based on the horse's posture and surface temperature when lunging with different head and neck positions. The study aims to determine if changes in a horse's posture correspond to increased metabolic activity, as indicated by body surface temperature. Thirteen horses included in the study were lunged with chambon (CH), rubber band (RB), and triangle side reins (TRs) as well as with a freely moving head (FMH). Images were taken in visible light and infrared. Principal Component Analysis (PCA) was used to analyze horse posture changes and a Pixel-Counting Protocol (PCP) was used to quantify surface temperature patterns. The horses' posture exhibited contrasting changes, reflected by a changing centroid shape (p < 0.0001) but not size (p > 0.05) when lunged with RB and TRs, but not CH. Different (p < 0.0001) surface temperature patterns were observed during lunging. FMH lunging resulted in lower temperatures over a larger surface, CH induced moderate temperatures on a smaller area, RB caused moderate to high temperatures across a broader surface, and TRs led to higher temperatures over a smaller region. The studied lunging cases returned different (p < 0.0001) surface temperature patterns. Lunging with FMH returned lower temperatures over a larger surface, CH moderate temperatures on a smaller area, RB moderate to high temperatures across a broader surface, and TRs higher temperatures over a smaller region. The proposed methods can be applied to evaluate the efficiency of lunging in horses.

Chitosan-based films with cannabis oil as a base material for wound dressing application.

This study focuses on obtaining and characterizing novel chitosan-based biomaterials containing cannabis oil to potentially promote wound healing. The primary active substance in cannabis oil is the non-psychoactive cannabidiol, which has many beneficial properties. In this study, three chitosan-based films containing different concentrations of cannabis oil were prepared. As the amount of oil increased, the obtained biomaterials became rougher as tested by atomic force microscopy. Such rough surfaces promote protein adsorption, confirmed by experiments assessing the interaction between human albumin with the obtained materials. Increased oil concentration also improved the films' mechanical parameters, swelling capacity, and hydrophilic properties, which were checked by the wetting angle measurement. On the other hand, higher oil content resulted in decreased water vapour permeability, which is essential in wound dressing. Furthermore, the prepared films were subjected to an acute toxicity test using a Microtox. Significantly, the film's increased cannabis oil content enhanced the antimicrobial effect against A. fischeri for films in direct contact with bacteria. More importantly, cell culture studies revealed that the obtained materials are biocompatible and, therefore, they might be potential candidates for application in wound dressing materials.

Application of Two-Dimensional Entropy Measures to Detect the Radiographic Signs of Tooth Resorption and Hypercementosis in an Equine Model.

Dental disorders are a serious health problem in equine medicine, their early recognition benefits the long-term general health of the horse. Most of the initial signs of Equine Odontoclastic Tooth Resorption and Hypercementosis (EOTRH) syndrome concern the alveolar aspect of the teeth, thus, the need for early recognition radiographic imaging. This study is aimed to evaluate the applicability of entropy measures to quantify the radiological signs of tooth resorption and hypercementosis as well as to enhance radiographic image quality in order to facilitate the identification of the signs of EOTRH syndrome. A detailed examination of the oral cavity was performed in eighty horses. Each evaluated incisor tooth was assigned to one of four grade-related EOTRH groups (0-3). Radiographs of the incisor teeth were taken and digitally processed. For each radiograph, two-dimensional sample (SampEn2D), fuzzy (FuzzEn2D), permutation (PermEn2D), dispersion (DispEn2D), and distribution (DistEn2D) entropies were measured after image filtering was performed using Normalize, Median, and LaplacianSharpening filters. Moreover, the similarities between entropy measures and selected Gray-Level Co-occurrence Matrix (GLCM) texture features were investigated. Among the 15 returned measures, DistEn2D was EOTRH grade-related. Moreover, DistEn2D extracted after Normalize filtering was the most informative. The EOTRH grade-related similarity between DistEn2D and Difference Entropy (GLCM) confirms the higher irregularity and complexity of incisor teeth radiographs in advanced EOTRH syndrome, demonstrating the greatest sensitivity (0.50) and specificity (0.95) of EOTRH 3 group detection. An application of DistEn2D to Normalize filtered incisor teeth radiographs enables the identification of the radiological signs of advanced EOTRH with higher accuracy than the previously used entropy-related GLCM texture features.

Photocatalytic Activity of Sulfanyl Porphyrazine/Titanium Dioxide Nanocomposites in Degradation of Organic Pollutants.

Magnesium(II) sulfanyl porphyrazine with peripheral morpholinethoxy substituents was embedded on the surface of titanium(IV) dioxide nanoparticles. The obtained nanocomposites were characterized with the use of particle size and distribution (NTA analysis), electron microscopy (SEM), thermal analysis (TGA), FTIR-ATR spectroscopy, and X-ray powder diffraction (XRD). The measured particle size of the obtained material was 327.4 ± 15.5 nm. Analysis with XRD showed no visible changes in the crystallinity of the material after deposition of porphyrazine on the TiO2 surface. However, SEM images revealed noticeable changes in the morphology of the obtained hybrid material: higher aggregation and less ordered structure of the aggregates. The TGA analysis revealed the lost 3.6% (0.4 mg) of the mass of obtained material in the range 250-550 °C. In the FTIR-ATR analysis, C-H stretching vibratins in the range of 3000-2800 cm-1, originating from porphyrazine moieties, were detected. The photocatalytic applicability of the nanomaterial was assessed in photodegradation studies of methylene blue and bisphenol A as reference environmental pollutants. In addition, the photocatalytic degradation of carbamazepine with porphyrazine/TiO2 hybrids as photocatalysts was studied, accompanied by an HPLC chromatography assessment of photodegradation. In total, 43% of the initial concentration was achieved in the case of bisphenol A, after 4 h of irradiation, whereas 57% was achieved in the case of carbamazepine. In each photodegradation reaction, the activity of the obtained photocatalytic nanomaterial was proved with almost linear degradation. The photodegradation reaction rate constants were calculated, and revealed 5.75 × 10-5 s-1 for bisphenol A and 5.66 × 10-5 s-1 for carbamazepine.

Chitosan-Gelatin Films Cross-Linked with Dialdehyde Cellulose Nanocrystals as Potential Materials for Wound Dressings.

In this study, thin chitosan-gelatin biofilms cross-linked with dialdehyde cellulose nanocrystals for dressing materials were received. Two types of dialdehyde cellulose nanocrystals from fiber (DNCL) and microcrystalline cellulose (DAMC) were obtained by periodate oxidation. An ATR-FTIR analysis confirmed the selective oxidation of cellulose nanocrystals with the creation of a carbonyl group at 1724 cm-1. A higher degree of cross-linking was obtained in chitosan-gelatin biofilms with DNCL than with DAMC. An increasing amount of added cross-linkers resulted in a decrease in the apparent density value. The chitosan-gelatin biofilms cross-linked with DNCL exhibited a higher value of roughness parameters and antioxidant activity compared with materials cross-linked with DAMC. The cross-linking process improved the oxygen permeability and anti-inflammatory properties of both measurement series. Two samples cross-linked with DNCL achieved an ideal water vapor transition rate for wound dressings, CS-Gel with 10% and 15% addition of DNCL-8.60 and 9.60 mg/cm2/h, respectively. The swelling ability and interaction with human serum albumin (HSA) were improved for biofilms cross-linked with DAMC and DNCL. Significantly, the films cross-linked with DAMC were characterized by lower toxicity. These results confirmed that chitosan-gelatin biofilms cross-linked with DNCL and DAMC had improved properties for possible use in wound dressings.

Application of the Two-Dimensional Entropy Measures in the Infrared Thermography-Based Detection of Rider: Horse Bodyweight Ratio in Horseback Riding.

As obesity is a serious problem in the human population, overloading of the horse's thoracolumbar region often affects sport and school horses. The advances in using infrared thermography (IRT) to assess the horse's back overload will shortly integrate the IRT-based rider-horse fit into everyday equine practice. This study aimed to evaluate the applicability of entropy measures to select the most informative measures and color components, and the accuracy of rider:horse bodyweight ratio detection. Twelve horses were ridden by each of the six riders assigned to the light, moderate, and heavy groups. Thermal images were taken pre- and post-exercise. For each thermal image, two-dimensional sample (SampEn), fuzzy (FuzzEn), permutation (PermEn), dispersion (DispEn), and distribution (DistEn) entropies were measured in the withers and the thoracic spine areas. Among 40 returned measures, 30 entropy measures were exercise-dependent, whereas 8 entropy measures were bodyweight ratio-dependent. Moreover, three entropy measures demonstrated similarities to entropy-related gray level co-occurrence matrix (GLCM) texture features, confirming the higher irregularity and complexity of thermal image texture when horses worked under heavy riders. An application of DispEn to red color components enables identification of the light and heavy rider groups with higher accuracy than the previously used entropy-related GLCM texture features.

Dialdehyde Starch Nanocrystals as a Novel Cross-Linker for Biomaterials Able to Interact with Human Serum Proteins.

In recent years, new cross-linkers from renewable resources have been sought to replace toxic synthetic compounds of this type. One of the most popular synthetic cross-linking agents used for biomedical applications is glutaraldehyde. However, the unreacted cross-linker can be released from the materials and cause cytotoxic effects. In the present work, dialdehyde starch nanocrystals (NDASs) were obtained from this polysaccharide nanocrystal form as an alternative to commonly used cross-linking agents. Then, 5-15% NDASs were used for chemical cross-linking of native chitosan (CS), gelatin (Gel), and a mixture of these two biopolymers (CS-Gel) via Schiff base reaction. The obtained materials, forming thin films, were characterized by ATR-FTIR, SEM, and XRD analysis. Thermal and mechanical properties were determined by TGA analysis and tensile testing. Moreover, all cross-linked biopolymers were also characterized by hydrophilic character, swelling ability, and protein absorption. The toxicity of obtained materials was tested using the Microtox test. Dialdehyde starch nanocrystals appear as a beneficial plant-derived cross-linking agent that allows obtaining cross-linked biopolymer materials with properties desirable for biomedical applications.