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Ouf, S. A., J. Almarashi, and A. - A. Mohamed, Characterization and Evaluation of Cold Atmospheric Plasma as Seedborne Fungal Disinfectant and Promoting Mediator for Physico-Chemical Characteristics of Moringa Oleifera Seedlings, : Research Square, 2022. Abstract

Large volume atmospheric pressure glow discharge plasma was developed and investigated to enhance the biological activities of M. oleifera seeds. The generated plasma was characterized optically, electrically, and spectroscopically. The large volume plasma size can reach 40 mm gape and 150 mm diameter. One hundred ns magnetic pulse compression high voltage system was used to generate the plasma where the current increases with increasing applied voltage at a constant frequency and discharge gap while the current decreases with increasing discharge gap at constant voltage and frequency. The plasma homogeneity was enhanced with higher applied voltage and frequency. Nitrogen second positive system bands dominated the plasma emission spectra, and its intensity increased with increasing applied voltage and frequency. There was a steady reduction in the total count of emerged seedborne fungi on the application of cold plasma with complete elimination of fungi at ≥ 13 A. The low doses of cold plasma (3.9 and 5.8 A) induced an increase in the seed germination, a significant increase in chlorophyll content (chl a and chl b ) and antioxidant activities of the seedlings emerged from soaked or wet seeds rather than dry seeds. There was a steady reduction in the total count of emerged fungi reaching complete decontamination of seedborne fungi at higher plasma doses (≥ 13 A). The plasma was harmful when applied at higher doses (≥ 13 A) and more harmful to the wet seeds compared to the dry ones, where the chlorophyll a content of the seedling developed from 1% H 2 O 2 soaked seeds.

Ouf, S. A., M. I. Ali, M. G. Haggag, D. O. Elsafty, and A. H. I. Faraag, "Enhancement of antidermatophytic activities of Citrullus colocynthis Schrad collected from different ecological habitats in Egypt using fluconazole", Phytomedicine Plus, vol. 2, no. 1, pp. 100178, 2022. AbstractWebsite

Background Dermatophytes may cause various and severe types of fungal infections in several communities. Herbal plant extracts may be inhibitory against cutaneous fungi related to Epidermophyton, Microsporum, and Trichophyton. The extract may be more efficient by its mixing with a low concentration of the antifungal agent. Purpose This study demonstrates the antidermatophytic activities of seed methanol extract of Citrullus colocynthis from different habitats in Egypt, namely Sinai desert, El-Wadi El-Gadid, and Wadi Qena. Methods A broth microdilution assay was used to determine the MIC and MFC of the extracts against the investigated fungal strains according to the Clinical and Laboratory Standards Institute (CLSI) M38-Ed3 guidelines. GC/MS was employed for the identification of the active constituents. Keratinase, lipase, phospholipase, and protease were measured spectrophotometrically or by determining the diameter of the reaction zone according to standard methods using a specific medium. Results C.colocynthis seed methanol extract from Sinai desert combined with 0.5 μg/ml of the antifungal drug fluconazole (FLU) was more effective than the extract or FLU, each on its own, in decreasing spore viability and reduction of fungal enzyme activities. The phytochemical analyses of the methanolic seed extract of the three cultivars show the presence of three common main compounds, namely, (9Z,12Z)-methyl octadeca-9,12-dienoate, (9Z,12E)-methyl octadeca-9,12-dienoate, and (9E,12E)-methyl octdeca-9,12-dienoate. The in-silico docking activity of the main compounds toward fungal enzymes showed a high binding affinity between the first compound and lipase enzymes of Microsporum canis and Trichophyton mentagrophytes. Conclusions The variation in the bioactivity of the plant extracts of a certain species may be varied due to the difference of the active constituents that are formed under specific conditions or stress. The inhibitory action of the extract to a certain pathogen could be improved when combined with lower drug concentration. Longer chain carbon compounds were frequently detected in the cultivar of Sinai compared to the compounds identified from the other localities suggesting that there was a variation in the active constituents according to the surveyed habitat although the main core of the constituents is maintained.

Zaki, S. A., S. A. Ouf, K. A. Abd-Elsalam, A. A. Asran, M. M. Hassan, A. Kalia, and F. M. Albarakaty, "Trichogenic Silver-Based Nanoparticles for Suppression of Fungi Involved in Damping-Off of Cotton Seedlings", Microorganisms, vol. 10, no. 2, 2022. AbstractWebsite

Mycogenic silver nanoparticles (AgNPs) produced by some biocontrol agents have shown the ability to inhibit the growth of numerous plant pathogenic fungi, which may be a unique method of disease management. This study describes the extracellular production of AgNPs by Trichoderma harzianum. The size, shape, charge, and composition of the AgNPs were subsequently studied by UV-visible spectroscopy, DLS, zeta potential, TEM, SEM, and EDX, among other methods. The AgNPs had sizes ranging from 6 to 15 nm. The antifungal activities of bio-synthesized AgNPs and two commercial fungicides (Moncut and Maxim XL) were tested against three soil-borne diseases (Fusarium fujikuroi, Rhizoctonia solani, and Macrophomina phaseolina). Cotton seedling illnesses were significantly reduced under greenhouse settings after significant in vitro antifungal activity was documented for the control of plant pathogenic fungi. The use of biocontrol agents such as T. harzianum, for example, may be a safe strategy for synthesizing AgNPs and using them to combat fungus in Egyptian cotton.

Ouf, S. A., A. M. F. Galal, H. S. Ibrahim, A. Z. Hassan, M. K. G. Mekhael, K. F. El-Yasergy, M. A. N. El-Ghany, M. A. Rizk, and A. G. Hanna, Phytochemical and antimicrobial investigation of the leaves of five Egyptian mango cultivars and evaluation of their essential oils as preservatives materials, , vol. 58, issue 8, pp. 3130 - 3142, 2021. AbstractWebsite

The sterols, hydrocarbons and fatty acids constituents of the leaves of five mango cultivars locally implanted in Egypt were identified. The effect of their essential oils (EOs) against food borne microorganisms was studied as preservative materials. The chemical constituents of the EOs isolated from mango leaves were identified by Gas Chromatography–Mass spectrometry (GC–MS) technique. Trans-caryophyllene, α–humulene and α–elemene were identified as terpene hydrocarbons, while 4-hydroxy-4-methyl-2-pentanone as oxygenated compounds were recorded in all tested cultivars with variable amounts. Results showed that Staphylococcus aureus and Escherichia coli were the most sensitive microorganisms tested for Alphonso EOs. On the other hand, Salmonella typhimrium was found to be less susceptible to the EOs of the studied cultivars. The EOs of different mango cultivars induced a steady decrease in the activity of amylase, protease and lipase at the minimum inhibitory concentration (MIC). The treatment of the tested bacteria with the EOs of mango cultivars caused a steady loss in enterotoxins even when applied at the sub-MIC. Bacteria-inoculated apple juice treated with minimum bactericidal concentration of Alphonso oil was free from the bacteria after 5 days of incubation at 25 °C. Eighteeen volatile compounds were found to reduce the activity of the amylase enzyme and the most active was cedrelanol (−7.6 kcal mol−1) followed by alpha-eudesmol (−7.3 kcal mol−1) and humulene oxide (−7 kcal mol−1). The binding mode of both of cedrelanol and alpha-eudesmol with amylase enzyme was illustrated.

Alhilal, M., Y. A. M. Sulaiman, S. Alhilal, S. M. Gomha, and S. A. Ouf, "Synthesis of Novel Acyclic Nucleoside Analogue Starting From 6-Aminouracil as Potent Antimicrobial Agent", Polycyclic Aromatic CompoundsPolycyclic Aromatic Compounds: Taylor & Francis, pp. 1 - 12, 2021. AbstractWebsite
Al-Surhanee, A. A., M. Afzal, N. A. Bouqellah, S. A. Ouf, S. Muhammad, M. Jan, S. Kaleem, M. Hashem, S. Alamri, A. A. H. A. Latef, et al., "The Antifungal Activity of Ag/CHI NPs against Rhizoctonia solani Linked with Tomato Plant Health", Plants, vol. 10, no. 11, 2021. AbstractWebsite

Pathogenic infestations are significant threats to vegetable yield, and have become an urgent problem to be solved. Rhizoctonia solani is one of the worst fungi affecting tomato crops, reducing yield in some regions. It is a known fact that plants have their own defense against such infestations; however, it is unclear whether any exogenous material can help plants against infestation. Therefore, we performed greenhouse experiments to evaluate the impacts of R. solani on 15- and 30-day old tomato plants after fungal infestation, and estimated the antifungal activity of nanoparticles (NPs) against the pathogen. We observed severe pathogenic impacts on the above-ground tissues of tomato plants which would affect plant physiology and crop production. Pathogenic infection reduced total chlorophyll and anthocyanin contents, which subsequently disturbed plant physiology. Further, total phenolic contents (TPC), total flavonoid contents (TFC), and malondialdehyde (MDA) contents were significantly increased in pathogen treatments. Constitutively, enhanced activities were estimated for catalase (CAT), superoxide dismutase (SOD), and ascorbate peroxidase (APX) in response to reactive oxygen species (ROS)in pathogen-treated plants. Moreover, pathogenesis-related genes, namely, chitinase, plant glutathione S-transferase (GST), phenylalanine ammonia-lyase (PAL1), pathogenesis-related protein (PR12), and pathogenesis-related protein (PR1) were evaluated, with significant differences between treated and control plants. In vitro and greenhouse antifungal activity of silver nanoparticles (Ag NPs), chitosan nanoparticles, and Ag NPs/CHI NPs composites and plant health was studied using transmission electron microscopy (TEM) and Fourier transform infrared (FTIR) spectrophotometry. We found astonishing results, namely, that Ag and CHI have antifungal activities against R. solani. Overall, plant health was much improved following treatment with Ag NPs/CHI NPs composites. In order to manage R. solani pathogenicity and improve tomato health, Ag/CHI NPs composites could be used infield as well as on commercial levels based on recommendations. However, there is an urgent need to first evaluate whether these NP composites have any secondary impacts on human health or the environment.

Alhilal, M., Y. A. M. Sulaiman, S. Alhilal, S. M. Gomha, and S. A. Ouf, "Antifungal Activity of New Diterpenoid Alkaloids Isolated by Different Chromatographic Methods from Delphinium peregrinum L. var. eriocarpum Boiss", Molecules, vol. 26, no. 5, 2021. AbstractWebsite

This paper aimed to investigate the potential antifungal influences of new alkaloids from Delphinium peregrinum L. var. eriocarpum Boiss. New Diterpenoid alkaloids Delcarpum (1), Hydrodavisine (4) and known alkaloids Peregrine (2), Delphitisine (3) were isolated by different chromatographic methods from the aerial parts of D. Peregrinum eriocarpum Boiss, which grows in Syria. The structures of alkaloids were proposed based on 1D NMR spectroscopy 1H-NMR, 13C-NMR, DEPT-135, DEPT-90, 2D NMR spectroscopy DQF-COSY, HMQC, EI-Ms mass spectrum, and IR spectroscopic measurements. The antifungal activity of the isolated alkaloids was evaluated against different dermatophyte fungal isolates compared with fluconazole. In the case of Peregrine (2) the minimum inhibitory concentrations(MICs) recorded 128–256, 32–64, and 32 for Epidermophyton floccosum, Microsporum canis, and Trichophyton rubrum, respectively, compared to 32–64, 16, and 32 μg/mL in the case of fluconazole, respectively. The MICs recorded on application of the four alkaloids mixture were 64, 32, and 16 in the case of E. floccosum, M. canis, and T. rubrum, respectively, which were significantly lower than that measured for each of the individual alkaloid and were compatible for fluconazole. In conclusion, MICs of the tested alkaloids showed a variable potential effect on the investigated fungal isolates. Peregrine (2) was the most effective alkaloid, however, the application of the mixture of alkaloids induced significant synergistic activity that was more pronounced than the application of individual ones.

Al-Surhanee, A. A., M. H. Soliman, and S. A. Ouf, "Chapter 10 - The role of soil microbes in the plant adaptation to stresses: current scenario and future perspective", Frontiers in Plant-Soil Interaction: Academic Press, pp. 237-258, 2021. Abstract

Environmental stresses hamper the growth and development of plants drastically, limiting their yield to considerable extent. Stresses alter normal metabolism and induce generation of toxic effects which ultimately downregulate growth and productivity. Environmental stresses hamper photosynthesis, mineral uptake and assimilation, and damage major cellular structures and their functioning. Soil microbes include a range of organisms that establish association with the plants and among the beneficial and more explored ones are arbuscular mycorrhizal fungi (AMF) and plant growth promoting rhizobacteria (PGPR). Both AMF and PGPR are believed to enhance plant growth by mediating several benefits to hosts. Mineral solubilization and availability, water content maintenance, modulation of root morphology and physiology, regulation of enzyme activity and sequestration of toxic ions, metals etc. are some of the key functions attributed to these beneficial microbes. The positive role of these microbes have been confirmed through molecular studies also. Their involvement in strengthening the tolerance mechanisms through upregulation of the antioxidant defense pathway, increased osmolyte accumulation, etc. have been well established. This chapter discusses the role of beneficial microbes, mainly AMF and PGPR, in improving the stress tolerance in plants focusing the relevant reports.

Ouf, S. A., and E. M. Ali, "Does the treatment of dried herbs with ozone as a fungal decontaminating agent affect the active constituents?", Environmental Pollution, vol. 277, pp. 116715, 2021. AbstractWebsite

Herbs and spices are food crops susceptible to contamination by toxigenic fungi. Ozone, as a decontamination approach in the industry, has attractive benefits over traditional food preservation practices. A contribution to the studying of ozone as an antifungal and anti-mycotoxigenic agent in herbs and spices storage processes is achieved in this research. Nine powdered sun-dried herbs and spices were analyzed for their fungal contamination. The results indicate that licorice root and peppermint leaves were found to have the highest population of fungi while black cumin and fennel record the lowest population. The most dominant fungal genera are Aspergillus, Penicillium, Fusarium, and Rhizopus. Ozone treatment was performed at a concentration of 3 ppm applied for exposure times of 0, 30, 90, 150, 210, and 280 min. After 280 min of exposure to ozone, the reduction of fungal count ranged from 96.39 to 98.26%. The maximum reduction in spore production was achieved in the case of A. humicola and Trichderma viride exposed for 210 min ozone gas. There was a remarkable reduction in the production of the total mycotoxin, reaching 24.15% in aflatoxins for the 150 min-treated inoculum in the case of A. flavus. The total volume of essential oil of chamomile and peppermint was reduced by 57.14 and 26.67%, respectively, when exposed to 3 ppm. For 280 min. In conclusion, fumigation with ozone gas can be used as a suitable method for achieving sanitation and decreasing microbial load in herbs and spices. Still, it is crucial to provide precautions on ozone’s effect on major active constituents before recommending this method for industrial application.

Mohamed, A. - A. H., A. H. Basher, J. Q. M. Almarashi, and S. A. Ouf, "Susceptibility of Staphylococcus epidermidis to Argon Cold Plasma Jet by Oxygen Admixture", Applied Sciences, vol. 11, no. 8, 2021. AbstractWebsite

Cold atmospheric pressure sterilization is one of the nominated and efficient techniques to prevent the spread of diseases. Reactive species such as O and OH and other radicals play a major role in the mechanism of plasma sterilization. Therefore, in this work, oxygen was mixed with different parentage from (0.2 to 1.2%) to argon to enhance the generation of the reactive species and increase the argon atmospheric pressure plasma sterilization efficacy. The emission spectra from the jet increase the radicle line intensities by increasing the percentage admixture of O2 with the argon gas to reach a maximum power at 0.8; then, it gradually decreases with a higher O2 percentage. The OH band intensity decreases with increasing the admixture of O2. The jet with different O2 percentages was tested against Gram-positive S. epidermidis, which is the causal agent of nosocomial infections. The maximum reduction in colony-forming units (CFU) was observed at 0.2% O2. No bacterial growth was observed at the later concentration applied for 8 min and the same case was detected at 0.4% O2 applied to 16 min.

Zaki, S. A., S. A. Ouf, F. M. Albarakaty, M. M. Habeb, A. A. Aly, and K. A. Abd-Elsalam, "Trichoderma harzianum-Mediated ZnO Nanoparticles: A Green Tool for Controlling Soil-Borne Pathogens in Cotton", Journal of Fungi, vol. 7, no. 11, 2021. AbstractWebsite

ZnO-based nanomaterials have high antifungal effects, such as inhibition of growth and reproduction of some pathogenic fungi, such as Fusarium sp., Rhizoctonia solani and Macrophomina phaseolina. Therefore, we report the extracellular synthesis of ZnONPs using a potential fungal antagonist (Trichoderma harzianum). ZnONPs were then characterized for their size, shape, charge and composition by visual analysis, UV–visible spectrometry, X-ray diffraction (XRD), Zeta potential, transmission electron microscopy (TEM), scanning electron microscopy (SEM) and energy-dispersive X-ray analysis (EDX). The TEM test confirmed that the size of the produced ZnONPs was 8–23 nm. The green synthesized ZnONPs were characterized by Fourier transform infrared spectroscopy (FTIR) studies to reveal the functional group attributed to the formation of ZnONPs. For the first time, trichogenic ZnONPs were shown to have fungicidal action against three soil–cotton pathogenic fungi in the laboratory and greenhouse. An antifungal examination was used to evaluate the bioactivity of the mycogenic ZnONPs in addition to two chemical fungicides (Moncut and Maxim XL) against three soil-borne pathogens, including Fusarium sp., Rhizoctonia solani and Macrophomina phaseolina. The findings of this study show a novel fungicidal activity in in vitro assay for complete inhibition of fungal growth of tested plant pathogenic fungi, as well as a considerable reduction in cotton seedling disease symptoms under greenhouse conditions. The formulation of a trichogenic ZnONPs form was found to increase its antifungal effect significantly. Finally, the utilization of biocontrol agents, such as T. harzianum, could be a safe strategy for the synthesis of a medium-scale of ZnONPs and employ it for fungal disease control in cotton.

Ouf, S. A., S. M. Gomha, M. Eweis, A. S. Ouf, I. A. A. Sharawy, and S. A. Alharbi, Antidermatophytic activity of some newly synthesized arylhydrazonothiazoles conjugated with monoclonal antibody, , vol. 10, issue 1, pp. 20863, 2020. AbstractWebsite

A new series of 5-arylhydrazonothiazole derivatives 5a–d has been synthesized, elucidated, and evaluated for their antidermatophytic activity. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of the newly synthesized products were investigated against 18 dermatophyte fungal isolates related to Epidermophyton floccosum, Microsporum canis, and Trichophyton rubrum. The morphological alterations induced by the synthesized derivatives singly or conjugated with the monoclonal antibody were examined on spores of T. rubrum using a scanning electron microscope. The efficacy of synthesized derivative 5a applied at its respective MFC alone or conjugated with anti-dermatophyte monoclonal antibody 0014 in skin infection treatment of guinea pigs due to inoculation with one of the examined dermatophytes, in comparison with fluconazole as standard reference drug was evaluated. In an in vivo experiment, the efficiency of 5a derivative conjugated with the antibody induced 100% healing after 45 days in the case of T. rubrum and M. canis-infected guinea pigs.

Abd-El-Raouf, R., S. A. Ouf, M. M. Gabr, M. M. Zakaria, K. F. El-Yasergy, and B. Ali-El-Dein, Escherichia coli foster bladder cancer cell line progression via epithelial mesenchymal transition, stemness and metabolic reprogramming, , vol. 10, issue 1, pp. 18024, 2020. AbstractWebsite

Bacteria is recognized as opportunistic tumor inhabitant, giving rise to an environmental stress that may alter tumor microenvironment, which directs cancer behavior. In vitro infection of the T24 cell line with E. coli was performed to study the bacterial impact on bladder cancer cells. EMT markers were assessed using immunohistochemistry, western blot and RT-PCR. Stemness characteristics were monitored using RT-PCR. Furthermore, the metabolic reprograming was investigated by detection of ROS and metabolic markers. A significant (p ≤ 0.001) upregulation of vimentin as well as downregulation of CK19 transcription and protein levels was reported. A significant increase (p ≤ 0.001) in the expression level of stemness markers (CD44, NANOG, SOX2 and OCT4) was reported. ROS level was elevated, that led to a significant increase (p ≤ 0.001) in UCP2. This enhanced a significant increase (p ≤ 0.001) in PDK1 to significantly downregulate PDH (p ≤ 0.001) in order to block oxidative phosphorylation in favor of glycolysis. This resulted in a significant decrease (p ≤ 0.001) of AMPK, and a significant elevation (p ≤ 0.001) of MCT1 to export the produced lactate to extracellular matrix. Thus, bacteria may induce alteration to the heterogonous tumor cell population through EMT, CSCs and metabolic reprogramming, which may improve cancer cell ability to migrate and self-renew.

Elaziz, D. A., M. Abdel-Ghany, S. Meshaal, R. E. Hawary, S. Lotfy, N. Galal, S. A. Ouf, and A. ELmarsafy, "Fungal infections in primary immunodeficiency diseases", Clinical Immunology, vol. 219, pp. 108553, 2020. AbstractWebsite

Primary immunodeficiency diseases (PID), encompass a heterogeneous group of diseases, with increased susceptibility to recurrent, severe infections. Invasive fungal infections raise a serious concern related to their morbidity and mortality. Herein, we describe various fungal infections among different PID patients. Twenty-eight PID patients diagnosed with fungal infections were included; fourteen patients with chronic granulomatous disease, two with Hyper Immunoglobulin E syndrome, one with LRBA deficiency and one with MHC class II defect, one with unclassified immune dysregulation, one with CD4 lymphopenia and one patient with Immune dysregulation Polyendocrinopathy Enteropathy X-linked syndrome. Aspergillus species were the most common isolated causative organisms in 78% of patients, Candida species were the causative organisms in 32%, Pneumocystis jirovecii caused infections in 7% followed by Malassezia furfur, Fusarium spp., Mucormycosis, and Penicillium chrysogenium 3.5% for each. The mortality rate among our patients was 10/28 (35.7%). PID patients are at high risk of developing fungal infections.

Ouf, S. A., K. F. El-Yasergy, H. A. Mohammed, A. El-Ghany, and N. Mohamed, "Efficacy of ozonized water for fungal decontamination of fresh fruit pieces decorating dessert cakes", Egyptian Journal of Botany, vol. 59, no. 3: National Information and Documentation Center (NIDOC), Academy of Scientific …, pp. 845–855, 2019. Abstract
Alananbeh, K. M., N. A. BouQellah, M. R. Al Harbi, and S. A. Ouf, "The efficacy of photosensitizers on mycelium growth, mycotoxin and enzyme activity of Alternaria spp.", Jordan Journal of Biological Sciences, vol. 11, no. 5, 2018. Abstract