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   2017| January-April  | Volume 16 | Issue 1  
    Online since May 8, 2017

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Statistical optimization of L-asparaginase production by using Fusarium solani
Abeer A El-Hadi, Heba A El-Refai, Mona S Shafei, Rania Zaki, Hanan Mostafa
January-April 2017, 16(1):16-23
Background and objective The enzyme l-asparaginase is important as a therapeutic agent in the treatment of acute lymphocytic leukemia. It has been observed that microorganisms such as yeast and filamentous fungi such as Aspergillus spp., Penicillium spp., and Fusarium spp. are commonly produced asparaginases with fewer adverse effects. Fusarium solani was selected to be the most potent microbial isolate for l-asparaginase production. The factors controlling l-asparaginase production, such as containing media, were implemented to increase the yield of l-asparaginase. Statistical experimental design such as the Plackett–Burman enables finding out the most effective factors that increase the yield of l-asparaginase. Materials and methods Aspergillus rubber, Aspergillus terreus, Epico niger, Penicillium cyclopium, and F. solani were chosen for screening the l-asparaginase activity using phenol red – a pH indicator. l-Asparagine and the modified Czapek Dox media were tested for the production of l-asparaginase from F. solani under different incubation times. To optimize l-asparaginase production by using F. solani, the combined effect of seven variables (l-asparagine, sucrose, NaNo3, K2HPO4, MgSO4, KCl, and time) were assessed using the central composite design. Results and conclusion All the four fungal strains screened for l-asparaginase activity showed positive results in the rapid plate assay method. The medium employed contained asparagine with phenol red, and after incubation, pink zones around the colonies were observed. F. solani was a potential source for a high yield of l-asparaginase enzyme and high substrate specificity. Modified Czapek medium II was the most suitable for l-asparaginase, showing an activity of 121 U/ml. The result on l-asparaginase activity according to the screening Plackett–Burman experiments gives a medium composed of the following (g/l): sucrose (1); l-asparagine (0.5); KH2PO4. (0.75), MgSO4 (0.7); H2O (0.72); KCl (0.72); (NH4)2SO4 (11.7) at an incubation time of 3 days. For further investigation, x1 K2HPO4, x2 sucrose, and x3 time were found to be the most significant variables affecting l-asparaginase activity. The second optimization step was to identify optimal values of the three factors that bring about maximum l-asparaginase activity, using the central composite designed experiment. The results showed that K2HPO4 (2.5 g/l), sucrose (4 g/l), and the time (8 days) were critical in the production of l-asparaginase.
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Protective effect of pumpkin seed extract against testicular toxicity induced by tramadol in adolescent and adult male albino rats: a light and electron microscopic study
Fatma M Minisy, Ahmed A Massoud, Enayat A Omara, Fatma G Metwally, Nabila S Hassan
January-April 2017, 16(1):43-52
Background Tramadol overdose is one of the most common causes of drug poisoning in Egypt, especially in young men with a history of substance abuse and mental disorders. Objective The aim of this study was to investigate the effect of tramadol on the testis and the possible protective role of pumpkin seed extract (PSE) in adolescent and adult albino rats. Materials and methods Twenty-four healthy growing male Wistar rats aged 3 weeks (adolescent) and another 24 adult rats aged 6 weeks were included in this study. Each group was divided into four equal groups and treated for 3 weeks; these four subgroups were the control group, the PSE alone-treated group (at 40 mg/kg body weight), the tramadol-treated group (at 20 mg/kg body weight/day for the first week and at 40 and 80 mg/kg for the second and third weeks, respectively) and the combination group treated with PSE (at 40 mg/kg body weight) and tramadol (at 20 mg/kg body weight/day in the first week and 40 and 80 mg/kg in the second and third weeks, respectively). At the end of the experiment, testicular tissue samples were collected and processed for histological and ultrastructural examination. Results and conclusion The histopathological examination of the tramadol-treated groups (adults and adolescents) indicated severe structural changes in germ cells in the form of degenerated apoptic cells, exfoliation of germ cells in the lumen of seminiferous tubules, hemorrhage, vacuolization, disorganization, and marked loss of cells. But PSE succeeded in protecting the testicular tissue and diminished the atrophic changes. It could be concluded that PSE has a prophylactic effect against tramadol-induced testicular damage and this action was more pronounced in adults than in adolescents.
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Evaluation of antidiabetic and antianxiety potential of Kalanchoe pinnata root standardized extracts
Varsha Tiwari, Anita Singh, Abhishek Tiwari
January-April 2017, 16(1):24-36
Background Kalanchoe pinnata is an important medicinal plant and possesses numerous medicinal values including antidiabetic potential. Aim The aim of this study was to explore the antidiabetic and antianxiety potential of K. pinnata root standardized extracts. Materials and methods Standardization of the extracts included the physicochemical and fingerprinting analysis of the extracts. Pharmacological studies included acute toxicity, antianxiety, the oral glucose tolerance test, and streptozotocin (STZ)-induced antidiabetic screening of standardized extracts. Diabetes was induced by a single intraperitoneal injection of STZ (65 mg/kg). Fasting blood glucose (FBG) levels were monitored in normal and diabetic rats at regular intervals of 0, 7, 14, and 21 days of the experiment. The antianxiety potential of ethyl acetate and ethanol extract at the doses of 100, 200, and 300 mg/kg was evaluated using an elevated plus maze model. Results Ethyl acetate extract at 300 mg/kg was found to have significant antianxiety potential, whereas ethanol extract at 100, 200, and 300 mg/kg failed to show a statistically significant antianxiety effect. Alcohol and aqueous extracts were found to show a statistically significant (P<0.001) decrease in FBG at 400 mg/kg, whereas both extracts at low doses failed to show a significant decrease in FBG on the oral glucose tolerance test. Treatment of STZ-induced diabetic rats with the alcohol and aqueous extract (200 and 400 mg/kg) led to a statistically significant (P<0.001) reduction in FBG of the diabetic rats after the 21st day of treatment. Conclusion The present study re-established the plant as an effective antidiabetic and antianxiety armament in fighting with the challenge of diabetes mellitus, expected to cover most of Europe and Asia in coming 20 years.
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Hydrogen sulfide donors or related derivatives are the future medicines of renal diseases
Marwan S.M. Al-Nimer
January-April 2017, 16(1):1-7
Hydrogen sulfide (H2S) is one of the three gasotransmitters that possess anti-inflammatory, antiapoptotic, and antioxidant properties. It maintains the function of the kidney through its effect on the glomeruli and the renal transport system. Literature review using PubMed, Excerpta Medica database (EMBASE), Google scholar, and Cochrane review revealed that H2S donors are introduced as exogenous H2S and have been found to target many organs in in-vitro and in-vivo studies. This review provides the main research that was performed on the H2S donors in the context of kidney disease. Exogenous H2S supplementation can be administered in different therapeutic areas promising therapeutic strategy in the setting of kidney diseases. Therefore, suitable pharmaceutical preparations of H2S donors are necessary to be launched in the markets for the prevention and treatment of acute/chronic renal diseases.
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Erratum: Assessment the microbiological and molecular aspects of soil isolated bacteria that suppress Pythium ultimum in Abha/KSA

January-April 2017, 16(1):69-69
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Effect of compounds identified in the active fraction of pericampylus glaucus on blood glucose and lipid profiles in streptozotocin-induced diabetic rats
Muhammad Kifayatullah, Mohd. Shahimi Mustapha, Md. Moklesur Rahman Sarker, Izharullah , Muhammad Amin
January-April 2017, 16(1):8-15
Objective The present study was designed to determine the effect of compounds identified in the active fraction of Pericampylus glaucus on blood glucose and lipid profiles in streptozotocin (STZ)-induced diabetic rats. Materials and methods Initially, the antidiabetic activity of petroleum ether, n-hexane, chloroform, and ethanolic extracts of P. glaucus was evaluated in STZ-induced diabetic rats at a dose of 400 mg/kg body weight. Then, the potential extract was fractionated by different solvent systems to obtain various fractions. Next, fractions were investigated again at a dose of 100 mg/kg body weight to find the active fraction. The active fraction was examined using STZ-induced diabetic rats for 21 days. The blood glucose levels were observed weekly and various biochemical parameters were determined on the day the rats were killed. Results The active fraction was subjected to gas chromatography mass spectrometry to find the compounds present in the active fraction of ethanolic extract of P. glaucus. The ethanolic extract was noted to have significant (P<0.001) antidiabetic activity in diabetic rats compared with the other extracts. Four fractions (FA, FB, FC, and FD) were collected from the active ethanolic extract. Among these, fraction B, which was collected from a mixture of petroleum ether and ethyl acetate, was found to have a high (P<0.001)-attenuation effect on blood glucose levels compared with the others, except for ‘fraction D’ (P<0.01), which was collected from ethanol. The data showed that the active fraction (fraction B) also induced significant (P<0.05, <0.01, <0.001) attenuation in the levels of triglyceride, total cholesterol, and LDL and a significant (P<0.01) improvement in the level of HDL. In gas chromatography mass spectrometry analysis, 10 peaks were noted, which suggests the presence of 10 compounds in the active fraction responsible for the blood glucose-lowering effect and biochemical parameters. Conclusion The present study confirmed the presence of compounds identified in P. glaucus responsible for the attenuation of blood glucose and lipid parameters.
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Possible role of a compound identified from Pericampylus glaucus plant in controlling blood glucose in experimental animals
Muhammad Kifayatullah, Mohammad S Mustapha, Mohammad M.R. Sarker, Kaleemullah , Muhammad Amin, Fawad Ali
January-April 2017, 16(1):37-42
Objective The aim of the present research work was to determine the possible mechanism of an active fraction identified from Pericampylus glaucus plant in controlling blood glucose in experimental animals. Materials and methods The effect of an active fraction from P. glaucus on intestinal absorption of glucose at a dose of 50 mg/kg (body weight) was investigated using an in-vivo method. The identification of the compound was carried out with gas chromatography mass spectrometry followed by 1H nuclear magnetic resonance. The active fraction and standard drug were given daily for 7 days. The animal groups were anesthetized through intraperitoneally injection with sodium pental at a dose of 40 mg/kg (body weight) after they were made to fast overnight to evaluate the effect of the active fraction on the absorption of glucose in comparison with the diabetic group. Results and discussions A single dose of 50 mg/kg (body weight) of the active fraction and standard drug acarbose produced significant (P<0.001) attenuations in the intestinal absorption of glucose as compared with the diabetic group. The amount of glucose absorbed was 94.05±0.9 mg/g tissue (weight) in the diabetic group, whereas that absorbed in the active fraction-treated group was 55.80±1.2 mg/g tissue (weight). However, the animals treated with standard drug acarbose showed a significant reduction in glucose absorption rate at 38.21±2.5 mg/g tissue (weight). The inhibition of glucose absorption by standard acarbose and active fraction was 59.37 and 40.66%, respectively, as compared with the diabetic control group. The gas chromatography mass spectrometry analysis indicated a single compound propanoic acid in plant sample. The molecular weight was 234, 1H nuclear magnetic resonance was 452 MHz, %area was 3.57, retention time was 2.990, and total area was 1 015 462. The molecular formula was C9H22O3Si at m/z 73 (high resolution). The presence of this compound in P. glaucus might be responsible for the inhibition of glucose absorption by blocking Na+, K+ ATPases activity. Conclusion The present experiment confirmed reduced glucose absorption rate and the antidiabetic activity is due to this possible mechanism.
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Assessment of some synthesized novel 9-substituted tetrahydroacridine derivatives in diabetic disease management in rats
Rehab M Abdel Megeed, Hassan Z Ghanem, Mohsen M Kamel, Maged F Bakeet, Abdel-Hamid Z Abdel-Hamid
January-April 2017, 16(1):53-61
Background and objective 9-Substituted 1,2,3,4-tetrahydroacridine derivatives, especially those bearing substituted aliphatic or aromatic amino groups at the position 9, have been widely used for treating some chronic diseases because of their role as acetylcholine esterase inhibitors. Therefore, some new substituted tetrahydroacridine derivatives were synthesized to investigate their efficiency as antidiabetic agents in vivo. Materials and methods Some new substituted tetrahydroacridine derivatives hybridized at their position 9 with nitrogen, oxygen, and/or sulfur heterocycles or sulfa drugs starting from the known intermediate compound 9-chlorotetrahydroacridine were synthesized. Biologically, these compounds were investigated for antidiabetic potentials using Wistar rats. Diabetes was induced using streptozotocin (45 mg/kg) and then biochemical assays and histopathological examinations were applied to assess the therapeutic efficiency and safety margins of these compounds. Results and discussion Biochemical and histopathological examinations demonstrated the efficiency and safety margins of these compounds in vivo. Different biochemical analyses and histopathological examinations were estimated in diabetic and treated groups as compared with the healthy one. Data listed in this study showed an acceptable improvement percentage in glucose level and α-amylase, liver function enzymes (glutamic pyruvic transaminase and glutamic oxaloacetic transaminase), and lipid profile (triglyceride, low-density lipoprotein, total cholesterol) parameters after treatment with new synthetic compounds. Histopathological examination showed regeneration of treated groups. 9-Sulfadiazine-tetrahydroacridine (9-SDTHA) derivative (2b) was the most safe and efficient compound. The superiority of this compound may be because of the presence of a pyrimidine ring, which is the main constituent of DNA and RNA structure. These compounds could be suggested as antidiabetic agents.
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Development and evaluation of nanosized aripiprazole-loaded bioflexy films using a biopolymer from Lagenaria siceraria for brain delivery through orosoft palatal mucosal platform
NV Satheesh Madhav, Vishakha Jaiswal, Abhijeet Ojha
January-April 2017, 16(1):62-68
Purpose The National Institute of Mental Health estimated the global cost of mental illness to be $6T by 2030. The purpose of the study was to deliver aripiprazole for brain targeting by suitably designing a bioflexy film for the treatment of different brain disorders. Materials and methods Bioflexy films were prepared by solvent casting technique using aripiprazole as a model drug, different concentrations of isolated biopolymer from Lagenaria siceraria, and a standard polymer. Results All formulations of aripiprazole were flexible, smooth, and transparent in nature, with a weight range from 23.73 to 34.48 mg, pH range from 7.27 to 7.39, and folding endurance of 94–116 times. The content uniformity was in the range of 97.2–98.6. Conclusion In-vitro and in-vivo release shows FL2 as the best formulation. On the basis in-vitro and in-vivo drug release, FL2 was found to be best formulation showing mucoadhesivity (36 h), t50% of 2.5 h, t80% of 23.4 h, and having R2 value of 0.9923, best-fit model Higuchi matrix analyzed by BIT-SOFT 1.12. Moreover, it was observed that significant amount of drug reaches to the brain through soft palatal route through neural pathway.
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