Archives of Advances in Biosciences

Summer Package  

Introduction: There is growing evidence revealing that genetic factors could be involved in the etiology of insulin resistance and diabetes. Recent studies now suggest that c-MYC oncogene amplification in beta-cells can cause downregulation of insulin gene expression and leading to diabetes. The present study was carried out to examine gene amplification level of c-MYC gene in healthy individuals compared to those with diabetes.

Materials and Methods : This case control study consisted of 70 subjects (34 diabetic patients and 36 healthy controls). The genomic DNA was extracted from blood samples using standard phenol-chloroform method, and Differential Quantitative PCR (DQ PCR) was accomplished. Subsequently, the mean Band Intensity Ratio (BIR) of c-MYC to γ-IFN was applied to determine the amplified products' molecular band intensity. The Real-time PCR was used to reconfirm the obtained results.

Results: The mean BIR of c-MYC to γ-IFN was 1.29 in diabetics, whereas in healthy individuals, the mean BIR was equal to 1.16.The findings indicate that the mean BIR in diabetics was almost 1.1 times higher than healthy controls; however, it was not statically different (P-value= .168).

Conclusions: Since no significant difference was found in terms of BIR of c-MYC to γ-IFN  in case and control groups, it can be concluded that c-MYC may not have a role to trigger the disease in this limited Iranian population. Further studies and larger statistical populations are needed to confirm our findings. This study showed that the DQ PCR technique could be reliable to screen gene amplification in large populations.

Introduction: Research has revealed that breast cancer is the second most common cancer among women and that the compounds in Artemisia plant have good anti-tumor potential. Nanomaterials significantly increase solubility, stability, and effective drug delivery. The aim of this study was to evaluate the efficacy of Nano-form of Artemisia essential oil in comparison with non-Nano-form in inhibiting MCF7 cells.

Materials and Methods:  After extracting essential oil from Artemisia Vulgaris plant, nanoparticles containing Artemisia Vulgaris essential oil were synthesized through homogenizer and sonication method. Different properties of nanoparticles including particle size, zeta potential, and morphology were measured. Finally, the toxicity effect of solid lipid nanoparticles containing essential oil on MCF7 cancer cell line was examined adopting MTT technique.Results: The results indicated that tDCS significantly reduced the scores on DDQ and OCDUS in the active tDCS group compared to the sham tDCS group (P<.05).

Results: The results of cellular effect of lipid nanoparticles containing Artemisia Vulgaris annua on MCF7 cells showed that increasing the concentration of lipid nanoparticles containing Artemisia Vulgaris essential oil, compared to purified essential oil, reduced the survival rate of MCF7 cells.

Conclusions: The findings show that lipid nanocarriers raise the release rate of essential oil compared to pure essential oil. Increase in concentration of lipid nanoparticles containing Artemisia essential oil reduces the survival rate of breast cancer cells. The IC50 obtained for unbaked essential oils and nanoparticles containing essential oils were 1105 and 262 μg/ml, respectively. Thus, it can be concluded that nanoparticle essential oil of this plant is effective in reducing the IC50 of the drug and increasing cytotoxicity.

Introduction: Multiple sclerosis (MS) is one of the most serious syndromes in human populations. Although the source of MS is unknown, some patterns have been discovered according which the genetic background and environmental factors are the key elements affecting its development. Since human samples cannot be repeatedly drawn from the spinal cord, animal models have been the best options available for studying MS so far. Experimental autoimmune encephalomyelitis (EAE) is the most recognized model in this regard. This study aimed to study EAE on rabbits to correlate symptoms and lesions, deducing whether this really can be a suitable model for predicting the algorithm lying behind the MS disease.

Materials and Methods:  For this purpose, 15 male two-month-old rabbits were divided into three groups, namely A, B, and C. The three groups were injected normal saline, complete Freund's adjuvant (CFA) + spinal cord homogenate, and normal saline + spinal cord homogenate via footpad and neck scruff, respectively. Then, they were submitted to laboratory and observed for histopathological changes.

Results: Group A did not show any signs or symptoms, while group B and C showed histopathological lesions. Moreover, group B was the only group showing clinical signs. There was also a significant difference between pathological and clinical signs in group A (p<.05), but not in group C (p>.05).

Conclusions: Considering the clinical and histopathological similarities between EAE and MS, the results suggest that EAE models are suitable to study MS.

Introduction: As one of the most common and invasive brain tumors, glioblastoma which originates in the nervous tissue of the brain has remained a therapeutic challenge given low success of conventional therapies. Small molecules including GW9508, due to their different roles in signaling and intracellular pathways and the production and increase of oxidative stress of mitochondrial origin, can cause cells to progress to apoptosis, also known as a cost-effective pharmacological factor. Therefore, in the present study, the anticancer and cytotoxic effects of GW9508 on A549 class lung cancer cells were investigated.

Materials and Methods:  In this experiment, the cell line (C118) was firstly cultured in DMEM culture medium containing 10% FBS and then treated with different concentrations of GW9508. MTT assay was used to determine IC50 and compare the viability of treated cells with different concentrations of GW9508 on days 1, 3, and 5 in the control group. To evaluate the effect, the qRT-PCR test was used with the IC50 concentration on the induction of apoptosis and expression of the P53 gene.

Results: The results showed that GW9508 significantly reduced the viability and proliferation of C118 cells in a dose- and time-dependent manner (P <.05). Morphological changes such as reduction of chromatin density and cell rotation were also observed in the cells. Also, molecular results showed that GW9508 was able to increase the expression of the P53 gene.

Conclusions: The GW9508 small molecule induces cell death in glioblastoma cancer cells by reducing cell viability and increasing P53 gene expression. As a result, it has therapeutic potential to induce cell death in cancer cells and to treat cancer.

Introduction: Non-obstructive azoospermia (NOA) is one of the reasons for infertility in men, and different factors including genetic factors are involved in its development. Since taking biopsies of the testicular tissue for assisted reproductive technologies (ARTs) is invasive and time-consuming, and the testicular tissue is heterogeneous, introducing a biomarker for predicting the possibility of the presence of sperm in the testicle can increase the ART efficiency. Accordingly, Cell-free seminal mRNA (CFs-mRNA), which is found in many fluids of the body including the seminal fluid of NOA individuals, can be employed as a biomarker for this purpose.

Materials and methods: This study was conducted on 15 men suffering NOA, candidates for testicular sperm extraction (TESE), along with 15 healthy men as control. The testicular tissue of 10 patients was examined using hematoxylin and eosin staining and then classified according to Johnsen scoring. RNA was extracted from the cell-free plasma of semen samples and cDNA was synthesized. The Expression level of TNP1 and DDX4 genes was investigated using real-time polymerase chain reaction (PCR).

Results: The expression of CFS-mRNA of the DDX4 gene was observed in only one sample of NOA individuals (10%), showing a score of 8. Further, the expression of CFS-mRNA of the TNP1 gene was observed only in two samples (20%) of NOA patients whose scores were 3 and 8.

Conclusion: Insufficiency or lack of expression of CFS-mRNA of TNP1 and DDX4 genes may be helpful in predicting the absence of sperm in the testicular tissue of NOA patients in terms of sperm retrieval for ART. Yet, further studies with more specific and sensitive techniques are required to achieve a more solid and precise conclusion.

Introduction: Researchers take a great interest in nanoparticles due to their unique properties and high level of performance. Yet, despite the functions of nanoparticles in various sciences and industries, their potential effects on human health especially fetal heart have not been fully investigated. The destructive effect of iron nanoparticles on the fetal heart is inevitable. Therefore, the aim of this study was to investigate the effect of iron oxide nanoparticles on fetal heart growth and development in vivo and in vitro on NMRI mice.

Materials and Methods In this study, mice were divided into three groups: 1- Control: (without the effect of iron oxide), 2- Sham: (injection of solvent iron oxide and distilled water on the 9th day of pregnancy) 3- Treatment: (under the influence of different concentration of nano-iron oxide particles (10, 30, and 50 µg/kg body weight) on the 9th day of pregnancy). On day 16 of pregnancy, fetuses were taken out and their heart was removed (in vivo method) and analyzed by morphological, histological, and statistical criteria. As for in vitro method, pregnant mice were anesthetized on day 15. The embryos were removed from the body. Their hearts were separated and cultured in a culture medium containing a certain dose of iron oxide nanoparticles. Then, morphological and histological changes were examined.

Results: Injection of iron nanoparticles at concentrations of 10, 30, and 50 g/kg caused a significant increase in fetal body weight and height. However, in the results of the examinations on the heart organs, no change in the diameter, weight, wall thickness of the ventricles and atria was observed both macroscopically and microscopically.

Conclusions: In the findings of our study, increase in body length and weight of fetuses can significantly indicate the possibility of increased cell division in the fetus and the ability of these nanoparticles to pass through the placenta and transfer from mother to fetus.

Introduction: Cancer immunotherapy has become a major player in modern oncology. In immunotherapy, immune cells selectively recognize and kill cancerous cells; this way, many side effects of chemotherapy and radiotherapy are reduced. One of the immunotherapy methods is the T cell therapy, which is based on the use of T cells with determined antigenic specificity for cancer cells. The search for unique tumor antigens which are specific to malignant cells and have the ability to stimulate cellular immunity is one of the main targets of malignancies treatment.

Materials and methods: D393-CD20 is an alternative splicing form of the CD20 surface receptor, which lacks 168 nucleotides in exons 3 to 7 of the CD20 sequence. D393-CD20 peptide is merely expressed on cancerous B cells, such as Burkitt’s lymphoma (BL), DLBCL and B-CLL. In this study we isolated and expanded a CD8+ T lymphocyte specific clone for a D393-CD20 antigen to examine the effect on B-CLL cell line. To this end, we evaluated the impact of cytotoxic T cells upon D393-CD20 antigen, expressed on malignant B cells. We also evaluated the ratio of apoptosis using flow cytometry and MTT.

Results: Results suggest that targeting D393-CD20 antigen with specific CD8+ T lymphocytes is very effective in preventing tumor cells growth.

Conclusion: Targeting D393-CD20 can be a proper choice for B-cell lymphoma immunotherapy.

Introduction: CMV is an infectious disease usually transfer from pregnant women to the fetus, or during breastfeeding transfer to the infant and have signs and symptoms like losing weight at birth, leukocytosis, diarrhea.

Case presentation:  In this case, the one year old girl does not has any sign and symptoms except leukocytosis and the infant mothers does not has any disease during pregnancy. The experiments were prescribed for diagnosing the reason of leukocytosis were CBC, Urine analysis, Immunoassay tests for EBV and CMV and serologic tests for Covid19, wright and Widal agglutination and 2ME tests for Brucella.

Conclusions: The results showed the rate of IgM and IgG Antibody were 27.688 and 228.761 respectively. It is noteworthy that there were no other signs and symptoms in this baby, which are commonly described for CMV infection, and other tests have been reported as usual.