Showing 3 results for Molaei Rad
Mojgan Farrokhi Karibozorg, Ali Farahnak, Mohammad Bagher Molaei Rad, Taghi Golmohammadi, Mohammad Reza Eshraghian,
Volume 2, Issue 2 (4-2014)
Abstract
Introduction: Hydatid cyst disease is caused by the protoscolices of Echinococcus granulosus. Alkaline phosphatase (ALP) enzyme is required for metabolism, physiology, immunology, and nutrients absorption in parasite. The aim of this study was to compare the level of ALP activity (as a pathological biomarker) in hydatid cyst protoscolices (HCP) with that of sheep liver tissue and to determine the effect of cystic infection on the enzyme activity. Methods: HCPs were collected from sheep livers with hydatid cysts at a local abattoir and washed 3 times with PBS buffer. HCP samples were freeze-thawed and sonicated, while the collected liver tissues were homogenized. Then, extract solutions were centrifuged and stored at -20°C. ALP activity was measured in the extract solutions of HCP and healthy and infected sheep liver tissue samples. The amounts and bands of protein samples were detected using Bradford method and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), respectively. To determine the significant difference between the two groups, independent two samples t-test was used. Results: The mean values of ALP-specific activity of healthy and infected livers and HCP were estimated 0.019, 0.175, and 1.28 U/ml/mg, respectively. Higher ALP activity level was observed in cystic liver compared to healthy liver (p<0.05). T-test analysis showed higher ALP enzyme activity for HCP compared to healthy liver (p<0.05). SDS-PAGE demonstrated a protein band with molecular weight of 59 kDa in HCP samples, which was identified as ALP. Conclusion: ALP activity in HCP and healthy liver indicates the importance of this enzyme in comparative biochemistry of liver and parasite. Higher level of ALP enzyme activity in cystic liver in comparison with healthy liver could be considered as a pathological biomarker for diagnosis of hydatid cyst disease with other hydatid disease parameters.
Ali Farahnak, Fariba Amni, Taghi Golmohammadi, Mohammad Reza Eshraghian, Mohammad Bagher Molaei Rad,
Volume 3, Issue 1 (1-2015)
Abstract
Introduction: To determine an indicator for Triclabendazole (TCBZ) efficacy, Alanine aminotransferase (ALT) activity in Fasciola hepatica (Iranian isolate) parasite in presence and absence of TCBZ was evaluated by an in vitro cultivation method. Also, ALT enzyme activity between none and parasitized-infected sheep liver tissues was assessed. Method: The sheep livers were collected and transferred immediately to the Department of Parasitology. Adult living parasites were recovered, washed and divided into two groups, treatment and control groups with 10 parasites in each. We added 15 μg TCBZ to the treatment group; then incubated both groups for 4 h at 37ºC. The parasites, infected and parasite free liver tissues were ground and homogenized by a Mortar and pestle, centrifuged, and supernatants were recovered. Protein concentration and ALT enzyme activity were measured in the supenatants. Results: The results of ALT enzyme activity assay showed 0.03 U/ml/mg protein for treated F. hepatica and 0.01 U/ml/mg protein for untreated samples, the mean values of difference was not significant (p>0.05). The difference between ALT activity in none and parasitized-infected liver was not significant (p>0.05). However, two-sample T-test analysis showed higher ALT activity in treated and untreated parasite in comparison with none and parasitized-infected liver specimens (p<0.05). In addition to ALT protein band for parasite and liver tissue, Cathepsin enzyme (proteases) was detected for parasite by SDS-PAGE analysis. Conclusion: ALT activity cannot be considered as a useful marker for TCBZ efficacy in F. hepatica treatment. However, ALT enzyme showed comparable activities in parasite and its host liver tissue.
Ali Farahnak, Leila Zaki, Mohamad Bagher Molaei Rad, Abolfazl Golestani, Mohammad Reza Eshraghiyan,
Volume 6, Issue 2 (4-2018)
Abstract
Introduction: Hydatidosis is an endemic parasitic disease of humans in Iran, and Albendazole (ABZ) is a drug of choice for treatment of this infection. As the Alkaline phosphatase (ALP) is necessary for the metabolism of parasites, this study was aimed to evaluate the effect of ABZ on ALP enzyme activity in hydatid cyst parasite as a marker for drug efficiency. Methods: In the present study, the ALP activity level was estimated in the extracts of the untreated parasite (Hydatid cyst protoscoleces) as well as the ABZ-treated samples with a final concentration of 100 µg. The protein concentration and the protein bands in the extracted samples were analyzed by Bradford and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) methods, respectively. Results: The results showed that the mean value of the ALP activity level of the treated samples (0.474 U/ml/mg) was significantly higher than that of untreated samples (0.205 U/ml/mg) (P<0.05). SDS-PAGE analysis demonstrated the higher intensity of the 59 kDa protein band in ABZ-treated samples, compared to the untreated sample. Conclusion: Considering the effect of the ABZ drug on ALP activity in the hydatid cyst protoscoleces, this enzyme might be regarded as an indicator for the effectivity of drug on this parasite.
