فهرست مطالب

Molecular and Biochemical Diagnosis - Volume:2 Issue:1, 2016
  • Volume:2 Issue:1, 2016
  • تاریخ انتشار: 1395/09/30
  • تعداد عناوین: 7
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  • Maryam Adelipour, Foad Abdollahpour, Abdolamir Allameh* Pages 1-16
    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.
    Keywords: Cancer stem cell, Biomarker, Tumor
  • Ragaa H. M. Salama, Yasser F. Abd, Elraheem, Khaled H. Mahmoud, Jalal A. Bilal, Almutlaq M. Abdullah, Aya A. A. Alsanory, Tasnim A. A. Alsanory Pages 17-25
    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.
    Keywords: Human islet cell antibody (ICA), Insulin auto antibodies (IAA), Antiglutamic acid decarboxylase (anti-GAD) antibodies
  • Mandana Salehizadeh *, Mitra Salehi, Mohamad Reza Falahiyan Pages 27-41
    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.
    Keywords: Antimicrobial, Mutacin, Streptococcus mutans, Gram-Positive Bacteria
  • Razieh Abdolvahabi *, Abdolfatah Sarrafnejad, Mohsen Nafar, Aliakbar Amirzargar Pages 43-50
    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
    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
    Keywords: Kidney transplantation, Acute rejection, SNP, TLR2, CD14
  • Syed Farhan Ahmad*, Zubair Anwar, Shahid Hussain, Maryam Jehangir, Irum Jehangir, Anwar Jamal, Jabar Zaman Khan Khattak, Ayaz Ali Khan Pages 51-64
    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.
    Keywords: Khyber Pakhtunkhwa, Hepatitis C, RT-PCR
  • Morteza Abkar, Abbas Sahebghadam Lotfi *, Jafar Amani, Mehdi Fasihi Ramandi Pages 65-78
    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.
    Keywords: Recombinant protein, Refolding, Inclusion Body
  • Shima Hadipour, Hossein Ghafoori *, Nooshin Sohrabi, Maryam Izaddoust Pages 79-94
    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.
    Keywords: ? - Amylase, Bacillus persicus, Q - Sepharose, Haloalkaline, Thermostable