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   2020| January-March  | Volume 19 | Issue 1  
    Online since March 24, 2020

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Optimizing the production of rice straw hydrolytic cellulase under solid-state fermentation using Aspergillus terreus RS2
Shaymaa A Ismail, Amira A Hassan
January-March 2020, 19(1):7-17
Background The enzymatic hydrolysis of lignocelluloses into fermentable sugars is the key step in biorefining, and cellulases are the key enzymes. The low titer of cellulase production and their high cost remain the most significant barriers to their industrial applications. The aim of this study was the economic production of cellulase by an Egyptian fungal isolate under solid-state fermentation by using rice straw as a carbon source. Additionally, the produced enzyme can be applied in the hydrolysis of rice straw and production of free sugars that can be used in several biotechnological industries. Materials and methods The isolated fungus was identified according to its cultural and morphological features followed by 18S rDNA sequencing. Optimization of the enzyme productivity was performed by applying Plackett–Burman and Box–Behnken designs. Finally, the hydrolysis of rice straw for the production of fermentable sugars using the produced enzyme was carried out, confirmed by scanning electron microscope and thin-layer chromatography analysis. Results and conclusion The optimum cellulase activity produced by the isolated fungus Aspergillus terreus RS2 was 124.94 U/g dry substrate. It was achieved at the optimum conditions that were conducted to be as follows: 3.75 g (1.5%w/v) of rice straw moistened with 11.25 ml (1 : 3 biomass to moistening agent) of modified Mandel’s medium [1% (NH4)2SO4 was the only constituent nitrogen source] of pH 7, incubated at 30°C for 8 days. Finally, the activity of the produced enzyme in the degradation of rice straw indicated the release of reducing sugars of 343.98 mg/g dry substrate with a saccharification percentage of 34.4% recorded after 4-h hydrolysis period.
  1,760 261 1
Prevalence of vitamin D deficiency and its relationship with epileptic severity in a pediatric department of tertiary care hospital
Chandana Kamili, Shivaram Prasad, Battula Harsha Vardhan, Kompelli Koushik, Nadigittu Akshita, Sanjana Yadav, Sharadha Radhakrishnan, Konde Abbulu, Venu Kola, Prasenjit Mondal
January-March 2020, 19(1):1-6
Aim To evaluate the prevalence of vitamin D deficiency in pediatric epileptic patients and to assess if there is any correlation between vitamin D level and disease severity in epileptic patients. Objective To estimate vitamin D deficiency and insufficiency as percentage of enrolled patients in pediatric department and to identify if there is any correlation between vitamin D deficiency and factors such as age, sex, BMI, total serum calcium levels, antiepileptic drugs, and types of seizures. Patients and methods A prospective interventional clinical study was carried out in a tertiary care hospital between September 2018 and February 2019. We included 30 pediatric epileptic patients of age group 1–14 years from whom the blood samples were withdrawn and were sent to the laboratory for the estimation of vitamin D levels. Results The data collected was analyzed using statistical method χ2 and analysis of variance in Prism 8.0 version. The prevalence of vitamin D deficiency and its correlation with epilepsy was determined. In this study, we found that there was a high prevalence of vitamin D deficiency in pediatric epileptic patients (23 patients out of 30–73%). The other risk factors for vitamin D deficiency found in this study are children of age group 5–10 years (43%), malnourished patients (40%), hypocalcemic patients (40%), and patients who use antiepileptic drugs for more than 2 years (57%). Conclusion Based on the aforementioned scientific analysis, it can be concluded that there is a high prevalence of vitamin D deficiency in pediatric patients with epilepsy. So, vitamin D3 supplements should be given for all epileptic children even before the initiation of antiepileptic therapy. Patients’ parents were educated on the importance of vitamin D.
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Optimization and comparative studies on activities of β-mannanase from newly isolated fungal and its mutant
Om K.H Khattab, Siham A Ismail, Nivien A Abosereh, Amany A Abo-Elnasr, Shaimaa A Nour, Amal M Hashem
January-March 2020, 19(1):29-46
Background and objective β-Mannanase has potential industrial applications in pharmaceutical field, bioethanol production, coffee extraction, food and feed technology, etc. So finding a new and promising enzyme source is a very important issue. The aim of this study was to improve the biosynthesis of β-mannanase by different techniques, such as mutation and optimization of the culture parameters. Materials and methods Five fungal isolates were tested for the production of β-mannanase. Enzyme activity, protein content, and specific activity were determined. The most potent isolated microorganism and its mutant were identified by using Transmission Electron Microscopy and 18SrDNA sequencing and phylogenetic analysis. Ultraviolet and gamma rays were used. Optimization studies were done to maximize enzyme production from the most potent microorganism and its highly productive and stable mutant, including culture conditions and medium compositions, and statistical optimization was also carried out. Primary characterization of β-mannanase was studied. Results and conclusion In our research, we found a stable mutant strain obtained by using gamma radiation at 150 GY. The first step of the fermentation, optimized by one-factor-at-a-time technique, increased the biosynthesis of β-mannanase for Penicillium citrinium 150 GY from 65.9 to 219 IU/ml compared with the wild strain, which increased from 16.82 to 26.5 IU/ml. Statistical optimization improved P. citrinium 150 GY β-mannanase from 219 to 296 IU/ml by applying Plackett–Burman design and increased the level of β-mannanase biosynthesis to 351 IU/ml. Primary characterization of β-mannanase produced by P. citrinium and P. citrinium 150 GY proved that they are almost the same, except in a little shift to higher value (5°C) in optimum temperature.
  1,094 176 1
Body composition changes in asthmatic children regarding steroid therapy duration
Rokia A El-Banna, Safenaz Y El Sherity, Maya G Aly
January-March 2020, 19(1):18-24
Background Childhood asthma is markedly increasing in developing countries. The first line of management according to national asthma guidelines is inhaled corticosteroids. Accurate body composition analysis, with persistent asthma control by steroid therapy, is essential in childhood, as the potential effects of the long-term and short-term treatments are still a matter of concern. Aim To assess the body composition (total and regional) distribution in Egyptian children with asthma receiving inhaled steroid with different duration therapy. Patients and methods Body composition was measured by dual-energy X-ray absorptiometry in a cross-section study of 160 Egyptian prepubertal school-aged children (4–12 years). There were 60 asthmatic children controlled by long-term steroid therapy and 60 asthmatics controlled by short-term steroid therapy, who were compared with 40 healthy children to assess the effect of duration of therapy. Results Asthmatic children controlled by long-term steroid therapy had significantly higher weight, BMI, and total and chest lean masses among both sexes within all different age groups (P<0.05). However, asthmatic female children controlled by long-term therapy had significantly higher total and pelvic fat masses rather than both short-term and healthy groups (P<0.05). Moreover, the body composition parameters (total and regional) had positive significant correlations with BMI. Conclusion The total lean mass, specifically chest lean mass, is increased in asthmatic children who are controlled by long-term steroid therapy, and evaluation of those children using dual-energy X-ray absorptiometry provides an accurate analysis of both total and regional body composition.
  977 142 -
Insights into the in-vitro hypocholesterolemic, antioxidant, antirotavirus, and anticolon cancer activities of the methanolic extracts of a Japanese lichen, Candelariella vitellina, and a Japanese mushroom, Ganoderma applanatum
Waill A Elkhateeb, Ghoson M Daba, Asmaa N El-Dein, Donia H Sheir, Walid Fayad, Mohamed N Shaheen, ELmahdy M Elmahdy, Ting-Chi Wen
January-March 2020, 19(1):67-73
Background and objective Medicinal mushrooms, as well as lichens are potential sources of bioactive compounds against many lethal diseases. Therefore, in-vitro evaluations of the biological activities of the 80% methanolic extracts of two Japanese originated samples, namely, the mushroom Ganoderma applanatum, and the lichen Candelariella vitellina have been performed. Materials and methods Extraction of C. vitellina and G. applanatum was conducted using 80% methanol. The resulted extracts were examined for their in-vitro hypocholesterolemic and antirotavirus activities, as well as antihuman colon cancer action. Results and conclusion It was revealed that both extracts have potent hypocholesterolemic effects. G. applanatum in concentration of 24 mg/ml could successfully reduce the cholesterol concentration to 100±0% after 96 h of incubation at room temperature, whereas reaching this degree of reduction in cholesterol concentration required using 32 mg/ml of C. vitellina extract. On the contrary, the extract of C. vitellina at concentration of 1.0 mg/ml exhibited a stronger DPPH scavenging activity (99.5±0.166%), in comparison with 66.24±0.43% in case of using the same concentration of the G. applanatum extract. C. vitellina extract showed antirotavirus in a therapeutic index of 11, whereas G. applanatum extract recorded 3.4 only. The antihuman colon cancer study elucidated that both extracts have a moderate activity toward HCT116 human colon carcinoma. C. vitellina extract achieved a cytotoxicity of 57.90±4.4%, whereas the extract of G. applanatum displayed only 43.10±0.9%. Studies on G. applanatum and C. vitellina introduce them as promising sources of biologically potent compounds that can be candidates for use in treatment of many diseases after further studies.
  890 129 2
Gene amplification and overexpression of Bacillus subtilis L-asparaginase
Wafaa K Hegazy, Mohamed S Abdel-Salam, Maysa E Moharam
January-March 2020, 19(1):25-28
Background and objectives The aim of this paper was to focus on improving production level of l-asparaginase by developing recombinant strains. Materials and methods Asp gene was cloned into the shuttle vector pNW33N, and the recombinant plasmid was used to transform Bacillus subtilis protoplast. Asp gene was expressed into both Escherichia coli and B. subtilis. Enzyme activity of the recombinant strains was measured as compared with wild-type strains. Results Asp gene was successfully subcloned into the recombinant plasmid named S-ASP-NRC-27. The gene was expressed efficiently in both host strains: E. coli and B. subtilis. The enzyme activity of the transformants was increased up to threefold under control of Lac Z promoter. Conclusion From the previous results, the shuttle vector pNW33N seemed to be a very useful plasmid as a cloning vector in a wide variety of the genus Bacillus. Both of Asp gene amplification and the control of Lac Z promoter had direct effects on producing the super Asp-expression strains.
  865 143 -
Investigation of lipoidal matter of Chenopodium quinoa seeds and its cytotoxicity potential against three human cancer cell lines
Shaymaa M Bata, Khaled A Shams, Ali Kh Khalil, Rasmia A Hassan, Ibrahim A Saleh, Ahmed R Hamed, Nahla S Abdel-Azim, Faiza M Hammouda
January-March 2020, 19(1):47-54
Background and objectives Quinoa (Chenopodium quinoa Willd) is a plant species that belongs to Chenopodiaceae family. It has a high nutritional value as it is rich in proteins, lipids, and fiber, and has an extraordinary balance of essential amino acids, vitamins, and minerals, in addition to a high number of health-beneficial phytochemicals, including saponins and phytosterols. The present study aims to investigate the lipoidal components of C. quinoa and assessment of C. quinoa lipoidal components and different fractions (methanol, chloroform, ethylacetate, and butanol) cytotoxicity potential against three human cancer cell lines using MTT reduction assay. Materials and methods The seeds of C. quinoa were extracted by different solvents and the extracts prepared were tested for their lipiodial constituents and cytotoxic activities in three cancer cell lines. Results and conclusion The results obtained from Gas Liquid Chromatography (GLC) analysis of nonsaponifiable fraction of C. quinoa seeds indicated that it consists mainly of a mixture of a series of n-alkanes and sterols (79.96 and 7.91%. respectively). The oil was also found to be rich in linoleic acid (52.7%) and oleic acid (25.33%). The investigation of the cytotoxic potential of the plant’s low polarity fractions, including CHCl3, and nonsaponifiable fraction against the studied cell lines revealed the sensitivity of A549 human lung cancer cells. Further studies are needed to uncover the mechanism and chemical constituents of the obtained activity of C. quinoa.
  807 144 -
The neuroprotective effects of natural antioxidant against brain injury induced by paracetamol in a rat model of protein malnutrition
Marwa A Masoud, Amr S Kotb, Ola M Abd El-Raouf, Ebtehal M Fikry
January-March 2020, 19(1):55-61
Backgroundand objective Apigenin (API), as flavonoid, is found in chamomile tea, parsley, celery, onions, lemon balm, and oranges. Therefore, in the present study, we investigated the neuroprotective role of API against oxidative stress, neuroinflammation, and neurotransmitter abnormality induced by cumulative dose of paracetamol (PA) in a rat model of protein malnutrition. Materials and methods A total of 30 male Wistar albino rats, weighing 150–200 g, were used in five groups. API (50 mg/kg, p.o., once daily for 1 week) was administered to low-protein-fed rats with PA (500 mg/kg, p.o, once)-induced brain injury. Results and conclusion API treatment obviously improved cerebral-reduced glutathione and malondialdehyde contents and also superoxide dismutase and myeloperoxidase activities. Additionally, it attenuated contents of serotonin, catecholamines, γ-aminobutyric acid, and cholinesterase activity. Moreover, API reduced the abnormal cerebral pathological lesions. Consequently, API has a protective effect on rat brain injury induced by both protein malnutrition and PA.
  747 116 -
Assessment of cytochrome P450 2E1 activity in Hausa/Fulani of northwest Nigeria using chlorzoxazone as a probe determination of polymorphism
Muhammad T Umar, Shaibu O Bello, Aminu Chika, Yakubu Abdulmumini
January-March 2020, 19(1):62-66
Background and objective High expression and activity of cytochrome P450 2E1 have been linked to non-alcoholic fatty liver disease; increased susceptibility to the gastric, nasopharyngeal, colorectal, urinary bladder and esophageal malignancies; and acetaminophen-induced hepatotoxicity. It plays key roles in activating procarcinogens to carcinogens, metabolism of xenobiotics, and hosts of endogenous compounds. This study aimed at determining the polymorphism of this highly polymorphic enzyme among Hausa/Fulani in northwest Nigeria. Materials and methods A total of 20 nonrelated Hausa/Fulani from Sokoto metropolis were selected by convenient sampling. A tablet of 250 mg chlorzoxazone was administered orally to them with 100 ml of distilled water after an overnight fast, and 3 h after dosing, urine was collected. HPLC equipped with a UV detector was performed for simultaneous estimation of chlorzoxazone and its metabolite 6-hydroxychlorzoxazone. Metabolic ratio index method was used for each participant The data generated were analyzed using Statistical Package for the Social Sciences version 20 (SPSS-20) by constructing frequency histogram and probit plots. A trend line was added to the probit plot, and polynomial equation obtained was resolved to get antimode. Participants with antimode greater than or equal to value of intercept on logMR were regarded as poor metabolizers, whereas those with less were extensive metabolizers. Result and conclusion Anti-mode was found to be −1.8, and only 7 of 20 participants were extensive metabolizers (35%, odds 0.54, 95% confidence interval: 0.22–1.3). Although convenience sampling was used, the findings are worrisome considering the highly polymorphic and the procarcinogenic nature of the enzyme.
  679 83 -
Coronary angiogenesis diabetic rats during consumption of Petroselinum crispum
Shiva Roshankhah, Amir Abdolmaleki, Arash Ziapoor, Mohammad Reza Salahshoor
January-March 2020, 19(1):74-80
Background and objectives Diabetes mellitus is one of the most prevalent metabolic disorders worldwide. Cardiac angiogenesis disruption occurs in diabetes. Petroselinum crispum has antioxidant properties leading to its therapeutic attributes. Materials and methods Sixty-four male Wistar rats were randomly sorted into eight groups: control, extract groups (50, 100, and 150 mg/kg of P. crispum), diabetic, and diabetic+extract groups. Diabetes was induced by intraperitoneal injection of streptozotocin (50 mg/kg), and different doses of extract were administered intraperitoneal for 30 days. The serum level of vascular endothelial growth factor was determined by enzyme-linked immunosorbent assay method; nitrite oxide was measured by Griess assay; and capillary density in the heart was evaluated by immunohistochemistry assay. Results and conclusion The values of all parameters were reduced significantly in the diabetic group compared with the control group (P<0.001). No significant modifications were observed in all extract groups compared with the control group as well as diabetic+extract groups compared with diabetic rats (P>0.05). The hydroalcoholic extract of P. crispum did not affect cardiac angiogenesis in normal and diabetic patients, which probably is due to the insufficient amounts of active ingredients, especially flavonoids available in P. crispum.
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