Tarbiat Modares UniversityMolecular and Biochemical Diagnosis Journal2383-05221320141001Production of biosimilar human like erythropoietin (EPO) by Human embryonic kidney (HEK293) cell line, its advantages and disadvantages to CHO cell line1431555777ENNadirAlipourDepartment of Biotechnology, Middle East Technical University, Ankara, Turkey.
Department of Medical Microbiology, Faculty of Medicine, Giresun University, Giresun, TurkeyNasrinGainiDepartment of Radiology, Faculty of Medicine, Trakya University, Edirne, TurkeyJournal Article19700101Background: Erythropoietin (EPO) is a glycoprotein hormone function to regulate the production of red blood cells. Deficiency of EPO is known to cause anemia in chronically infected renal patients and they require regular blood transfusion. Availability of recombinant EPO has eliminated the need for blood transfusion and now it is extensively used for the treatment of anemia. Glycosylation of erythropoietin is essential for its secretion, stability, protein conformation and biological activity. However, maintenance of human like glycosylation pattern during manufacturing of EPO is a major challenge in biotechnology. Currently, Chinese hamster ovary (CHO) cell line is used for the commercial production of erythropoietin but this cell line does not maintain glycosylation resembling human system. With the trend to eliminate non-human constituent from biopharmaceutical products, as a preliminary approach, we have investigated the potential of human emberyo kidney cell line (HEK293) to produce recombinant EPO. Methods: Initially, the secretory signal and Kozak sequences was added before the EPO mature protein sequence using overlap extension PCR technique. PCR-amplified cDNA fragments of EPO was inserted into mammalian expression vector under the control of the cytomegalovirus (CMV) promoter and transiently expressed in CHO and HEK293 cell lines. After RT-PCR analysis, ELISA and Western blotting was performed to verify the immunochemical properties of secreted EPO. Results: Addition of secretory signal and Kozak sequence facilitated the extra-cellular secretion and enhanced the expression of EPO protein. Significant expression (P < 0.05) of EPO was observed in the medium from HEK293 cell line. Conclusions: HEK293 cell line has a great potential to produce glycosylated EPO, suggesting the use of this cell line to produce glycoproteins of the therapeutic importance resembling to the natural human system.https://mbd.modares.ac.ir/article_5777_89abe98de6071178edb1b28901a8f459.pdfTarbiat Modares UniversityMolecular and Biochemical Diagnosis Journal2383-05221320141001SOD1 C6W mutation and exon deletion in an amyotrophic lateral sclerosis patient1571695778ENParisaGhiasiDepartment of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, IranSamanHosseinkhaniDepartment of Neurology, Tehran University of Medical Sciences, Tehran, IranShahriarNafissiDepartment of Neurology, Tehran University of Medical Sciences, Tehran, IranKhosroKhajehDepartment of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, IranJournal Article19700101Background: Despite the genetic heterogeneity reported in familial ALS (FALS), SOD1 gene mutations are the most frequent cause of FALS, accounting for around 20% of familial cases (ALS1) and isolated sporadic cases. Mutant forms of SOD1 exhibit toxicity that promotes the death of motor neurons. It is well documented that FALS produces protein aggregates in the motor neurons of FALS patients, which have been found to be associated to mitochondria.
Methods: In this study, we cloned the SOD1 gene, using reverse transcriptase-polymerase chain reaction (RT-PCR) method, from both a healthy control and a living 79 -year-old man with diagnosis of sporadic form of ALS who had shown unusual rapid progression of disease. RNA samples were available from lymphocytes of them. pET28a expression system and BL21 chemically competent Escherichia coli strain as host were used for protein expression.
Results: DNA Sequencing data showed both heterozygosis C to G transition at nucleotide position 21 leading to a C6W changing at protein level and a deletion at nucleotides position 73 to 169 leading to complete deletion of exon two.https://mbd.modares.ac.ir/article_5778_ca7be8306ecc3f5fa30ff2c41e64fa7b.pdfTarbiat Modares UniversityMolecular and Biochemical Diagnosis Journal2383-05221320141001Studies on the relationship of CTLA-4 +49A/G gene with Recurrent Miscarriage in Northwest of Iran1711765779ENMortazaBonyadiCenter of Excellence for Biodiversity, Faculty of Natural Sciences, University of Tabriz, Tabriz, Iran.
Liver and Gastrointestinal Research Center, Tabriz University of Medical Science, Tabriz, IranSaraParsaCenter of Excellence for Biodiversity, Faculty of Natural Sciences, University of Tabriz, Tabriz, IranSiminTaghaviAlzahra Hospital, Tabriz, Iran.
Maternal Fetal Medicine, Obstetrics and Gynecology Unit, University of Medical Sciences, Eastern Azarbaijan, IranNargesZeinalzadehCenter of Excellence for Biodiversity, Faculty of Natural Sciences, University of Tabriz, Tabriz, IranJournal Article19700101Background: Immunological factors are important in pregnancy loss because of the interaction between mother and fetus. T-regulatory cells as the component of humeral immune response play important role in the fetu-maternal interface. One of the regulatory mechanisms for these cells is mediated by antigen independent co-stimulatory signals and interaction of Cytotoxic T-Lymphocyte Antigen 4 (B7/CTLA-4) is one of these signals. The CTLA-4 which down regulates the activation and proliferation of T-cells occurs in a competitive interaction with CD28 to bind to B7. The aim of this study was to find out the relationship of CTLA-4 +49A/G gene with Recurrent Miscarriage in a group of Iranian women.
Methods: In the present study, 60 women with the history of two or more pregnancy loss were selected and considered as the case group. A group of women (n=60) with at least two live births without any previous history of pregnancy loss and autoimmune diseases were taken as control group. Genomic DNA was extracted from whole blood using standard protocols. The CTLA-4 +49 A/G were detected using polymerase chain reaction-restriction fragment length polymorphisms assay.
Results: The results showed that CTLA-4 +49 A/G polymorphisms were not significantly different in women with the history of two or more pregnancy loss compared to normal individuals. The frequency of G-allele polymorphism was 39.16% and 35.83% in patients and controls respectively.
Conclusions: The data presented may suggest that the CTLA-4 is not associated with recurrent miscarriage in an Iranian population in Northwest region.https://mbd.modares.ac.ir/article_5779_7d3d5bcad324d3edc08e40738e663554.pdfTarbiat Modares UniversityMolecular and Biochemical Diagnosis Journal2383-05221320141001Relationship between idiopathic female infertility with KISS1receptor gene mutation in northern Iranian women1771835780ENHamidrezaVaziriDepartment of Biology, Faculty of Sciences, University of Guilan, IranZahraSayarDepartment of Biology, Faculty of Sciences, University of Guilan, IranRoyaFarajiFaculty of Medicine, Guilan University of Medical Sciences, Rasht, IranJournal Article19700101Background: The kiss peptins and its receptor G protein coupled receptor (GPR54) or KISS1 receptor system are being described as key signaling molecules for reproductive function in animal models and humans. They play essential roles in regulation of the hypothalamic- pituitary- gonadal (HPG) axis and the onset of puberty and fertility. Objective: This study was performed to delineate the association of T305C (Leu 102 Pro) KISS1 receptor gene mutation with idiopathic female infertility in Iranian women. Methods: In this study, 140 healthy women with at least one child and no history of infertility and abortion and 130 idiopathic infertile women were recruited for this study. By using allele specific PCR (AS-PCR) method, the allele and genotype frequencies among infertile and healthy women were determined. Results: The gene frequencies of the 305 T and C allele of the KISS1 receptor were 45% and 54% among infertile women and 50% and 50% among healthy controls, respectively. The distribution of genotype frequencies in the patients and controls was as follows: TT (Leu/Leu) was 15% and 0%, TC (Leu/Proline) was 60% and 100% and CC (Pro/Pro) was 24% and 0% respectively. Structural analysis was performed using the MedCalc program (version 12). Our results suggests that significant association were not observed in genotype (P=0.8) and allelic (P=0.6) distribution between cases and controls. Conclusions: The data presented show that mutant allele C is not a risk factor for infertility, suggesting that the presence of KISS1 receptor T305C mutation is probably not associated with idiopathic female infertility in this population (P>0.05).https://mbd.modares.ac.ir/article_5780_294a8ed24b1ad22ec2e7efea049b8737.pdfTarbiat Modares UniversityMolecular and Biochemical Diagnosis Journal2383-05221320141001The effects of aqueous extract of chicory root on steatosis, lipid profile and liver damage enzyme markers in tamoxifen-treated rats1851945781ENAbolfazlNasiriDepartment of Clinical Biochemistry, School of Medicine, Hamadan University of Medical Sciences, Hamadan, IranNasrinZiamajidiDepartment of Clinical Biochemistry, School of Medicine, Hamadan University of Medical Sciences, Hamadan, IranHamidBehroujDepartment of Clinical Biochemistry, School of Medicine, Hamadan University of Medical Sciences, Hamadan, IranRoghayehAbbasalipourkabirDepartment of Clinical Biochemistry, School of Medicine, Hamadan University of Medical Sciences, Hamadan, IranArashDehghanDepartment of Pathology, School of Medicine, Hamadan University of Medical Sciences, Hamadan, IranJournal Article19700101Background: Accumulation of triglycerides in the liver i.e. steatosis, is a well-known side-effect of tamoxifen administration to patients suffering from breast cancer. Cichoriumintybus (chicory) is a plant used as traditional medicine for curing liver disorders. In this study, the effects of extract prepared from chicory roots on tamoxifen-induced liver steatosis and related biochemical factors in animal model using rats has been investigated. Methods: Female rats of Wistar strain were divided into four groups and treated as follows; 1-Control: received vehicle; 2- Chicory root-extract treated: rats were given by gavage the aqueous chicory root extract (1 g/kg body weight/day for 14 days).3- Tamoxifen-treated: rats received tamoxifen (1 mg/kg body weight/day, for 7 days). 4- Tamoxifen+chicory-group: animals received tamoxifen (1 mg/kg body weight/day for 7 days) followed by chicory extract given by gavage (1 g/kg body weight/day for 14 days). After treatment, blood was collected by cardiac puncher, plasma was separated and plasma levels of glucose, total protein, triglyceride, cholesterol,LDL-C, HDL-C and activities of ALT, AST and ALP were measured. Liver tissues were homogenized used for measuring tissue triglyceride and histological examinations. Results: The data show that tamoxifen treatment caused a significant decrease in the level of serum cholesterol, HDL-C and total protein. However, serum ALT level was increased in tamoxifen-treated rats compared to controls. Increased serum ALT in tamoxifen-treated rats was recovered in rats treated with plant extract (tamoxifen+chicory-group). HDL-C and total protein levels were unaffected in rats fed chicory extracts. Tamaxifen-treated animals showed signs of liver steatosis as shown by histological examination and accumulation liver triglyceride. The steatosis markers such as accumulated triglyceride in liver was significantly reduced due to the plant extract treatments when compared to tamoxifen-group. Conclusions: Dietary extract prepared from chicory roots is effective in modulation of tamoxifen-induced liver damage and steatosis.https://mbd.modares.ac.ir/article_5781_8e2c381d4dd04f1c55093f22c59c3a08.pdfTarbiat Modares UniversityMolecular and Biochemical Diagnosis Journal2383-05221320141001Hepatic ethoxyresorufin O-deethylase activity of liver microsomal cytochrome P-450 (CYP1A1) in young and adult rats treated with paracetamol and β-naphthoflavone1952045782ENMohammadRahmati-YamchiDepartment of Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, I.R. Iran.
Department of Biochemistry, Faculty of Medicine, Tabriz University of Medical Science, Tabriz, I.R. IranYousefRasmiDepartment of Biochemistry, Faculty of Medicine, Urumia University of Medical Science, Urumia, I.R. IranAbdolamirAllamehDepartment of Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, I.R. IranJournal Article19700101Background: Age-related differences in the ethoxyresorufin O-deethylase (EROD) activity of CYP1A1 and its inducibility in rats may determine the toxic potential of acetaminophen. This study was carried out to compare the effects of acetaminophen (APAP) and β-naphthoflavone (βNF) on CYP1A1 activity in young and adult rats.
Methods: For this purpose, young and adult rats (n = four / group) were treated with different doses of APAP. Likewise groups of young and adult rats were treated with a single dose of β-naphthoflavone (βNF, 67 mg / kg b.w). EROD was measured in microsomal fraction using resorufin as the substrate.
Results:The results showed that a single i. p. injection of APAP (25 mg / kg B.W.) failed to alter liver microsomal EROD in young and adults. Whereas, in adults treated with 250 and 450 mg APAP / kg B.W, liver CYP1A1 was elevated to about 45 and 60% respectively. The rate of CYP1A1 induction in young rats with single dose of APAP (450 mg/kg B.W) was approximately 32%. Induction in CYP1A1 was noticed 4 h after APAP injection and returned to normal levels in 24 h. The inducibility of CYP1A1 in rats treated with a toxic dose of APAP was comparable to the data obtained from animals treated βNF, 67 mg / kg b.w.
Conclusion: These results together with our previous reports indicate a similar pattern of changes in CYP1A1 in both the age-groups treated with toxic doses of APAP may suggest that the inducible CYP1A1 can equally contribute to protection against liver damage in young and adult rats.https://mbd.modares.ac.ir/article_5782_cf9dc5e4e194fc21f397b4cac9cc3ae9.pdfTarbiat Modares UniversityMolecular and Biochemical Diagnosis Journal2383-05221320141001Identification of the Propionibacterium acnes using Polymerase Chain Reaction (PCR) in the Acne vulgaris lesions2052125783ENMasoodGhaneDepartment of Microbiology, Faculty of Biological Sciences, Islamic Azad University, Tonekabon Branch, Tonekabon, IranNeginNaghdiDepartment of Microbiology, Faculty of Biological Sciences, Islamic Azad University, Tonekabon Branch, Tonekabon, IranJournal Article19700101Background: Acne vulgaris is an inflammatory chronic disease of pilosebaceous unit. One of the most important factors playing a role in occurrence of acne is presence of <em>Propionibacterium acnes</em>. With the aim of molecular identification of the P. acnes from the acne vulgaris lesions, current research was carried out. Methods: In this study, contents within the lesions was collected from 70 patients. The presence of the <em>P. acnes</em> was examined by a specific PCR technique. Results: Of 70 samples, 58 samples (82.85%) were determined to be positive in terms of presence of <em>P. acnes</em>. No significant relationship was observed between presence of <em>P. acnes</em> and each one of the studied demographic factors, including gender, age, disease period, family background and treatment background. Conclusions: The adopted molecular technique has obviated the limitations associated with the culture method for identification of the bacteria. To overcome the problems with conventional culture techniques for <em>P. acne</em>, this PCR method is promising for better identification of this bacterium.https://mbd.modares.ac.ir/article_5783_634841a6831464b64c072c8510c7f35c.pdf