Nova Biologica Reperta

Analyzing the association of MMP-3 promoter polymorphism and its serum concentration with the outcome of in vitro fertilization and embryo transfer

Farhad Mashayekhi

This study aimed to investigate the polymorphism of matrix metalloproteinase -3 (MMP-3) gene and its expression in the serum of infertile female patients received in vitro fertilization and embryo transfer (IVF-ET). To do so, 100 women with unsuccessful IVF-ET (IVF^–) and 100 women with successful IVF-ET procedure and clinical pregnancy (IVF⁺) were included. Genetic polymorphism and serum concentration of MMP3 were investigated by ARMS-PCR and ELISA, respectively. The results showed no significant association between MMP-3 gene polymorphism and IVF-ET outcome among the two groups studied. However, a significant decrease in the concentration of MMP-3 serum in the IVF^– group was observed in comparison with the IVF⁺group (P=0.000002). Moreover, we showed that the serum MMP-3 levels in CC, AC and AA genotypes in the IVF^– group were 33, 65.33 and 86 ng/ml, respectively. In conclusion, while there is no significant difference between MMP-3 promoter polymorphism and IVF-ET outcome between the IVF⁺ and IVF^- groups, a significant decrease in MMP-3 serum levels in IVF^- group was seen as compared with the IVF⁺ group. It could be also suggested that the CC genotype is associated with a decreased level of MMP-3 serum concentration and may be associated with IVF-ET failure.

Study of Cisplatin effects in comparison with Curcumin on Cdc42 gene expression in human Calu-6 lung carcinoma cell line

Mohammad Nabiuni

Lung carcinoma is the second most common type of cancer. Inefficiency of the current treatments and the undesirable side effects of chemotherapy drugs made the know-how of the treatment important. The purpose of this study is to investigate the synergic effect of curcumin and Cisplatin in comparison with the sole application of each treatment on Calu-6 cell line, an epithelial cell line of human lung carcinoma, and the expression of Cdc42 gene. The viability of Calu-6 was examined after 24- or 48-hour treatment with doses of 0.5 to 8 µg/ml of curcumin, 0.1 to 50 µg/ml of cisplatin and combined doses of curcumin and Cisplatin by MTT assay. To measure apoptosis and the expression of Cdc42 gene, flow cytometry and Real-Time PCR were utilized. Decrease of cell viability and induction of cell death were observed in the cells treated with 0.67 µg/ml of curcumin and 1.7 µg/ml of cisplatin (the lowest effective dose) and the combined treatment with the same doses of each drug after 24-hour treatments. The maximum rates of early and late apoptosis were related to treatment with curcumin and the combined treatment. The gene expression analysis results indicated that both Curcumin and Cisplatin decrease the expression of Cdc42 gene, moreover, their co-administration showed synergic effects. Therefore, Curcumin could be an appropriate option for complementary administration with other chemotherapy agents in order to reduce their efficient dose, and to reduce their side effects.

The effect of cerebrospinal fluid-derived exosomes on neural differentiation of adipose mesenchymal stem cells in alginate hydrogel scaffold

Javad Baharara

Nowadays, researchers have made extensive efforts to find new treatments for nerve damage. Meanwhile, the role of exosomes in cell-cell communication is considered to be a new mechanism. Exosomes can act as suitable differentiating agents. The aim of this study was to investigate the differentiating effect of cerebrospinal fluid-derived exosomes on adipose mesenchymal stem cells in alginate hydrogel. Exosomes were extracted from the cerebrospinal fluid by ultracentrifugation and were then identified by atomic force microscopy (AFM), SEM and DLS technique. In addition, Adipose Mesenchymal Stem cells in alginate hydrogel were treated with different concentrations of exosomes. Cell survival was assessed by MTT and Acridine Orange/Ethidium Bromide methods. Cell differentiation was processed by immunocytochemistry and Real-Time PCR. Examinations confirmed the presence of exosomes with an approximate size of 70 nm. Cell survival results indicate that he ability of cells to survive and proliferate during 14 days. Also, the expression of MAP2 proteins (microtubule-associated protein 2) and Nestin (intermediate filament protein) was confirmed by immunocytochemistry. The results of Real Time - PCR showed that during the seventh and fourteenth days the expression level of MAP2 gene increased and the expression of Nestin gene showed a significant decrease compared to the control group. This study showed that exosomes extracted from cerebrospinal fluid can cause neuronal differentiation of Adipose mesenchymal stem cells in alginate hydrogel scaffolds.

Kinetic study of peroxidase enzyme in the presence of purine alkaloids

Tayyebeh Rahmati darvazi

Peroxidase catalyzes different oxidation of substrates using hydrogen peroxide, a reactive oxygen specie (ROS). ROS, at low concentrations, act as messenger to regulate intracellular signaling, whereas, at high concentrations, they can overcome the immune system by creating oxidative stress. Some common beverages such as coffee, tea and soft drinks contain high levels of xanthine alkaloids including theophylline and theobromine. In this study, the effect of theophylline and theobromine on peroxidase activity was kinetically studied by measuring the absorption of 4-aminoantipyrine, oxidized in the presence and absence of theophylline and theobromine at 510 nm for 3 minutes. The results showed that theobromine and theophylline acted as inhibitors with IC₅₀ of 0.50 and 0.55 mM, respectively. K_m and V_max values showed that both compounds are non-competitive inhibitors. The values of K_i were calculated as 0.03 and 0.045 mM for theobromine and theophylline, respectively. Lower values of K_i and IC₅₀ for theobromine compared to theophylline indicates that theobromine has a higher inhibition strength and binding tendency to the enzyme-substrate complex. Hence, it is concluded that theobromine has a stronger inhibitory effect on POD activity.

Validation of two sympatric fish species of Urmia chub, Petroleuciscus ulanus and Urmia bleak, Alburnus atropatenae, based on morphologic characters in Mahabad-Chai River

Soheil Eagderi

Atta Mouludi-Saleh¹, Soheil Eagderi¹, Keyvan Abbasi² & Manoochehr Nasri³
¹Department of Fisheries, Faculty of Natural Resources, University of Tehran, Karaj, Iran;²Inland Waters Aquaculture Research Center, Iranian Fisheries Sciences Research Institute, Agricultural Research, Education and Extension Organization, Bandar Anzali, Iran;³Department of Fisheries Science and Engineering, Faculty of Agriculture and Natural Resources, Lorestan University, Khorramabad, Iran
Correspondent author: Soheil Eagderi, soheil.eagderi@ut.ac.ir

Abstract: Due to the presence of taxonomic ambiguity regarding Petroleuciscus ulanus and Alburnus atropatenae and the tendency to treat them as synonyms of each other, this study was conducted to validate these two species by comparing their morphometric and meristic characters in the Mahabad-Chai River, Urmia Lake basin. A total of 64 specimens of P. ulanus and 68 specimens of A. atropatenae were sampled and were then investigated for 17 morphometric and 7 meristic characters. Measurements were standardized to eliminate the effects of size due to allometric growth pattern, then their normality and abnormality were examined. Standardized data were analyzed using t-test, Mann-Whitney, principal component analysis (PCA) and discriminative function analysis based on P-value of Hotelling's t-test (DFA/Hotelling's T-test) methods. The results showed that the two species were significantly different in 11 morphometric and 2 meristic traits (P<0.05). DFA analysis also showed significance differences between the two studied species. Our results refute the synonymy hypothesis of Petroleuciscus ulanus and Alburnus atropatenae on the basis of morphometric and meristic traits. Therefore, based on the results of this study, the taxonomic identities of these two species are valid.

Evaluation of phytochemical characterizations and antifungal potency of aqueous and methanolic extracts of Tulipa biflora

Vida Tafakori

The purpose of this study was to evaluate phytochemical characterizations and the in vitro antifungal capacity of the aqueous and methanolic extracts of the flower of Tulipa biflora Pall., against various yeast and mold species. For this purpose, fresh flowers were grinded and then macerated in methanol and water overnight, the resulted extracts were then evaluated by various tests. The results indicated that the extracts had different phytochemical components such as terpenoids, tannins, flavonoids, alkaloids, phenols, and phlobatannins. After the evaporation of solvents, antifungal activities of the concentrated extracts were evaluated by the well-diffusion method on Candida albicans, Trichosporon asahii, Yarrowia lipolytica, Aspergilus niger and Fusarium oxysporum. The results showed that the extracts were effective on yeasts and mold species studied except for Aspergilus niger. In order to determine the minimum inhibitory concentration (MIC) and minimum biocidal concentration (MBC) of the extracts, anti-microbial tests were performed in micro-plates. The methanolic extract had more antifungal effectsthan that in the aqueous extract. Therefore, the extract of Tulipa biflora could be introduced as a source for antifungal agents.