Cell-free seminal mRNA of DDX4 and TNP1 Genes as Potential Biomarkers of the Presence of Sperm in the Testicular Tissue

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

Ineffectiveness of Methylation in Rgulation of VHL, ECAD, and RUNX3 Genes in Erythroid Cells Differentiated by Erythropoietin

Background: Vast variety of intermediate factors including cell cycle regulators, growth factors, transcription factors, and signaling pathways are involved in hematopoietic stem cell (HSC) commitment and differentiation into distinct lineages. VHL, Ecad, and RUNX3 are among these. Epigenetics is currently introduced as a potential mechanism to control the gene regulation. The aim of this study is to reveal the correlation between the expression level and methylation pattern of mentioned genes after in vitro differentiation of cord blood HSCs into erythroid lineage mediated by erythropoietin.Materials and Methods: After isolation and expansion, the CD34+ cord blood stem cells were divided into two parts. The first part was used to extract the DNA and RNA and the second to differentiate into erythroid lineage. Methylation specific PCR (MSP) and Real-time PCR were used to determine the methylation status and expression levels of the genes, respectively.Results: Although the significant upregulation observed for VHL and Ecad genes and a down-regulation for RUNX3 gene after differentiation, no remarkable changes were seen in methylation pattern compared with cord blood HSCs by MSP technique.Conclusion: It is appearing that methylation pattern in promoter region has not an effective role in expression of VHL, Ecad, and RUNX3. Moreover, considering the inability of MSP method to detect subtle differences in methylation level a more sensitive method is needed to distinguish the methylation levels of these genes before and after erythroid differentiation

Challenges in the treatment of Iranian patients with leukemia in comparison with developed countries from the perspective of specialists

     Evaluation of the factors associated with treatment process of leukemia and comparison with current related approaches in developed countries can present a good indicator to assess the weak and strong points in healthcare system of our country in leukemia treatment. The objective of this research is general and specific description of the challenges and shortcomings in Iranian healthcare system and monitoring of hematologic malignancies as well as comparison with developed countries. Our study is a descriptive-cross-sectional study. 100 hemato-oncologist , pathologists, and faculty members throughout the country were selected by random cluster sampling. Data collected using questionnaires with Cronbach's alpha coefficient of 0.76 . SPSS and Chi-square test were used for data analysis. According to the specialists, lack of advanced diagnostic facilities as well as cell and BM banks together with high treatment expenses are the main factors contributing to poor treatment processes in Iran, which are far from worldwide standards.The use of novel currently methods used in developed countries for leukemia treatment, financial and psychological support of patients under treatment , making underprivileged provinces well-equipped ,balanced  specialist service distribution relative to capital city either in diagnosis or treatment are factors which makes system standardized. Moreover, integrated institutional work in relation to leukemia incidence and statistical analysis of mortality and morbidity rate can pave the way for reducing and eliminating the problems in diagnosis and treatment of leukemia patients

Correlation between Methylation and Expression Level of P15 and P16 Genes during Differentiation of Cord Blood Stem Cells into Erythroid Lineage Mediated by Erythropoietin

Background: Several influential factors such as transcription factors and intracellular signaling components are involved in differentiation of stem cells into a specific lineage. P15 and p16 proteins are among these factors. Accumulating evidences has introduced the epigenetic as a master regulator of these factors during lineage specification. The main objective of this study is to determine the correlation between the expression level and methylation pattern of P15 and P16 genes in erythroid lineage after in vitro differentiation by erythropoietin (EPO).Materials and Methods: The purified and expanded CD34+ cord blood stem cells were differentiated into erythroid lineage in the presence of EPO. DNA was isolated from both cord blood stem cells and differentiated cells. The Real-Time PCR performed using cDNA and the isolated DNA was used in methylation Specific PCR (MSP) reaction for methylation pattern analysis in both pre and post differentiation stages.Results: The study demonstrated that P15 and P16 genes have partial methylation after erythroid differentiation by EPO. The Expression of P15 gene was higher after differentiation and the expression of P16 gene had a slightly decreased level in post differentiation stage.Conclusion: Significant increase in P15 gene expression after differentiation to erythroid lineage, suggests the remarkable efficacy of this gene in erythroid function. According to upregulation of P15 gene after differentiation despite unchanged methylation status and slight down regulation of P16 gene with slight hyper-methylation of the gene it can be suggested that although the methylation can affects the expression level of P16 gene, the P15 gene is not affected by this mechanism during erythroid differentiation mediated by EPO

Ineffectiveness of Methylation in Rgulation of VHL, ECAD and RUNX3 genes in Erythroid Cells Diffrentiated by Erythropoieten

Background: Vast variety of intermediate factors including cell cycle regulators, growth factors, transcription factors, and signaling pathways are involved in hematopoietic stem cell (HSC) commitment and differentiation into distinct lineages. VHL, Ecad, and RUNX3 are among these. Epigenetics is currently introduced as a potential mechanism to control the gene regulation. The aim of this study is to reveal the correlation between the expression level and methylation pattern of mentioned genes after in vitro differentiation of cord blood HSCs into erythroid lineage mediated by erythropoietin. Materials and Methods: After isolation and expansion, the CD34+ cord blood stem cells were divided into two parts. The first part was used to extract the DNA and RNA and the second to differentiate into erythroid lineage. Methylation specific PCR (MSP) and Real-time PCR were used to determine the methylation status and expression levels of the genes, respectively. Results: Although the significant upregulation observed for VHL and Ecad genes and a down-regulation for RUNX3 gene after differentiation, no remarkable changes were seen in methylation pattern compared with cord blood HSCs by MSP technique. Conclusion: It is appearing that methylation pattern in promoter region has not an effective role in expression of VHL, Ecad, and RUNX3. Moreover, considering the inability of MSP method to detect subtle differences in methylation level a more sensitive method is needed to distinguish the methylation levels of these genes before and after erythroid differentiation

Evaluation of Coagulase-Positive Staphylococcus Aureus Contamination in Lighvan Cheese on Retail Stores

Background: Staphylococcus aureus is a Gram-positive, non-sporulated, lack of capsule, immovable, aerobic and anaerobic and able to tolerate high levels of salt (up to 15 percent). Therefore, foods containing salt provides an ideal environment for bacterial growth. It is also one of the most important bacterial born of spoilable food products, especially in dairy products. Lighvan traditional cheese is consumed in many regions of Iran, especially consumption of these cheeses is very high in Tehran which in the manufacture of such cheeses, the unpasteurized and raw milk is used and the sanitary conditions during processing and subsequent storage are low. So, considering the importance of this issue in the release of poisoning, the coagulase-positive Staphylococcus aureus contamination in Lighvan cheese on Retail Stores was evaluated. Materials and Methods: In order to perform this study, 22 samples of traditional Lighvan cheese were collected randomly from Tehran stores and according to Iran national standards and by using of bird-parker, Cook Meat cultures and coagulase test were evaluated. Results: Total of 22 samples, 14 samples (63/6%) were contaminated by S. aureus over standard and 5 (22/7%) were contaminated with Staphylococcus aureus coagulase positive. Conclusion: This study demonstrated that sanitary conditions should be observed during the production and Use of pasteurized milk in cheese production

Survey the Bacteriological Quality of Juice in the Juice Shop of Shahrood City and its Relationship with Food Safety Knowledge and Performance of Vendors in 2013-2014: A Case Study

Backgrounds and Objective: Juice drinks have high nutritional value, antioxidants, vitamins and minerals which are widely consumed due to therapeutic properties. The aim of this study was to survey the bacteriological quality of juices in Shahrood city. Materials and Methods: This descriptive cross-sectional study was done in the spring and summer seasons of 2014-2015, on juices sold in the Shahrood city study. A self-report questionnaire confirmed by the ministry of health, with certain modifications (as needed), was applied to determine the bacteriological quality of juices and the relationship between food safety knowledge and practices of vendors. In this study, all juice shops (9 units) were considered and the research was carried out by visiting and completing the questionnaires. In order to determine the bacteriological quality of fruit juices, sampling was done from juice shops. The samples the assessed for the presence of Staphylococcus, E. coli, total coliform, yeast and mold counts by using standard methods. The results were compared with the standards of the Institute of Standards and Industrial Research of Iran. Then, obtained data were analyzed by Excel and SPSS software. Result: About 16.12 % of the samples contaminated to Staphylococcus aureus, 22.58% to Escherichia coli, 12.9% to total coliform, 38.7% to yeast and 9.67% to mold. Also, a significant correlation between parameters such as level of literacy (Education) of vendors, amount of sales, site and refrigerator hygiene, personal hygiene, food hygiene, and sanitation was existed (P ≤ 0.001). Conclusion: The results showed that all biological parameters were higher than the standard level, which was due to the lack of principles and rules of personal hygiene, food hygiene and sanitation. Hence, management, monitoring and purposeful education was much more needed for food safety supervision and training to prevent contamination of food with pathogenic agents, and greater emphasis on compliance with regulations is Article 13

A novel de novo canonical splice site mutation in the PTCH1 gene in a male patient with mild psychomotor retardation and autistic traits: a case report

Abstract Basal cell nevus syndrome (BCNS), or Gorlin syndrome, is a rare autosomal dominant disorder caused by mutations in the tumor suppressor gene PTCH1 with complete penetrance and variable expressivity characterized by a broad spectrum of developmental anomalies and a predisposition to neoplasms. Herein, we report a novel de novo splice site mutation in the PTCH1 gene related to mild developmental delay and autistic traits in a 4-year-old male patient

Synthesis and Characterize of Zeolite- Zero Valent Iron/ Silver Nanocomposite (Z-Fe0 /Ag) for Removal Fluoride in Aqueous Solutions (Study of Isotherms, Kinetics and Thermodynamics)

Background: Fluoride is a pollutant that entered to the environment through Glass, Fertilizer and Semiconductor Material industries and cause problems such as Alzheimer's and brain injuries. Adsorption with using inexpensive materials like zeolite is the best options to remove this pollutant. The main problem of absorption method is its low speed. So the aim of present study is investigated the effect of Fe0/Ag nanoparticles as catalyst in performance of zeolite for removal of fluoride. Methods: In this study were used reduction method for converting Fe2+ to Fe0 and co-precipitation method for coating of Fe0 on Zeolite. For depositing silver on nano zero valent iron the rapid mixing along with the high temperature were used. Absorbent characteristics were analyzed with using SEM, TEM and XRD techniques. Then effect of pH, contact time, stirring speed, temperature, amount of adsorbent and Fluoride concentration were evaluated and optimized. Results: pH=3, 60 min contact time, 200 rpm stirring speed and adsorbent concentration equal to 10g/l were obtained as the optimum conditions for Fluoride removal. Investigating the isotherms and kinetics relationships showed that the experimental data correlate to the Langmuir adsorption isotherm model (R2>0.991) and pseudo-second order kinetics (R2>0.933). The reaction thermodynamic study also expressed that process is endothermic and non-spontaneous Conclusion: According on the obtained results can be stated This Absorbent has appropriate efficiency in fluoride removal, But the catalytic properties of zero valent iron and silver was rejected in relation to removal this contaminants

Immobilization of Laccase Enzyme on Silica-Coated Iron Oxide Nanoparticles and Comparison of Stability and Activity of Free and Immobilized Laccase

Background: Laccase enzyme is capable of oxidizing many resistant and non-biodegradable environmental pollutants, so it has been studied frequently in recent years and is widely used in biodegradation of contaminants. Despite its abundant applicability, due to its short life span, nonrecovery, thermal instability and instability in organic environments, its widespread use is very limited. The present study aimed to increase the stability of laccase by immobilizing it on silica coated iron oxide nanoparticles. Materials and Methods: Fe3O4 nanoparticles were synthesized based on the co-precipitation method and after coating with silica, their surface was modified by amine groups. The enzyme was then immobilized by covalent binding using glutaraldehyde. Specifications of synthesized nanoparticles and immobilized enzyme were investigated using X-ray diffraction (XRD), Field Emission Scanning Electron Microscopy (FESEM), and Energy-dispersive X-ray spectroscopy (EDX). Results: Results of successful laccase immobilization on nanoparticles showed that laccase immobilization significantly increased storage and thermal stability, maintaining activity in a wider range of temperature and pH than free laccase. Conclusion: The immobilization of laccase on silica-coated iron oxide nanoparticles can reduce the barriers and challenges of various enzymes by increasing its efficiency and stability