فهرست مطالب

Iranian Biomedical Journal - Volume:24 Issue: 6, 2020
  • Volume:24 Issue: 6, 2020
  • تاریخ انتشار: 1399/06/16
  • تعداد عناوین: 8
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  • Fatemeh Masaebi, Mehdi Azizmohammad Looha, Mohammad Rostami-Nejad*, Mohamad Amin Pourhoseingholi, Navid Mohseni, Gabriel Samasca, Iulia Lupan, Mostafa Rezaei-Tavirani, Mohammad Reza Zali Pages 335-341
    Background

    It has been established that the level of some inflammatory cytokines increases in celiac disease (CD) and non-celiac gluten sensitivity (NCGS) in comparison with healthy subjects. Therefore, the primary interest in our research was proposing an accurate tool to diagnose patients with CD and NCGS from healthy individuals in an Iranian population.

    Methods

    The serum samples were examined in 171 participants, including 110 CD patients, 46 healthy individuals, and 15 NCGS. The commercial ELISA kits were used to detect the level of the following cytokines: IL-1, IL-6, IL-8, IL-15, and IFN-γ. The receiver operating characteristic (ROC) curve analysis was applied to determine the optimal thresholds for high sensitivity, specificity, positive and negative predictive values of cytokines, as the indicators of CD, NCGS, and healthy control groups.

    Results

    In NCGS group, the values of area under the ROC curve for IL-1, IL-8, and IFN-γ were 71%, 78%, and 70%, respectively. To differentiate the CD and NCGS groups from the control group, IL-15 had the highest sensitivity (82.70%), specificity (56.50%), positive predictive value (81.98%), and negative predictive value (57.78%), followed by IL-8 with the highest sensitivity of 74.50%, specificity of 73.30%, and positive and negative predictive values of 95.35% and 30.21%, respectively.

    Conclusion

    The obtained results demonstrate that IL-15 and IL-8 could be proposed as potential markers in their optimal cut-off points for distinguishing CD from the NCGS and the healthy control. Based on our findings, the evaluation of cytokine levels can be recommended as a useful tool for the diagnosis of CD and NCGS in a clinical practice.

    Keywords: Celiac disease, Cytokines, Sensitivity, specificity
  • Elham Seifali, Gholamreza Hassanzadeh, Marzieh Mahdavipour, Keywan Mortezaee, Ashraf Moini, Leila Satarian, Faezeh Shekari, Abdoreza Nazari, Shabnam Movassaghi, Mohammad Akbari* Pages 342-355
    Background

     Ischemic stroke, as a health problem caused by the reduced blood supply to the brain, can lead to the neuronal death. The number of reliable therapies for stroke is limited. Mesenchymal stem cells (MSCs) exhibit therapeutic achievement. A major limitation of MSC application in cell therapy is the short survival span. MSCs affect target tissues through the secretion of many paracrine agents including extracellular vesicles (EVs). This study aimed to investigate the effect of human umbilical cord perivascular cells (HUCPVCs)-derived EVs on apoptosis, functional recovery, and neuroprotection.

    Methods

    Ischemia was induced by middle cerebral artery occlusion (MCAO) in male Wistar rats. Animals were classified into sham, MCAO, MCAO + HUCPVC, and MCAO + EV groups. Treatments began at two hours after ischemia. Expressions of apoptotic-related proteins (BAX/BCl-2 [B-cell lymphoma-2] and caspase-3 and -9), the amount of terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)-positive cells, neuronal density (microtubule-associated protein 2 [MAP2]), and dead neurons (Nissl staining) were assessed on day seven post MCAO.

    Results

    Administration of EVs improved the sensorimotor function (p < 0.001) and reduced the apoptotic rate of Bax/Bcl-2 ratio (p < 0.001), as well as caspases and TUNEL-positive cells (p < 0.001) in comparison to the MCAO group. EV treatment also reduced the number of dead neurons and increased the number of MAP2+ cells in the ischemic boundary zone (p < 0.001), as compared to the MCAO group.

    Conclusion

      Our findings showed that HUCPVCs-derived EVs are more effective than their mother’s cells in improving neural function, possibly via the regulation of apoptosis in the ischemic rats. The strategy of cell-free extracts is, thus, helpful in removing the predicaments surrounding cell therapy in targeting brain diseases.

    Keywords: Apoptosis, Extracellular vesicles, Ischemia, Middle cerebral artery
  • Shiva Irani*, Mahsa Paknejad, Masoud Soleimani, Azam Soleimani Pages 356-364
    Background

    A growing body of literature has revealed the effective role of miR-34a, as a tumor suppressor and regulator of expression of multiple targets in tumorigenesis and cancer progression. This study aimed at evaluating the potential effects of miR-34a alone or in combination with paclitaxel on breast cancer cells.

    Methods

    After miR-34a transduction by lentiviral vectors in two MCF-7 and MDA-MB-231 cell lines of breast cancer, effects of the elevated expression of miR-34a in the cell viability and the cell proliferation were determined using MTT assay  in treated and untreated cells with paclitaxel. The mRNA level of the CCND1 (Cyclin D1)gene was then measured in the two cell lines using the qRT-PCR assay. Finally, the influence of miR-34a and paclitaxel on apoptosis and cell cycle progression were examined by flow cytometry.

    Results

    The CCND1 mRNA expression levels were significantly down-regulated by overexpressed lentiviral miR-34a in MCF-7 and MDA-MB-231 cells. Combined treatment by miR-34a and paclitaxel reduced the cell viability and proliferation compared to single-drug treatment. In addition, the cell cycle arrest appeared at two phases by the combination of miR-34a and paclitaxel in MDA-MB-231 cells.

    Conclusion

    Our results suggest that miR34a, in combination with paclitaxel, has a potential for decreasing the cell viability and proliferation. Moreover, it can reduce the expression of CCND1 mRNA independent of the paclitaxel effect.

    Keywords: Breast cancer, Cyclin D1, Drug resistance, Paclitaxel, miR-34a
  • Arash Poursheikhani*, Hassan Yousefi, Javad Tavakoli-Bazzaz, Seyed H. Ghaffari Pages 365-373
    Background

    Epithelial ovarian cancer (EOC) is one of the most lethal gynecological malignancy worldwide. Although the majority of EOC patients achieve clinical remission after induction therapy, over 80% relapse and succumb to the chemoresistant disease. Previous investigations have demonstrated the association of epidermal growth factor receptor (EGFR) with resistance to cytotoxic chemotherapies, hormone therapy, and radiotherapy in the cancers. These studies have highlighted the role of EGFR as an attractive therapeutic target in cisplatin-resistant EOC cells.

    Methods

    The human ovarian cell lines (SKOV3 and OVCAR3) were cultured according to ATCC recommendations. The MTT assay was used to determine the chemosensitivity of the cell lines in exposure to cisplatin and erlotinib. The qRT-PCR was applied to analyze the mRNA expression of the desired genes.

    Results

    Erlotinib in combination with cisplatin reduced the cell proliferation in the chemoresistant EOC cells in comparison to monotherapy of the drugs (p < 0.05). Moreover, erlotinib/cisplatin combination synergistically decreased the expression of anti-apoptotic and also increased pro-apoptotic genes expression (p < 0.05). Cisplatin alone could increase the expression of multi-drug resistant genes. The data suggested that EGFR and cisplatin drive chemoresistance in the EOC cells through MEKK signal transduction as well as through EGFR/MEKK pathways in the cells, respectively.

    Conclusion

     Our findings propose that EGFR is an attractive therapeutic target in chemoresistant EOC to be exploited in translational oncology, and erlotinib/cisplatin combination treatment is a potential anti-cancer approach to overcome chemoresistance and inhibit the proliferation of the EOC cells.

    Keywords: Cisplatin, Epidermal growth factor receptor, Ovarian cancer
  • Shima Mehrabadi, Seyed Morteza Karimiyan*, Ghorbangol Ashabi, Khadijeh Moradbeygi, Marjan Hoseini Pages 374-380
    Background

    Tolerance and dependence to anti-nociceptive effect of morphine restricted its use. Nowadays co-administration of morphine and other drugs suggests diminishing this tolerance. Baclofen is one of the drugs that may be beneficial in the attenuation of tolerance to morphine. Studies have shown that changes in transient receptor potential vanilloid type 1 (TRPV-1) expression during administration of morphine have a pivotal role in developing morphine tolerance. Therefore, the effect of baclofen on TRPV-1 expression during chronic administration of morphine was investigated in this study.

    Methods

    A total of 48 rats were divided into four groups of control, morphine single injection, morphine tolerance, and morphine tolerance + baclofen. To induce morphine tolerance in rats, animals received 10 mg/kg of i.p. morphine sulfate once a day for 10 days. In the treatment group, baclofen (0.5 mg/kg) was injected for 10 days, before morphine injection. Finally, to evaluate baclofen treatment on morphine analgesia and hyperalgesia, thermal hyperalgesia and formalin test were used. TRPV-1 and protein kinase C (PKC) expression and protein production in DRG of spinal cord were then evaluated by real-time PCR and Western blot.

    Results

    In baclofen treatment group, thermal hyperalgesia and formalin test improved in comparison with morphine tolerance group. In morphine tolerance group, both TRPV-1/PKC gene expression and protein levels  increased in comparison with the control group. However, following the baclofen treatment, the TRPV-1 and PKC levels  decreased.

    Conclusion

    Baclofen can enhance anti-nociceptive effect of morphine by modulating TRPV-1 channel and PKC activity.

    Keywords: Baclofen, Morphine, Protein kinase C, Spinal cord
  • Ashraf I Khasawneh*, Nisreen Himsawi, Jumana Abu-Raideh, Muna A. Salameh, Mohammad Al-Tamimi, Sameer Al Haj Mahmoud, Tareq Saleh Pages 381-393
    Introduction

    Biofilm formation in Staphylococcus aureus is a major virulence factor. Both methicillin-sensitive Staphylococcus aureus (MSSA) and methicillin-resistant Staphylococcus aureus (MRSA) are common causes of community- and hospital-acquired infections and are associated with biofilm formation. The status of biofilm-forming genes has not been explored in Jordanian nasal carriers of S. aureus. This study investigates antibiotic resistance patterns and the prevalence of biofilm-forming genes between MSSA and MRSA in two distinct populations in Jordan.

    Methods

    A total of 35 MSSA and 22 MRSA isolates were recovered from hospitalized patients and medical students at Prince Hamzah Hospital, Jordan. Antibiotic susceptibility was tested using disk diffusion method and Vitek 2 system. The phenotypic biofilm formation was tested using Congo red agar and microtiter plate assays. The prevalence of the biofilm-forming genes was determined using multiplex PCR.

    Results

    Among 57 S. aureus isolates, 22 (38.6%) isolates were MRSA and were highly resistant against benzylpenicillin, oxacillin, and imipenem. The frequencies of the icaADBC were 77.1%, 97.1%, 94.3%, and 97.1% respectively in MSSA compared to 86.4%, 100%, 100%, and 100% in MRSA isolates. On the other hand, the frequency of the fnbA, fnbB, clfA, fib, clfB, ebps, eno, and cna genes was 81.8%, 90.9%, 95.5%, 90.9%, 86.4%, 100%, 100%, and 40.9%, respectively in the MRSA isolates.

    Conclusion

    In both groups, MRSA isolates, in comparison to MSSA, were significantly more resistant to cefoxitin, oxacillin, imipenem, tetracycline, clindamycin, and trimethoprim-sulfamethoxazole. Unexpectedly, biofilm formation and gene prevalence between MRSA and MSSA isolates showed no significant difference, suggesting other potential virulence mechanisms.

    Keywords: Biofilms, Methicillin-resistant Staphylococcus aureus, Methicillin-sensitive Staphylococcus aureus
  • Seyed Dawood Mousavi-Nasab, Farzaneh Sabahi, Hooman Kaghazian, Mahdi Paryan, Siamak Mirab Samiee, Mostafa Ghaderi, Fatemeh Zali, Manoochehr Makvandi* Pages 394-399
    Background

    HRV is the causative agent of severe gastroenteritis in children and responsible for two million hospitalizations and more than a half-million deaths annually. Sequence characteristics of the gene segments encoding the VP7 and VP4 proteins are used for the genotype classification of rotavirus. A wide variety of molecular methods are available, mainly based on reverse transcription PCR for rapid, specific and sensitive genotyping of rotaviruses. This study describes an alternative real-time PCR assay for genotyping of rotavirus.

    Methods

    The samples of stools studied in this research have been collected from patients referred to Childrenchr('39')s Medical Centers, Tehran, Iran. Rotavirus detection and genotyping were performed using the RT-PCR and semi-nested RT-PCR, respectively. Samples were then genotyped with a new real-time PCR.

    Results

    The real-time PCR was able to genotype all positive samples with a mean Ct of 28.2. Besides, a concordance rate of 100% was detected between real-time PCR and semi-nested RT-PCR.

    Conclusion

    In this study, the genotyping of rotavirus with real-time PCR showed that this method can provide several favorable features, including high sensitivity and specificity, and a wide dynamic range for rotavirus genotyping.

    Keywords: Gastroenteritis, Genotype, Real-time polymerase chain reaction, Rotavirus
  • Rasoul Alizadeh, Sanaz Jamshidi, Mohammad Keramatipour, Parisa Moeinian, Rozita Hosseini, Hasan Otukesh, Saeed Talebi* Pages 400-403
    Background

    Nephronophthisis (NPHP) is a progressive tubulointestinal kidney condition that demonstrates an AR inheritance pattern. Up to now, more than 20 various genes have been detected for NPHP, with NPHP1 as the first one detected. X-prolyl aminopeptidase 3 (XPNPEP3) mutation is related to NPHP-like 1 nephropathy and late onset NPHP.

    Methods

    The proband (index patient) had polyuria, polydipsia and chronic kidney disease and was clinically suspected of NPHP. After the collection of blood sample from proband and her parents, whole exome sequencing (WES) was performed to identify the possible variants in the proband from a consanguineous marriage. The functional importance of variants was estimated by bioinformatic analysis. In the affected proband and her parents, Sanger sequencing was conducted for variants’ confirmation and segregation analysis.

    Results

    Clinical and paraclinical investigations of the patient was not informative. Using WES, we could detect a novel homozygous frameshift mutation in XPNPEP3 (NM_022098.2: c.719_720insA; p. Q241Tfs*13), and by Sanger sequencing, we demonstrated an insertion in XPNPEP3.

    Conclusion

    The homozygous genotype of the novel p.Q241Tfs*31 variant in XPNPEP3 may cause NPHP in the early childhood age.

    Keywords: Nephronophthisis, Whole exome sequencing, XPNPEP3