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   2020| July-September  | Volume 19 | Issue 3  
    Online since September 30, 2020

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Lichens − masters of extraordinary symbiosis with potent pharmaceuticals
Waill A Elkhateeb, Ghoson M Daba
July-September 2020, 19(3):197-201
Lichens are an excellent example of a symbiotic relationship between members belonging to two unrelated separate kingdoms (fungi and algae), which results in the collection of secondary metabolites. These metabolites can be fungal originated, algal originated, or unique compounds not produced by either fungi or algae individually. Although involved since centuries in traditional folk medicine, lichens have attracted extra attention of scientists owing to the emergence of new diseases, which has required screening for novel compounds capable of curing or supporting currently used compounds. This review highlights the nature, importance, nutritional and pharmaceutical uses, and applications of these enigmatic dual organisms.
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Production and one-step purification of serratiopeptidase enzyme from Serratia marcescens with potent anti-inflammatory and antioxidant power
Mostafa A El-Abd, Eman A Ibrahim
July-September 2020, 19(3):238-243
Background and objective Inflammation is a huge problem facing the world. NSAIDs are the most popular and widespread drugs used for lowering the impaired symptoms accompanied with inflammation. This group has plentiful disorders to the gastrointestinal tract reaching to gastric ulcer. The objective of this study was to produce a natural product having an anti-inflammatory power without the side effects concomitant with the used drugs. The serratiopeptidase enzyme has a potent anti-inflammatory action without any abnormal disorders. Materials and methods Different media and cultural conditions were evaluated for the optimal production of the serratiopeptidase enzyme from a local bacterial strain. Partial purification was employed using the ammonium sulfate precipitation technique. Results and conclusion Serratiopeptidase enzyme was produced by the bacterial strain Serratia marcescens and its anti-inflammatory and antioxidant power were declared. The best medium was chosen for the optimal productivity and the optimum pH was 6 that recorded the highest anti-inflammatory and anti-oxidant activity at 67.5 and 71.8%, respectively, which is powerful than any chemical drugs used. Serratiopeptidase enzyme produced by this strain was partially purified by ammonium sulfate precipitation revealed an increase in specific activity by two times than the crude one with 14.7 recovery. The authors conclude the use of serratiopeptidase enzyme for treating the inflammation instead of the traditional ways.
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Occurrence of terpenes, polyketides, and tannins in some Japanese lichens and green mosses
Waill A Elkhateeb, Ghoson M Daba
July-September 2020, 19(3):216-223
Terpenes, polyketides, and tannins are valuable chemical classes that are famous for their varied biological activities such as being antioxidant, anticancer, antibiotic, and immunosuppressant agents. On the other hand, lichens and mosses are rich sources of biologically active compounds. However, rare studies have described neither the chemical analyses nor the biological activities of such samples. In this review, the chemical constitutions of five Japanese originated lichens and moss samples belonging to the species Candelariella vitellina, Lepraria incana, Dirinaria applanata, Brachythecium velutinum, and Brachythecium rutabulum were described with focus on three chemical classes which are terpenes such as Dechloromycorrhizin A, Hericenone A, demethoxyviridin, Scytalidic acid, Ovellin B, Ceriporic acid B, Ganodermatriol, Fomefficinic acid A, Ganoderol A, Ganoderol F; polyketides such as Chaetoquadrin A, Comazaphilone C, Hormothamnione, Arnottianamide, Avermutin, 4′-Hydroxyphlebiarubrone, 4′-Hydroxyphlebiarubrone, Citropone A, Atrovenetin; and finally secondary metabolites (tannins) such as 5′ methoxydehydrodiconiferyl alcohol, ellagic acid 3,3′-di-O-methyl ether, 5,5′-dehydrodiferulic acid, 3,3′,4-tri-O-methylellagic acid, and 5′-methoxydehydrodiconiferyl alcohol.
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Egyptian red sea seagrass as a source of biologically active secondary metabolites
Abdel-Hamid A Hamdy, Nabaweya M El-Fiky, Ahmed A El-Beih, Magdy M.D Mohammed, Walaa S.A Mettwally
July-September 2020, 19(3):224-237
Background and objective The Red Sea seagrass Halophila stipulacea (Forsskål) Ascherson and Thalassodendron ciliatum (Forsskål) den Hartog were poorly investigated either for their biological activities or for chemical constituents. This study aims to investigate the phytochemical constituent of both grass, along with studying the different biological activities (osteoclastogenesis, antioxidant activity, and anticancer activity) of the crude extract as well as purified compounds. Materials and methods The present study used three different in-vitro bioassay methods to screen the fractions and/or isolated compounds of both seagrass, to assess their possible biological activity. Osteoclastogenesis assay, antioxidant activity, and anticancer activity were carried out using different assays of the different anticancer mechanism of action. Results and conclusion Ten secondary metabolites were isolated and identified for the first time from Red Sea seagrass H. stipulacea (Forsskål) Ascherson: (1) p-hydroxybenzoic acid, (2) bis(2-ethyl hexyl) phthalate, (3) benzoic acid, (4) p-hydroxybenzaldehyde, (5) thymidine, (6) stigmasterol, (7) oleic acid, (8) linoleic acid, (9) linoleic methyl ester, and (10) apigenin. Furthermore, a new dihydrochalcone and a known flavonoid were isolated and identified from Red Sea seagrass T. ciliatum (Forsskål) den Hartog: (11) 2,4′ dihydroxy-4-methoxy-6-glucosyl dihydrochalcone (Thalassodendron B) and (12) rutin. Fractions of both seagrass showed promising biological activities.
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Implementation of nanotechnology in drug analysis as a new solution for old problems: a review
Wafaa A Zaghary, Nermeen A Abdallah, Ayman A Abdelrahim, Marwa A Zanoun, Emily T Hanna, Tamer M Sakr
July-September 2020, 19(3):202-215
There is an increasing interest in nanoparticles nowadays. In this review, we will enlighten the use of nanoparticles for detection and quantification of some drugs, antibiotics, antivirals, and amino acids using different analytical methods.
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Production of dextranase by aspergillus fumigatus NRC-F103 and its application in cane juice treatment and enhancing ethanol production from sugarcane molasses
Mohamed Fadel, Fatima H Motawe, Amira M Roshdy
July-September 2020, 19(3):283-290
Background and objectives One of the important applications of dextranase enzyme is preventing dextran accumulation in sugarcane juice as well as, consequently, for enhancing the resultant fermentable reducing sugars and ethanol yield in the fermentation of sugarcane molasses by yeasts. Materials and methods Different materials and methods were used for fungal strains [screening, mutation ultraviolet (UV), inoculum preparation, cultivation type ‘solid-state fermentation,’, culture substrates], dextranase (production, assay, soluble protein determination), purification, characterization, application in ethanol production by fermentation from sugarcane molasses, dextran estimation, and determination of reducing sugars. Results and conclusion Six fungal strains (namely Aspergillus oryzae FK-923, Aspergillus niger F-93, A. niger F-258, Aspergillus awamori NRC-F18, Aspergillus fumigatus NRC-F103, and Trichoderma viride NRC-F107) were screened on sorghum, sugar beet pulp, wheat bran, and orange peels using the solid-state fermentation technique to produce dextranase enzyme. The fungus A. fumigatus NRC-F103 cultivated on orange peels showed promising enzyme yield than other tested fungal strains. Then, optimization of culture conditions for dextranase production was carried out. Moisture content, initial pH value, incubation temperature, and incubation period were optimized to be 2 : 1 (v/w), 5.0, 35°C and 96 h, respectively. Five inorganic nitrogen sources were trailed at equivalent levels as sole nitrogen in the fermentation medium did not result in any increase in enzyme activity. Subjecting the fungal to UV resulted in a 75% increase in enzyme activity corresponding to the mother strain before subjecting to UV. Under the above conditions, 118 U/g original substrate was obtained. Isopropanol 1 : 1 (v/v) was applied for precipitation enzyme protein, as 32% of total protein involving 68% of total enzyme activity was obtained and specific activity was 42.94 U/mg protein compared with 16.4 U/mg protein in the culture supernatant. A study on obtained dextranase showed that it has an optimum that pH ranged from 4.5 to 5.5 as well as it gave the highest activity when incubated between 35 and 40°C. Promising results were obtained when the enzyme was applied in cane juice to prevent accumulation of dextran. Enzyme supplementation to diluted sugarcane molasses (26%, w/v) resulted in an increase in reducing sugars by 2.64%. Ethanol was increased by about 2.36% (v/v) in the fermentation medium supplemented with an enzyme compared with the unsupplemented medium and the fermentation efficiency increased from 89 to 92.5%.
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Methanolic extracts and different fractions of whole plants of Leucas zeylanica show promising analgesic and antioxidative activities
Molla M Islam, Ela S Parvin, Raihan S Zahir, Mahmud Z Al
July-September 2020, 19(3):260-265
Background and objective Leucas zeylanica is a medicinal plant used traditionally in tropical Asian countries including Bangladesh. This study was conducted to assess the pharmacological activities of whole plants of L. zeylanica. Materials and methods Methanolic crude extracts (MCEs) and petroleum ether-soluble fractions (PESFs), chloroform-soluble fractions (CSFs), and ethyl acetate-soluble fractions (EASFs) of whole plants were studied for the probable peripheral analgesic, central analgesic, and antioxidant and antimicrobial activities by acetic-acid-induced writhing and radiant heat tail-flick tests in mice, 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging, and disk diffusion method, respectively. Results and conclusion The MCEs showed the highest peripheral analgesic activity among the test samples with an inhibition of pain sensation of 40% (at 200 mg/kg body weight dose, MCE200) and 32% (at 100 mg/kg body weight dose, MCE100) in comparison to diclofenac sodium (66.67%, P<0.001). In central analgesic activity test, elongation of flicking time was calculated at 30, 60, and 90 min of administration of test samples where morphine was the positive control. All extracts and fractionates showed significant increase in tail-flicking time. However, methanolic extract showed the highest activity (at 60 min) among all test samples (15.44±0.256, P<0.001). In antioxidant tests, butylated hydroxytoluene (BHT) was used as the positive control where methanolic extracts exhibited the highest antioxidant potentials among the test samples. IC50 (μg/ml) values were 19.61 (BHT), 28.46 (MCE), 65.61 (EASF), 84.75 (PESF), and 97.09 (chloroform-soluble fractions). Additionally, whole plant extracts showed weak antimicrobial activity. This ethnopharmacological investigation suggests that methanolic extracts and different fractions from L. zeylanica have strong analgesic and antioxidant potential and can be a significant source of natural medicine.
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Use of echocardiography and glutathione S-transferase to detect heart complications in β-thalassemic patients
Assem M Abo-Shanab, Naglaa Kholoussi, Dawoud Habib, Iman A Helwa, Gehan Hussin, Naglaa Omar, Ghada El-Kamah, Mohamed El-Dessouk, Abdelgawad Fahmy
July-September 2020, 19(3):244-251
Background and objective β-Thalassemia major (TM) is an inherited disorder of hemoglobin synthesis and characterized by defective hemoglobin synthesis, resulting in ineffective erythropoiesis, severe anemia, increased erythrocyte turnover, and excessive iron absorption. Accordingly, iron overload develops and may accumulate in the liver, heart, and endocrine organs. Several gene polymorphisms have been studied as protective or predisposing factors for cardiac dysfunction in patients with TM. Moreover, echocardiographic left ventricular (LV) diastolic evaluation is used to detect early myocardial dysfunction secondary to iron overload. This study aimed at determining some diastolic and tissue Doppler echo indices to predict iron load. Materials and methods This study included 42 β-thalassemic patients, among whom, 16 proved to have cardiac complications after clinical evaluation. Their age ranged from 3 to 25 years. Participants were subjected to clinical evaluation, molecular analysis to detect glutathione S-transferase M1 (GSTM1) gene polymorphism, and transthoracic color Doppler echocardiography to detect early myocardial dysfunction. Results Seven (43.7%) patients had the functional wild-type allele (GSTM1 non-null genotypes), whereas nine (56.2%) patients were homozygous for the GSTM1 null allele. There was a statistically significant increase regarding both right and LV E/A in GSTM1 null genotype when compared with GSTM1 non-null genotype in β-thalassemic patients with cardiac complications. Moreover, LV and right ventricular diastolic function has been significantly affected in participants with GSTM1 null genotype in β-thalassemic patients with cardiac complications, and particularly, LV diastolic function has been significantly impaired in cases experiencing frequent blood transfusion in β-thalassemic patients with cardiac complications. Conclusion Follow-up of patients with β-thalassemia and evaluating echocardiographic changes may permit better assessment of patients and early recognition of cardiac affection before disease progression.
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Synthesis and characterization of microbial poly3-hydroxybutyric acid nanocarrier for curcumin as an antibreast cancer agent
Mostafa A El-Abd, Mona M Agwa
July-September 2020, 19(3):266-273
Background and objective In a previous study, the authors produced, optimized, characterized, and purified the poly3-hydroxybutyric acid (PHB) using the locally isolated Bacillus flexus bacterial strain. The development of a biodegradable drug carrier as an efficient delivery system has received great interest over the past few decades. The objective of this study was to produce a nano-PHB carrier for curcumin to be more effective in tumor fighting. Materials and methods PHB was produced on the optimized medium by B. flexus strain. The nano-PHB form was produced using the nanoprecipitation technique. The size, shape, and characteristics of loaded curcumin nano-PHB particles were performed using zeta-potential, scanning electron microscopy, and Fourier transform infrared spectroscopy techniques. The antitumor effect of the nano-PHB loaded curcumin was performed using human breast adenocarcinoma MCF-7 cell line. Results and conclusion In this study, a naturally developed, biodegradable, and biocompatible nanosized carrier for curcumin-targeted delivery in breast-cancer cells with higher encapsulation efficiency (95.5%) was formulated. The size range of both free PHB and curcumin-loaded PHB was 237 and 260 nm respectively. The nanoparticles exhibited a spherical shape with no aggregation which is confirmed by electron microscopy, indicating a higher colloidal stability. The curcumin-loaded PHB nanocarrier showed a sustained drug release behavior. In-vitro anticancer assays showed the superiority of curcumin-loaded PHB nanocarrier over free curcumin for fighting breast cancer. These results show that the PHB biopolymer acts as an efficient carrier vehicle for the curcumin.
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Co-milling of oxcarbazepine with Soluplus for the enhancement of solubility and dissolution rate
Amjad Hussain, Muhammad S Arshad, Sidra Noreen, Javaria Khalid, Nasir Abbas, Jahanzeb Mudassir
July-September 2020, 19(3):291-296
Background and objective Poor solubility and dissolution rates affect the bioavailability of drugs. The aim of this study was to improve the solubility and dissolution rate of a poorly soluble drug, oxcarbazepine by its mechanochemical activation via the co-milling technique. Materials and methods The drug and Soluplus (in two different ratios) were co-milled in a planetary ball bill. The bulk properties, solubility, and dissolution rate were determined and differential scanning calorimetry, powder X-ray diffraction, Fourier-transform infrared spectroscopy (FTIR), and laser diffraction (for particle size determination) techniques were used to characterize drug and co-milled formulations. Results and discussion The results have shown good compressibility and excellent flow of co-milled mixtures as compared with the drug. The solubility of the drug (0.448±2 mg/ml) was increased by 2–3-fold in co-milled mixtures while the dissolution rate of oxcarbazepine was increased up to 2.5–3 times. Both differential scanning calorimetry and powder X-ray diffraction results have shown a reduction of crystallinity while the Fourier-transform infrared spectroscopy spectra indicated no interaction. Laser diffraction studies have shown ∼5 times reduction in mean particle size. Conclusion The study concludes that co-milling is effective in enhancing solubility and dissolution of poor soluble drugs.
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Evaluation of enzyme production and biocontrol agent of zinc nanoparticles from Gonatorrhodiella parasitica
Sherien M.M Atalla, Nadia G El Gamal, Khalil M.S.A.
July-September 2020, 19(3):252-259
Background and objectives Biosynthesis of zinc nanoparticles (Zn NPs) is considered one of the new approaches for enzyme production and good biofertilizer and biocontrol agent for pathogenic disease of plant. The selected fungal strain Gonatorrhodiella parasitica was used for biosynthesis of Zn NPs. Determination of enzyme activity in fermentation medium containing olive mill wastes (OMW) as substrate for production of protease, pectinase, and chitinase at different pH values (6) was done to produce detailed information on the progress of reduction of zinc nitrate on the nanosecond times (6.0, 8.0, and 10.0). Absorbance ultraviolet–visible spectra of Zn NPs at pH 6.0 were at λmax 280 nm. Transmission electron spectroscopy measurements indicate that extracellular biosynthesis of Zn NPs by G. parasitica was in the size of 16 nm. Composting of OMW inoculated with G. parasitica for 15–30 days led to decrease in total phenol, flavonoids, 2,2 diphenyl-1-picrylhydrazyl (DPPH), and aflatoxins found in OMW. Comparison of G. parasitica free and nano zinc particles as biocontrol agent of soil-borne fungi causing preemergence and postemergence damping off of green bean plants (seed treatment) under field conditions measured from 7 to 60 days and its effect of growth parameters confirm the use of Zn NPs for enzymes production and biocontrol application. Materials and methods The local fungal strain G. parasitica used in this study was isolated from soil and identified at the Department of Chemistry of Natural and Microbial Products at the National Research Center. Determination of enzymes activity (pectinase, chitinase and protease) was done. Determination of major phytochemicals in prepared extracts (total phenolic content, total flavonoid content, DPPH scavenging activity, and aflatoxin) was done. A field experiment was conducted during the 2018 season at Kafer-Eldawar (Behira) Governorate, in a heavily naturally infested field, to study the effect of free and Zn NPs of G. parasitica on soil-borne fungi causing preemergence and postemergence damping off of green bean plants (seed treatment) under field conditions. Results and conclusion Composting of OMW inoculated with G. parasitica for 15–30 days led to decrease in total phenol, flavonoids, DPPH, and aflatoxins found in OMW. Moreover, suppressive soil-borne fungi caused preemergence and postemergence root rot of green bean plants (seed treatment) under field conditions.
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Mesenchymal stem cells’ therapeutic potential for endotoxin-induced brain and spleen injuries in rats
Marwa A Masoud, Wedad A Hassan, Nahla A Hassaan, Hanaa A Mansour
July-September 2020, 19(3):297-306
Background and objective Inhalation of bacterial endotoxin induces an acute inflammation in various organs, especially the brain and spleen. This study examined the therapeutic effects of bone marrow mesenchymal stem cells (BM-MSCs) in lipopolysaccharide (LPS)-induced brain-spleen injuries in rats as compared with dexamethasone. Materials and methods A total of 32 male Wistar albino rats, weighing 180–200 g, were used in the study and were divided into four groups. Group 1 (normal) rats received 20 μl of saline in each nostril for two consecutive days. Group 2 animals received LPS (20 μl of LPS of Escherichia coli in each nostril for 2 consecutive days) that induced brain-spleen injuries and served as a positive control group. Group 3 animals were injected with dexamethasone (2 mg/kg, once, intraperitoneal). Group 4 animals received (1×106) BM-MSCs in 500 μl PBS/rat via intraperitoneal injection once before acute injury induction with LPS. At the end of the experiment, the authors studied the sickness behavior by assessing open field behavior and measured oxidative and inflammatory parameters. Results and conclusion LPS-induced open field behavior impairments (decreased locomotion and rearing and increased immobility), with elevation of a number of inflammatory cells, especially neutrophils. Moreover, LPS-induced elevation of lipid peroxidation along with reduction of both reduced glutathione and superoxide dismutase in brain and spleen tissues and increased interleukin-1β and myeloperoxidase contents in rats, compared with normal group. These harmful effects were hindered after treatment with MSCs. In conclusion, MSCs prevented both sickness and depressive-like behavior via neuroinflammatory pathway and could be a novel approach to therapy for LPS-induced serious injuries in rats but needs further clinical studies.
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Biological evaluation of ethyl acetate extract of Chaetomium cupreum against Ehrlich ascites carcinoma cells in Swiss albino mice
Nazir A Wani, Waseem I Khanday, Sheihk A Ali, Sharmila Tirumale
July-September 2020, 19(3):274-282
Background Chaetomium genus is a natural source of different types of secondary metabolites or pigments. These secondary metabolites display a broad spectrum of biological properties including antimicrobial, antioxidant, anti-inflammatory, and anticancer activity. Objective The objective of the study was to evaluate the anticancer activity of ethyl acetate extract of Chaetomium cupreum against Ehrlich ascites carcinoma (EAC) cells in Swiss albino mice. Materials and methods Methods involved are evaluation of acute toxicity study, tumor induction using EAC cells, estimation of various hematobiochemical parameters, and evaluation of antioxidant enzymes and markers of oxidative stress. Results The ethyl acetate extract of C. cupreum-treated EAC-bearing mice at the concentration of 200 mg/kg body weight (bwt) reduced ascitic fluid volume (1.65±0.70 ml) and ascitic fluid weight (1.32±0.69 g) as compared with ascitic fluid volume (4.79±0.52 ml) and ascitic fluid weight (3.93±0.57 g) in EAC control group. Similarly, the cell apoptosis was higher in EAC-bearing mice treated with standard 5-fluorouracil at 50 mg/kg bwt (96.04%) as compared with treatment with ethyl acetate extract at 50 mg/kg bwt (21.92%) followed by 100 mg/kg bwt (36.63%) and increased further at 200 mg/kg bwt (47.48%) in treated groups. In hematological estimation, the EAC-bearing mice treated with ethyl acetate extract at 200 mg/kg bwt showed increased red blood cell count (3.78±0.07×106/μl) and hemoglobin content (6.02±01 g/dl) and decreased white blood cells count (5.45±0.01×103/μl). In biochemical estimation, ethyl acetate extract treatment in EAC-bearing mice at 200 mg/kg bwt decreased aspartate aminotransferase activity (64.10±0.07 U/l), alanine aminotransferase (55.71±0.65 U/l), alkaline phosphatase (107.04±0.02 U/l), cholesterol (124.38±0.04 mg/dl), and triglycerides (155.38±0.04 mg/dl). Similarly, in enzymatic antioxidants and oxidative stress, the ethyl acetate extract-treated EAC-bearing mice at 200 mg/kg bwt increased superoxide dismutase (27.10±0.03 U/mg protein), catalase (20.20±0.02 U/mg protein), and reduced glutathione (24.04±0.03 U/mg protein), whereas decreased glutathione peroxidase (38.04±0.07 U/g hemoglobin) and malondialdehyde content (170.50±0.06 nmol/mg protein) significantly. Conclusion The results of the present finding showed that ethyl acetate extract of C. cupreum possesses significant anticancer potential.
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