International Journal of Medical Laboratory

1-6

Authors: Homayoon Bashiri 1,2 M.D., Mahsa Madani 3 M.D., Lotfollah Asgari 2,3 M.D., Arezoo Bozorgomid 1* Ph.D.

Background and Aims: Limited information is available on the frequency of advanced adenomas in diminutive colon polyps. Thus, this study aimed to investigate the pathological characteristics and the frequency of high-grade dysplasia in diminutive colorectal polyps in individuals referred to colonoscopy examination in Kermanshah, Iran. Materials and Methods: Demographics characteristics, location and diameter of the polyp, histological assessment of the polyps, grade, and others were retrieved from colonoscopy reports. Results and Conclusions: During the study period, 250 diminutive colorectal polyps were detected. The histological assessment showed that 36.4% were adenomatous, 32.8% were hyperplastic, and 30.8% were inflammatory polyps. Only two diminutive polyps (0.8%) had high-grade dysplasia, and the frequency of adenocarcinoma in our study was 0.4%. Besides, the frequency of adenomatous polyps was higher in the proximal versus the distal colon. These findings emphasize the urgent need for a colorectal cancer screening plan in the Iranian population to improve therapeutic outcomes.

23-34

Authors: Yasaman Issazadeh 1 M.Sc., Mohadeseh Farnaghizad 1 M.Sc., Sarvenaz Falsafi 2 Ph.D., Hoora Mazaheri 1 Ph.D., Shokoofeh Ghazi 1 Ph.D., Ava Behrouzi 1* Ph.D.

Background and Aims: Hospital infections and their antibiotic resistance have become a global concern recently. One of the most prominent factors in hospital infections is Pseudomonas aeruginosa (P. aeruginosa), which can become resistant to many antibiotics due to its ability to form biofilms. Recently, scientists have tried to replace antibiotic therapy with alternative therapies such as probiotics which can reduce or eliminate the pathogenic bacteria's ability to form biofilms. Therefore, the present study revealed that some genes, such as algD and PpyR, were involved in biofilm formation in P. aeruginosa. Furthermore, the inhibitory effect of the supernatant of lactobacillus agilis on the biofilm formation of P. aeruginosa was evaluated in the current study. Materials and Methods: In this study, the effect of the supernatant of probiotic Lactobacillus agilis on the biofilm formation of P. aeruginosa and also the expression of two genes effective in biofilm formation (algD and PpyR) were investigated. Antibiograms were performed to detect the most resistant bacteria since there is a link between biofilm formation and antibiotic resistance. Further, the effects of probiotics on the expression of PpyR and algD genes were discussed. Results: Results showed that the biofilm formation of P. aeruginosa was significantly reduced in the presence of lactobacillus agilis. Conclusions: According to the current study, it could be concluded that because of antibiotics resistance and their associated mechanisms, probiotics could be used as a replacement for antibiotics in many treatments.

46-56

Authors: Mohammad Shokouhian 1 M.Sc., Hossein Mozdarani 2 Ph.D., Masoud Soleimani 3 Ph.D., Majid Safa 1 Ph.D., Mohammad Reza Rezvany 1, 4, 5* Ph.D.

Background and Aims: The occurrence of single and double-strand breaks of DNA damage is the major obstacle for proliferation under various environmental factors and, if not repaired, can result in many consequences, including mutation, cell death, and others. So, the present study was conducted to evaluate the damage of DNA and the expression status of DNA repair system genes before and after stem cell proliferation. Materials and Methods: The MACS method isolated the umbilical cord blood hematopoietic stem cells (UCB-HSCs). In order to investigate cell death, the study of Annexin V/PI was done by flow cytometry. Comet assay made observation and identification of DNA breaks, and the expression of genes normally involved in the repair of DNA breaks was evaluated by real-time polymerase chain reaction. Results: The average number of stem cells increased by 1.9-fold after three days of proliferation. The apoptotic percentage of cells was negligible (less than 0.2%), and the purity of the CD34+ cells was reduced by about one-third in three days (67%). By examining the expression of DNA repair genes, including KU70, KU80, RAD51, and XRCC1, their increased fold change was not significant. In a microscopic examination of stem cells in the comet assay, there was no significant difference between DNA damage before (1.33% ± 0.31) and after (2.08% ± 0.92) replication. Conclusion: In our investigation, neither DNA damage nor changes in the DNA break repair were observed. However, further studies are required to clarify the DNA break repair by recruiting more UCB-HSCs samples.

67-74

Authors: Erfan Zaker 1, 2 M.Sc., Fateme Zare 3,4 Ph.D., Seyed Hossein Hejazi 5 Ph.D., Hossein Khanahmad 2 MD., Ph.D., Seyed Mehdi Kalantar 1* Ph.D.

Background and Aims: A monoclonal antibody (mAb) can unambiguously identify, quantify, and purify an antigen or particular epitope at a large scale. The superiority of these antibodies lies in their specificity for the antigenic determinant. So, this study aims to prepare mouse mAb-secreting hybridoma against human gamma interferon (IFN-γ) and determine the produced antibody's characters. Materials and Methods: Mouse splenic B lymphocytes immunized with recombinant human IFN-γ were fused with mouse SP2/0 cells. The hybridized cells were selected by hypoxanthine-aminopterin-thymidine and hypoxanthine-thymidine media to obtain monoclonal antibody-producing hybridoma cells. Finally, indirect enzyme-linked immunosorbent assay (ELISA), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and western blot were used to confirm the creation of antibody-secreting hybridoma cells. Results: mAb against IFN-γ were produced by fusing SP2/0 mouse non-secretory myeloma cell line with the spleen cells of immunized mice. This antibody's indirect ELISA optical density was 2.055 on average, and the desired antibody bands were confirmed in SDS-PAGE compared to Septicol® (commercial antibody). Also, in the western blot, the desired antibody could bind to the antigen. IFN-γ transferred on nitrocellulose membrane. In ELISA and western blot tests, anti-mouse IgG conjugated antibodies were used; therefore, the mAb IgG isotype was taken into consideration. Conclusion: In this study, a mouse mAb was obtained by immunization of Balb/C mice and fusion of spleen cells of these mice with the SP2/0 cells, which can specifically bind to recombinant human IFN-γ and can be used to detect IFN-γ secretion in all types of intracellular infections, including latent tuberculosis.