Mohajerani, Mr, Sarikhani, A, Gandomani, M, Eslamirad, Z, Mosayebi, M, Didehdar, M,
Volume 8, Issue 5 (winter[PERSIAN] 2015)
Abstract
Abstract Background and Objective: Malassezia that is a part of normal flora is lipophilic yeast involved in a variety of skin diseases such as seborrheic dermatitis, pityriasis versicolor, atopic dermatitis and psoriasis. Seborrheic dermatitis affects most often the sebaceous-gland-rich areas of skin such as face, scalp, and parts of the upper trunk. Dandruff is a mild variant of seborrheic dermatitis characterized by scaling. In this study, Malassezia species causing dandruff were identified. Material and Methods: In this descriptive study, the samples (n= 60) from participants with dandruff were examined under a microscope using 10% KOH solution and cultured in Leeming and Notman ager medium. DNA Extraction was performed from colonies by glass bead and the Malassezia genus, and species were detected by CfoI enzyme using PCR-RFLP method Results: Of 60, 40 (66.6%) were positive for Malassezia yeast. The positive samples in direct examination grew in culture medium. Malassezia species isolated were Malassezia globosa (25 cases), Malassezia restricta (10 cases), Malassezia furfur (3 cases) and Malassezia sympodialis (2 cases). Conclusions: In most studies, the Malassezia species were identified as the agents causing seborrheic dermatitis. In our study, Malassezia globosa was isolated as a dominant species. Keywords: Seborrheic Dermatitis, Malassezia SPP, Arak
Parisa Bakhshi , Massoud Saidijam, Delavar Shahbazzadeh, Nazanin Mohajerani, Hassan Mirzahoseini,
Volume 9, Issue 5 (Nov,Dec-2015 2015)
Abstract
Abstract
Background and Objective: Hirudin is an anticoagulant polypeptide secreted from the salivary glands of leeches. Recombinant hirudin is a strong anticoagulant agent in arterial and venous thrombosis. The aim of this study was to evaluate the effect of inserting protein A signal peptide sequence of pEZZ18 plasmid on expression and secretion of the recombinant hirudin in E.coli.
Methods: the synthetic hirudin gene was amplified by PCR using specific primers. First, the gene was purified and cloned into PTG19-T cloning vector, and then it was subcloned into pEZZ18 expression vector by SalI / SacI enzymatic digestion and finally transformed into E.coli JM107. After the expression of recombinant hirudin protein, different cellular fractions were isolated and analyzed on SDS-PAGE and further confirmed by Western blotting.
Results: PCR product (522 bp) was first subcloned into the T-Vector (replicating vector) and then successfully subcloned into the pEZZ18 (expression vector). Cloning and subclonig were confirmed by enzymatic digestion and Colony PCR. After the expression and isolation of fractions, the presence of hirudin (about 29 kDa) in different cell fractions due to the effects of signal peptide was observed in SDS-PAGE and finally confirmed by Western blotting.
Conclusion: The gene of anticoagulant hirudin protein (desirudin) was cloned into the pEZZ18 vector containing Protein A signal peptide sequence and later transformed into E.coli JM107. The recombinant hirudin protein expression in the extracellular space was approved.
Keywords: Hirudin; Desirudin; Protein Sorting Signals.
Maryam Mohajerani , Afsane Aghaei ,
Volume 10, Issue 1 (Jan,Feb 2016 2016)
Abstract
Abstract
Background and objective: Peroxidases catalyze protein oxidation and lipid peroxidation. The activity of these enzymes in nerve cells is involved in causing disorders such as Alzheimer's and Parkinson's disease. This study investigated the effect of Citrus aurantium, Foeniculum vulgare and Rosmarinus officinalis essential oils on activity of peroxidase enzyme.
Methods: All three medicinal plants were dried at room temperature. Their essential oil was extracted by steam distillation using a Clevenger apparatus. Optimal reaction conditions were determined in the presence of hydrogen peroxide and guaiacol as substrate and hydrogen donor, respectively. Enzyme kinetics of zucchini peroxidase were evaluated by increasing the amount of essential oils in optimal reaction conditions. Enzyme reaction rate for each of the essential oils and the Km and Vmax values were determined.
Results: The results indicated concentration-dependent effect of the extracted essential oils on enzyme kinetics at optimum temperature of 50 °C and optimal pH of 6.5. The essential oil of Citrus aurantium had non-competitive inhibitory effects on the enzyme with Km of 6.25 mM, while the enzyme’s Vmax significantly reduced by increasing the concentration. Foeniculum vulgare showed mixed inhibition effect with Km of 7.14 mmol per 20 μl of the essential oil, but had a decreasing effect on the Vmax in smaller amounts. Finally, Rosmarinus officinalis showed activating effects by reducing the Km to 4-5.88 mM.
Conclusion: The essential oils of Citrus aurantium and Foeniculum vulgare are inhibitors of the peroxidase enzyme and can be further studied as natural herbal medicines.
