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   2019| April-June  | Volume 18 | Issue 2  
    Online since July 5, 2019

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Optimization and characterization of l-asparaginase production by a novel isolated streptomyces spp. strain
Abeer A El-Hadi, Hanan Mostafa Ahmed, Rania A Hamzawy
April-June 2019, 18(2):111-122
Introduction and purpose Microbial l-asparaginase (l-asparagine amino hydrolase, E.C. has been applied as the most important chemotherapeutic agent in the treatment of certain human cancers, especially in acute lymphoblastic leukemia. Actinomycetes are recognized as a comparatively less explored source for l-asparaginase and therefore act as candidates for the production of l-asparaginase. The possibility of using novel actinomycete as a source of production of some industrially important microbial l-asparaginase was investigated in this study. Materials and methods Genomic DNA of the actinomycetes isolate from soil samples was extracted using the Gene JET Genomic DNA purification kit (Thermo scientific #k0721). The actinomycete isolate was identified by 16S rDNA. The identified actinomycete isolate was inoculated on starch casein agar slants and incubated for 7–10 days at 28°C, and then it was maintained at 4°C until further use. Inoculum was prepared from a 7-day old culture of the strain. Production of l-asparaginase was initially tested in four different media. The actinomycete strain was used further for the optimization of cultural conditions, namely, l-asparagine substrate concentrations, pH, temperature, and incubation conditions. The pH of the medium was varied from pH 3.0 to pH 9.0; the incubation temperature was varied from 30 to 50°C. The effect of carbon source and nitrogen source on l-asparaginase production was studied. Modified Czapex Dox broth was supplemented with different carbon and nitrogen sources such as starch, mannitol, mannose, sucrose, cellulose, and fructose at a concentration of 1% (w/v) and ammonium sulfate, beef extract, yeast extract, and peptone at a concentration of 0.2% (w/v), respectively, keeping other components constant. The properties of Streptomyces spp. l-asparaginase were also studied such as pH, assay temperature, and thermal stability. Results and discussion Genotypic characterization of the most promising unknown actinomycete isolate showing the maximum production was identified by 16S rDNA. PCR amplified the 16 s rDNA region using primers. Genotypic characterization of the most promising unknown actinomycete isolate showing the maximum production was identified by16S rDNA. The 16 s rDNA region was amplified by PCR (about 1000 bp) using primers. According to sequencing similarities and multiple alignments, the isolate was found to be closely related to Streptomyces griseoplanus strain NRRL-ISP 5009 16 s ribosomal RNA gene with 85% identity. Higher enzyme activity was observed in modified Czapex Dox broth as compared with other media used. Modified Czapex Dox broth was supplemented with different concentrations of l-asparagine; the enzyme production was maximum at 1.5% l-asparagine (126.20 U/ml). Analysis of the culture supernatant showed that the enzyme activity rise from 126.20 U/ml on the fifth day to 141.11 U/ml on the 10th day which its peak enzyme production activity. Different carbon sources such as starch, mannitol, lactose, sucrose, and glucose were amended in asparagine-modified Czapek Dox broth to determine their impact on l-asparaginase production. Biosynthesis of l-asparaginase by S. griseoplanus strain has been reported to be higher when the basal medium was supplemented with starch. For the production of l-asparaginase by S. griseoplanus strain, yeast extract has been reported as a good nitrogen source. According to the properties of the enzyme, the maximum activity was achieved at 45°C. The half-lives of the free enzyme were calculated to be 521 min (8.5 h) at 50°C, 312.6 min (5.2 h) at 55°C, and 195.2 min (3.25 h), at 60°C.
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Phytochemical investigation of Corchorus olitorius and Corchorus capsularis (Family Tiliaceae) that grow in Egypt
Amal Z Hassan, Maged K.G Mekhael, Atef G Hanna, András Simon, Gábor Tóth, Helmut Duddeck
April-June 2019, 18(2):123-134
Background and objective Corchorus (Family Tiliaceae) is a genus of annual herbs. Nearly 40 species are known to occur in nature and distributed in the tropics of both hemispheres. Because of the wide medicinal applications of compounds isolated thereof, the present investigation deals with the isolation and structure elucidation of some phytochemicals from Corchorus olitorius (molokheya) and Corchorus capsularis that grow in Egypt. Materials and methods Phytochemical investigation of the seeds and different plant organs of both C. olitorius and C. capsularis was achieved applying different separation techniques. Petroleum ether extraction followed by saponification of the extract led to the isolation of phytosterols, hydrocarbons and fatty acids. Essential oils were obtained from the leaves by extraction with methylene chloride. Methanolic extraction led to the isolation of cardiac glycosides. Identification of isolated compounds was realized through Rf values, shift reagents and spectroscopic tools such as ultraviolet and nuclear magnetic resonance. The fatty acids were identified using gas liquid chromatography. Results and conclusion A study of the lipid contents (fatty acids, phytosterols and hydrocarbon components) of seeds, roots, leaves and stems of C. olitorius as well as the seeds and vegetative part of C. capsularis, which grow locally in Egypt, was carried out. The identification of the lipid content was achieved by comparing the retention time of their peaks in gas liquid chromatography with those of authentic samples. Gas chromatography/mass spectrometry study of the chemical constituents of the essential oils of the leaves of C. olitorius and C. capsularis led to the identification of 11 and 21 compounds with a total concentration of 24.7 and 62.9%, respectively. Cedrane-5-one (17.7%) and γ-terpinene (12.1%) represent the major compounds in each plant, respectively. Phytochemical investigation of C. olitorius led to the isolation of raffinose I, coroloside II, glucoevatromonoside III, erysimoside IV and olitoriside V and gluco-olitoriside VI. Meanwhile, the study of the vegetative parts of C. capsularis led to the isolation of 3-O-glucopyranosyl-β-sitosterol VII. The isolated compounds were identified by spectral tools (hydrogen-1, carbon-13-nuclear magnetic resonance, electron ionization mass spectrometer).
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Erratum: Production, characterization, and antioxidant activities of bacterial exopolysaccharides extracted from petroleum oil water

April-June 2019, 18(2):178-178
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GC-MS analysis and in-vitro hypocholesterolemic, anti-rotavirus, anti-human colon carcinoma activities of the crude extract of a Japanese Ganoderma spp
Waill A Elkhateeb, Ghoson M Daba, Donia Sheir, Asmaa Negm El-Dein, Walid Fayad, ELmahdy M Elmahdy, Mohamed N.F Shaheen, Paul W Thomas, Ting-Chi Wen
April-June 2019, 18(2):102-110
Background and objective Medicinal mushrooms are mines of various biologically active compounds. Therefore, chemical analysis and in-vitro evaluation of some biological activities of the Japanese originated mushroom Ganoderma spp. were conducted. Materials and methods Extraction of the fruiting bodies of Ganoderma spp. was accomplished using 80% methanol. This extract was investigated for its in-vitro cholesterol-lowering activity, anti-rotavirus effect, and anti-human colon cancer influence. Moreover, a gas chromatography–mass spectrometry analysis for this extract was performed. Results and conclusion The gas chromatography–mass spectrometry analysis resulted in the detection of 39 compounds, which were generally saturated and unsaturated fatty acids, and alkenes. The crude extract exhibited a promising in-vitro cholesterol-lowering activity (100±0%) after 96 h of incubation at room temperature. The same crude extract showed a moderate anti-rotavirus SA-11 strain effect with a therapeutic index of 9.3. Moreover, Ganoderma spp. extract displayed a strong activity toward HCT116 human colon carcinoma cell line, resulting in a cytotoxicity of 84.03±0.93% on HCT116 cell line monolayers. Ganoderma spp. crude extract represents a promising source of biologically active compounds that could by further investigations represent support and/or alternative to the currently used drugs.
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Contrast computed tomography versus PET/CT in the assessment of bronchogenic carcinoma
Omar Hussein, Safenaz El Sherity, Yehia Omar
April-June 2019, 18(2):135-140
Background In the past decades, diagnostic imaging modalities of bronchogenic carcinoma were chest radiography and computed tomography (CT) to determine the tumor size and mediastinal lymph nodes involved, as well as liver and adrenal metastases. Now PET/CT has become a routine procedure for the primary assessment (initial staging) in the detection of functional tumor activity (viable cells). Objective The objective of this study was to compare between contrast CT and PET/CT in the assessment of bronchogenic carcinoma (initial staging) and impact of weight and BMI on it. Patients and methods This was a cross-sectional study that involved 100 patients for initial staging of newly diagnosed bronchogenic carcinoma examined by contrast CT and PET/CT. In addition to anthropometry measures, weight and height were taken and BMI was calculated. Results The initial staging of bronchogenic carcinoma showed significant differences between PET/CT versus contrast CT (P=0.001). Upstaging done by PET/CT in six patients (stages I and II), as well as an agreement between CT and PET/CT in stage III and IV was detected. The evaluation showed a sensitivity and specificity of 75.2–89.4% and 78.2–90.0% for CT and 97.2–100% and 98.5–100% for PET/CT, respectively. There is a significant positive correlation between tumor size and its metabolic activity measured by the maximum standardized uptake value. However, there was no significant correlation between BMI and maximum standardized uptake value; moreover not any significant association between BMI and metastatic deposits was detected. Conclusion PET/CT is a powerful imaging modality for the assessment of functional behavior of tumor cells to avoid false results depending on the morphology only as contrast CT, which leads to change the decision taken for the management of bronchogenic carcinomas.
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Effect of nonionizing rays on growth, chemical constituents, and molecular aspects of Catharanthus roseus plant
Sami A Metwally, Rasha S Tawfik, Bedour H Abo-Leila, Tarek A.A El-Tayeb
April-June 2019, 18(2):141-148
Background and objective Catharanthus roseus (L.) is a perennial tropical plant and is considered one of the important ornamental and medicinal plants of the family Apocynaceae. It is used as an anticancer and antidiabetic agent. Catharanthus leaves are used for menorrhagia and rheumatism. The laser rays are used as a physical method to improve the germination, the plant growth, and the vigor of the seeds. The study aimed to evaluate vegetative growth, flowering, and marker gene on Catharanthus leaves and to investigate its chemical constituents under the effect of laser rays. Materials and methods The pot experimental study was carried out at greenhouse of National Research Centre, Giza, Egypt, on C. roseus. Two types of laser [helium cadmium (He-Cd) and argon (Ar) laser] were used at different exposure times (0, 4, 8, and 12 min), and various morphological, flowering, chemical constituents were determined. Inter-simple sequence repeat (ISSR) marker was used to illustrate the effect of He-Cd and Ar laser on C. roseus at different exposure time. Results Generally, the highest values of number of leaves, plant height, root length, fresh and dry weight of leaves, and number of days to flower, photosynthetic pigments, proline %, carbohydrate, vinblastine, and vincristine were obtained with 8 and 12 min of He-Cd laser treatments. Conversely, 4-min He-Cd and 8-min Ar laser time exposure recorded decrease in photosynthetic pigments, proline %, and carbohydrate concentration. ISSR showed that mean polymorphic percentage was 25%. ISSR investigation demonstrated that the control, He-Cd laser exposure for 8 min, and Ar laser exposure for 12 min of Catharanthus plant produced unique positive markers, which were found to be mutant specific. Conclusion It was concluded that laser rays with different time exposure had potential effect on growth, flowering, chemical constituents, and the production of Catharanthus plant.
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Quantification of fosamprenavir in spiked human plasma using liquid chromatography–electrospray ionization–tandem mass spectrophotometry–application to pharmacokinetic study
Manish K Thimmaraju, Prasenjit Mondal, Kola Venu, Gummadi Sridhar Babu, Gaju Rajkumar, Bookya Padmaja
April-June 2019, 18(2):149-159
Background Fosamprenavir (FSV) is used for the treatment of HIV infections. It is a prodrug of the protease inhibitor and antiretroviral drug amprenavir. Aim This research work described about the estimation of FSV in spiked human plasma using electrospray ionization, LC-MS/MS technique, and its application to pharmacokinetic study in rabbits. Materials and methods Liquid–liquid extraction technique was used for the extraction of FSV in spiked human plasma. The separation was achieved using ZORBAX SB-C18 column with 4.6 mm internal diameter with 5 μm particle size using acetonitrile: 5 mmol/l ammonium acetate in water (85 : 15, v/v) as a mobile phase. Positive ion mode was selected for the product ion mass spectra, m/z 585.6–418.2 for FSV and m/z 589.2–469.1 for FSV-deuterated (internal standard), The US Food and Drug Administration guidelines were adopted for successful validation of the developed method. Results The retention time of FSV was found to be 1.51 min, for FSV-deuterated it was 1.62 min, with a runtime of 2.5 min. The present method exhibits excellent intraday and interday accuracy with %nominal 95–98.4% and precision percentage coefficient variation up to 3% in all quality control (QC) levels. The developed method demonstrated excellent matrix and analyte selectivity (%interference=0), matrix effect (matrix factor 2.09 at lower quantitation limit and 1.14 at high QC level) and satisfactory stability study results in all types (%nominal 94.03–100.80%). The linearity range was found to be 0.510–200.185 ng/ml with a correlation coefficient (r2) of 0.998. The calculated accuracy and precision values in the ruggedness study were within 15–20% in all QC levels. The percentage coefficient variation of the pharmacokinetic study on rabbit plasma samples was also conducted and the parameters of FSV showed Tmax of 2 h and the mean Cmax, AUC0→t and AUC0→α for test formulation were 98.6, 351.3, and 354.9, respectively. Conclusion This method was successfully optimized, validated, and applied favorable for the pharmacokinetic study of marketed formulation in rabbit blood samples in a single oral human-equivalent dose. The applicability of the developed method undoubtedly can further extend during preclinical and clinical trials.
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Influence of two extraction methods on essential oils of some Apiaceae family plants
Mohammed Sayed Aly Mohammed, Mohamed Salah Hussein Tawfik, Ahmed El-Gohary Ibrahim
April-June 2019, 18(2):160-164
Background Renewed interest in natural materials as food flavors and preservatives has led to the search for suitable essential oils. One factor that influences the essential oil content (%) is the extraction method used. Carum carvi (caraway), Anethum graveolens L. (anise), and Pimpinella anisum L. (dill) are well known plants from Apiaceae family widely spread in Egypt, where they have good climatic and soil conditions for high yield and good quality. Essential oil content is the main criteria for determining the quality of the fruits of these plant species. The aim of the study was to choose the best method for essential oil extraction. Objective The present research was conducted to evaluate the possible impacts of two types of distillation methods − hydrodistillation and hydro-steam distillation on essential oil content (%) and its main constituents of caraway, anise, and dill fruits. Materials and methods Seeds of the three species were subjected to two types of distillation methods − hydrodistillation and hydro-steam distillation. The essential oil content (%) of the three plants were determined and gas chromatography-mass spectrometry analyses was carried out to identify the chemical constituents of the oil samples and their percentage were calculated in order to clear the effect of the two extraction methods applied. Results and conclusion It was established that while hydrodistillation gave higher essential oil yields for caraway and dill seeds (3.14 and 2.36%, respectively), hydro-steam distillation gave the maximum mean values of essential oil content of anise seeds (0.76%). The maximum values of the main components such as carvone (54.45%), transanethole (98.97%), and carvone (57.71%) were obtained as a result of hydrodistillation method for caraway, dill, and anise seeds, respectively.
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Optimization of parameters for lipase production by Aspergillus niger NRRL-599 using response surface methodology
Rania A. Abd El Aal, Yousseria M Shetaia, Mona S Shafei, Sanaa K Gomaa, Hassaan A El Menoufy, Heba A El-Refai
April-June 2019, 18(2):165-171
Background and objective Lipases are characterized to catalyze precise chemical transformation. That is they have widespread applications in detergents, cosmetics, food, organics synthesis, and pharmaceutical industries. In recent years, the study and development of lipase production in solid state fermentation, also using sequential statistical strategy as optimization for some vital factors are gaining more attention. This may be attributed to several advantages using techniques such as simplicity of fermentation media, no need for complex machinery. Equipment and control system, compactness of fermentation vessel due to lower water volume, high yields, less energy needed, and lower capital. Materials and methods Aspergillus niger NRRL-599 strain was tested for lipase production using different agroindustrial wastes including wheat bran, wheat germ cake oil, jojoba cake oil, almond cake oil, and olive oil as substrates under solid state fermentation. For further optimization, lipase activity was studied using two sequential optimization. Results and conclusion Olive oil waste was the most suitable substrate for lipase production (125 U/ml). Using one variable at a time, maximum lipase activity (200 U/ml) and specific enzyme activity (357.1 U/mg) were recorded in the presence of 5% w/v olive oil, 48 h inoculum age, 4% v/v inoculum size after 14 days fermentation at 30°C. The screening of the seven physiological factors using Plackett–Burman design showed that only three variables; that is tween 80, moisture content, and inoculum age affected significantly lipase production. Optimization by Box–Behnken design resulted in the highest lipase activity (420 U/ml) in which the most effective variables were inoculum size, inoculum age, moisture content, and tween 80 1%.
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Chemical and biological evaluation of olive leaves as a waste by-product of olive oil industry
Mona T.M Ghanem, Wafaa A Tawfik, El-Sayed M Mahdy, Mohamed Essameldin Abdelgawad, Nahla S Abdel-Azim, Moustafa M El-Missiry
April-June 2019, 18(2):172-177
Background and objectives Finding new uses for by-products of cultivated plants is of great value economically and to the environment. Leaves represent about 10% of the total weight of olives yield. It is worth to obtain value-added products from this material. In this article, the leaves were evaluated chemically and biologically for phenolics and flavonoids as well as for microelements and macroelements and fatty acids. Also, antioxidant and antimicrobial activities were carried out. Materials and methods Air-dried powdered olive leaves were defatted with hexane and the marc was then soaked in 80% methanol and successively extracted with CH2Cl2, EtOAc, and n-BuOH. Total phenolic and flavonoid contents were determined as chlorogenic acid and rutin equivalents, respectively. Microelements and macroelements were detected in addition to fatty acids. The antioxidant effect was determined in vitro using 1,1-diphenyl-2-picrylhydrazyl and antimicrobial activity was carried out using in-vitro agar well diffusion method. Results and conclusion Total phenolics were found to be highest in the 80% methanolic extract and the lowest in water and ethyl acetate fractions. 1,1-diphenyl-2-picrylhydrazyl-free radical scavenging activity of olive leaf extracts were in this order: 80% methanolic extract, water extract, ethyl acetate fraction and butanol fraction. Also, the calcium : potassium value was 15: 1. Fatty acid profile revealed that linolenic acid was the major fatty acid in terms of percent (49.45%). Ethyl acetate fraction showed positive antibacterial activity and negative antifungal activity whereas water, 80% methanol, and butanol fractions have positive antifungal and negative antibacterial activity. Conclusion Olive leaves could be considered as a potential inexpensive source for food supplements for human health.
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Association of human microbiome with health and disease states
Hanan A Al-Ashaal
April-June 2019, 18(2):75-87
This review deals with microorganisms that are associated with the human body, their types and diversity. The survey also covers the symbiotic and adverse effects of the located flora and how could friendly microbial communities change to vigorous pathogenic enemies. Besides, the study includes special oversight on the natural compounds that may help to overcome serious diseases that are caused by or that result from microbiome alteration or dysbiosis.
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Medicinal mushrooms as a new source of natural therapeutic bioactive compounds
Waill A Elkhateeb, Ghoson M Daba, Paul W Thomas, Ting-Chi Wen
April-June 2019, 18(2):88-101
In the ancient books of traditional medicines, medicinal mushrooms were occupying the headlines, and the main topics were confirming to their miraculous therapeutic powers. The presence of various phenolic compounds, polysaccharides, and terpenoids and other compounds, is the reason for their potent biological activities as anticancer, antioxidant, antimicrobial, antiaging, hepatic protective, hypoglycemic, hypocholesterolemic, and much more biological activities are discovered every day. Many mushroom genera are famous for their promising therapeutic capabilities. One of the mushrooms genera attracting attention is Cordyceps which has long been used in Asian countries for maintaining long and healthy life. Numerous studies on different metabolic activities of Cordyceps have been performed both in vitro and in vivo. This review describes the importance of medicinal mushrooms with focus on Cordyceps as an example of globally commercialized mushrooms.
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