Comparative study between zinc oxide nanoparticles synthesis by biogenic and wet chemical methods in vivo and in vitro against Staphylococcus aureus.

ZnO nanoparticles (ZnO-NPs) can be used as nano medicine for Staphylococcus aureus infection, which causes deleterious effects on liver, kidney and lung tissue, as it causes catarrhal bronchitis, peri-bronchial oedema, lymphocytic granulomas, oedematous fluid and haemorrhage inside the bronchi, and interstitial pneumonia. In this research ZnO nanoparticle (ZnO-NPs) synthesis by biogenic method using green alga Ulva fasciata and by wet chemical method. Both of them tested in vitro and in vivo against Staphylococcus aureus. The characterization of ZnO-NPs was detected by U.V spectroscopy, Fourier-transform infrared (FTIR), Energy dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD), Scanning Electron Microscope (SEM) and Transmission Electron Microscope (TEM). In vivo assessment eight groups, each group contain of five rats and the treatment as follow (1) an uninfected control group; (2) an infected group; groups (3), (4), and (5) were injected with biogenic or chemical ZnO-NPs or zinc acetate, as the bulk group, respectively; and groups (6), (7) and (8) were infected and then treated in the same manner as groups (3), (4), and (5), respectively. The blood profile, biochemical parameters, phagocytic activity and histological assessment of liver, kidney and lung tissue of each rat was investigated after 20 days. The rats treated with 5 mg/1 kg natural ZnO-NPs showed improved lung characteristics, and the number of platelets in the infected groups treated with ZnO-NPs from chemical and natural sources (G6 and G7) was close to those in the control group. However, the trend was reversed for regarding lymphocytes, which remained at higher levels in uninfected animals treated with synthetic ZnO-NPs (G4) than in infected rats treated with synthetic ZnO-NPs (G7). Moreover, a significant difference in phagocytic activity was found among all groups compared to that of controls. Compared to control group rats (G1), uninfected rats injected with only natural ZnO-NPs (G3) showed a significant (P < 0.05) improvement in the phagocytic index. We propose that ZnO-NPs produced from natural sources are preferable to those produced from chemical sources for use as nano medicine for the treatment of S. aureus infection in albino rats.

The Expression of Triticum aestivum Cysteine-Rich Receptor-like Protein Kinase Genes during Leaf Rust Fungal Infection.

Understanding the role of cysteine-rich receptor-like kinases (CRKs) in plant defense mechanisms is crucial for enhancing wheat resistance to leaf rust fungus infection. Here, we identified and verified 164 members of the CRK gene family using the Triticum aestivum reference version 2 collected from the international wheat genome sequencing consortium (IWGSC). The proteins exhibited characteristic features of CRKs, including the presence of signal peptides, cysteine-rich/stress antifungal/DUF26 domains, transmembrane domains, and Pkinase domains. Phylogenetic analysis revealed extensive diversification within the wheat CRK gene family, indicating the development of distinct specific functional roles to wheat plants. When studying the expression of the CRK gene family in near-isogenic lines (NILs) carrying Lr57- and Lr14a-resistant genes, Puccinia triticina, the causal agent of leaf rust fungus, triggered temporal gene expression dynamics. The upregulation of specific CRK genes in the resistant interaction indicated their potential role in enhancing wheat resistance to leaf rust, while contrasting gene expression patterns in the susceptible interaction highlighted potential susceptibility associated CRK genes. The study uncovered certain CRK genes that exhibited expression upregulation upon leaf rust infection and the Lr14a-resistant gene. The findings suggest that targeting CRKs may present a promising strategy for improving wheat resistance to rust diseases.

Essential oil from Lavandula angustifolia elicits expression of three SbWRKY transcription factors and defense-related genes against sorghum damping-off.

Sorghum damping-off, caused by Fusarium solani (Mart.) Sacc., is a serious disease which causes economic loss in sorghum production. In this study, antagonistic activity of lavender essential oil (EO) at 0.5, 0.75, 1.0, 1.25, 1.5, and 1.6% against F. solani was studied in vitro. Their effects on regulation of three SbWRKY transcription factors, the response factor JERF3 and eight defense-related genes, which mediate different signaling pathways, in sorghum were investigated. Effects of application under greenhouse conditions were also evaluated. The results showed that lavender EO possesses potent antifungal activity against F. solani. A complete inhibition in the fungal growth was recorded for lavender EO at 1.6%. Gas chromatography-mass spectrometric analysis revealed that EO antifungal activity is most likely attributed to linalyl anthranilate, α-terpineol, eucalyptol, α-Pinene, and limonene. Observations using transmission electron microscopy revealed many abnormalities in the ultrastructures of the fungal mycelium as a response to treating with lavender EO, indicating that multi-mechanisms contributed to their antagonistic behavior. Results obtained from Real-time PCR investigations demonstrated that the genes studied were overexpressed, to varying extents in response to lavender EO. However, SbWRKY1 was the highest differentially expressed gene followed by JERF3, which suggest they play primary role(s) in synchronously organizing the transcription-regulatory-networks enhancing the plant resistance. Under greenhouse conditions, treating of sorghum grains with lavender EO at 1.5% prior to infection significantly reduced disease severity. Moreover, the growth parameters evaluated, the activities of antioxidant enzymes, and total phenolic and flavonoid contents were all enhanced. In contrast, lipid peroxidation was highly reduced. Results obtained from this study support the possibility of using lavender EO for control of sorghum damping-off. However, field evaluation is highly needed prior to any usage recommendation.

Thiolation of Myco-Synthesized Fe3O4-NPs: A Novel Promising Tool for Penicillium expansium Laccase Immobilization to Decolorize Textile Dyes and as an Application for Anticancer Agent.

Environmental pollution due to the continuous uncontrolled discharge of toxic dyes into the water bodies provides insight into the need to eliminate pollutants prior to discharge is significantly needed. Recently, the combination of conventional chemotherapeutic agents and nanoparticles has attracted considerable attention. Herein, the magnetic nanoparticles (Fe3O4-NPs) were synthesized using metabolites of Aspergillus niger. Further, the surfaces of Fe3O4-NPs were functionalized using 3-mercaptoproionic acid as confirmed by XRD, TEM, and SEM analyses. A purified P. expansum laccase was immobilized onto Fe3O4/3-MPA-SH and then the developed immobilized laccase (Fe3O4/3-MPA-S-S-laccase) was applied to achieve redox-mediated degradation of different dyes. The Fe3O4/3-MPA-S-S-laccase exhibited notably improved stability toward pH, temperature, organic solvents, and storage periods. The Fe3O4/3-MPA-S-S-laccase exhibited appropriate operational stability while retaining 84.34% of its initial activity after 10 cycles. The catalytic affinity (Kcat/Km) of the immobilized biocatalyst was increased above 10-fold. The experimental data showed remarkable improvement in the dyes' decolorization using the immobilized biocatalyst in the presence of a redox mediator in seven successive cycles. Thus, the prepared novel nanocomposite-laccase can be applied as an alternative promising strategy for bioremediation of textile wastewater. The cytotoxic level of carboplatin and Fe3O4-NPs singly or in combination on various cell lines was concentration-dependent.

Synergistic antibacterial activity of compact silver/magnetite core-shell nanoparticles core shell against Gram-negative foodborne pathogens.

The development of innovative antibacterial drugs against foodborne pathogens has led to an interest in novel materials such as nanomaterials. The unique features of nanomaterial qualify it for use as an antibacterial treatment. Noble metals and metal oxide nanoparticles, such as silver and magnetite nanoparticles, have been shown to be effective antibacterial medications against a range of microorganisms. In this work, Ag@Fe3O4 -NPs were fabricated by using a wet chemical reduction and modified co-precipitation techniques. The antibacterial efficiency of the Ag/Fe3O4 core shell nanoparticles was investigated by applying various techniques, such as the Kirby-Bauer Disk Diffusion test, minimum inhibitory concentration (MIC) and bactericidal concentration (MBC), Colony Forming Unit (CFU), and kill time assay. The toxicity mechanism of Ag@Fe3O4 -NPs against Salmonella typhimurium and Escherichia coli was studied by apoptosis and reactive oxygen species (ROS) assays. The data revealed that a cubic core was surrounded by a silver shell, which indicated the regular morphology of silver magnetite core shell nanoparticles without any aggregation. Furthermore, Ag@Fe3O4 -NPs is more toxic against S. typhimurium and E. coli than Ag-NPs and Fe3O4 NPs. The MIC values for Ag/Fe3O4 NPs against S. typhimurium and E. coli were 3.1 and 5.4 µg/ml, respectively, whereas the MIC values for Ag-NPs and MNPs against S. typhimurium and E. coli were 4.1 and 8.2 µg/ml for Ag-NPs and 6.9 and 10.3 µg/ml for MNPs. The results showed the ability of Ag@Fe3O4 -NPs to induce apoptosis by generating ROS. Also, the ability of Ag@Fe3O4 -NPs to liberate free Ag+ and generate ROS via the Haber-Weiss cycle may be a plausible mechanism to explain the toxicity of Ag@Fe3O4 -NPs - NPs.

Innovative Artificial-Intelligence- Based Approach for the Biodegradation of Feather Keratin by Bacillus paramycoides, and Cytotoxicity of the Resulting Amino Acids.

The current study reported a new keratinolytic bacterium, which was characterized as Bacillus paramycoides and identified by 16S rRNA, and the sequence was then deposited in the GenBank (MW876249). The bacterium was able to degrade the insoluble chicken feather keratin (CFK) into amino acids (AA) through the keratinase system. The statistical optimization of the biodegradation process into AA was performed based on the Plackett-Burman design and rotatable central composite design (RCCD) on a simple solid-state fermentation medium. The optimum conditions were temperature, 37°C, 0.547 mg KH2PO4, 1.438 mg NH4Cl, and 11.61 days of incubation. Innovatively, the degradation of the CFK process was modeled using the artificial neural network (ANN), which was better than RCCD in modeling the biodegradation process. Differentiation of the AA by high-performance liquid chromatography (HPLC) revealed the presence of 14 AA including essential and non-essential ones; proline and aspartic acids were the most dominant. The toxicity test of AA on the HepG2 cell line did not show any negative effect either on the cell line or on the morphological alteration. B. paramycoides ZW-5 is a new eco-friendly tool for CFK degradation that could be optimized by ANN. However, additional nutritional trials are encouraged on animal models.

Production and immobilization of ß-glucanase from Aspergillus niger with its applications in bioethanol production and biocontrol of phytopathogenic fungi.

ß-Glucanase has received great attention in recent years regarding their potential biotechnological applications and antifungal activities. Herein, the specific objectives of the present study were to purify, characterize and immobilize ß-glucanase from Aspergillus niger using covalent binding and cross linking techniques. The evaluation of ß-glucanase in hydrolysis of different lignocellulosic wastes with subsequent bioethanol production and its capability in biocontrol of pathogenic fungi was investigated. Upon nutritional bioprocessing, ß-glucanase production from A. niger EG-RE (MW390925.1) preferred ammonium nitrate and CMC as the best nitrogen and carbon sources, respectively. The soluble enzyme was purified by (NH4)2SO4, DEAE-Cellulose and Sephadex G200 with 10.33-fold and specific activity of 379.1 U/mg protein. Tyrosyl, sulfhydryl, tryptophanyl and arginyl were essential residues for enzyme catalysis. The purified ß-glucanase was immobilized on carrageenan and chitosan with appreciable yield. However, the cross-linked enzyme exhibited superior activity along with remarkable improved thermostability and operational stability. Remarkably, the application of the above biocatalyst proved to be a promising candidate in liberating the associate lignocellulosic reducing sugars, which was utilized for ethanol production by Saccharomyces cerevisiae. The purified ß-glucanase revealed an inhibitory effect on the growth of two tested phytopathogens Fusarium oxysporum and Penicillium digitatum.

Alleviation of Lead Stress on Sage Plant by 5-Aminolevulinic Acid (ALA).

Oxidative stress is imparted by a varying range of environmental factors involving heavy metal stress. Thus, the mechanisms of antioxidant resistance may advance a policy to improve metal tolerance. Lead as a toxic heavy metal negatively affects the metabolic activities and growth of medicinal and aromatic plants. This investigation aimed to assess the function of 5-aminolevulinic acid (ALA) in the alleviation of Pb stress in sage plants (Salvia officinalis L.) grown either hydroponically or in pots. Various concentrations of Pb (0, 100, 200, and 400 µM) and different concentrations of ALA (0, 10, and 20 mg L-1) were tested. This investigation showed that Pb altered the physiological parameters. Pb stress differentially reduced germination percentage and protein content compared to control plants. However, lead stress promoted malondialdehyde (MDA) and H2O2 contents in the treated plants. Also, lead stress enhanced the anti-oxidative enzyme activities; ascorbate peroxidase superoxide, dismutase, glutathione peroxidase, and glutathione reductase in Salvia plants. ALA application enhanced the germination percentage and protein content compared to their corresponding controls. Whereas, under ALA application MDA and H2O2 contents, as well as the activities of SOD, APX, GPX, and GR, were lowered. These findings suggest that ALA at the 20 mgL-1 level protects the Salvia plant from Pb stress. Therefore, the results recommend ALA application to alleviate Pb stress.

Emerging mutations in envelope protein of SARS-CoV-2 and their effect on thermodynamic properties.

Structural proteins of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are potential drug targets due to their role in the virus life cycle. The envelope (E) protein is one of the structural proteins; plays a critical role in virulency. However, the emergence of mutations oftenly leads to drug resistance and may also play a vital role in virus stabilization and evolution. In this study, we aimed to identify mutations in E proteins that affect the protein stability. About 0.3 million complete whole genome sequences were analyzed to screen mutations in E protein. All these mutations were subjected to stability prediction using the DynaMut server. The most common mutations that were detected at the C-terminal domain, Ser68Phe, Pro71Ser, and Leu73Phe, were examined through molecular dynamics (MD) simulations for a 100ns period. The sequence analysis shows the existence of 259 mutations in E protein. Interestingly, 16 of them were detected in the DFLV amino acid (aa) motif (aa72-aa75) that binds the host PALS1 protein. The results of root mean square deviation, fluctuations, radius of gyration, and free energy landscape show that Ser68Phe, Pro71Ser, and Leu73Phe are exhibiting a more stabilizing effect. However, a more comprehensive experimental study may be required to see the effect on virus pathogenicity. Potential antiviral drugs, and vaccines may be developed used after screening the genomic variations for better management of SARS-CoV-2 infections.

Emerging Mutations in Nsp1 of SARS-CoV-2 and Their Effect on the Structural Stability.

The genome of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) encodes 16 non-structural (Nsp) and 4 structural proteins. Among the Nsps, Nsp1 inhibits host gene expression and also evades the immune system. This protein has been proposed as a target for vaccine development and also for drug design. Owing to its important role, the current study aimed to identify mutations in Nsp1 and their effect on protein stability and flexibility. This is the first comprehensive study in which 295,000 complete genomes have been screened for mutations after alignment with the Wuhan-Hu-1 reference genome (Accession NC_045512), using the CoVsurver app. The sequences harbored 933 mutations in the entire coding region of Nsp1. The most frequently occurring mutation in the 180-amino-acid Nsp1 protein was R24C (n = 1122), followed by D75E (n = 890), D48G (n = 881), H110Y (n = 860), and D144A (n = 648). Among the 933 non-synonymous mutations, 529 exhibited a destabilizing effect. Similarly, a gain in flexibility was observed in 542 mutations. The majority of the most frequent mutations were detected in the loop regions. These findings imply that Nsp1 mutations might be useful to exploit SARS-CoV-2's pathogenicity. Genomic sequencing of SARS-CoV-2 on a regular basis will further assist in analyzing variations among the drug targets and to test the diagnostic accuracy. This wide range of mutations and their effect on Nsp1's stability may have some consequences for the host's innate immune response to SARS-CoV-2 infection and also for the vaccines' efficacy. Based on this mutational information, geographically strain-specific drugs, vaccines, and antibody combinations could be a useful strategy against SARS-CoV-2 infection.

Overexpression of Terpenoid Biosynthesis Genes From Garden Sage (Salvia officinalis) Modulates Rhizobia Interaction and Nodulation in Soybean.

In legumes, many endogenous and environmental factors affect root nodule formation through several key genes, and the regulation details of the nodulation signaling pathway are yet to be fully understood. This study investigated the potential roles of terpenoids and terpene biosynthesis genes on root nodule formation in Glycine max. We characterized six terpenoid synthesis genes from Salvia officinalis by overexpressing SoTPS6, SoNEOD, SoLINS, SoSABS, SoGPS, and SoCINS in soybean hairy roots and evaluating root growth and nodulation, and the expression of strigolactone (SL) biosynthesis and early nodulation genes. Interestingly, overexpression of some of the terpenoid and terpene genes increased nodule numbers, nodule and root fresh weight, and root length, while others inhibited these phenotypes. These results suggest the potential effects of terpenoids and terpene synthesis genes on soybean root growth and nodulation. This study provides novel insights into epistatic interactions between terpenoids, root development, and nodulation in soybean root biology and open new avenues for soybean research.