فهرست مطالب

Cell Journal (Yakhteh)
Volume:16 Issue: 2, Summer 2014

  • تاریخ انتشار: 1393/03/16
  • تعداد عناوین: 15
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  • Najmaldin Saki, Saeid Abroun*, Saeideh Hajizamani, Fakher Rahim, Mohammad Shahjahani Page 99
    Multiple myeloma (MM), is the second most common blood cancer after non-Hodgkin’s lymphoma. Genetic changes, structural and numerical chromosome anomalies, are involved in pathogenesis of MM, and are among the most important prognostic factors of disease-associated patient survival. MicroRNAs (miRNAs) are small 19-22 nucleotide single-stranded RNAs involved in important cellular processes. Cytogenetic changes in plasma cells alter miRNA expression and function. MiRNAs act as tumor suppressors and oncogenes by affecting intracellular signaling pathways. MiRNA expression is associated with a specific genetic change and may assist with diagnosis and disease prognosis. This study aims to evaluate recent findings in MM-associated cytogenetic changes and their relationship with changes in the expression of miRNAs. We have determined that MM-associated cytogenetic changes are related to changes in the expression of miRNAs and CD markers (cluster of differentiation) are associated with disease survival. Information about these changes can be used for therapeutic purposes and disease prognosis.
    Keywords: Multiple Myeloma, Prognosis, MicroRNAs (miRNA), Tumor Suppressor
  • Malihe Moshiri, Fatemeh Todehdehghan*, Abdolhossein Shiravi Page 111
    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.
    Keywords: Sperm, Basic Reproductive Rate, Snake, Viper, Iran
  • Shiva Nemati, Reza Jabbari, Mostafa Hajinasrollah, Nargess Zare Mehrjerdi, Hossein Azizi, Katayoun Hemmesi, Reza Moghiminasr, Zahra Azhdari, Ardeshir Talebi, Soroush Mohitmafi, Ahmad Vosough Taqi Dizaj, Giuve Sharifi, Hossein Baharvand, Omidvar Rezaee, Sahar Kiani* Page 117
    Objective
    Currently, cellular transplantation for spinal cord injuries (SCI) is the subject of numerous preclinical studies. Among the many cell types in the adult brain, there is a unique subpopulation of neural stem cells (NSC) that can self-renew and differentiate into neurons. The study aims, therefore, to explore the efficacy of adult monkey NSC (mNSC) in a primate SCI model.
    Materials And Methods
    In this experimental study, isolated mNSCs were analyzed by flow cytometry, immunocytochemistry, and RT-PCR. Next, BrdU-labeled cells were transplanted into a SCI model. The SCI animal model was confirmed by magnetic resonance imaging (MRI) and histological analysis. Animals were clinically observed for 6 months.
    Results
    Analysis confirmed homing of mNSCs into the injury site. Transplanted cells expressed neuronal markers (TubIII). Hind limb performance improved in transplanted animals based on Tarlov’s scale and our established behavioral tests for monkeys.
    Conclusion
    Our findings have indicated that mNSCs can facilitate recovery in contusion SCI models in rhesus macaque monkeys. Additional studies are necessary to determine the improvement mechanisms after cell transplantation.
    Keywords: Neural Stem Cell, Spinal Cord Injury, Primates, Transplantation
  • Hassan Azhdari, Zarmehri, Saeed Semnanian*, Yaghoub Fathollahi, Firouz Ghaderi Pakdel Page 131
    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.
    Keywords: Orexin, A, Tail Flick, Pain
  • Masoud Hajia, Mohammad Farzanehkhah*, Bahareh Hajihashemi, Alireza Dolatyar, Mohsen Imani, Roghieh Saburian, Marjan Rahnamaye Farzami, Mohammad Rahbar Page 141
    Objective
    In-time diagnosis of Streptococcus pneumoniae (S. pneumonia) can play a significant role in decreasing morbidity and mortality rate. Applying molecular methods has gained popularity due to the existing limits of routine diagnostic methods. Examining the expression of different genes of this bacterium through different molecular methods suggests that lytA gene has a higher sensitivity and specificity in diagnosis of Streptococcus pneumoniae. The aim of this study was to evalutate lytA gene expression in diagnosis of invasive S. pneumonia in culture positive specimens by real-time polymerase chain reaction (PCR).
    Materials And Methods
    IIn this a descriptive study, All received specimens were isolated to identify S. pneumoniae. DNA was then extracted and after optimizing the test and determining the detection limit, samples were tested by real-time PCR using lytA gene primers.
    Results
    Twenty seven isolates were diagnosed as S. pneumoniae. In all, the extracted DNA was positive in real-time method. The electrophoresis of the products also confirmed the presence of single product b along with the 53 base pair fragment. The detection limit of the test was less 6 colony forming unit (CFU).
    Conclusion
    Real-Time PCR seems to provide reliable and rapid results. We suggest that this test should be conducted on the preliminary isolated specimens, since applying various biochemical tests need one extra working day.
    Keywords: Streptococcus pneumoniae, lytA Gene, Real Time PCR
  • Mahban Rahimifard, Mona Navaei, Nigjeh, Neda Mahroui, Sanaz Mirzaei, Zahra Siahpoosh, Amir Nili, Ahmadabadi, Azadeh Mohammadirad, Maryam Baeeri, Reza Hajiaghaie, Mohammad Abdollahi* Page 147
    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.
    Keywords: Islets of Langerhans, Oxidative Stress, Medicine, Iranian Traditional, In Vitro
  • Mozhgan Torabi, Mahnaz Kesmati*, Hooman Eshagh Harooni, Hosein Najafzadeh Varzi Page 163
    Objective
    Nano components are today’s new wonder material. However, the safety or toxicity of these components in humans is not yet clear. In a previous study we indicated that nano ZnO (nZnO) has a stronger anxiolytic effect compared to the conventional ZnO (cZnO). The present study was designed to evaluate the intraperitoneal administration of an opioidergic receptor agonist and antagonist of as well as the intra CA1 administration of an opioidergic receptor antagonist on the anxiolytic properties of nano and conventional ZnO in adult male Wistar rats.
    Materials And Methods
    In this experimental study, rats received drugs via two modes of injection; intraperitoneal (IP.) and intra CA1 (intra hippocampus, CA1 area). Firstly, nZnO (5, 10, 20 mg/kg), cZnO (5, 10, 20 mg/kg), morphine 6 mg/kg, and naloxone 1 mg/kg were injected IP and naloxone 1μg/rat was injected intra CA1. Subsequently, morphine and naloxone (IP and intra CA1) were co-injected with the effective dose of nZnO and cZnO. An elevated plus maze was used to evaluate anxiety related behavior and anxiety parameters 30 minutes after the second injection.
    Results
    The results indicated that the anxiolytic effects of nZnO 5 mg/kg and cZnO 10 mg/kg were equal. When injected intraperitoneally, naloxone increased anxiety but did not inhibit the anxiolytic effect of nZnO and cZnO. The anxiolytic effects of morphine potentiated the anxiolytic effects of ZnO, particularly nZno. When introduced via intra CA1 injection naloxone alone had no effect on anxiety behaviors and did not inhibit the anxiolytic effect of nZnO.
    Conclusion
    It seems that the opioidergic system activity involved in the anxiolytic effect of nano and conventional ZnO may operate through shared and unshared pathways.
    Keywords: Nanoparticls, Zinc Oxide, Anxiety, Opioid, Hippocampus
  • Mahdi Zamani *, Fatemeh Karami, Fariba Shirvani, Laleh Kia, Lashaki, Bizhan Shahbazkhani Page 171
    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.
    Keywords: CTLA4, CD14, Celiac disease
  • Ahmadreza Assareh, Habib Haybar *, Hosein Malekzadeh, Leila Yazdanpanah, Mohammadreza Bozorgmanesh Page 179
    Objective
    Β2-microglobulin (β2M) associated amyloidosis is an inevitable complication of chronic kidney disease (CKD). Testing β2M in the blood is invasive and expensive. On the other hand, oral fluid is a perfect medium to be explored for public health and disease surveillance. However, it has never been studied if salivary concentration of β2M reflects its concentration in the serum. The current study; therefore, aimed to examine the relationship between salivary and serum β2M in a sample of adult diabetic men with CKD.
    Materials And Methods
    Among diabetic patients referred to the Nephrology Department of The Golestan Hospital of Ahvaz due to CKD, 40 men not requiring renal replacement therapy were consecutively recruited for this cross-sectional study. Patients were excluded if they had any disease or were using any drugs that might affect the oral mucosa or saliva. The concentration of β2M was measured in both serum and saliva. The correlation between serum and salivary β2M was measured by calculating spearman’s ρ.
    Results
    The Spearman’s ρ for correlation between serum and salivary β2M was -0.017 (p=0.917), indicating lack of correlation. Serum and salivary creatinine (Spearman’s ρ=0.54; p value<0.001) as well as serum and salivary urea nitrogen levels (Spearman’s ρ=0.39; p value=0.014) were correlated.
    Conclusion
    Salivary β2M levels poorly agreed with serum β2M levels, and thus may not be used as a surrogate for serum β2M in CKD patients who did not require replacement therapy.
    Keywords: ?2, Microglobulin, Chronic Kidney Disease, Saliva
  • Fereshteh Nejad, Dehbashi, Mahmoud Hashemitabar, Mahmoud Orazizadeh, Somaieh Bahramzadeh, Elham Shahhosseini Pourshoushtary, Layasadat Khorsandi* Page 187
    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.
    Keywords: Exendin, 4, Mesenchymal Stem Cells, Insulin, producing Cells, PDX, 1, GLUT, 2
  • Malek Soleimani Mehranjani *, Milad Falahat Chian Page 195
    Objective
    Mesenchymal stem cells (MSCs) can differentiate into various cell types. Since cysteine has structural similarities to neuronal inducers β-mercaptoethanol and glutathione, we examined its effect on neural induction of rat bone marrow MSCs.
    Materials And Methods
    In this experimental study, cells were treated in a medium containing 1mM cysteine for 24 hours prior to treatment with neuron inducing medium containing 10 mM cysteine for 1, 2 and 3 hours. Cell viability and morphology were assessed by 3-(4,5-dimethylthiazol-2-Yl)-2,5-diphenyltetrazolium bromide (MTT) assay and, Hoechst, propidium iodide and acridine orange staining respectively. Expression of nestin and β-Tubulin III genes, as neural cell-specific markers, was studied reverse transcription polymerase chain reaction (RT-PCR). The data was statistically analyzed using One-Way ANOVA and Tukey’s test and p<0.05 was considered significant.
    Results
    After 3 hours of treatment, neuron like morphology with a considerable expression of nestin and β-Tubulin III genes was apparent. The mean cell viability was not significantly different at 1, 2 and 3 hours following induction, compared with the control cells.
    Conclusion
    Cysteine can induce neural features in rat bone marrow MSCs without reducing cell viability. Therefore, it can be considered as a safer alternative to toxic neural inducer agents such as β-mercaptoethanol.
    Keywords: Mesenchymal Stem Cell, Neural Induction, Cysteine, Nestin, ? Tubulin III
  • Elahe Safari, Ahmad Zavaran Hosseini *, Zuhair Hassan, Khosro Khajeh, Mehdi Shafiee Ardestani, Behzad Baradaran Page 203
    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.
    Keywords: VEGFR2, Pseudomonas Exotoxin, Immunotoxin
  • Faiza Sharif *, Merijn A.G. De Bakker, Michael K. Richardson Page 211
    Objective
    Genes involved in bone and tissue remodelling in the vertebrates includematrix metalloproteinase-9 (mmp-9), receptor activator of necrosis factor κ-β (rank), cathepsin- k (Ctsk) and tartrate-resistant acid phosphatase (TRAcP). We examine whether these markers are expressed in cells of zebrafish embryos of 1-5 days post fertilization. We also examine adult scales, which are known to contain mature osteoclasts, for comparison.
    Materials And Methods
    In this experimental study, in situ hybrdisation, histochemistry and serial plastic and paraffin sectioning were used to analyse marker expression.
    Results
    We found that mmp-9 mRNA, TRAcP enzyme and Ctsk YFP protein were expressed in haematopoietic tissues and in the cells scattered sparsely in the embryo. Ctsk and rank mRNA were both expressed in the branchial skeleton and in the developing pectoral fin. In these skeletal structures, histology showed that the expressing cells were located around the developing cartilage elements, in the parachondral tissue. In a transgenic zebrafish line with YFP coupled to Ctsk promoter, Ctsk expressing cells were found around pharyngeal skeletal elements. To see whether we could activate osteoclasts, we exposed prim-6 zebrafish embryos to a mixture of 1 μM dexamethasone and 1 μM vitaminutes D3. These compounds, which are known to trigger osteoclastogenensis in cell cultures, lead to an increase in intensity of Ctsk YFP expression around the skeletal elements.
    Conclusion
    Our findings show that cells expressing a range of osteoclast markers are present in early larvae and can be activated by the addition of osteoclastogenic compounds.
    Keywords: Zebrafish, Osteoclasts, TRAcP, Ctsk, mmp, 9
  • Fatemeh Taghizade Mortezaee, Mohammad Amin Tabatabaiefar, Morteza Hashemzadeh Chaleshtori, Sepideh Miraj * Page 225
    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 geneticpolymorphisms 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 betweenthe 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.
    Keywords: ESR1, CYP1A1, Uterine Leiomyoma, Polymorphism
  • Sima Shahabi, Seyed Gholam Ali Jorsaraei *, Ali Akbar Moghadamnia, Effat Barghi, Ebrahim Zabihi, Masoumeh Golsorkhtabar Amiri, Ghorban Maliji, Alieh Sohan Faraji, Maryam Abdi Boora, Neda Ghazinejad, Hajar Shamsai Page 231
    In some traditional therapies, it has been claimed that camphor (a crystalline ketone obtained from cinnamomum camphora) would be a suppressor of sexual behaviors and sex hormones. This study evaluated the effects of camphor on sex hormones, like luteinizing hormone (LH), follicle-stimulating hormone (FSH) and testosterone. In this experimental study, 56 male rats were divided into 5 groups, including control (n=12), sham (n=11) and three treatment groups (n=11) in three different doses. The sham groups received daily intra peritoneal (IP) injections of the vehicle (ethanol 10%) for 30 days. Three treatment groups received different daily IP injections of the camphor (1, 2 and 5 mg/Kg) for 30 days and the control groups didn’t received anything. Serums were used for assaying LH, FSH and testosterone. The level of LH significantly increased in all doses of camphor among the treatment groups as compared to the control (p<0.05), but camphor in doses 2 and 5 mg/Kg significantly reduced the FSH level as compared to control group (p<0.05). No significant changes were seen in testosterone levels. Camphor increased level of LH, decreased level of FSH, whereas it failed to change level of testosterone. The claim of inhibitory effect of camphor on sexual activity could not be confirmed by this study. More investigations in this field are suggested.
    Keywords: Camphor, FSH, LH, Testosterone