Unraveling Extremely Damaging IRAK4 Variants and Their Potential Implications for IRAK4 Inhibitor Efficacy.

Interleukin-1-receptor-associated kinase 4 (IRAK4) possesses a crucial function in the toll-like receptor (TLR) signaling pathway, and the dysfunction of this molecule could lead to various infectious and immune-related diseases in addition to cancers. IRAK4 genetic variants have been linked to various types of diseases. Therefore, we conducted a comprehensive analysis to recognize the missense variants with the most damaging impacts on IRAK4 with the employment of diverse bioinformatics tools to study single-nucleotide polymorphisms' effects on function, stability, secondary structures, and 3D structure. The residues' location on the protein domain and their conservation status were investigated as well. Moreover, docking tools along with structural biology were engaged in analyzing the SNPs' effects on one of the developed IRAK4 inhibitors. By analyzing IRAK4 gene SNPs, the analysis distinguished ten variants as the most detrimental missense variants. All variants were situated in highly conserved positions on an important protein domain. L318S and L318F mutations were linked to changes in IRAK4 secondary structures. Eight SNPs were revealed to have a decreasing effect on the stability of IRAK4 via both I-Mutant 2.0 and Mu-Pro tools, while Mu-Pro tool identified a decreasing effect for the G198E SNP. In addition, detrimental effects on the 3D structure of IRAK4 were also discovered for the selected variants. Molecular modeling studies highlighted the detrimental impact of these identified SNP mutant residues on the druggability of the IRAK4 ATP-binding site towards the known target inhibitor, HG-12-6, as compared to the native protein. The loss of important ligand residue-wise contacts, altered protein global flexibility, increased steric clashes, and even electronic penalties at the ligand-binding site interfaces were all suggested to be associated with SNP models for hampering the HG-12-6 affinity towards IRAK4 target protein. This given model lays the foundation for the better prediction of various disorders relevant to IRAK4 malfunction and sheds light on the impact of deleterious IRAK4 variants on IRAK4 inhibitor efficacy.

Detoxification gene expression, genotoxicity, and hepatorenal damage induced by subacute exposure to the new pyrethroid, imiprothrin, in rats.

The pyrethroid imiprothrin is widely used worldwide for control of insects in the agriculture and public health sectors. No sufficient information is however available concerning detoxification gene expression, i.e., cytochrome P450 1A2 (CYP1A2) and metallothionein 1a gene, oxidative stress, lipid peroxidation, DNA damage, cytotoxicity, genotoxicity, and organ injury induced by imiprothrin in mammals. This study is designed to explain the mechanism of imiprothrin induced detoxification gene expression, DNA damage, cytotoxicity, genotoxicity, and organ toxicity in male rats. The benchmark dose (BMD) was calculated to find the best sensitive markers to imiprothrin toxicity. Imiprothrin was injected intraperitoneally (i.p.) into male rats once a day for 5 days with doses of 19, 38, and 75 mg/kg body weight (b.wt.). Imiprothrin caused a significant increase in lipid peroxidation and changes in oxidative stress biomarkers in treated rats. Significant dose-dependent changes in the liver and kidney biomarkers were observed. Histopathological alterations were seen in the liver and kidney tissue of male rats. Imiprothrin also significantly increased chromosomal aberrations (CA) and micronuclei in bone-marrow cells, and induced lipid peroxidation, oxidative stress, cytotoxicity, and liver and kidney dysfunction, and damage. Imiprothrin induced DNA damage and over detoxification gene expression of CYP1A2 and metallothionein 1a gene in hepatocytes of male rats. Imiprothrin thus shows clastogenic and genotoxic potential. The mechanism for hepatorenal toxicity and injury, genotoxicity/cytotoxicity of imiprothrin might be due to enhanced lipid peroxidation, and oxidative stress associated with overproduction of free radicals, especially reactive oxygen species, and an imbalance in redox status. From the BMD models, aspartate aminotransferase (AST), total protein, uric acid, superoxide dismutase (SOD), and micronuclei (MPEs) were very sensitive markers to imiprothrin toxicity.

Biological applications study of bio-nanocomposites based on chitosan/TiO2 nanoparticles polymeric films modified by oleic acid.

The aim of the present study was to prepare and characterize nanocomposite films to improve the treatment of skin wounds by applying the film as a bandage. To modify chitosan (Cs) and to prepare nanocomposites, a mixture between titanium dioxide nanoparticles (TiO2 NPs) was performed at different concentrations (2, 5, 10 and 15 wt%) and oleic acid (OA). The thin nanocomposite films were prepared by using casting method. The prepared films (Cs, Cs/TiO2 NPs, Cs/OA and Cs/OA/TiO2 NPs) were described by water absorption (swelling study) and biological degradation. Physico-chemical characterizations of Cs, Cs/OA, Cs/TiO2 NPs and Cs/OA/TiO2 NPs (with only 15 wt% TiO2 NPs) films were determined by X-ray diffraction, transmission high-resolution electron microscopy, field emission scanning electron microscopy, thermal analysis and Fourier transform infrared spectroscopy as well as their mechanical properties. Antimicrobial activity against microorganisms has been studied to assess activity against bacteria. The prepared nanocomposite films showed good antimicrobial activity for both Gram-positive and Gram-negative bacteria. The therapeutic effects of Cs-TiO2 NPs-oleic acid nanocomposites on healing excision wounds were studied in rat animal model. The data obtained revealed that groups treated with nanocomposites showed enhancement wound closure and speed up wound healing time.

Thymoquinone-PLGA-PVA Nanoparticles Ameliorate Bleomycin-Induced Pulmonary Fibrosis in Rats via Regulation of Inflammatory Cytokines and iNOS Signaling.

Pulmonary fibrosis is considered one of the most chronic interstitial illnesses which are not easily treated. thymoquinone's (TQ) benefits are still partly problematic due to poor water solubility; therefore, it was loaded onto PLGA-PVA carriers. This study aimed to evaluate the potential effect of TQ-PLGA-PVA nanoparticles (TQ-PLGA-PVA-NPs) on pulmonary fibrosis induced by bleomycin in albino rats. Forty male rats were randomized into four groups. The first group served as the control group; the second and the third groups received bleomycin intratracheally, whereas the third group received TQ-PLGA-PVA-NPs after 4 weeks from bleomycin administration. The fourth group was administrated TQ-PLGA-PVA-NPs alone. The designed nanoparticles appeared around 20 nm size (10-30 nm), had a spherical shape, and had 80% encapsulation efficiency. The histological examination of rats simultaneously treated with TQ-PLGA-PVA-NPs and bleomycin revealed reduction in the thickness of the alveolar septa and improvement of the other lung structures, with the presence of lymphocytes admixed with exfoliated epithelium in a few lumina remaining. Ultrastructural findings revealed marked collagenolysis and the release of nanoparticles from ruptured pneumocytes within the alveolar septa after 14 days from TQ-PLGA-PVA-NPs administration. Very active pneumocyte types II were seen in the TQ-PLGA-PVANP group. Additionally, immunohistochemical expression of inducible nitric oxide (iNOS) and estimation of inflammatory cytokines in lung tissues including interleukin 10 (IL 10) and transforming growth factor-beta (TGF-ß1) confirmed the antioxidant and anti-inflammatory effects of TQ-PLGA-PVANPs. The study concluded that TQ-PLGA-PVA-NPs could attenuate the bleomycin-induced pulmonary fibrosis, through the inhibition of lung inflammation and the suppression of bleomycin- induced oxidative stress.

Thymoquinone ameliorates pulmonary vascular damage induced byEscherichia coli-derived lipopolysaccharide via cytokine downregulation in rats.

Our study investigated the ameliorative effects of thymoquinone (TQ) on the pulmonary blood vessels which were injured after intratracheal administration of Escherichia coli-derived lipopolysaccharide (LPS) in a rat model. Forty rats (150 ± 50 g) were randomly divided into four groups equally. The first group was intratracheally administered LPS (Escherichia coli O55:B5) at a dose 200 µg. The second group was co-administered intraperitoneal injection of TQ and LPS daily for one week. The third group was provided intraperitoneal injection of 1 mg of TQ. The fourth group was administered normal saline intratracheally at the rate of 200 µl. The results revealed that cytokine level of interleukin-1 beta (IL-1ß) and tumor necrosis factor alpha (TNFα) in serum was reduced in TQ-treated rats. Immunohistochemical study showed that expression of NF-kB was countered in the lung tissue by the application of TQ. In addition, the lesion score for various pathological aberrations were checked when rats were treated with TQ. From the results of the present study, it was concluded that TQ has an ameliorative effect on the pulmonary blood vascular damage via rearrangement of the cytokines in response to LPS injury in the rat model.