International Journal of Medical Laboratory

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.

Background and Aims: One of the major subjects for improving in vitro fertilization (IVF) outcome is the quantity and quality of retrieved oocytes. In vitro maturation (IVM) provides an opportunity for using immature oocytes routinely discarded in clinics. This study aimed at evaluating the quality of embryos derived from in vivo and rescue in vitro matured oocytes.
Materials and Methods: Totally, 462 immature oocytes as cases and 466 mature (MII) oocytes as controls were included for study of their developmental competence. Oocytes underwent intracytoplasmic sperm injection insemination and then denuded oocytes were microscopically assessed regarding cytoplasmic and nuclear maturity and quality.
Results: The morphological assessments showed fertilization rate of 60.9 and 61.4%, the embryo formation rate of 86.7% and 90.9% and arresting rate of 27.3% and 25.6% for the case and control oocytes, respectively. Evaluating embryo quality in the cleavage stage indicated that 63% of the embryos in the case group and 68% of the embryos in the control group were of good quality. There was no significant difference between fertility rate and arresting rate of oocytes matured in both groups, although the embryo formation rate and the quality of embryos differed significantly.
Conclusions: Our findings suggest that IVM is a valuable and practical option for patients who had to cancel IVF treatment cycles because of severe responses or resistance to routine hormonal therapies or those with low functional ovarian reserve.

Background and Aims: Mercury, with its oxidative activity, causes damage to the antioxidant enzymes thus resulting in physiological disorders. Sodium selenide is an antioxidant that protects antioxidant enzymes. The aim of this study was to investigate the protective effect of sodium selenide on renal toxicity induced by mercuric chloride in rats.
Materials and Methods: Animals were divided into 6 groups of 6 each. Control group was treated with mercuric chloride (2.5 mg/kg) subcutaneously. Groups 2, 3 and 4 of the rats were treated with 2.5 mg/kg of mercuric chloride and sodium selenide subcutaneously administered at doses of 0.02, 0.04 and 0.08 mg/kg, respectively. The fifth group received 0.04 mg/kg sodium selenide subcutaneously after 25 days of mercuric chloride (2.5 mg/kg) treatment. The sixth group was treated with 2.5 mg/kg of mercuric chloride subcutaneously and 100 mg/kg of vitamin E by gavage.
Results: After data analysis of the activity of the enzyme catalase, superoxide dismutase, glutathione peroxide, creatinine and urea, there was a significant difference in the activity of these factors compared with the control group in blood and kidney tissue (p<0.05).
Conclusions: This study indicated that mercuric chloride has toxic effects in blood and kidney and sodium selenide may be able to reduce its’ blood and renal toxicity.

Background and Aims: One of the neurotransmitters in the brain is Histamine which acts as several biological mechanism regulators like inflammation, gastric acid secretion, and neuromodulation. Inactivation of Histamine occurs by histamine N-methyltransferase (HNMT) enzyme. The HNMT transfers a methyl group from S-adenosyl-L-methionine to Histamine and is the main process for the termination of neurotransmission actions of Histamine in the mammalian central nervous system.
Materials and Methods: In this case, a family was referred to the genetic clinic to diagnose the cause of their disorder. The clinical form, pedigree, and questionnaire were completed for the family, and the parents gave their written consent for all tests and photographs publication. Both siblings have moderate learning and intellectual disability. Whole exome sequencing was performed and Sanger sequencing for co-segregation was used.
Results: Bioinformatics analysis revealed a homozygous missense variant in HNMT (c.623T>C p.Leu208Pro) which causes non-syndromic autosomal recessive intellectual disability in this consanguineous family. Analysis of segregation confirmed this mutation. P.Leu208Pro mutation reduces the stability of the protein, which reduces the inactivation of Histamine.
Conclusion: HNMT should be considered an important gene in the genetic evaluation of consanguineous families with intellectual disability. 

Background and Aims: Hypertyrosinemia type 3 (HT3) is an inherited error in tyrosine metabolism caused by a mutation in the 4-hydroxyphenylpyruvate dioxygenase (HPD) gene. Here we report a one and half-year-old girl infant who was diagnosed based on increased serum tyrosine levels and increased urinary excretion of p-hydroxyphenyl derivatives.
Materials and Methods: The proband was one and half-year-old Iranian girl who was diagnosed based on increased serum tyrosine levels and increased urinary excretion of p-hydroxyphenyl derivatives. In this study, we used Whole-Exome Sequencing to identify the genetic reason for the disease and the funded mutation confirmed by Sanger sequencing.
Results and Conclusion: Through whole-exome sequencing screening of heterozygotes c.413C>T (p.T138M) and c.75G.A (p.W25Ter) in the HPD gene and genetically confirmed by Sanger sequencing. There were heterozygous conditions c.413C>T (p.T138M) and c.75G.A (p.W25Ter) in father and mother respectively. This mutation in her parents was also confirmed by Sanger sequencing.

Background and Aims: Polycystic ovary syndrome (PCOS) has a largely unknown etiology and is a common heterogeneous endocrine disorder in women of reproductive age, characterized by polycystic ovaries, hyperandrogenism, insulin resistance, and chronic anovulation. MicroRNAs (miRNAs) are small, noncoding RNA sequences that negatively regulate gene expression at the post-transcriptional level. miRNA-103 has been associated with metabolic disorders such as obesity and diabetes, which are also associated with PCOS. This study aims to describe the different expressions of circulating miR-103 in the plasma of PCOS women.
Materials and Methods: This case-control study includes 25 women with PCOS and 25 healthy women. Plasma total RNA was isolated and, after polyadenylation, converted to total cDNA. Then, the expression level of microRNA-103 was analyzed by quantitative reverse transcription-polymerase chain reaction, and SNORD miRNA was used as the reference gene.
Results: Our results indicate that the expression level of miRNA-103 significantly decreased in the PCOS group compared to healthy women, which can be related to the etiology and progression of PCOS (p < 0.05).
Conclusions: Our findings recommend further studies in the larger statistical population and more accurate techniques to confirm microRNA-103 as a noninvasive diagnosis biomarker.

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.