Cell Journal (Yakhteh)

111-116

Authors: Malihe Moshiri , Fatemeh Todehdehghan , Abdolhossein Shiravi

Objective: Zanjani viper (Vipera albicornuta) is an endemic venomous snake in East Azerbaijan Province, Iran which is medically important due to its application for antivenin production in the laboratory. We need to produce this snake in captivity. This study was conducted to characterize mature male Zanjani viper and to evaluate its sperm reproductive parameters. Materials and Methods: This applied- descriptive study was conducted on twenty Zanjani viper samples collected from Ag Dag Mountain in East Azarbaijan Province, Iran, between September and October 2010. After the snakes were anesthetized and sacrificed humanly, their morphometric specifications and sperm reproductive parameters, including concentration, motility, vitality, morphology, and survival time, were measured. Results: Morphometric specifications and evaluation of sperms of the snake showed the following information: Zanjani male viper, body length of 73.65 ± 4.35 cm, tail length of 5.465 ± 0.48 cm, and mature snakes with testicular volumes of 0.61 ± 0.81 ml (right) and of 0.46 ± 0.17 ml (left). Our findings revealed average sperm concentration of 0.47 ± 0.1 ×106 ml-1, motility of 49 -55 %, vitality of 46.11 ± 9.63 %, normal morphology of 61.71 ± 5.3%, and survival time of 6 ± 2 hours at the laboratory temperature. Statistical analyses were performed using Student’s t test for comparison of two values, and one-way ANOVA was applied where three values were compared. Conclusion: Results suggest that mature Zanjani male viper with mature sperms in its vas deferens is present in late summer and early autumn seasons in Bostanabad County, Iran.

131-140

Authors: Hassan Azhdari-Zarmehri , Saeed Semnanian , Yaghoub Fathollahi , Firouz Ghaderi Pakdel

Objective: It is well known that intracerebroventricular (ICV) and supraspinal injections of orexin-A elicit analgesia, but the mechanism(s) of action remains unidentified. This study aims to characterize the effect of orexin-A on rostral ventromedial medulla (RVM) neurons which are involved in the descending nociception modulating pathway. Materials and Methods: In this experimental study, we injected orexin-A or vehicle directly into rats’ ICV while the tail flick (TF) latencies were measured and the on- and off-cell firing activities were monitored for more than 60 minutes. Results: In response to noxious stimuli on the tail, we observed increased firing rate of on-cells and a decreased association with the firing rate of off-cells and in neutral cells the firing rate was unchanged just prior to tail flicking. ICV injection of orexin-A decreased the spontaneous firing rate of on-cells (the type of RVM neurons believed to have facilitatory action on nociception). Furthermore, orexin-A increased firing rate of off-cells (the type of RVM neurons believed to have an inhibitory action on nociception). Orexin-A reduced the TF-related responses of on-cells and TF-related pause duration of off-cells. Conclusion: These results have shown that the analgesic effect produced by orexin-A may be induced by brainstem neurons. It is suggested that the orexinergic system from the hypothalamus to the RVM may have a potential role in modulation of nociceptive transmission.

147-162

Authors: Mahban Rahimifard , Mona Navaei-Nigjeh , Neda Mahroui , Sanaz Mirzaei , Zahra Siahpoosh , Amir Nili-Ahmadabadi , Azadeh Mohammadirad , Maryam Baeeri , Reza Hajiaghaie , Mohammad Abdollahi

Objective: Pancreatic islets have fewer antioxidant enzymes than other tissues and thus are vulnerable to oxidative stress. In the present study, the effects of nine specifically selected Iranian medical plants on the mitochondria function and survival of isolated rat islets were examined. Materials and Methods: In this experimental study, following laparotomy, pancreases of rats were removed and the islets isolated and incubated in vitro for 24 hours. Logarithmic doses of plant materials were added to the islets and incubated for an additional 24 hours after which the viability of the cells and production of reactive oxygen species (ROS) were measured. Levels of insulin production in relation to static and stimulated glucose concentrations were also determined. Results: The tested compounds markedly increased survival of the islet cells, their mitochondrial activity, and insulin levels at the same time as reducing production of ROS. Greatest effects were observed in the following order: Peganum harmala, Glycyrrhiza glabra, Satureja hortensis, Rosmarinus officinalis, Teucrium scordium, Aloe vera, Zingiber officinale, Silybum marianum, and Hypericum perforatum at doses of 10, 103, 104, 10, 102, 102, 10-1, 10 and 103 μgmL-1, respectively. Conclusion: Based on these results, we suggest that pretreatment with these selected Iranian medical plants can improve the outcomes of pancreas transplants and grafts through the control of oxidative stress damage.

171-178

Authors: Mahdi Zamani , Fatemeh Karami , Fariba Shirvani , Laleh Kia-Lashaki , Bizhan Shahbazkhani

Objective: Celiac disease (CD) is developed via autoimmune reactions against gluten which is mainly found in grains. Although HLA DQB1 locus is the most important genetic susceptibility to CD, some other variants such as A49G and G1359T of CTLA4 and CD14 genes respectively have been proposed as CD predisposing genetic factors in many various studies. We aimed to assess possible roles of A49G and G1359T polymorphisms in CD susceptibility in the Iranian population. Materials and Methods: In this case-control, one hundred CD patients and 100 healthy matched controls with average age of 30-33 years were selected. They were genotyped for both A49G and G1359T polymorphisms using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results: There was no association between genotypes of A49G variant of CTLA4 and risk of CD (p<0.05). The G1359T polymorphism of CD14 gene also did not show any significant association with risk of CD among the studied population. However, patients with CD14 T/T genotype were more classified in the severe form (Marsh III) of CD, showing border line significance (p<0.05). Conclusion: No association was identified between the combination of 1359T and A49G alleles with risk of CD. These lacks of association could be due to small sample size and considering further studies in various populations and ethnicities seems to be required.

187-194

Authors: Fereshteh Nejad-Dehbashi , Elham Shahhosseini Pourshoushtary , Layasadat Khorsandi , Mahmoud Hashemitabar , Mahmoud Orazizadeh , Somaieh Bahramzadeh

Objective: The aim of this study was to evaluate the effect of exendin-4 (EX-4) on differentiation of insulin-producing cells (IPCs) from rat bone marrow-derived mesenchymal stem cells (RAT-BM-MSCs). Materials and Methods: In this experimental study, RAT-BM-MSCs were cultured and the cells characterized by flow cytometry analysis of cell surface markers. RAT-BM-MSCs were subsequently treated with induction media with or without EX-4. After induction, the presence of IPCs was demonstrated with dithizone (DTZ) staining and gene expression profiles for pancreatic cell differentiation markers (PDX-1, GLUT-2, insulin) were assessed using reverse transcription polymerase chain reaction (RT-PCR). Insulin excreted from differentiated cells was analyzed with radioimmunoassay (RIA). The two-tailed student’s t-test was used for comparison of the obtained values. Results: The percentage of DTZ-positive cells significantly increased in EX-4 treated cells (p<0.05). Expression of the islet-associated genes PDX-1, GLUT-2 and insulin genes in EX-4 treated cells was markedly higher than in the cells exposed to differentiation media without EX-4. RIA analysis demonstrated significant release of insulin with the glucose challenge test in EX-4 treated cells compared to EX-4 untreated cells. Conclusion: The results of this study have demonstrated that EX-4 can enhance differentiation of IPCs from RAT-BM-MSCs.

203-210

Authors: Elahe Safari , Ahmad Zavaran Hosseini , Zuhair Hassan , Khosro Khajeh , Mehdi Shafiee Ardestani , Behzad Baradaran

Objective: Immunotoxins (ITs) have been developed for the treatment of cancer, and comprise of antibodies linked to toxins. Also vascular endothelial growth factor (VEGF) plays a key role in tumor angiogenesis, and the blockade of VEGF receptor-2 (VEGFR2) inhibits angiogenesis and tumor growth. The aim of this study was to produce anti-VEGFR2/rPE (Pseudomonas exotoxin) 38 IT to test its cytotoxic activity and mechanism of action. Materials and Methods: In this basic research and experimental study, at first, DNA that encodes recombinant PE38 protein was inductively expressed in Escherichia coli (E.coli) and purified by nickel-sepharose chromatography and further analyzed by western blot. Then, for production of IT, rPE38 was chemically conjugated to anti-VEGFR2. The cytotoxicity response of IT treatment was evaluated by 3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide (MTT) test in Human Umbilical Vein Endothelial Cell (HUVEC) and Michigan Cancer Foundation-7 (MCF-7) (VEGFR2+) cell lines. The mechanism of IT cytotoxicity was observed by Annexin V staining and flow cytometry. Continuous variables were compared with the analysis of variance (ANOVA; for all groups). P values less than 0.05 were considered statistically significant. Results: SDS-PAGE showed 98% purity of rPE38 and IT. In vitro dose-dependent cytotoxicity assay demonstrated that anti-VEGFR2/PE38 is toxic to VEGFR2-positive cells. IT treatment significantly inhibited proliferation of HUVEC and MCF-7 in a VEGFR2-specific manner as compared with the control groups (p<0.05). Flow cytometry showed that the mechanism of IT induced cell death is mediated by apoptosis. Conclusion: IT treatment also caused remarkable synergistic cytotoxicity characterized by decreased cell viability, and an increased apoptotic index by both anti-VEGFR2 and PE38. Thus these results raise the possibility of using anti-VEGFR2/PE38 IT for cancer therapy because nearly all tumors induce local angiogenesis with high VEGFR expression.

225-230

Authors: Fatemeh Taghizade Mortezaee , Mohammad Amin Tabatabaiefar , Morteza Hashemzadeh Chaleshtori , Sepideh Miraj

Uterine leiomyoma (UL) is the most common benign smooth muscle cell tumor with as yet unknown etiology and pathogenesis. This study was carried out to investigate the association of ESR1-351 A>G, ESR1 -397 T>C and CYP1A1 (Ile462Val) polymorphisms with UL in female patients of Iranian origin. In this case-control study, 276 patients with UL and 156 healthy women were recruited. The genetic polymorphisms ESR1-351 A>G, ESR1-397 T>C and CYP1A1 (Ile462Val) were genotyped by polymerase chain reaction- restriction fragment length polymorphism (PCR-RFLP). No significant difference were found in frequencies of both genotypes and alleles of ESR1-351 A>G, ESR1-397 T>C and CYP1A1 (Ile462Val) polymorphisms between the two groups (p>0.05). Our findings indicated that these ESR1 and CYP1A1 polymorphisms were not associated with the development of UL in the cases reported here.