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Table of Contents
October-December 2019
Volume 18 | Issue 4
Page Nos. 285-435
Online since Tuesday, January 28, 2020
Accessed 41,005 times.
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REVIEW ARTICLES
Fomitopsis officinalis mushroom: ancient gold mine of functional components and biological activities for modern medicine
p. 285
Waill A Elkhateeb, Ghoson M Daba, Marwa O Elnahas, Paul W Thomas
DOI
:10.4103/epj.epj_46_19
Polypore mushrooms have been used medicinally for thousands of years. Agarikon (
Fomitopsis officinalis
) is a medicinal polypore mushroom containing a host of pharmacologically active compounds that beneficially affect human health. Agarikon is known for its capability of producing various biologically active compounds with medical applications such as antiviral, antibacterial, anticancer, and anti-inflammatory agents. This review describes the importance of medicinal mushrooms, with a specific focus on Agarikon as an example of a globally commercialized medicinal mushroom.
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Metabolomics applications in disease diagnosis, treatment, and drug discovery
p. 290
Hassan Z Ghanem, Mai O Kadry, Rehab M Abdel-Megeed, Abdel-Hamid Z Abdel-Hamid
DOI
:10.4103/epj.epj_10_19
Metabolomics-based discovery has an important role in disease diagnosis, discovery of drug, and treatment expansion. This tool is a novel biomarker that can provide a biochemical insight into disease stages and could estimate the activity of certain drugs; it can observe the preclinical and clinical cases, leading to discovery of more efficient methods for treatment approach. This review discussed the potential of metabolomics technology as a very important approach in disease diagnosis, therapy, as well as new drug discovery.
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ORIGINAL ARTICLES
Effect of lemon balm (
Melissa officinalis
) aqueous extract on streptozotocin-induced diabetic rats
p. 296
Mahitab I EL-Kassaby, Abeer A.A Salama, Hagar H Mourad, Khaled G Abdel-Wahhab
DOI
:10.4103/epj.epj_14_19
Background
Lemon balm (
Melissa officinalis
) has a significant role in curing diseases and maintaining health through its antioxidant capacity. The aim of this study was to evaluate antidiabetic effect of lemon balm aqueous extract (LBAE) on streptozotocin (STZ)-induced diabetic rats.
Materials and methods
The extract was administered to STZ-induced diabetic rats in low and high doses (200 and 400 mg/kg body weight/day, respectively) for 4 weeks. Serum insulin, glucose, lipid profiles, alkaline phosphatase, serum alanine aminotransferase, aspartate transaminase, creatinine and urea levels were determined, whereas total antioxidant capacity, malondialdehyde, nitric oxide, Na
+
/K
+
ATPase activity (ATPase), tumor necrosis factor-α, and cluster of differentiation 4 levels were evaluated in liver and kidney tissue homogenates.
Results and conclusion
Oral administration of LBAE significantly decreased glucose, total cholesterol, triglycerides, low-density lipoprotein cholesterol, malondialdehyde, nitric oxide, tumor necrosis factor-α, and cluster of differentiation 4 levels. However, insulin, high-density lipoprotein cholesterol, and total antioxidant capacity levels significantly increased with respect to diabetic control group. These findings revealed that LBAE possesses antihyperglycemic and antihyperlipidemic effects against STZ-induced disorders in diabetic rats. Hence, it can be used in the management of diabetes mellitus.
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Comparative antibacterial study between bioactive glasses and vancomycin hydrochloride against
Staphylococcus aureus
,
Escherichia coli
, and
Pseudomonas aeruginosa
p. 304
Dina A Maany, Zainab M Alrashidy, Nabil A Abdel Ghany, Wafa I Abdel-Fattah
DOI
:10.4103/epj.epj_15_19
Background
This work targets the comparison of the antibacterial activity of different bioactive glasses as particles and those coating the surface of 316 l stainless steel sheet, with that of vancomycin hydrochloride antibiotic, to determine the best efficiency of the aforementioned materials for medical and surgical purposes.
Materials and methods
Different bioactive glass composites (borate, B, S, and B5), composed of different ratios of oxides, such as SiO
2
, Na
2
O, CaO, B
2
O
3
, P
2
O
5,
and MgO, were prepared. The antimicrobial activity of different synthesized glasses as well as vancomycin hydrochloride antibiotic was carried out against various Gram-negative and Gram-positive pathogens. The different bioactive glasses (0.05 g) were placed each in wells (1 cm in diameter) of pathogen-seeded nutrient agar, as particles or coated on 316 l stainless steel 1.0×1.5 cm sheets for agar diffusion method. The antibacterial test of vancomycin hydrochloride in different concentrations (25, 50, 75, and 100 mg/ml in distilled H
2
O) was carried out. The pathogen cell viability in presence and absence of glass composite was investigated using electron microscopy and cell count method. Nutrient broth (50 ml) was inoculated with
Staphylococcus aureus
along with 0.05 g of borate particles, incubated at 37°C and 150 rpm for 6 h. Then, the samples were examined under electron microscope, and the final pH was measured. A volume of 0.1 ml of each sample was further inoculated on solid nutrient agar, incubated at 37°C for 24 h, and then colony count was carried out.
Results and discussion
The borate bioactive glass was effective either as particles or coated on 316 l stainless steel. The other types of bioactive glasses coating the stainless steel produced a better antibacterial activity than the particles. The transmission electron microscope, showed the damaged bacterial cells of
S. aureus
after incubation with borate bioactive glass. The colony count of
S. aureus
after bioglass treatment was 18×10
2
, whereas in the control sample was 25×10
6
; the final pH was 10.4.
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Anti-urolithiatic activity of
Solanum nigrum
hydroalcoholic extract in ethylene glycol-induced urolithiasis in rats
p. 311
Abeer A.A Salama, Mahitab I El-Kassaby, Azza Hassan
DOI
:10.4103/epj.epj_21_19
Background and objectives
Urolithiasis is a growing public health problem. Asymptomatic kidney stones are kept under observation. The aim of this study was to explore the anti-urolithiatic activity of the hydroalcoholic extract of
Solanum nigrum
fruit in ethylene glycol-induced urolithiasis in rats.
Materials and methods
Urolithiasis was induced by oral administration of ammonium chloride 1% and ethylene glycol (0.75% v/v) in drinking water for 28 days. Hydroalcoholic extract of
Solanum nigrum
fruit (200 and 400 mg/kg) and cystone (750 mg/kg) were administered orally from the 15th day as a curative regimen.
Results and conclusion
Administration of ethylene glycol caused an elevation of serum creatinine, urea, calcium, and malondialdehyde and a reduction of magnesium and glutathione. In addition, renal content of tumor necrosis factor alpha was elevated and adiponectin renal content was reduced in urolithiatic control. Histopathological examination revealed tubular degeneration, dilatation, presence of calcium oxalate crystals in the lumen of renal tubules, and intense interstitial mononuclear cell infiltration in the lithiatic control group. Treatment with both doses of
Solanum nigrum
reversed all biochemical parameters and histopathological alterations. The results demonstrate that the hydroalcoholic extract of
Solanum nigrum
has
potent anti-urolithiatic activity against calcium oxalate urolithiasis induced by ethylene glycol through tumor necrosis factor alpha inhibition and adiponectin stimulation as well as in maintaining balance between stone promoter (calcium) and inhibitor (magnesium).
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Biosynthesis of biologically active chitinase utilizing some Egyptian chitinaceous wastes and the properties of the synthesized enzyme
p. 320
Heba M Shalaby, Salah A Abo-Sdera, Saadia M Easa, Abdel-Mohsen S Ismail
DOI
:10.4103/epj.epj_23_19
Background and objective
Chitin-degrading enzymes have an utmost practical importance in many fields such as medicine, agriculture, and industry. These enzymes are used as effective antibacterial, antifungal, antihypertensive, and antioxidant agents and also as excellent food quality enhancers. The objective of the present article was to formulate the production medium and to pinpoint the proper growth conditions for the chosen microorganism producing highly active chitinase enzymes. The general properties of the crude enzyme preparation were determined to define the proper conditions for enzyme action. Under the specified conditions, the capability of the enzyme preparation for antimicrobial and antioxidant activities were decided.
Materials and methods
Eighteen recommended microbial strains were screened for biologically active chitinolytic enzymes productivity. Chitinase enzyme was determined, and also the important properties of the crude chitinase were duly pinpointed. Finally, biological activities of the crude enzyme were studied.
Results and conclusion
and conclusion Among all the 18 organisms,
Streptomyces halstedii
H2 was the most potent producer of an influential chitinase enzyme. The maximum chitinase activity of 49.5 U/ml was obtained from medium contains glucose 6 g/l, ammonium nitrate (0.9 g/l), and urea (0.64 g/l) at 30°C and pH 9.0. The important properties of the streptomycetal chitinase were duly pinpointed as follows: optimum enzyme and substrate concentrations were 1.6 mg/ml and 1.4% (w/v), respectively, and optimum reaction pH and temperature were 7.2 and 45°C, respectively. The crude preparation was stable for 60 min at pH 7.2 and 30°C and retained 92.6% of the original activity. Under the specified conditions, at varying concentrations, the enzyme preparation exhibited considerable 2, 2-diphenyl-1-picrylhydrazyl radical scavenging activity accompanied with low antimicrobial activity, pointing out the partial purification necessity of the crude enzyme preparation.
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Purification and characterization of alkaline protease produced by
Streptomyces flavogriseus
and its application as a biocontrol agent for plant pathogens
p. 332
Elsayed E Mostafa, Moataza M Saad, Helmy M Hassan, Elsayed Hussien Zeidan
DOI
:10.4103/epj.epj_1_19
Background
A protease is an enzyme that helps proteolysis: protein catabolism by hydrolysis of peptide bonds. Proteases were the first enzymes to be commercialized, partly on the account of the history of availability and on account of need.
Aim
The aim of the present work is studying the properties of the partially purified enzyme and some application of the enzyme.
Materials and methods
In this study, alkaline protease was produced by
Streptomyces flavogriseus
using the shaking method. The enzyme was partially purified by ammonium sulfate precipitation. The most active fraction was evaluated on Sephadex G-100 column chromatography. The activity of the enzyme was determined at different temperatures and different pH values using various buffers.
Results and conclusion
In this study, alkaline protease produced by
S. flavogriseus
was partially purified by ammonium sulfate precipitation at 75% w/v saturation and pooled on Sephadex G-100 with a yield of 60.85% and specific activity of 85.14 U/mg protein. The enzyme was optimally active at 45°C in 0.5 mol/l potassium phosphate buffer (pH 8.0) after 30 min of incubation and was broadly stable at 45°C for 60 min and pH 6–8. The alkaline protease relative activity was increased in the presence Mn
2+
, Co
2+
, K
+
, Na
+
, Fe
2+
, and Mg
2+
at concentration 1 mmol/l, respectively.
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Genetic identification and optimization of novel β-glucosidase-producing
Lysinibacillus sphaericus
QS6 strain isolated from the Egyptian environment
p. 341
Ahmed F El-Sayed, Nivien A Abo-Sereih, Abeer E Mahmoud, Tahany M El-Kawokgy, Abbas A El-Ghamery
DOI
:10.4103/epj.epj_51_19
Background and objective
β-Glucosidase-producing bacteria are potential sources for biotransformation of lignocellulose biomass and agricultural wastes into biofuels. The aim was the isolation, screening, molecular identification, and optimization of highly efficient β-glucosidase-producing bacteria under different growth conditions.
Materials and methods
Cellulose-degrading bacteria were isolated and screened for β-glucosidase enzymes. Then, they were identified by phenotypic and genotypic identification. Optimization for β-glucosidase production was studied under different culture conditions.
Results and conclusion
Highly efficient β-glucosidase-producing strain QS6 was selected and identified morphologically and biochemically as Lysinibacillus sp. using 16 s rDNA gene sequencing approach and bioinformatics analysis. Strain QS6 was most similar to
Lysinibacillus sphaericus
, with similarity of 98%. Phylogenetic analysis was done to determine the relationship of strain QS6 with different strains of genus Lysinibacillus sp. It indicated that the suitable culture conditions of producing β-glucosidase were the culture temperature of 35°C, the initial pH of 7.0, the incubation time of 24 h, and 1% inoculum size. While studying the effect of carbon sources on β-glucosidase production, it was found that cellobiose (1%w/v) was the best carbon source for inducing β-glucosidase production. Moreover, the nitrogen source peptone at 0.5% w/v was optimum for β-glucosidase production by this bacterium.
L. sphaericus
QS6 was found to be sensitive to antibiotics (amoxicillin, streptomycin, tetracycline, cefadroxil, kanamycin, chloramphenicol, ampicillin, erythromycin, and tobramycin). Moreover, in-vitro antibacterial bioassay of the most potent β-glucosidase-producing strain (QS6) showed high antimicrobial activity against
Escherichia coli
(1.9 cm) and
Pseudomonas aeruginosa
1.0 cm). A promising Lysinibacillus sp. completely identified as
L. sphaericus
QS6 (GenBank MN493725.1) is an efficient source of β-glucosidase production.
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Foliar application of selenium and humic acid changes yield, essential oil, and chemical composition of
Plectranthus amboinicus
(Lour.) plant and its antimicrobial effects
p. 356
Ahmed E El-Gohary, Heba M Amer, Salah H Salem, Mohamed S Hussein
DOI
:10.4103/epj.epj_22_19
Background and objective
Plectranthus amboinicus
is an indigenous vegetable that can be freshly eaten. This plant is used for medicine to cure common illnesses such as cough, stomachache, headache, and skin infection.
Materials and methods
This study was conducted to study the effect of both selenium (2, 4, 8, 12, and 16 g/l) and humic acid (1.5 and 3.00 g/l), in addition to control, which was sprayed with water.
Results and conclusion
Generally, mass production of
P. amboinicus
(Lour.) plants has significantly increased as a result of application of different levels of selenium and humic acid treatments, compared with the control treatment. Essential oil percentage and yield (ml/plant) increased significantly as a result of selenium and humic acid treatments compared with control (S0H0). For essential oil constituents, the results clear that carvacrol (5.96–15.45%) is the first main compound followed by γ-Terpinene (6.74–11.80 %). The third main component is Limonene (3.23–11.32%), whereas the fourth one is α-Muurolene. Moreover, these treatments had a positive effect on selenium, total carbohydrates, photosynthetic pigments, and total phenolic content. Based on scavenging the stable ATBS [2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)] radical, all treatments increased significantly inhibition % especially S4H2 compared with untreated plants. Antibacterial and antifungal activities of
P. amboinicus
were studied.
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Pomegranate peels ameliorate renal nitric oxide synthase, interleukin-1β, and kidney injury molecule-1 in nephrotoxicity induced by acrylamide in rats
p. 368
Mohamed A.M Kandeil, Kamel M.A Hassanin, Mahmoud M Arafa, Hebatullah A Abdulgawad, Ghada M Safwat
DOI
:10.4103/epj.epj_25_19
Background and objectives
Acrylamide (AA) is considered a toxic intermediate product of the Millard reaction and liberated in high-carbohydrate foods exposed to high temperatures. AA formed during baking, frying, roasting, or grilling of such food. Various studies have recorded the toxic effects of AA on many biological functions. The aim of our study is to determine such toxic effect on the kidney and the prophylactic role of pomegranate peels (PP) as a waste portion of the fruit.
Materials and methods
In this study, 60 male albino rats were administered 40 mg/kg body weight AA orally for 17 consecutive days. To evaluate the nephrotoxic effects of AA, some biochemical parameters were measured. Also, 200 mg/kg body weight PP were administered orally as a prophylaxis for 31 consecutive days.
Results and conclusion
In the AA group, alterations in renal function were observed, which was evident from significantly high levels of urea, creatinine, and uric acid. Estimation of serum and urine electrolytes (Na
+
, K
+
, and Cl
−
) showed electrolyte imbalance as well. AA-induced renal oxidative stress was expressed as low levels of renal antioxidants (glutathione, glutathione peroxidase, and superoxide dismutase) and high levels of renal oxidants (malondialdehyde and nitric oxide). To clarify the Pathogenesis of AA nephrotoxicity, estimation of renal nitric oxide synthase and interleukin-1β is carried out showing high significant level. Direct damage in renal tissue is resembled in high level of renal kidney injury molecule-1. As stated before, the administration of AA resulted in acute nephrotoxicity, whereas PP played a vital role in reducing this toxicity. Lower levels of urea, creatinine, and uric acid were observed in the AA+PP group and electrolyte balance was achieved, indicating the prophylactic effect of PP. PP showed antioxidant activity as higher levels of glutathione, glutathione peroxidase, and superoxide dismutase recorded and also lower levels of nitric oxide and malondialdehyde controlling oxidative stress of AA. The levels of kidney injury molecule-1, interleukin-1β, and nitric oxide synthase improved significantly. Finally, we can state that PP could ameliorate the nephrotoxic effect of AA.
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Euphorbia lathyris
L.-modulated hyperlipidemia induced by high-fat diet in an experimental animal model
p. 377
Eman A Younis, Khaled Mahmoud, Hanan F Aly
DOI
:10.4103/epj.epj_28_19
Background and objectives
Hyperlipidemia has been embroiled in atherosclerosis, which is the primary factor of heart disease and stroke. An important aim in the pivotal restrain of atherosclerosis awaits an affordable and safe drug that will fundamentally confirm to have a great effect where treatment is decisive. Humans have been served by a variety of drugs extracted from a natural source, and the most of these cures were provided by higher plants.
Materials and methods
The aim of this study is to investigate and assess the hypolipidemic effect of
Euphorbia lathyris
L. extracts on high-fat diet-induced hyperlipidemia in rats, and it shows a scientific proof for expansion of
E. lathyris
L. as a prospect naturalistic oral hypolipidemic drug. Rats were administrated orally with cholesterol in a dose of 100 mg/1 ml olive oil/1 kg five times a week for 12 consecutive weeks. This was accomplished by assessing the activities of particular biochemical parameters associated with liver functions in tissue of high-fat-diet group when compared with those of control one. Liver function tests, included aspartate aminotransferases; alanine and alkaline phosphatases; antioxidants and oxidative stress biomarkers, such as glutathione, catalase, and glutathione-S-transferase; total antioxidant capacity; nitric oxide; and lipid peroxide. In addition, lipid profile (cholesterol, total lipid, high-density lipoprotein-cholesterol, and triglyceride) were also evaluated. Moreover, histological examination of liver, heart, and kidney was done to prove the biochemical outcomes.
Results and conclusion
The present findings strongly clarify defects in all biochemical measurements, such as increase in the malondialdehyde level and liver function enzyme activities. However, reductions in enzymatic and nonenzymatic antioxidant in liver of hyperlipidemic rats were marked. On the contrary, treatment of hyperlipidemic rats with low and high dose of methanol and petroleum ether extract of
E. lathyris
natural product proved to have ameliorating effects in all biochemical parameters under investigation. The previous biochemical observations were also accompanied by histopathological examinations of the heart, liver, and kidney sections, which showed significant recovery of hepatocytes and amelioration in histological architectures of heart and kidney. It could be concluded that the two extracts of
E. lathyris
showed improvement in liver function enzyme activities, oxidant–antioxidant biomarkers, and lipid profile activities in liver of high-fat-diet rats.
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Optimization and purification of cellulase produced by
Penicillium decumbens
and its application
p. 391
Nehad E.A., Yoness M.F., Reem A.A.
DOI
:10.4103/epj.epj_31_19
Background and objectives
Cellulases are an enzyme group based on catalytic action. They include endocellulase, exocellulase, beta glucosidase, cellulose phosphorylases, and oxidative cellulases. This work was aimed at the production of cellulase by fungal strains from
Penicillium
sp. Selection of the best organism that gives the highest productivity of the enzyme, examination of the cellulase production under the optimum conditions, purification of cellulase, identification by high-performance liquid chromatography, and its application in clarification and yield increase of apple juice were also studied.
Materials and methods
Three strains of
Penicillium
sp. were examined using the method of Congo red for cellulase production. The factors affecting cellulase production by fungus
Penicillium decumbens
were identified. Cellulase produced by
P. decumben
s was purified using ammonium sulfate precipitate (80% saturation) followed by ion exchange chromatography by Sephadex G-200. High-performance liquid chromatography technique was used to measure the purity of cellulase produced from
P. decumbens
. The cellulase enzyme was used to increase the yield of apple juice and apple juice clarification, as examples of its application in the food industry.
Results and conclusion
The
P. decumbens
colony proved to have the largest decolorization zone, and the cellulase produced was a large amount (21.5 U/ml). The highest activity of cellulase was seen in the media containing 50% carboxymethylcellulose and 50% dates molasses waste (Dibs) as carbon source after incubation for 6 days, and the optimum pH and temperature for the production cellulase were pH 4.0 at 30°C. Utilization of 80% ammonium sulfate gave pure enzyme cellulase (38.25 U/ml) and has a high degree of specific activity (25.5 U/mg protein). Cellulase activity of 42 U/ml and the degree of specific activity of 46.6 U/mg protein, with a 4.3-fold purification of cellulase, with 42% recovery from the crude cellulase, was obtained with Sephadex G-200. The results revealed an increase in the quantity of produced apple juice treated by cellulase enzyme.
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Genetic improvement of fungal β-mannanase and its molecular differentiation
p. 403
Nivien A Abosereh, Siham A Ismail, Om K.H Khattab, Shaimaa A Nour, Amany A Abo-Elnasr, Amal M Hashem
DOI
:10.4103/epj.epj_32_19
Background
and objective β-Mannanase is an enzyme that has great potential in many industrial application including feed, food, pharmaceutical, cosmetics, production of mannan and manooligosaccharides, pulp and paper, bioethanol and biodiesel productions, and oil and textile industries. The aim of this study was to describe the potential of gamma and ultraviolet (UV) rays to optimize the production of industrially important β-mannanase enzyme by subjecting
Penicillium citrinium
Egy5LC368457 to these rays.
Materials and methods
Various doses and times of UV and gamma irradiation were used. Genetic diversity was resolved by mistreatment with the Random Amplified Polymorphic Polymer (RAPD-PCR) technique. Ten RAPD oligonucleotide primers amplifying DNA of β-mannanase showed reproducible banding patterns.
Results and conclusion
The results of this study revealed the highest β-mannanase activity was produced by gamma ray 150 Gy (37.42 IU/ml) with 2.27-fold higher than the wild type. A total of 64 bands were obtained from nine of these markers with 44% polymorphic bands. The size of the amplified bands ranged between ∼75 and 3000 bp. The genetic polymorphism value of each primer was determined, which ranged between 2 and 9 bands. The primer efficiency of amplification ranged between 3.13 and 23.44%, and the discriminatory power ranged from 4.5 to 25. In conclusion, UV and gamma ray irradiation can induct mutations, which can be carefully acclimatized and commercially propagated under suitable condition. RAPD technique could be successfully applied to the newly β-mannanase and can differentiate mutants.
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The potential of cultivated milk thistle by-products as cancer chemopreventive and anti-inflammatory drugs
p. 411
Sherin K Ali, Nahla S Abdel-Azim, Ali K Khalil, Mohamed-Elamir F Hegazy, Tarik A Mohamed, Ahmed R Hamed, Khaled A Shams, Faiza M Hammouda
DOI
:10.4103/epj.epj_34_19
Background and objectives
Seeds of
Silybum marianum
(milk thistle), growing wild in Egypt, have been used since ancient times in traditional medicine. This study aims at identifying chemical and bioactivity properties of the oil of the seeds of cultivated
S. marianum
, obtained by cold pressing, as well as the extracts of the leaves. The prepared extracts were tested for their cancer chemoprevention and anti-inflammatory activities.
Materials and methods
The phytochemical constituents of cold-pressed seed oil and extracts of the leave were determined using gas chromatography–mass spectrometry and high-performance liquid chromatography (HPLC), respectively. The prepared extracts were tested for their cancer chemopreventive and anti-inflammatory activities.
Results and conclusion
HPLC profiling of leaves extract indicated that gallic acid and naringenin are the major phenolic acid and flavonoid, respectively. Additionally, HPLC analyses indicated the presence of seven main active components of silymarin in seeds. The total extract from leaves caused a moderate NAD(P)H-quinone oxidoreductase 1 protein induction and inducible nitric oxide synthase protein expression inhibition.
Conclusion
Cultivated
S. marianum
(milk thistle) by-products (oil and leaves) could have possible applications in food and pharmaceutical industry
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Molecular characterization of alkaline protease-coding gene from
Bacillus licheniformis
MK90 mutants with biofilm inhibitory activity
p. 419
Ahmed A Hamed, Mohamed Khedr, Mohamed Abdelraof
DOI
:10.4103/epj.epj_47_19
Background
Enzymes are organic materials that accelerate biochemical processes without themselves undergoing change. They can be produced by plant, animal, fungi, and bacteria. Bacterial proteases are much favorable than any other sources, because bacteria can grow quickly and can be easily cultivated in laboratory.
Objective
To isolate and screen bacteria from soil samples for their ability to produce alkaline protease, and to improve the alkaline protease production followed by evaluation of its antimicrobials and antibiofilm activity.
Materials and methods
Sample collection was carried out from different locations in Egypt. Isolation of bacteria from soil samples was done using serial dilution method on skim milk agar. All isolated bacteria were screened for their ability to produce protease enzyme. The bacterial isolate showing maximum alkaline protease activity was identified using 16 S rRNA genetic identification. To induce mutations, ultraviolet (UV) irradiation was used. The most active mutant strains were selected for production, purification, and characterization of alkaline protease followed by evaluation of alkaline protease antimicrobial and antibiofilm activity.
Results and conclusion
Three UV mutants (MT2, MT4, and MT26) out of 48 displayed proteolytic activity more than other mutants and wild type (WT).
Bacillus
alkaline extracellular protease gene was genetically characterized through isolation of the genomic DNA of
Bacillus licheniformis
MK90 WT, and the best protease-producing UV mutants were followed by amplification, sequencing, and analyses. WT strain and best protease-producing mutants were compared at proteomic level through sodium dodecyl sulfate polyacrylamide gel electrophoresis for total cellular proteins. Then protease enzyme of WT and mutants was purified and characterized. This study reports that the
B. licheniformis
protease was active at an alkaline pH and wide range of temperatures (40–60°C), reflecting its potential application in detergent and laundry industries. On the contrary, the antibiofilm activity of the protease enzymes was evaluated toward four pathogenic bacterial strains, i.e.,
Staphylococcus aureus
,
Pseudomonas aeruginosa
,
Escherichia coli
, and
Bacillus subtilis,
and the results showed that proteases from
B. licheniformis
MK90 may be useful for controlling biofilm formation by some pathogenic bacteria.
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ERRATUM
Erratum: Potential effects of silver nanoparticles, synthesized from Streptomyces clavuligerus, for controlling of wilt disease caused by Fusarium oxysporum
p. 434
DOI
:10.4103/1687-4315.277082
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