International Journal of Medical Laboratory

84-98

Authors: Fatemeh Montazeri , Ali Mohammad Foroughmand , Seyed Mehdi Kalantar , Abbas Aflatoonian , Mohammad Ali Khalilli

As numerical and structural defects in chromosomes are an inevitable consequence of IVF, Pre-implantation genetic diagnosis and screening (PGD/PGS) methods are used for detecting abnormalities in embryos before implantation to the uterus to increase the successful rate of IVF. Pre-implantation genetic diagnosis and screening approaches can be achieved by different techniques such as NGS, CGH and FISH. Among these approaches, FISH-based PGD/PGS is challenging in that it requires experience and skill to increase its facility and validity. Therefore, based on literature review and our experiences obtained from genetic laboratory of Yazd Reproductive Sciences Institute (Yazd, Iran), we were ditermined to discuss these challenges. After reviewing the available protocols and articles, we compared results of different methods for performing pre- and post-examination FISH process. Required samples in each section were obtained from embryo in cleavage or blastocyst stage. According to our team's experience, we recommend the cleavage stage biopsy and our modified fixation method. Also, we do not recommend more than two round hybridization on the same cell. Many studies have shown that FISH-based PGD is an efficient method for decreasing IVF failure in infertile patients. This paper introduces the best biopsy and fixation method and, includes some useful tips and tricks on type and number of probe, removing the cytoplasm, denaturation and hybridization, data evaluation and scoring criteria

103-112

Authors: Maryam Ahmadi , Farzaneh Tafvizi , Elham Moslemi

Background and Aims: Prostate cancer (PCa) is one of the most common cancers among men in Iran. Since changes in the regulation of proto-oncogenes expression are the main causes of most human cancers, including PCa, evaluating the expression of marker genes can be helpful for early diagnosis of cancer and better understanding of its etiology. The present study compared c-Myc expression level in prostatic adenocarcinoma and benign prostatic hyperplasia (BPH). Material and Methods: Paraffin-embedded prostatic tissues from patients with prostate adenocarcinoma (n=38) and BPH (n=38) were selected. The samples were included only if the patients underwent radical prostatectomy and had no history of hormone therapy, chemotherapy, or radiotherapy. After RNA extraction and cDNA synthesis, c-Myc expression in the samples was compared using SYBR green-based real-time polymerase chain reaction. Results: Significantly higher c-Myc mRNA expression was observed in adenocarcinoma samples than in BPH group (p=0.001). No significant correlation was observed between c-Myc expression and Gleason Score (p>0.05). There were no significant correlations between c-Myc expression and prostate-specific antigen levels and age (p>0.05). Conclusions: The c-Myc mRNA expression increased in the PCa samples compared with the BPH group. It seems that c-Myc expression can be introduced as a prognostic marker for determination of the invasive potential of tumor cells. Further tests and studies conducted with larger sample sizes may help to use this marker in differentiating malignant from benign samples.

123-132

Authors: Shiva Aghaei , Seyedhossein Hekmatimoghaddam , Mehdi Kalantar , Mohammad Hassan Sheikha , Mohammad Sobhan , Ali Jebali

Volume 5, Issue 2 (May 2018) IJML 2018, 5(2): 123-132 | Back to browse issues page XML Print Download citation: BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks Send citation to: Mendeley Zotero RefWorks Aghaei S, Hekmatimoghaddam S, Kalantar M, Sheikha M H, Sobhan M, Jebali A. Fe3O4@Ag Nanoprobe for Detection of Ovarian Cancer Cell Line Using Magnetic Resonance Imaging. IJML. 2018; 5 (2) :123-132 URL: http://ijml.ssu.ac.ir/article-1-212-en.html Fe3O4@Ag Nanoprobe for Detection of Ovarian Cancer Cell Line Using Magnetic Resonance Imaging Shiva Aghaei , Seyedhossein Hekmatimoghaddam , Mehdi Kalantar , Mohammad Hassan Sheikha , Mohammad Sobhan , Ali Jebali Abstract: (40 Views) Background and Aims: Magnetic resonance imaging (MRI) plays an essential role in molecular imaging by delivering the contrast agent into targeted cells. The aim of this study was to evaluate the use of magnetic nanoparticles containing iron oxide and silver (Fe3O4@Ag core-shell nanoprobe) as a contrast agent for the detection of ovarian cancer cell line ovcar-3. Materials and Methods: Co-precipitation method was used for synthesis of Fe3O4Ag nanoprobe which was then characterized by Fourier transform infrared spectroscopy (FTIR), dynamic light scattering (DLS) and scanning electron microscope (SEM). To evaluate the ability of this nanoprobe in detection of the ovcar-3 cell line by MRI, the cells were exposed to different (5 to 50 µg/mL) concentrations of Fe3O4@Ag nanoprobe before contrast intensity calculation by MRI. Results: SEM images revealed that Fe3O4@Ag nanoparticles are spherical, about 100 nm. FTIR showed strong absorbance picks belonging to the stretching vibration of Ag and Fe-O. It was found that contrast intensity of Fe3O4@Ag nanoprobe decreases as concentration decreases. Statistical analysis revealed significant difference in concentrations of 30, 40 and 50 μg/mL, compared to control (p<0.05). In the presence of Ovcar-3 cells, higher concentrations (10, 20, and 30 μg/mL) of nanoparticles also significantly increased contrast intensity in comparison with control (p<0.05). Conclusions: This novel magnetic nanoparticle can be used as an effective contrast agent for improving MRI in detection of ovarian cancer cells. The sensitivity of this contrast agent may be improved by binding to targeting molecules such as antibody and aptamer.

141-149

Authors: Zakieh Javidan , Nasrin Ghasemi , Hamid Reza Ashrafzade

Background and Aims: The cytochrome P450 1A1 (CYP1A1) plays a curial role in phase I metabolism of polycyclic aromatic hydrocarbons to their ultimate biologically active intermediates that have potential reproductive toxicity in men. Reproductive functions in men may be impaired by many environmental, physiologic, and genetic factors. The majority of the environmental factors are xenobiotics. Metabolic active xenobiotics exert adverse effects via covalent interactions between intermediate metabolites and cellular macromolecules such as DNA or protein. Materials and Methods: Genotyping two polymorphisms, CYP1A1*2A and CYP1A1*2C, was done using polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) assay in a case–control study including 105 infertile men and104 healthy fertile subjects. Results: The results showed that frequency of CYP1A1*2A was significantly different between the patients and the controls (p = 0.036). Analysis indicated that CYP1A1*2A CC genotype was significantly associated with an increased risk of male infertility (OR = 5.4, CI=1.12-26.04; 95%) compared with the AA genotype. No significant association was detected between CYP1A1*2C polymorphism and male infertility. Conclusions: The CYP1A1*2A single nucleotide polymorphism can be considered as an effective agent in azoospermia.