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Background: Trimethyltin (TMT) is a toxic agent that causes oxidative stress, a laboratory model for inducing hippocampal injuries. Pentoxifylline (PTX) inhibits phosphodiesterase, inflammation and oxidative stress. This study evaluated the neuroprotective effects of PTX on injuries induced by TMT in the hippocampus. Methods: Sixty male Wistar rats were divided into five groups of 12 each. Group 1 received normal saline while Group 2 received a single dose of TMT (8 mg/kg). The other four groups received TMT at 8 mg/kg plus 60, 100 or 120 mg/kg PTX twice daily for six consecutive days. The rats’ working and reference memory were investigated, using radial arm maze tasks. At the end of the experiments, the rats’ brains were removed and processed for histological study of the hippocampus. Results: The TMT treatment prolonged the four baited arms tasks (P

Background: Chronic lymphocytic leukemia (CLL) is one of the most common hematologic malignancy in adults worldwide. This cancer has a poor prognosis at different stages. So, the identification of new biomarkers is important for diagnosis of B-CLL. Considering the oncogenic role of APRIL molecule in this leukemia as well as the regulatory role of miRNAs in different signaling pathways, the present study evaluated the miRNAs targeting APRIL gene in B-CLL. Methods: The miRNAs were predicted and selected using bioinformatics algorithms. A total of 80 plasma samples were subjected to RNA extraction and synthesis of cDNA. The expressions levels of predicted miRNAs and APRIL gene in plasma of B-CLL patients and healthy individuals were assessed by Real time PCR analysis. ROC analysis was performed to investigate the role predicted miRNAs as novel biomarkers in diagnosis of B-CLL. Results: The results of the prediction showed that miR-145-5p and miR-185-5p target the APRIL gene. The expression level of APRIL gene was strikingly higher in plasma of B-CLL patients than in the healthy individuals (102, P= 0.001). On the other hand, expression levels of miR-145-5p and miR-185-5p were strikingly lower in B-CLL patients than in the healthy individuals (0.07, P= 0.001) (0.29, P= 0.001). Also, ROC curve analyses demonstrated that miR-145-5p and miR-185-5p are specific and sensitive and may serve as new biomarkers for the detection of B-CLL. (AUC; 0.95, sensitivity; %90) (AUC; 0.87, sensitivity; %63). Conclusion: These data suggest that miR-145-5p and miR-185-5p target the APRIL gene and might have a role in diagnosis of B-CLL. Therefore, these two miRNAs can be served as a novel and potential biomarker for detection of B-CLL.

Recently, members of the semaphorin family have received major attention in various medical fields, especially autoimmunity. In this study, we selected semaphorin-3A (Sema3A), semaphorin-7A (Sema7A), and their receptors to determine the possible relationship between these molecules and multiple sclerosis (MS).We measured the gene expression of Sema3A, Sema7A, neuropilin-1 (NP-1), plexin-C1, and β1 integrin in the blood samples of relapsing-remitting multiple sclerosis (RRMS) patients, treated with high-dose interferon-β1a (IFN-β1a), low-dose IFN-β1a, IFN-β1b, and glatiramer acetate (GA) via quantitative real-time polymerase chain reaction (qRT-PCR) assay, and then, compared the results of treatment-naive patients with the healthy controls.The gene expression of Sema3A (P = 0.02), NP-1 (P 0.001), and plexin-C1 (P 0.01) significantly decreased in the treatment-naive group, compared to the healthy controls. Sema3A significantly increased in all treated patients, compared to the treatment-naive patients (P 0.001). However, expression of NP-1 (P 0.001), plexin-C1 (P 0.001), and β1 integrin (P 0.05) only increased in patients receiving high-dose IFN-β1a, IFN-β1b, and GA. Expression of Sema7A increased in only two groups of patients treated with IFN-β1b (P 0.001) and GA (P = 0.018), without any significant decrease in the treatment-naive group, compared to the healthy controls (P 0.05).Our findings confirm that the presence of Sema3A, Sema7A, and their receptors can play critical roles in the treatment of MS patients. Therefore, they can be potential target molecules for MS treatment in the future.

Background T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive and malignant neoplasm that arises from the hematopoietic T-cell precursors. Inactivation of FBXW7 gene is frequently observed in T-cell acute lymphoblastic leukemia, suggesting a significant tumor-suppressive role for FBXW7 in the pathobiology of this leukemia. Considering the role of microRNAs in cell proliferation and regulation of apoptosis, the aim of this study was to identify novel oncogenic microRNAs that suppress FBXW7 in patients with T-ALL. Patients and Methods The expression levels of two bioinformatically predicted microRNAs - miR-32 and miR-107 were compared in patients with T-ALL and a control group. A total of 80 plasma samples were subjected to RNA extraction, and the microRNA expression profiles were assessed by the RT-qPCR. The expression level of miR-103 was used as the endogenous reference for normalization of quantitative data. Results The plasma levels of miR-32 and miR-107 in patients with T-ALL were significantly higher (5.65, P < 0.001) and lower (0.432, P = 0.002), respectively. On the other hand, the expression levels of FBXW7 gene were significantly downregulated by -76.9 fold in T-ALL patients (P < 0.001). The results of the ROC curve analysis indicated that overexpression of miR-32 might be used to distinguish T-ALL patients with reasonable sensitivity and specificity. Conclusion miR-32 is considered as a novel oncomir that targets FBXW7 and might have a role in the etiology or progression of T-ALL. Furthermore, miR-32 can potentially serve as a non-invasive biomarker for detection of T-ALL.

Background Myeloproliferative neoplasms (MPN) are heterogeneous diseases that classified by the presence of Philadelphia chromosome into Philadelphia chromosome negative (Ph-neg) and positive (Ph-pos) myeloproliferative neoplasms. In ph-neg group A somatic point mutation (c.1849G>T) in the JAK2 gene, part of the JAK2-STAT signal-transduction pathway, causes substitution of phenylalanine for valine (V617F) in the JAK2 protein and has been identified. This mutation was seen in PV by 65% to 97% and ET (30–57%) and primary myelofibrosis (35–95%). Highly sensitive methods have been used to determine the presence of the JAK2V617F mutation instead of direct sequencing. We aimed to assess JAK2 exon14 mutations by high-resolution melting (HRM) analysis, which allows variation screening in compare to other method for detecting mutation. Methods The mutation analysis included 45 individuals who were subjected for diagnosis of ph-neg MPN. Genomic DNA was isolated and different methods are performed. Results PCR RFLP, ARMS PCR and HRM method has a detection sensitivity comparable with conventional methods (Qiagen) to identify the mutations and sequencing. Conclusions For HRM analysis is cost-effective and beside that it is enzyme independence method also this method able to show amount of the mutant allele carried in samples and it’s helpful for treatments follow-up and determining MRD for them.

Background: Timely and sensitive diagnosis of gastric cancer is crucial for efficient treatment and survival of the patients. microRNAs have been considered as diagnostic biomarkers in different type of cancers including gastric cancer. In the present study, the expression profile of four microRNAs, miR-103, miR-107, miR-194 and miR-210 were evaluated in patients with intestinal-type of gastric cancer (IGC) in order to assess their diagnosis utility as noninvasive biomarkers. Methods: A total number of 100 plasma samples from patients with gastric cancer and healthy controls were obtained and total RNA was extracted using a commercial monophasic solution of phenol and guanidium thiocyanate. Reverse transcription (RT) reactions were performed by specific stem-loop RT primers and M-MuLV RT-enzyme. The expression patterns of microRNAs were assessed using reverse transcription quantitative real-time PCR (RT-qPCR) method and the expression of SNORD47 RNA was used as the reference for normalization. Results: The results indicate that the plasma levels of miR-107, miR-194, and miR-210 were significantly lower in patients. Receiver operating characteristic (ROC) curve analysis showed that the patients could be distinguished from healthy individuals at the cutoff levels of 0.504, 0.266, and 0.394 of miR-107, miR-194, and miR-210, respectively. On the other hand, the expression levels of these miRNAs were not significantly different in different clinicopathological stages of the disease. Conclusion: These findings suggest that the plasma levels of miR-107, miR-194 and miR-210 were downregulated in patients with ICG and propose these molecules as potential non-invasive biomarkers for detection of IGC.

Background: Coronavirus disease 2019 (COVID-19) is an infectious disease caused by a newly discovered coronavirus. Understanding the spreading routes of SARS-CoV-2 is crucial for patient management and defining biosafety strategies for public and health care workers. Objectives: In the current study, the virus shedding in the upper respiratory as well as blood, stool, and urine specimens of infected patients, was examined using quantitative real-time PCR assay (qRT-PCR). Methods: The samples of 50 positive patients with high, moderate, and low virus titers in their respiratory specimens were collected. All specimens were subjected to RNA extraction and RT-qPCR assay using two different internally-controlled test systems. Results: The results indicated that no positive results were observed in the urine samples of all patients. The viral genome was diagnosed in 5% of blood and 3.3% of rectal swab samples. The Cq (Cycle of quantification)-values of positive results on the stool and blood samples were always higher than the Cq-value of the respiratory specimen of the same patient. Conclusions: SARS-CoV-2 is mainly detected in the respiratory samples, and the virus is not detectable in the urine. The importance of viremia and the existence of the virus in feces in virus spread in the human population needs further investigation.

There has recently been increasing interest in the transgenerational effects of opioids. Herein, transgenerational consequences of maternal chronic morphine consumption before gestation were investigated at the behavioral and molecular levels of next two generations. Twelve female Wistar rats were randomly divided into two groups: pregestation morphine-consuming and control mothers. Morphine-consuming mothers had access to morphine solution ad libitum for 2 months, whereas the control mothers received only tap water. One month after stopping morphine consumption, rats were mated. After parturition, male and female offspring and later grandoffspring of morphine-consuming and control mothers were divided and used as the study groups. Behavioral testing comprised spatial memory assessment using Morris water maze. Hippocampal expressions of Mecp2 and Hdac2 were investigated through real-time PCR. Spatial memory was significantly diminished in male but not female offspring and grandoffspring of morphine-consuming mothers versus control (P

Introduction: Multiple sclerosis (MS) is a neuroinflammatory autoimmune disease which involves the central nervous ­system. Although the primary cause of MS is obscure, effects of some cytokine and chemokine patterns in both innate and adaptive immune systems have been described. ­Objectives: Since limited studies have examined the role of interleukin (IL)-11 and chemokine CCL27 in MS, we aimed to identify changes in IL-11 and CCL27 gene expression and serum levels in relapsing-remitting MS (RRMS) patients, treated with interferon (IFN)-β and glatiramer acetate (GA). Methods: The serum level and gene expression of IL-11 and CCL27 were measured and compared between treatment-naïve MS patients and RRMS patients who were treated with high-dose IFN-β1a, low-dose IFN-β1a, IFN-β1b, and GA via enzyme-linked immunosorbent assay (ELISA) and quantitative real-time polymerase chain reaction. Results: A significant decrease was observed in the serum level of CCL27 in treatment-naïve patients and IFN-β1b-treated patients compared to the healthy controls. On the other hand, a significant increase was found in the protein level of CCL27 in low-dose and high-dose IFN-β1a groups compared to the treatment-naïve group. In addition, CCL27 gene expression was higher in patients treated with GA than in the treatment-naïve group. There were no significant changes in the gene expression or protein level of IL-11 in all experimental groups. Additionally, a positive correlation was found between IL-11 and CCL-27. Conclusion: Our results suggest the inflammatory role of CCL27 in MS patients, while IFN-β1a seems to play a compensatory role for this chemokine.

Hydatidosisis a parasitic disease caused by the larval stage of Echinococcus granulosus with different genotypes, and major complications in vital organs such as liver, lungs and, brain. Also, this parasite can infect animals and cause economic damages. Recently, some investigations indicated that the genetic variation of the parasite affects the antigenic, immunogenic and pathogenic features. Therefore, present study conducted to genotyping of the E. granulosus larva based on mitochondrial cox1 gene in livestock in the endemic areas of Markazi province, Iran. In this study, 49 hydatid cysts samples collected from 36 sheep, 11 goats and 2 cattle from different slaughterhouses of Markazi province in central part of Iran, 2017. The mitochondrial cox1 gene was amplified and genotyping were accomplished using sequence analysis. The sequencing analysis indicated that the main genotype G1 (61%) and G3 (37%) were identified. Also, one of the samples shows similarity with the G2 (2%) genotype. The results showed the statistically significant differences between the genotypes in different livestock (P

Background: Chronic infection with hepatitis B virus (HBV) is related to more than 50% of all cases of hepatocellular carcinoma (HCC). The HBx protein of the virus plays an important role in carcinogenesis through different mechanisms, including interaction with the Notch1 signaling pathway. Several microRNAs have also been shown to play essential roles in the carcinogenesis of HCC, and the molecules can be considered the novel biomarkers for the diagnosis of different types of cancer. Methods: In the present study, the expression levels of four bioinformatically predicted microRNAs, including miR-214, miR-6510, miR-5193, and miR-34a, were compared in individuals with HBV-associated HCC and controls in order to assess the diagnostic utility of these microRNAs as noninvasive biomarkers. Seventy three plasma samples were subjected to RNA extraction, and the microRNA expression profiles were assessed by RT-qPCR. The expression of miR-103 was used as the endogenous reference for the normalization of quantitative data. Results: The plasma expression of all miRNAs was significantly lower in the HCC group, but the downregulation was most marked for the newly described molecule, miR-5193 (-18.86 folds and -5.71 folds compared to healthy individuals and chronic HBV patients). Another novel microRNA, miR-6510, was downregulated by -4.23 folds (P = 0.001). The results of the ROC curve analysis indicated that the differential expression of miR-5193 could distinguish HCC patients with high sensitivity and specificity. Conclusions: The study microRNAs may have a role in HCC development and progression. In addition, miR-5193 can be potentially used as a non-invasive biomarker for the detection of HBV-induced HCC.

Accurate identification of dermatophyte species is important both for epidemiological studies and for implementing antifungal treatment strategies. Although nucleic acid amplification-based assays have several advantages over conventional mycological methods, a major disadvantage is their high cost. The aim of this study was to develop a rapid and accurate real-time PCR-based high-resolution melting (HRM) assay for differentiation of the most common dermatophyte species. The oligonucleotide primers were designed to amplify highly conserved regions of the dermatophyte ribosomal DNA. Analysis of a panel containing potentially interfering fungi demonstrated no cross reactivity with the assay. To evaluate the performance characteristics of the method, a total of 250 clinical isolates were tested in comparison with the long-established PCR-RFLP method and the results were reassessed using DNA sequencing, as the reference standard method. The assay is able to type dermatophytes using normalised melting peak, difference plot analysis or electrophoresis on agarose gel methods. The results showed that, in comparison to PCR-RFLP, the developed HRM assay was able to differentiate at least 10 common dermatophytes species with a higher speed, throughput and accuracy. These results indicate that the HRM assay will be a useful sensitive, high throughput and cost-effective method for differentiating the most common dermatophyte species.

Background Cell-mediated immunity including T-cells (T helper and cytotoxic) plays an essential role in efficient antiviral responses against coronavirus disease-2019 (COVID-19). Therefore, in this study, we evaluated the ratio and expression of CD4 and CD8 markers in COVID-19 patients to clarify the immune characterizations of CD4 and CD8 T-cells in COVID-19 patients. Methods Peripheral blood samples of 25 COVID-19 patients and 25 normal individuals with similar age and sex as the control group were collected. White blood cells, platelets, and lymphocytes were counted and CD4 and CD8 T lymphocytes were evaluated by flow cytometry. Results The number of white blood cells, lymphocytes, and platelets were reduced significantly in COVID-19 patients (P 0.05); however, the CD8 MFI increased significantly in COVID-19 infected patients (P

Quantitative real-time PCR (qPCR) assay is accepted as the method of choice for monitoring human cytomegalovirus (HCMV) infection in hematopoietic stem cell transplant recipients, but the high cost of commercial kits has hampered its use in many developing countries. In this study, an affordable in-house qPCR was used to manage HCMV infection in pediatric patients and the diagnostic value of this method was compared with the conventional pp65 antigenemia assay. A total number of 1179 samples from 82 recipients were used in this study, and the effect of some potential risk factors on HCMV reactivation was evaluated. The qPCR was able to detect HCMV reactivation earlier and with higher sensitivity than antigenemia assay. Forty-six episodes of reactivation were detected in 39 patients, of which all were detected by the qPCR assay, while only 21 episodes were diagnosed by antigenemia. The DNAemia level of 1284 IU/ml plasma was defined as the optimal cutoff value for starting pre-emptive therapy. It was shown that the acute GVHD severity and the relationship of donor and recipient are the most significant risk factors for HCMV reactivation. The data suggest that the antigenemia method for monitoring HCMV reactivation could be substituted by the qPCR assay.

Characterization of clinically important dermatophytes in North of Iran using PCR-RFLP on ITS region. Didehdar M1, Shokohi T2, Khansarinejad B3, Ali Asghar Sefidgar S4, Abastabar M5, Haghani I5, Amirrajab N5, Mondanizadeh M6. Abstract OBJECTIVE: Dermatophytosis is considered as one of the major public health problems in the world. Accurate identification of the dermatophytes species is important for recognition of the environment and animal sources of infection. In this survey, 1500 clinical specimens taken from outpatients suspected of dermatophytosis and they were examined by direct microscopy and culture. MATERIALS AND METHODS: In total, three hundred and twelve dermatophytes were isolates recovered in culture, they were identified using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method based on the ribosomal DNA internal transcribed spacer (ITS) regions. RESULTS: Tinea pedis was the most common clinical form (32.1%) followed by tinea cruris (24.4%), tinea manuum (16.4%), tinea unguium (8.3%), tinea corporis (7.3%), tinea faciei (6.4%), and tinea capitis (5.1%). Trichophyton interdigitale was the most frequent isolate (38.2%), followed by Trichophyton rubrum (29.8%), Epidermophyton floccosum (16.6%), Trichophyton tonsurans (14.8%) and Microsporum canis (0.6%). The frequency of dermatophytosis was higher in males than in females and in the age-group of 21-30 years. CONCLUSION: Our finding indicated that the incidence of dermatophytosis caused by anthropophilic dermatophytes in Mazandaran province is increasing. Also, this study provides valuable data for the prevention and control of dermatophytosis in the southern coast of the Caspian Sea. KEYWORDS: Dermatophytes; Identification moleculaire; Molecular identification; Nord de l’Iran; North of Iran

High-level Expression and Purification of Active Human FGF-2 inEscherichia coli by Codon and Culture Condition Optimization Mohammad Reza Soleyman 1 ; Mostafa Khalili 1 ; Behzad Khansarinejad 2 ; and Maryam Baazm 3, * 1 Department of Biotechnology, School of Medicine, Arak University of Medical Sciences, Arak, IR Iran 2 Department of Microbilogy and Immunology, School of Medicine, Arak University of Medical Sciences, Arak, IR Iran 3 Department of Anatomy, School of Medicine, Arak University of Medical Sciences, Arak, IR Iran *Corresponding Author: Maryam Baazm, Department of Anatomy, School of Medicine, Arak University of Medical Sciences, Arak, IR Iran. Tel: +98-9166621131, Fax: +98-8634173526, E-mail: Dr.Baazm@arakmu.ac.ir. Abstract Background: Basic fibroblast growth factor (bFGF) is a member of a highly conserved superfamily of proteins that are involved in cell proliferation, differentiation, and migration. Objectives: The objective of this study was to overexpress and purify the high-level active human bFGF in Escherichia coli (E. coli). Materials and Methods: This experimental study was conducted in the Islamic Republic of Iran. After codon optimization and gene synthesis, the optimized FGF-2 gene was subcloned into plasmid pET-32a. pET32-FGF-2 was transformed into E. coli BL21 for expression. The cultivation parameters were optimized to produce a high yield of FGF-2. Results: The optimal conditions were determined as follows: cultivation at 37°C in TB medium, with 1 mM isopropyl-β-D-thiogalactopyranoside (IPTG), followed by post-induction expression for 6 h. Under the abovementioned conditions, the expression volumetric productivity of FGF-2 reached 1.48 g/L. Conclusions: A fusion tag from the pET32 expression plasmid permits the recovery of the recombinant fusion FGF-2 from E. coli,without affecting its biological activity. Keywords: Fibroblast Growth Factor 2; Codon; Protein; E. coli

INTRODUCTION: Prevalence of influenza A virus (Flu-A), respiratory syncytial virus (RSV), and human metapneumovirus (hMPV) was assessed in children with acute respiratory infections (ARIs). METHODS: Nasopharyngeal aspirates and throat swabs were subjected to real-time polymerase chain reaction (PCR) to detect RSV and Flu-A and to conventional PCR to detect hMPV. RESULTS: Of the 156 children assessed, 93 (59.6%) carried at least one virus, with 35.9% positive for RSV, 14.1% for hMPV, and 9.6% for Flu-A. The prevalence of co-infections was 2.6%. CONCLUSIONS: The high detection rate may reflect increased sensitivity of real-time PCR compared to traditional PCR and viral culture.