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Fifteen azo dyes were subjected to microbial degradation by four fungal strains and their mixtures as well as five bacterial strains and their mixtures. The biodegradation efficiency was determined by LC/MS/MS analysis. The most active bacterial strain was B. subtilis, where it showed the highest biodegradation capacity (71.8% to 100%) of eight azo dyes. Bacterial strain B. brevis came next to it. However, the consortium of the five bacterial strains gave lower degradation percentages. On the other hand, the fungal strains and their consortium were more potent in biodegradation of all tested azo dyes, where ten azo dyes were completely (100%) degraded by the consortium. Two of widely used azo dyes, direct violet and methyl red, were further studied in relation to the intermediate biodegradation products by each of the tested fungi and bacteria as well as the bacterial and fungal consortia using GC/MS/MS. The major biodegradation product of methyl red was 2-amino benzoic acid by all of the tested bacterial strains and A. niger, while the major biodegradation products of direct violet by both bacteria and fungi were ethanol, 2(2-butyxyethoxy), followed by 4- methyl benzoic acid and phenol, 2,4-bis (1,1-dimethyl ethyl). The results of this study suggest the successful use of the four fungal consortium for the biodegradation of the azo dyes. These findings are important to design bioremediation technology for treating the azo dye residues.

Wnt5a is a representative protein ligand that activates a beta catenin independent pathway in Wnt signaling in cancer progression and spread in the palmitoylated form. So that its synthesis is augmented through promotion of its gene transcription in the malignant cells. Cisplatin is an important chemotherapeutic drug used in treatment of different types of advanced cancer. However, its usage is limited due to severe dose-limiting side effects, due to the lack of targeting therapy of cisplatin. On the other hand, the cancer cells developed resistance from cisplatin that is solved partially by C-glycosylation of cisplatin. Through this review article, we strongly suggest that the Wnt5a protein can be used as drug delivery molecule for cisplatin to target the malignant cells. This can be performed through the following steps: Firstly: use of Wnt5a in the depalmitoylated form to avoid its effect on cancer progression with improvement of its binding efficacy by two suggested modifications: 1) Glycosylation with sialic acid-containing glycoside to increase the binding affinity with the sialic acid on the surface of the malignant cells; 2) GPI anchoring modification: This can be performed through the Serine 282 in the Wnt5a molecule. Secondly, binding of the C-glycosylated cisplatin molecule with the modified Wnt5a molecule through the C-glycosylation bond. Since one Wnt5a molecule contains four asparagine molecules, it can bind with four molecules of C-glycosylated cisplatin molecules, hence higher concentration of cisplatin will attack one malignant cell.

Many cancer cells in solid tumors are hypoxic or pseudohypoxic and create acidic environment for malignancy progression. Under low oxygen conditions (hypoxia), hypoxia-inducible factors (HIFs) play pathological roles in cancer cell survival and spreading. HIF regulates several genes such as genes of glucose transporters that enhance anaerobic glycolysis, angiogenesis, erythropoiesis and carbonic anhydrase IX (CA-IX). CA-IX is a cell-surface glycoprotein that catalyzes the hydration of CO2 to protons and bicarbonate ions (respiratory acidification). This process is involved in adaptation to acidosis and implicated in cancer progression. Therefore, CA-IX inhibitors (such as sulfonamide-based compounds) showed hoping results in reduction malignancy progression. The article aims to reversal the malignant hypoxic environment in solid tumors to create a condition of weakness within the cancer for further focused cisplatin potency. This article suggests the use of modified synthesized HIF as a drug delivery molecule for both carbonic anhydrase IX inhibitor and glycosylated cisplatin that damages the DNA of malignant cell. HIF molecule has high affinity to bind with CA IX-expressing malignant cells, which is followed by cell entrance via endocytosis. Once the HIF-Cisplatin-CA-inhibitor complex enters the cell, the carbonic anhydrase inhibitor will improve the cellular pH that makes the environment unsuitable for HIF 1α function and it may be ubiquitinated. So, the raise in target genes transcription will be arrested. On the other hand, once the synthetized HIF is degraded, the cisplatin molecules will be released inside the malignant cell and start to damage its DNA. This approach may be a good solution for many solid tumors.

The increasing demand on new antimicrobial compounds is very important nowadays because the microbes become more resistant to the old antimicrobial compounds. Hence, this study was designed to isolate and identify antimicrobial compounds from Celery seeds (Apium graveolens). The seeds were extracted by hexane, methylene chloride and methanol (sequentially). Purification was achieved by column chromatography. 1-phenyl-1-hepten-3-ol was isolated from hexane extract and the chemical structure was identified by spectroscopic studies (IR, proton NMR and GC-MS). The hexane extract showed the highest value as antimicrobial activity against Bacillus cereus, Salmonella sp., Staphylococcus sp. and Penicillium sp, Fusarium oxysporum, Alterneria solani, and Aspergllus niger. However, the extracts (hexane, methylene chloride and methanol) didn’t show any inhibition zone against Pseudomonas sp .

Candida albicans is a common human yeast that infect several epithelial tissues including vagina. The increase of drug-resisting C. albicans encouraged the researchers to find alternative treatment. Honey medical signatures such as bactericidal, antifungal and anti-candida made it a possible candidate for disease treatment. In addition, rose essential oil possesses a wide range of biochemical activities in folkloric medicine including anti-microbial activities. The present research utilizes honey alone or in conjunction with Taifi rose (Rosa damascena) oil as anti-candida agent to treat vaginal candidiasis. Three local monof oral honeys from different flower sources and/or geographic origins were tested with four concentrations (50, 80 and 95%), while two concentrations of the Taifi rose oil (1 and 2%). anti-candida activity of honey alone or in conjunction with Taifi rose oil was determined as well as phenolic and flavonoids contents were determined. Also, GC-MS analysis of volatile oils and alkaloids were evaluated. The results of this study indicated that acidity is within the allowed range for commercialization and long-lasting storage. All honeys tested inhibited completely the C. albicans growth at concentrations 80% and 95% either incubation after 48 or 72 h. Also, only Markh and Manuka honeys were completely inhibited C. albicans growth at 50% concentration. Also, C. albicans growth inhibited completely at 2% Taifi rose oil after the incubation periods of 48 and 72 h. The phenolic compounds and flavonoids were analysed by mass spectrometry analysis which revealed the Markh honey showed the presence of gallic acid and quercetin that proved to have antifungal activity. It could be concluded that mixed Markh honey and Taifi rose oil treatment was capable to inhibit C. albicans growth completely. Further research is required to determine the anti-candida activity of the mixture of Markh honey and Taifi rose oil in the human body as a new therapeutic drug to treat vaginal candidiasis.

Most scientific studies on Calotropis procera refer to the plant as an important source of pharmaceutical compounds and its valuable benefits in medicine. One of the most important substances in this plant is the potential immunostimulant β-sitosterol (BS) that acts in improving human health. This study focused on the effects of lighting before and after irrigation on the BS accumulation pathway namely steroid biosynthesis. Studying the enzymes in BS biosynthetic pathway indicated the upregulation at dawn and predusk of the SMT2 and SMO2 genes encoding sterol methyltransferase 2 and methylsterol monooxygenase, two key enzymes in BS accumulation in C. procera. The results almost indicated no regulation at the different time points of the CYP710A gene encoding sterol 22-desaturase, an enzyme that acts in depleting β-sitosterol towards the biosynthesis of stigmasterol. RNA-Seq data was validated via quantitative RT-PCR and results were positive. The data of ultra-performance liquid chromatography-tandem mass spectrometry analysis with regard to BS accumulation also aligned with those of RNA-Seq analysis. We focused on the effects of light before and after watering on BS accumulation in C. procera. Our results show that BS accumulation is high at dawn in both dehydrated and well-watered condition. While, the BS was dramatically decrease at midday in well-watered plants. This increase/decrease in BS content is correlated with rates of expression of SMT 2 gene. This gene is a key convertor between the different branches in the cardiac glycoside biosynthesis. Accordingly, it could be suggested that BS (or one of the descendent product) may play an important role in C. procera tolerance to drought/light intensity conditions.

Fusarium oxysporum f. sp. lycopersici attacks tomato plants and causes wilt disease. Fusarium Pathogenicity is including pectinases enzymes which enable the Fungal penetration into host cell wall. The present study is focused on using Computational tools such as Auto- Dock program for screening of inhibitors of endo and exopolygalacturonase enzymes.It based on Lamarckian Genetic Algorithm (LGA) that estimate the binding energy and inhibition constant as parameters to select the best binding.The binding energy and amino acids interactions for the selected inhibitors were compared with that of the enzyme substrate (polygalacturonic acid) Allium species such as onion plant have been used widely as antimicrobial and antifungal plants.It contains (up to 5% ) of non-protein sulfur amino-acids, including S-E-Prop-1-enyl-L-cysteine S-oxide, S-3-Allylsulphinyl-L-alanine and S-Methylcysteine sulfoxide have satisfactory binding interactions and inhibition constant with endo and exopolygalacturonase. In the present study, these compounds were extracted from white onion bulb Giza 20 and detected in the onion extract LC/MS analysis. The Inhibitory effect of these compounds for exopolygalacturonase enzyme was confirmed experimentally by determination of the enzyme activity in the presence and the absence of these compounds. White onion extract 45% inhibition percentage of the exopolygalacturonase activity.The enzyme kinetic study showed increase in the Km value with stable V-max value in presence of 7µg/µL of the onion extract.Also, In-vitro experiment of inhibition of F. oxysporum growth in presence 20% and 40% of onion extract showed inhibition percentages of 47% and 53% respectively The results concluded that onion extract inhibits Fusarium growth through inhibition of exo and endo polygalacturonase. The inhibitory effect of onion extract could be due to its contents of S-E-Prop-1-enyl-L-cysteine S-oxide, S-3-Allylsulphinyl-L-alanine and S-Methylcysteinesulphoxide, these compounds have excellent binding interactions and inhibition effects on both exo- and endo-polygalacturonases enzymes of Fusarium oxysporum f. sp. lycopersici Keywords: Chemoinformatic, Auto- Dock program,Fusarium oxysporum, Pectinases, exopolygalacturonase; Onion extract.

Wastewater from textile industries contains azo dye residues that negatively affect most environmental systems. The biological treatment of these wastes is the best option due to safety and cost concerns. Here we isolated and identified 19 azo dye-degrading fungi and optimized conditions resulting in enhanced degradation. The fungi belonged to five species of Aspergillus and a single Lichtheimia sp. All fungi were evaluated for their ability to decolorize 20 azo dyes. While the most easily transformable azo dye was direct violet (decolorization ranged from 71.1 to 93.3%), the most resistant to decolorization was fast green azo dye. The greatest degradation potential of azo dyes (direct violet and methyl red) was optimized using the most promising four fungal strains and changing media glucose concentration, nitrogen source, and micronutrients. Biomass, lignin peroxidase, and laccases production were also determined in the optimization studies. The decolorization of both azo dyes by the four fungal strains was greatly enhanced by glucose supplementation. The fungal strains were not able to produce lignin peroxidases in the absence of organic nitrogen source. Both yeast extract and casamino acid supplementation enhanced decolorization of direct violet and methyl red dyes and production of lignin peroxidase by the fungal strains. In contrast, the laccases were absent in the similar medium enriched with the same organic nitrogen sources.

Lipase producing fungus was isolated and identified as a strain of Fusarium solani based on its 18s rDNA sequencet. The enzyme it produces was purified by diethyl amino ethyl sephadex (DEAE-sephadex) column chromatography. The specific activity of the pure enzyme was 1.98 U/mg protein. The kinetics study showed that Km and Vmax values were 0.63µM and 29.4µM/min/mg protein, respectively. The MW was 95.27 kDa. Effects of pH, incubation temperature and organic solvents on the lipase activity were studied. The maximum enzyme activity was obtained at pH 8.5 and incubation temperature 35°C. Hexane and butanol inhibited enzyme activity by 51% and 72.6 %, respectively, while DMSO stimulated the activity by 47.8%. The lipase was immobilized by fusion to chitosan-coated iron oxide magnetic nanoparticles and cross-linked by glutaraldehyde. The reusability and storage period of the immobilized enzyme showed that the enzyme retained 80% of its activity after 15 reuse cycles and retained 97% of activity after 30 days of storage at 4°C. The immobilized lipase was tested for synthesis of sugars-oleate esters and the ester products were analyzed by liquid chromatography tandem-mass spectrometry (LC/MS/MS). This investigation identified the potential for use of the obtained F. solani lipase in industrial applications that utilize organic solvents or alkaline pH values, such as detergent industry.

Silymarin, a Silybum marianum seed extract containing a mixture of flavonolignans including silybin, is being used as an antihepatotoxic therapy for liver diseases. In this study, the enhancing effect of gamma irradiation on plant growth parameters of S. marianum under salt stress was investigated. The effect of gamma irradiation, either as a single elicitor or coupled with salinity, on chalcone synthase (CHS) gene expression and silybin A + B yield was also evaluated. The silybin A + B content in S. marianum fruits was estimated by liquid chromatography-mass spectrometry (LC-MS/MS). An increase in silybin content was accompanied by up-regulation of the CHS1, CHS2 and CHS3 genes, which are involved in the silybin biosynthetic pathway. The highest silybin A + B production (0.77 g/100 g plant DW) and transcript levels of the three studied genes (100.2-, 91.9-, and 24.3-fold increase, respectively) were obtained with 100GY gamma irradiation and 4000 ppm salty water. The CHS2 and CHS3 genes were partially sequenced and submitted to the NCBI database under the accession numbers KT252908.1 and KT252909.1, respectively. Developing new approaches to stimulate silybin biosynthetic pathways could be a useful tool to potentiate the use of plants as renewable resources of medicinal compounds.

The fungus Lichtheimia corymbifera was isolated on mineral salt medium (MSM) supplemented with direct green azo dye as a sole carbon source and identified by sequencing the ITS1-ITS4 gene region. This fungus showed lignin peroxidase activity when grown in liquid MSM supplemented with methyl red azo dye as a sole carbon source. The lignin peroxidase was isolated by ammonium sulfate precipitation, and purified by DEAE-SephadexA50 column chromatography and gradient SDS-PAGE separation. The enzyme bands on gels were identified by reaction with pyrogallol and hydrogen peroxide, picked, and hydrolyzed by tryps in followed by amino acids sequencing by LC/MS/MS. Two different peroxidases were identified by homology to entries in the PEAKS database and Uni Prot repository. The first (accession number A0A068RV79) was comprised of 360 AA residues with MW 40.16KD. The second (accession number A0A068S1N8) was comprised of 271 AA with a MW 30.91KD. A kinetic study of the most abundant peroxidase (MW 30.91 KD) showed a maximum enzyme activity (Vmax) (10.23 mg/ml/20 s) and the lowest Km value (0.33 μM hydrogen peroxide) at 0.8 μM of hydrogen peroxide and 32 μM of pyrogallol. The kinetics of this enzyme was reported for the first time in this work; furthermore, no previous articles reported the ability of L. corymbifera to degrade the azo dyes.

Thymoquinone (TQ) is a major constituent of Nigella sativa oil with reported anti-oxidative activity and anti-inflammatory activity in animal cells. It also inhibits proliferation and induces programmed cell death (apoptosis) in human skin cancer cells. The present study sought to detect the influence of TQ on dividing cells of three plant systems and on expression of Bcl2-associated athanogene-like (BAG -like) genes that might be involved during the process of cell death. BAG genes are known for the regulation of diverse physiological processes in animals, including apoptosis, tumorigenesis, stress responses, and cell division. Synthetic TQ at 0.1 mg/mL greatly reduced wheat seed germination rate, whereas 0.2 mg/mL completely inhibited germination. An Evans blue assay revealed moderate cell death in the meristematic zone of Glycine max roots after 1 h of TQ treatment (0.2 mg/mL), with severe cell death occurring in this zone after 2 h of treatment. Light microscopy of TQ-treated (0.2 mg/mL) onion hairy root tips for 1 h revealed anti-mitotic activity and also cell death-associated changes, including nuclear membrane disruption and nuclear fragmentation. Transmission electron microscopy of TQ-treated cells (0.2 mg/mL) for 1 h revealed shrinkage of the plasma membrane , leakage of cell lysate, degradation of cell walls, enlargement of vacuoles and condensation of nuclei. Expression of one BAG-like gene, previously associated with cell death, was induced 20 min after TQ treatment in Glycine max root tip cells. Thus, TQ has multiple effects, including cell death, on dividing plant cells and plants may serve as a useful system to further investigate the mechanisms underlying the response of eukaryotic cells to TQ.

Disposal of the automotive lubricating oil after usage can cause environmental hazards. This study aimed to isolate a microbial isolate is that able to biodegrade lubricating oil effectively. A new yeast strain, identified by 18S-rRNA gene sequencing as Candida viswanathii KA-201l, was isolated from used lubricating oil, showed high biodegradation efficiency for different used lubricating oils. Capability of this isolate to degrade different high and low molecular weight hydrocarbons, castor oil, diesel oil and grease was tested. It showed high degradation efficiency for most of the tested compounds. The biodegradation products were analyzed by gas chromatography-mass spectroscopy (GC-MS) in most cases. The biodegradation efficiency under high osmotic pressure was studied. It effectively biodegrade lubricating and diesel oils (58.6 and 93.9%, respectively) at 6% salt concentration after four days only. C. viswanathii KA-2011 can be effectively used for removal of lubricants, diesel or vegetative oils pollution from soil, wastewater and sea water. Use of C. viswanathii KA-2011 in the bioremediation of lubricant or gasoline contaminated sea-water save the aquaculture from these pollutants, as well as it open new horizons in using of contaminated soil and wastewater in agriculture. Keywords: Lubricating oils, diesel oil, biodegradation, Candida viswanathii

Thirty actinomycete isolates were isolated from soil and tested against Candida albicans in vitro. The active isolate was identified by 16s-rRNA gene sequencing method as Streptomyces toxytricini. The antifungal compound was extracted with ethyl acetate followed by diethyl ether. Both HPLC and GC–MS analysis confirmed presence of one pure compound in the diethyl ether extract. The compound is a yellow liquid has a maximum absorbance at 240 nm in methanol. The chemical structure was elucidated by 1D and 2D-NMR and IR analyses. The elucidated molecular formula was C36H54O14. The compound is a polyacetal tricyclononane derivative, composed of a tricyclononane ring attached from the carbon atom number four with an oligo-acetal chain (six acetal groups in chain) and from the carbon atom number seven with a methoxy carbonyl benzene-1,3-dicarboxylic acid. The purposed name is: 4- {[tricycle(3.2.1.11,3)non-8-yl] methoxy carbonyl benzene-1,3-dicarboxylic acid} (2,4,5,6,7,8,9 heptaoxa, 3-ethoxy, 5,6,7,9-tetramethyl unidecane).

Several human pathogenic fungi can acquire resistance against the available antifungal compounds or need prolonged time of therapy, which may cause toxicity. Thymol, thymoquinone (TQ) and thymohydroquinone (THQ) are principle constituents of the essential oil of Nigella sativa seeds. TQ and THQ were organically synthesized from thymol in our laboratory. The chemical structure and purity were confirmed by using GC-MS analysis. The three compounds were primary tested at concentration of 1mg/ ml for their preliminary antifungal effect against thirty human pathogens isolated from patients. The tested pathogens were eleven dermatophyte, eight yeast and eleven mold isolates. All of the three compounds showed 100% inhibition for the thirty pathogens at concentration of 1mg/ ml. The minimum inhibitory concentration (MIC) test of each compound was studied on eight dermatophyte, five yeast and five mold isolates. Thymoquinone was the best antifungal compound against all of the tested dermatophytes and yeasts, followed by thymohydroquinone and thymol respectively. Thymol was the best antifungal against molds followed by TQ and THQ respectively. Keywords: Nigella sativa, thymol, thymoquinone, thymohydroquinone, antifungal, dermatophytes.