Tarbiat Modares University Molecular and Biochemical Diagnosis Journal 2383-0522 2 1 2016 03 01 Cancer stem cell markers 1 16 5784 EN Maryam Adelipour Department of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran Foad Abdollahpour Department of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran Abdolamir Allameh Department of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran Journal Article Cancer stem cells (CSC) are the tumor-associated cells existed within tumors or hematological cancers which share characteristics similar to normal stem cells. The common characteristics of a normal stem cell and a CSC are their differentiation capacity and self-renewal in tumors. The expression pattern of CSC markers differs depending on the type and location of cancers. CD molecules are probably the most common biomarkers for CSCs. CD molecules such as CD133, CD24, CD44, CD138 and similar CD molecules are well known markers for identification of CSCs. In addition, ATP-Binding Cassette (ABC) transporters such as ABCG2 and ABCB5 as well as EpCAM, ALDH1 and CXCR4 have been used to identify certain CSCs. Therefore these markers may be considered specific for better identification and diagnosis of a specific tumor. Currently studies are in progress to find new cell surface markers which can distinguish specific markers from other markers for isolation and characterization of CSCs. The future of this area of research is promising in developing novel prognostic assays and therapeutic approaches based on cellular and signaling functions of these markers.

https://mbd.modares.ac.ir/article_5784_6157966f9b9e2f35d2266675bad8b7f8.pdf

Tarbiat Modares University Molecular and Biochemical Diagnosis Journal 2383-0522 2 1 2016 03 01 Biochemical markers could predict type-1 diabetes mellitus 17 25 5785 EN Ragaa H. M. Salama Department of Medical Biochemistry, Faculty of Medicine, Assiut University, Assiut, Egypt Yasser F. Abd-Elraheem Pediatric, College of Medicine, Qassim Universitym, KSA Khaled H. Mahmoud Pediatric, College of Medicine, Qassim Universitym, KSA Jalal A. Bilal Pediatric, College of Medicine, Qassim Universitym, KSA Almutlaq M. Abdullah Maternal Childhood Hospital, Qassim, KSA Aya A. A. Alsanory Medical Students, Assiut University, Egypt Tasnim A. A. Alsanory Pharmacy Students, Assiut University, Egypt Journal Article Background : Type 1 diabetes mellitus (T1DM) is an autoimmune disease characterized by selective destruction of pancreatic beta cells. Methods: The study included 80 children, 20 of them have T1DM, 40 children were selected from first degree relatives to the same child and 20 healthy children serve as control. Body mass index (BMI) was calculated, random blood glucose and glycosylated hemoglobin A1c (GHbA1c) were measured. The following biochemical markers were measured in sera of all subjects by ELISA kits: Human insulin ,C-peptide, human islet cell antibody (ICA), insulin auto antibodies (IAA) and antiglutamic acid decarboxylase (anti-GAD) antibodies. Results : This study showed that diabetic children had high level of ICA (65%), IAA (55%), anti-GAD antibodies (50%) and decrease in C-peptide (60%). Whereas the relatives showed high level of anti-GAD antibodies (30%), IAA(25%), ICA(2.5) and decrease in C-peptide (30%). Anti-GAD antibodies were significantly higher among the relatives of the diabetics compared to the healthy controls. Conclusions : The strongest predictors of diabetes were C- peptide and islets cell antibodies, which had odd ratio 4.7 and 3.1, respectively. Autoantibodies could distinguish T1DM patients from healthy control subjects and they may also identify individuals at high risk during progression from pre-diabetes to overt disease.

https://mbd.modares.ac.ir/article_5785_51425b752a0b402ed3effc83fc4bbb74.pdf

Tarbiat Modares University Molecular and Biochemical Diagnosis Journal 2383-0522 2 1 2016 03 01 Frequency and expression of mutacin and putative bacteriocin genes in isolated Streptococcus mutans and its antimicrobial activity against gram-positive and gram-negative bacteria 27 41 5786 EN Mandana Salehizadeh Department of Microbiology, Islamic Azad University (Tehran Olom Tahghighat), Tehran, IR Iran Mitra Salehi Department of Microbiology, Islamic Azad University, North Tehran Branch, Tehran, IR Iran Mohamad Reza Falahiyan Department of Microbiology, Islamic Azad University, North Tehran Branch, Tehran, IR Iran Journal Article Abstract Background: Streptococcus mutans in the oral cavities sable to produce mutacin (bacteriocin-like substances) with antibiotic properties. The aim of this study was to investigate the frequency and expression of genes encoding mutacins typeI, II, III and IV and also two of 8 genes in a cluster encoding the putative bacteriocins, the designated bsm 283, bsm 299, bsm 423, bsm 1889c, bsm 1892c, bsm 1896, bsm 1906c and bsm 1914, were also screened by PCR and specific primers for each type of mutacin biosynthesis gene and then mutacin activity against the indicator strains determined. Methods : In this study, dental clinic samples were collocated; Streptococcus mutans was detected using biochemical tests and molecular methods (PCR). Frequency of mutacin biosynthesis genes types I, II, III and IV, bsm299 and bsm1899 were measured by PCR, using specific primers for each type of mutacin biosynthesis gene. Furthermore, the antimicrobial spectra of Streptococcus mutans isolates against other indicators, including Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Salmonella typhi, Pseudomonas, Escherichia coli were evaluated using well diffusion, disk diffusion and the minimal inhibitory concentrations (MICs) methods. Results : Out of 56 samples collected from patients referred to Milad Hospital dental clinic on October 2011 and three private dental clinics on November 2011, 24 strains of Streptococcus mutans produced mutacins. 67.52% of the strains had a wide antimicrobial spectrum and 37.5% of 67.5% had a high frequency of genes with more inhibitory activity against, Staphylococcus epidermidis, Staphylococcus aureus and Enterococcus faecalis respectively that are more related to putative bacteriocins. The expression frequency of the bsm gene (putative bacteriocins) was higher than that of the characterized mutacins types (I–IV). The lowest dilution rate mutacin was found against Staphylococcus epidermidis (0.0625 unit/mL). Conclusion : These findings suggested that all putative bacteriocins may represent a large repertoire of inhibitory substances produced by Streptococcus mutans. Therefore, the high diversity of mutacin-producing phenotypes, associated with high frequency of expression of the biosynthesis genes screened and wide antimicrobial activity against Staphylococcus epidermidis could be used as safe antimicrobial agents in treatment of superficial infections such as, removing the pimple caused by Staphylococcus epidermidis.

https://mbd.modares.ac.ir/article_5786_77431ca7981f1f1483ae8a58bcbb6e0e.pdf

Tarbiat Modares University Molecular and Biochemical Diagnosis Journal 2383-0522 2 1 2016 03 01 Single nucleotide polymorphisms of innate immune receptors in patients with renal rejection 43 50 5787 EN Razieh Abdolvahabi Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Abdolfatah Sarrafnejad Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Mohsen Nafar Chronic Kidney Disease Research Center, Department of Nephrology, Shahid Labbafinejad Medical Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran Aliakbar Amirzargar Department of immunology, School of medicine, Tehran University of Medical Sciences, Tehran, Iran and Molecular Immunology Research Center, Tehran University of Medical Sciences, Tehran, Iran Journal Article Background: The innate immunity plays an important role in the host response to transplantation by Toll-like receptors and results in development of acute allograft rejection. The aim of this study was to evaluate the association of TLR2 and CD14 (co-receptor) gene polymorphisms with acute rejection in kidney transplant recipients. Methods : The study was conducted in a population of 239 subjects consisting of 71 patients with acute rejection, 71 patients without acute rejection (SGF) and 97 Healthy Control (HC). The allele and genotype frequencies of TLR2 (R753Q, rs5743708) and CD14 (-159 C>T, rs2569190) polymorphisms were genotyped by Real-time PCR in the study groups. Results : Genotype distribution of CD14 -159 polymorphism was significantly different in AR vs. SGF and HC. CD14 -159 TT genotype was more prevalent in rejection than SGF and HC (P<0.0001, P<0.007, respectively). Also Graft loss, defiened as need of dialysis after acute rejection, was occurred in 24 patients (33.8%) from AR group. The frequencies of three genotype in CD14 (TT, CT, CC) in rejection With Graft loss were 75.0%, 20.8% and 4.1% respectively, While 25.5%, 31.9% and 42.5% in rejection without Graft loss (P<0.0001 for TT vs. CT, CC). Many recipients with AR were involved with graft loss had CD14 -159 TT genotype, whereas only a few recipients without graft loss had TT genotype (p<0.0001). Conclusion : Therefore, due to the importance of CD14 polymorphism (-159 C/T, rs2569190) in disease progression and also as a biomarker, could be considered as a crucial therapeutic target in early prognosis of acute rejection

https://mbd.modares.ac.ir/article_5787_a32d7eeaae19821fd9ce317f3ce952a7.pdf

Tarbiat Modares University Molecular and Biochemical Diagnosis Journal 2383-0522 2 1 2016 03 01 A detailed report on the specific area based percentage and diagnostic methods of hepatitis C in Khyber Pakhtunkhwa, Pakistan 51 64 5788 EN Syed Farhan Ahmad Department of Biotechnology and Bioinformatics, Islamic International University Islamabad, Pakistan Zubair Anwar Department of Virology and Immunology, NIH, Islamabad, Pakistan Shahid Hussain Department of Bioinformatics, Capital University of Science & Technology, Islamabad, Pakistan Maryam Jehangir Department of Bioinformatics, Shaheed Benazir Bhutto Woman University, Peshawer, Pakistan Irum Jehangir Department of Microbiology, Khyber Medical University, Peshawar, Pakistan Anwar Jamal Department of Medicine, Khyber Medical College, Peshawar, Pakistan Jabar Zaman Khan Khattak Department of Biotechnology and Bioinformatics, Islamic International University Islamabad, Pakistan Ayaz Ali Khan Department of Biotechnology, University of Malakand, Chakdara, Lower Dir, Pakistan Journal Article Background: The prevalence of hepatitis C virus (HCV) varies tremendously in different parts of the world. This study reviews the percentage and molecular diagnosis of Hepatitis C in the persons from Khyber Pakhtunkhwa, Pakistan that visited to a particular laboratory. Methods : The method includes the diagnostic procedure steps by Real Time PCR. A Total numbers of 1050 Persons were screened during four months i.e. January-April, 2014. The collected data was evaluated for prevalence rate, age wise prevalence, gender wise prevalence and comparison of RT-PCR and ICT. Results : Overall percentage was 64.85 which is an overestimation of a true prevalence because of the specific sampling method applied to current study. Middle age persons were more affected. The percentage was higher in male (56.9) as compared to female (43.02). The RT-PCR diagnostic test was found to be more sensitive for the detection of HCV comparative to ICT. Conclusion : It is recommended that government should establish such laboratories equipped with RT-PCR for timely and accurate detection of HCV. Moreover, awareness programs are required to decrease the burden of HCV in the Pakistani population. https://mbd.modares.ac.ir/article_5788_967edfdcdfbcc3b2d253fac24326e5b5.pdf

Tarbiat Modares University Molecular and Biochemical Diagnosis Journal 2383-0522 2 1 2016 03 01 Optimization of a method for refolding of bacterial recombinant proteins 65 78 5789 EN Morteza Abkar Department of Molecular Genetics, Faculty of Basic Sciences, Tarbiat Modares University, Tehran, Iran Abbas Sahebghadam Lotfi Department of Clinical Biochemistry, Faculty of Medicine, Tarbiat Modares University, Tehran, Iran Jafar Amani Applied Microbiology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran Mehdi Fasihi Ramandi Molecular Biology Research Center, Baqiyatallah University of Medical Science, Tehran, Iran Journal Article Background: The over-expression of recombinant proteins in large amount is important for production of therapeutic proteins and structural study. There are several systems for expression of recombinant proteins. One of the most relevant expression systems is Escherichia coli (E. coli). Although this organism has many advantages, most of recombinant proteins expressed in E. coli hosts form inclusion bodies. For gaining biological activities, these structures should be refolded. Many techniques have been developed for in vitro protein refolding. Methods: In this study, a method was designed for inclusion body solubilization and protein refolding. IBs were solubilized in the solution containing 2M urea. This is a mild solubilization method without creating random coil structures in the protein. Results : Inclusion bodies undergo mild solubilization with maintain native-like secondary structures. Solubilized proteins were refolded on chromatography column by using native buffer conditions. The results showed the recombinant proteins were purified with high efficiency without aggregation. Conclusions : The results suggest that this method is easy, efficient, cheap procedure and usable for obtaining refolded recombinant proteins. In addition, purified protein with the method can be used in diagnosis and/or treatment of diseases.

https://mbd.modares.ac.ir/article_5789_f0efb5f6cb4ce54821a9c5c6e1dff052.pdf

Tarbiat Modares University Molecular and Biochemical Diagnosis Journal 2383-0522 2 1 2016 03 01 Purification and characterization of an extracellular thermostable alkaline α-amylase from the moderately halophilic bacterium, Bacillus persicus 79 94 5790 EN Shima Hadipour Department of Biology, Payam-e-Noor University, Tehran Branch, Pardis, Iran Hossein Ghafoori Department of Biology, University of Guilan, Rasht, Iran Nooshin Sohrabi Department of Biology, Payam-e-Noor University, Tehran Branch, Pardis, Iran Maryam Izaddoust Department of Biology, University of Guilan, Rasht, Iran Journal Article Background: Today a large number of bacterial amylases are available commercially in industry. The goal of the present study was purification and biochemical characterization of an extracellular thermostable alkaline α-amylase from the novel moderately halophilic, Bacillus persicus from the Aran-Bidgol, Iran. Methods : Purification of enzyme, was carried out by acetone precipitation, ultrafiltration and Q-Sepharose cation exchange chromatography. Results : The purified native enzyme showed a molecular mass of 53 kDa composed of a monomer by SDS–PAGE. The optimum pH, temperature and NaCl concentration were 10, 45 ∘C and 0.85 M respectively. It retained 50% of activity at 1.25 M NaCl and about 73% of activity at highly alkaline pH of 10.5, therefore it was a moderately halophilic and also can activate by divalent metal ions especially Ca2+ and Mg2+. The apparent values of Km and Vmax were obtained 1.053 mg/ml and 356μ/min respectively. Conclusion : In the present study we report the purification and characterization of a moderately halophilic α-amylase from a newly isolated Bacillus persicus. The purified enzyme shows interesting properties useful for industrial and biotechnological applications. The molecular cloning and structural studies of this α-amylase are in progress in our laboratory. https://mbd.modares.ac.ir/article_5790_8763d72bba4a7ade23f9ae1f09f4efc7.pdf