International Journal of Medical Laboratory

239-250

Authors: Amir Ghorbanihaghjo Ph.D., Shahin Alizadeh Fanalou Ph.D. Navid Farahmandian Ph.D., Elham Bahreini* Ph.D.

Background and Aims: Age-related macular degeneration (ARMD) is a degenerative retinal disorder that causes progressive loss of central vision in older adults. The study aimed to determine the effect of asymmetric dimethylarginine (ADMA) as oxidizing metabolite and paraoxonase (PON1) activity within its phenotypes as an antioxidant agent in the development of such multifactorial disease. Materials and methods: Forty-five exudative ARMD (E-ARMD) patients and 45 healthy controls were enrolled for this case-control study. Serum PON1 activity was measured using paraoxon and phenylacetate as substrates. PON1 phenotype was determined using the double-substrate method. The ADMA and oxidized LDL (OX-LDL) levels were determined by enzyme-linked immunosorbent assay method. Blood lipid profile was measured, and nontraditional lipid indexes were calculated. Results: Three phenotypes were determined for PON1 among the participants in the study, including AA, AB, and BB phenotypes with low, moderate, and high activity, respectively. AA phenotype was more frequent among E-ARMD, while AB and BB phenotypes were more common among healthy subjects. The mean ADMA and OX-LDL levels were significantly higher in the patients comparing to the controls (p=0.02 and p=0.01, respectively). No significant differences were found in ADMA and OX-LDL levels between phenotypes in each group and also among similar phenotypes. LDL, cholesterol, and even all comprehensive lipid indexes except (atherogenic index of plasma) were higher in E-ARMD patients compared with the healthy group. Conclusions: It was concluded that high-risk individuals could be identified by evaluating the blood factors involved in oxidative stress, and antioxidant therapies may reduce the incidence and progression of the disease.

258-266

Authors: Parisa Bagheri1 M.Sc., Farhad Zaker2 Ph.D., Mohammad Hadi Sadeghian3 M.D., Ph.D., Soodeh Namjoo2 Ph.D., Saeedeh Hajebi Khaniki4 Ph.D., Zahra Chehreghani1 M.Sc., Maryam Sheikhi3 M.Sc., Hossein Ayatollahi3* M.D., Ph.D.

Background and Aims: DNA methyltransferase3A (DNMT3A) is necessary for the adjustment of gene expression, and the mutations in the DNMT3A gene are reported in a variety of leukemia cases. DNMT3A mutations are during cancer progression and cause poor prognosis in many leukemias. Thus, this gene can be a target for new treatments. This study aimed to examine the distribution of DNMT3A mutations in Iranian acute leukemia patients. Materials and Methods: In this study, diagnostic samples from 45 patients with de novo acute leukemia, including 22 acute myeloid leukemia (AML) patients, and 23 acute leukemia lymphoblastic (ALL) patients were screened, from April 2017 to March 2018 for the incidence of DNMT3A mutations by polymerase chain reaction and direct sequencing. Results: A total of 2 (9.1%) AML cases and 1 (4.34%) ALL cases were found to have the DNMT3A R882H mutation. It was found that a total of 22.7% and 21.7% of patients with AML and ALL had polymorphism rs368516543, respectively. DNMT3A mutations were considerably associated with higher age in AML patients. Conclusions: The findings suggest that the DNMT3A mutations are probably a new biomarker in the early examination and treatment of acute leukemia, even though further studies are needed.

280-288

Authors: Mahtab Teimouri1 Ph.D., Batool Hashemibeni2* Ph.D., Mohammad Mardani2 Ph.D., Muhammadhosein Moradi3 M.D. Student

Background and Aims: The current study was conducted to compare the expression levels of collagen type Π and X during chondrogenesis of human adipose-derived mesenchymal stem cells (hADMSCs) pellet and micromass cultures. Materials and Methods: Extracted hADMSCs were cultured until three passages and then transferred to pellet and micromass cultures in the experimental groups of day 7 and day14. For pellet and micromass cultures, aliquots of 5×105 cells/ml were centrifuged and respectively cultured in the conical tubes and droplets (12.5 μl) of the 24-well plates containing chondrogenic medium. Realtime-polymerase chain reaction technique was performed for gene expression levels. Results: Increased expression of collagen type Π was shown in micromass day14 compared to micromass day 7, pellet day 14 (p<0.01) and pellet day 7 (p<0.001). Also, an increased expression of collagen type Π was seen in micromass day 7 and pellet day 14 compared to pellet day 7 (p< 0.05). Expression of collagen type X increased in pellet day 14 compared to micromass on days 7 and 14 (p<0.001, p<0.01) and pellet day14 compared to pellet day7 (p< 0.05). An increased expression of collagen type X was shown in pellet day 7 compared to micromass on days 7 and 14 (p<0.05). Conclusions: According to the results, higher expression of collagen type Π and lower expression of collagen type X in micromass cultures that are prepared by cell suspension play a better role during cellular condensation that leads to the formation of large nodules exhibiting cartilage-like morphology, suggests a higher efficiency for micromass cultures.

312-320

Authors: Nima Rahmati1 M.D., Ph.D., Fatemeh Hajighasemi1* Ph.D.

Background and Aims: Nitric oxide (NO) has an essential role in inflammation and has been related to pathogenesis and the progress of numerous inflammatory-based diseases, including some cancers. Peganum harmala (P. harmala) is a medicinal plant used for the treatment of numerous diseases such as several infections. Also, anti-inflammatory effects of P. harmala extracts and its derivatives (harmaline and harmine) by suppressing myeloperoxidase, NO, and other mediators have been demonstrated in vivo. In this study, the effect of P. harmala seeds aqueous extract on NO production in U937 monocytic cells and peritoneal macrophages has been evaluated in vitro. Materials and Methods: U937 and mice peritoneal macrophages were cultured in Roswell Park Memorial institute-1640 with 10% fetal calf serum. Then, the cells at the logarithmic growth phase were incubated with different concentrations of aqueous extract of P. harmala seeds (0.1-1 mg/ml) for 24 hours. Next, NO production was assessed by the Griess method in the culture medium. Results: P. harmala seeds aqueous extract did not significantly affect lipopolysaccharide-induced NO production in U937 cells and peritoneal macrophages after 24 hours incubation time compared with untreated control cells. Conclusions: These results suggest that the anti-inflammatory effects of P. harmala may be mediated through NO-independent mechanism(s). However, further studies are warranted to define the P. harmala aqueous extract impact on NO expression in other related normal and cancerous cells.