Monthly Publishing Peer Reviewed Journal

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1. Microwave-Assisted Synthesis of Methyl Esters of Alginic Acids as Potential Drug Carrier
Dilyana Murdzheva, Nadezhda Petkova, Mina Todorova, Ivelina Vasileva, Ivan Ivanov, Panteley Denev
Eco-friendly synthesis of methyl esters of two alginic acids with different ratio between M-block and G-blocks were performed by microwave irradiation. The alginates methyl esters have been characterized by degree of esterification (DE), HPLC-SEC, IR-FT and NMR spectroscopy and thermal analysis (TGA/DTA). The influence of reaction time on DE of alginic acids esters were investigated and compared with conventional and ultrasound–assisted synthesis. The highest DE (45 %) of methylated alginic acid (M/G ratio 61/39) was obtained by microwave-assisted synthesis (MAS) for 5 min. In addition, MAS accelerate esterification process as significantly reduced the reaction time from 60 min to 5 minutes. The microwave-assisted esterification did not degrade the alginic acid polymer chains as the coefficient of polydispersity remained constant 1.09 for alginic acid (M/G ratio 61/39) and 1.17 for alginic acid (M/G ratio 37/63), respectively. The ability of complexation of methyl esters of alginic acids with Ca2+ ions was investigated and the promising results were obtained for future design of modified alginate-based micro particles as potential drug carriers.

2. Ethnomedicinal Plants: Study on Antifungal Activity of Essential oil of Pistacia khinjuk (Combined with the Dominance γ-Terpinene) Against Candida albicans.
Reza Tahvilian, Rohallah Moradi, Hossein Zhale, Mohammad Mahdi Zangeneh, Akram Zangeneh, Hossein Yazdani, Majid Hajialiani
Medicinal plants are considered modern resources for producing agents that could act as alternatives to antifungal drugs in demeanor of antifungal drug-resistance fungi. Pistacia khinjuk is a native plant in Iran, which this plant has been used as an indigestion, tonic, toothache, antiflammatory, antipyretic and astringent in Iran. The aim of the current study was to determine chemical composition of P. khinjuk essential oil and evaluate its antifungal activity against common pathogens (Candida albicans) with broth macro-dilution and agar well and disk diffusion methods. The chemical composition of the essential oil was identified using gas chromatography coupled with mass spectrometer detector (GC-MS). The antifungal activity of P. khinjuk essential oil was evaluated by micro-dilution method in Sabouraud-Dextrose broth medium and agar well and disk diffusion assay. According to results of GC-MS analysis, γ-terpinene (81.14%) (w/w), β-Pinene (3.93%) (w/w), α-Terpinolene (2.38%) (w/w) were the abundant components of the essential oil. The results revealed that the essential oil exhibited strong levels of antifungal activity against this tested microorganism. Regarding the MIC and MFC values C. albicans was very high sensitivity to the essential oil. Our findings indicated that P. khinjuk essential oil had a potential to be applied as antifungal agent.

3. Ameliorative Effect of Zinc on Oxidative Stress Induced by Electromagnetic Fields Emitted from Computers
Heba Mahdy-Abdallah, Khadiga Salah Ibrahim, Noha Mohamed Hegazy, Ahmed Ibrahim Amin, Hamdy A A Hammouda, Eman Essam Shaban
Background: A personal computer (PC) is a device with wide existence and use in modern societies. A PC user is exposed to a wide range of electromagnetic fields (EMFs). Objectives: This work aimed to investigate the possible effects of radiation emitted from computer monitor on the oxidant/antioxidant status of workers. Potential ameliorative effects of zinc (Zn) on antioxidant status was considered. Three groups were included in this work. Group B comprised 42 computer workers. The same workers were given zn tablets in a dose of 25 mg/day orally. They were termed group A. A control group composed of sixty-three subjects were included. They were matched for age and socioeconomic status. The study lasted for fifty-six days. Malondialdehyde (MDA) and reduced glutathione (GSH) levels and superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) activities were measured in serum of all participants. We found that serum levels of MDA were significantly increased while zinc concentration, GSH level and (CAT, SOD, and GPx) activities decreased significantly in group B compared with group C. In group A, all parameters were improved when compared with group B. Conclusion: Our results demonstrated that chronic exposure to EMRs emitted from computer induced oxidative stress. Zn supplementation can safeguard against resultant oxidative stress.

4. Molecular Docking of Bioactive Compounds from Piper Plants Against Secreted Aspartyl Proteinase of Candida albicans Causing Oral Candidiasis
P Mani, Muniraj Menakha, Mohammad Saleh Al- Aboody, Wael Alturaiki, Rajendran Vijaya kumar
Oral candidiasis is a common opportunistic fungal infection of the oral cavity caused by Candidaalbicans. A the most serious level, mortality rates of candidiasis are high. Resistance to antimicrobial agents among bacteria and fungi is a persistent problem complicating the management of critically ill patients. In recent years, drug resistance to human pathogenic fungus has been commonly reported from all over the world. Even though pharmaceuticalsectors have produced a number of new antibiotics in the last three decades, resistance to these drugs by microorganisms has increased. Hence, the present study was carried out to evaluate the effect of Piper nigrumand Piper betle against opportunistic pathogen Candida albicans causing oral candidiasis. Totally 55 and 25 compounds were identified from P.nigrum and P.betle respectively. The identified compounds were tested against oral candidiasis causing enzymes by C. albicans through molecular docking and observed Piperine has great potential. The selected compounds from these plants could be utilizedfor thedevelopment of novel drugs against oral candidiasis.

5. The Protective Efficacy of Quercetin on Mefenamic Acid-Induced Gastric Mucosal Damage
Ika Kustiyah Oktaviyanti, Iskandar Thalib, Eko Suhartono
The objectives of this study were to determine the protective effect of quercetin on NSAIDs-induced gastric mucosal damage by measuring the level of superoxide radical, hydrogen peroxide (H2O2), protein carbonylation (PC), and advanced oxidation protein products (AOPPs). In this experiment, a gastric mucosa was taken from male rats (Rattus novergicus). Samples then homogenized and divided into three groups with; T1 served as control which contains gastric mucosa homogenate only; T2 which contains gastric mucosa homogenate + 500 mg of mefenamic acid; and T3 which contains gastric mucosa homogenate + 500 mg of mefenamic acid + 250 mg/L of quercetin. After treatment, every 15-minute superoxide radical, H2O2, PC, and AOPPs levels were estimated. The results revealed that mefenamic acid increased the rate of superoxide radical, H2O2, PC, and AOPPs formation, while quercetin decreases the rate of all parameters. These results indicated that mefenamic acid induce gastric mucosal damage through oxidative stress and inflammation mechanisms as can be seen from the increasing of superoxide radical, H2O2, PC, and AOPPs levels. Also, the results indicated that quercetin have a protective effect to gastric mucosa as can be seen from the decreasing of all parameters.

6. Formulation Development and Evaluation of Hydrogel Based Gastroretentive Drug Delivery System of Antihypertensive Drug
Parveen Kumar Sharma, Gyati Shilakari Asthana, Abhay Asthana
Site specific drug delivery has become the important aspect of research. In present study focused has been made to formulate gastroretentive drug delivery (GRDDS) of drugs having narrow absorption window or having good absorption at proximal part of gastrointestinal tract (GIT) to deliver at stomach site. Hydrogel based gastroretentive formulation of ramipril were prepared by gas blowing technique with the help of using monomers like acrylamide and acrylic acid; bisacrylamide (BIS) as cross linker; pluronic F-127 as foam stabilizer; ammonium per sulphate (APS) as redox initiator; N, N, N’, N’-tetramethylethylenediamine (TEMED) as catalyst and sodium bicarbonate (NaHCO3) as foaming agent. Chitosan was taken as secondary polymer to enhance the mechanical properties of formulation. Hydrogel were characterized with respect to various parameters such as swelling ratio, mechanical strength, apparent density, porosity, void fraction, drug entrapment and in vitro drug release. Further scanning electron microscopy (SEM) and stability studies of formulation were carried out. Study showed the influence of chitosan various concentration on mechanical strength and also on swelling properties. The drug release from chitosan based hydrogel showed for prolonged period of time. Stability studies showed no change in physical appearance and insignificant difference in entrapment efficiency. Thus prepared formulation could be a better targeting for gastroretentive drug delivery.

7. Qualitative Phytochemicals Analysis and Larvicidal Activity of Hydro-Ethanolic Extract of Moroccan Mentha pulegium Against Larvae Mosquito Culex pipiens (Diptera: Culicidae) Vector of Infectious Diseases
Fouad El-Akhal, Yassine Ez zoubi, Khadija Essafi, Abdelhakim El Ouali Lalami
The mosquitos are vectors of many tropical diseases and subtropical. The main targeted mosquito vector in this study, Culex pipiens, causes serious clinical human diseases. In this work, which has never been realized before in the North center of Morocco, the phytochemical screening of the extract of Mentha pulegium and its insecticidal activity on larval stages 3 and 4 of Culex pipiens were studied. The phytochemical constituents were obtained using a simple qualitative analysis method and biological tests were realized according to a methodology inspired from standard World Health Organization protocol. The percent yield of the hydro-ethanolic extract from aerial part of Mentha pulegium was 8.7±0.2. The phytochemical screening of extract of Mentha pulegium grown in North center of Morocco revealed a presence of tanins, gallic tannins, flavonoids, sterols and terpenes, and mucilage components. However, catechic tannins, glycosides and coumarins were not detected. The highest larvicidal activity was observed in the hydro-ethanolic extract of Mentha pulegium against Culex pipiens with LC50 and CL90 values in the order of 0.38 (0.35 – 0.49) and 0.64 (0.50- 0.69) mg/ml respectively. From the results, it can be concluded that hydro-ethanolic extract had excellent potential for controlling mosquito larvae in Morocco.

8. Assessment of Vitamin D and Iron Levels in Childhood Asthma
Hanan M Hamed, Gamal Abdel Naser Yamamah, Nabil Abd El-Aziz Ibrahim, Nevine Elsaid Mohammed Elhelaly, Eman A Wadalla, Mahmoud Mohamed Shahat
Associations between an unhealthy diet and asthma are reported.  A recent attention has focused on the possible relationship between vitamin D and iron and the presence of asthma. The aim: To assess the levels of serum Vitamin-D and iron in children complaining of asthma to demonstrate their possible effect on asthma and pulmonary function. Methods: A cross sectional, case control study was designed. It included sixty patients with partly controlled asthma and uncontrolled asthma according to GINA criteria,2010 were recruited from Asthma Clinic, New Children’s Hospital, Cairo University from 20 September 2013 until 30 Mai 2014.Thirty healthy age and sex matched are included as control. All children were subjected to full history and clinical examination. Serum vitamin D and iron were measured. pulmonary function was done. Results: 33.3% and 25% of control and asthmatics patients have insufficient vitamin D levels (20-30 ng/ml). The frequency of asthmatics patients with iron deficiency is significantly higher than control (46.7% Vs 16.7%) (p= 0.004).  In uncontrolled group, vitamin D was correlated positively with absolute eosinophilic count (p= 0.002 &r = 0.5) and negatively with FEV1(p = 0.01&r = – 0.5). Conclusion: High prevalence of vitamin D insufficiency in both asthmatic and healthy children. Vitamin D may play a role in asthma pathogenesis. Serum iron of asthmatics was lower than the healthy group. Recommendation Further prospective studies on large scale aiming the use of an inexpensive intervention in a substantial number of asthmatic children.

9. Medium Optimization by Artificial Neural Networks for Maximizing the Triglycerides-rich Lipids from Biomass of Chlorella vulgaris
Mohammad Hossein Morowvat, Younes Ghasemi
Artificial neural networks (ANN) have been widely accepted for different microalgal biotechnology aspects due to their versatile applications and advantages. A naturally isolated strain of Chlorella vulgaris, was investigated to assess the influence of three major culture medium constituents including glucose (6-18-1 g L), nitrate (0.5-4.5 g L-1) and phosphate (0.01-0.08 g L-1) sources on lipid production using artificial neural network (ANN) method. A feed forward method was used. The most significant parameter for lipid production was found to be carbon concentration. The highest amounts of lipid production (1.944 g L-1) and biomass productivity (0.31 g L-1 d-1) was observed in the optimized medium condition composed of glucose (15 g L-1), nitrate (1.04 g L-1) and phosphate (0.005 g L-1). The suitability and robustness of using ANN methods for optimization process in microalgal biodiesel production assessment was indicated. Moreover, palmitic acid (C16:0) and linoleic acid (C18:2) were the most abundant fatty acids in the studied C. vulgaris strain under mixotrophic mode of cultivation.

10. Hydrogel Advancement: A New Approach for Gastroretentive Drug Delivery
Sharma P K, Asthana G S, Asthana A
Oral administration of dosage form is the priority for the patients as per its easy administration, economical formulation and various other reasons. Drugs having good absorption in proximal gastrointestinal tract (GIT) part possess bioavailability problem due to less gastric residence time of drug in the stomach. Thus superporous hydrogel (SPH) is a supreme approach to deliver the drug in upper part of GIT. SPH are prepared by using polymers which are hydrophilic in nature and can absorb considerable amount of water and swell. These SPH increase their dimension due to high swelling and resist entry from pylorus to small intestine. Thus increase gastric residence time. There are three generations of SPH based on their swelling index and mechanical strength. First generation called Conventional SPH (CSPH); second generation called SPH composite (SPHC) and third generation called SPH hybrid (SPHH). Third generation SPH possess good mechanical properties and are elastic in nature. Thus these are more suitable for prolonged gastric residence time. Beside their utility in gastroretentive drug delivery, SPH possess various pharmaceutical applications such as peptide drug delivery, fast dissolving tablet, tissue engineering, appetite suppressant, aneurysum and chemoembolisation. This article describes a brief discussion of SPH and its various aspects related with its method of preparation; characterization; various generations of SPH and its applications.

11. Developing a Robust Method for Quantification of β-carotene in Dunaliella salina Biomass Using HPLC Method
Mohammad Hossein Morowvat, Younes Ghasemi
Current analytical methods to determine β-carotene, a naturally occurring carotenoid pigment, such as spectrophotometry or color assay lack the appropriate selectivity and sensitivity and sometimes with significant fluctuations in the obtained results. In this study, a robust reverse-phase high-performance liquid chromatographic (RP-HPLC) method was developed from the photosynthetic unicellular green microalga Dunaliella salina. β-carotene extraction was performed using acetone and chloroform method (2:1 v/v). The extracted β-carotene specimens were chromatographed using a C18 chromatography column with methanol: chloroform (85:15 v/v) and 2.0 mL min-1 flow rate. Two distinct standard curves with different concentrations (0.2-50 µg mL-1 and 100-500 µg mL-1) were provided for β-carotene determination. The observed accuracy, precision and limits of detection and quantitation were quite within the satisfactory levels. The limit of detection (LOD) and quantitation (LOQ) of method were found to be 0.1 μg mL-1 and 0.2 μg mL-1, respectively. The obtained values for β-carotene determination through HPLC method were in agreement with the conventional colorimetric routes (R2=0.9587). The suggested analytical method might be magnificently exploited for β-carotene quantification in D. salina and also other related microalgal strains.

12. Germination of Glycine max Seeds Potentiates its Antidiabetic Effect in Streptozotocin Induced Diabetic Rats
Maha H Mahmoud, Mona M Taha, Eman M Shahy
Glycine max was a native legume to East Asia and now it is grown worldwide. It contains phytoestrogens, antioxidants and many bioactive compounds. It is used for stimulation of immune system and as adjuvant therapy in diabetes, liver and kidney disorders and some cancers. Objectives: The present study aims to evaluate the impact of germination on the antidiabetic property of Glycine max in diabetic rats. Methods: Glycine max seeds were germinated and dried then total polyphenol, flavonoid and the antioxidant power were estimated for non-germinated and germinated Glycine max. The feeding experiment was done on 24 albino rats for 5 weeks. Diabetes was induced by Streptozotocin. Nutritional assessment was done. Biochemical parameters were determined including hemoglobin, glucose, urea, creatinine, AST, ALT, lipid profile, G6PDH, total antioxidant activity and lipid peroxidation. Results: Results showed that germination increased total polyphenol, flavonoids and antioxidant power. Biochemical analysis revealed that germinated Glycine max was more effective to improve blood glucose, deterioration in body weight, alternation of HDL-C and LDL-C and restored antioxidant parameters to some extent. Conclusion: Consequently, it can be concluded that germination of Glycine max potentiates its hypoglycemic effect in diabetic rats and prevents or even reduces the complications of diabetes mellitus.

13. Potential Therapeutic Target of Tramadol in Management of Epilepsy
Azadi A, Barati M
Epilepsy is a neurologic disease that caused by genetic or acquired etiologies. Patients with this disorder usually take pain killer beside antiepileptic agents. Tramadol is an opiate-like analgesic that binds the μ-receptors of opioids and inhibits the re-uptake of monoamines and serotonin in the central nervous system. The mammalian target of rapamycin (mTOR) is a serine-threonine kinase that regulates many physiological functions. Many preclinical and some clinical data have represented that excessive activation of mTOR signaling can be responsible for epilepsy syndromes. Tramadol can inhibit the phosphorylation level of phosphatidylinositol 3′ -kinase (PI3K), increase the protein expression level of tumor suppressor gene (PTEN) and also increase c-Jun N-terminal kinase (JNK) phosphorylation. Because of these three different pathways, although acute and high dose consumption of tramadol can cause seizure, but it can inhibit mTOR signaling and be effective in epilepsy treatment in chronic use. In this article we introduced the probable signaling pathways of tramadol in epilepsy treatment and also the reason of causing epileptic seizures.

14. Oxidative and Chlorinative Stress Biomarkers in Liver Cells of Rats Exposed to Cadmium In-vitro
Iwan Aflanie, Nila Nirmala Sari, Eko Suhartono
Cyanide is one of the rapid acting poisons. Exposure to cyanide can cause several health problems, including liver cells damaged. However, the mechanism of liver cells damage by cyanide still not clear. In this present study, we proposed that cyanide could induced liver cells damaged via oxidative and chlorinative stress pathway. Thus, our study aimed to investigated the effect of cyanide on liver cells via oxidative and chlorinative stress pathway by measuring the levels of hydrogen peroxide (H2O2) malondialdehyde (MDA), Advanced Oxidation Protein Products (AOPPs), the myeloperoxidase (MPO) acivity, and the chlorinative index (CI). The results revealed that cyanide exposure positively correlated with H2O2, MDA, AOPPs level, and CI, and negatively correlated with MPO activity in liver cells homogenate. These results indicated that cyanide exposure induce liver cells damage through the oxidative and chlorinative stress pathway which can be seen from the correlation between cyanide exposure and the levels of H2O2, MDA, and AOPPs, the activity of MPO, and the CI.

15. A Study to Compare Three Types of Assisted Pelvic Floor Muscle Training Programmers in Women with Pelvic Floor Muscle Weakness
Jenifer Augustina, P Ponmathi, V P R Sivakumar
Objective: To compare the effects of three types of Assisted Pelvic floor rehabilitation programmes over Pelvic floor muscle strength in subjects with Pelvic floor muscle weakness. Study Design: Experimental study design. Subjects: Thirty subjects with pelvic floor weakness were taken in the study , age group between 38 to 50 years and pelvic floor muscle strength 3-6mmhg according to Pfx2, and divided into 3 groups(10-group A,10-groupB,10groupC) Interventions: 10 subjects  in the group A were trained to contract pelvic floor musculature along with transversus abdominis,10 subjects in the group B were trained to contract pelvic floor musculature along with theraband exercises and 10 subjects in the group C were trained to contract pelvic floor musculature along with ball squeezes. Outcome Measure: Pfx2 Results: statistical analysis was done by using ANNOVA test. Results showed no significant difference in Pfx2 measurements between three groups in the study, but clinically group C showed improvement. Conclusion: It is concluded that there is no statistically significant improvement of three types of assisted pelvic floor rehabilitation programmes over pelvic floor muscle strength in subjects with pelvic floor muscle weakness, but clinically group C showed improvement. Studies with Large sample size and digital Periton perineometer has to be carried out for better clinical outcome.

16. Sterols and Lipids from Agaricus bisporus
Consolacion Y Ragasa, Jo Madeleine Ann Reyes, Maria Carmen S Tan, Robert Brkljača, Sylvia Urban
Chemical investigation of the dichloromethane extract of Agaricus bisporus led to the isolation of ergosterol (1), brassicasterol (2), trilinolein (3) and linoleic acid (4).  The structures of 1 – 4 were identified by NMR spectroscopy.

17. Platelet Count and Platelet Indices Used as Potential Markers for First Malaria Infection Diagnosis
Orathai Tangvarasittichai, Manop Srikong, Surapon Tangvarasittichai
Hematological changes, especially thrombocytopenia associated with malaria infection are well recognized. We aim to determine platelet count and platelet indices as risk markers for first screening of malaria infection diagnosis. A total of 100 patients with malaria infection attended at the Clinical Laboratory Unit, Thong Pha Phum Hospital. After parasitological confirmation of exclusive infection by malaria, platelet counts and platelet indices were determined. Receiver operating characteristic (ROC) analysis is used for calculation of the area under the curve (AUC) and estimation the sensitivity and specificity of these markers. Platelet count, mean platelet volume (MPV), platelet distribution width (PDW), and plateletcrit (PCT) were significantly altered in patients with malaria infection. A significant inverse relationship was observed between PCT and parasitemia (%Malaria). Platelet count is the highest AUC while the combination of platelet counts and PCT is the best risk marker for first malaria infection diagnosis with 99.0% sensitivity and 95% specificity, respectively. Therefore, platelet count and PCT indices are the effectiveness markers for malaria infection and may using for severity (%Malaria) demonstration in patients with malaria infection, especially in endemic area.