The Antimicrobial Effects of Ciprofloxacin Combined with Green Synthesized Glutathione-coated Silver Nanoparticles on Biofilm Formation of Pseudomonas Aeruginosa

Introdaction:Nowadays, antibiotic resistance is rising at an alarming rate. Essentially, one of the important ways for bacteria such as P. aeruginosa to survive in the presence of antibiotics is biofilm formation. In the current report, we have focused on inhibiting the microbial biofilm formation of P. aeruginosa through combining glutathione (GSH) coated silver nanoparticles (AgNPs) and Ciprofloxacin (Cip).Materials and Methods: AgNPs were biosynthesized using Eucalyptus Camaldulensis leaf extracts and surface modification of AgNPs was done, using glutathione.  Synthesized nanoparticles were characterized by FTIR, XRD, DLS, SEM, and CHN tests. Then, 50 isolates of P. aeruginosa were originated from different samples of hospitalized patients from Sina hospital and 24 isolates were selected as a strong biofilm producer using microtiter plate method for further studies. Finally, the synergistic effect of GSH-coated AgNPs and Cip was investigated on biofilm formation of P. aeruginosa. Result:The images of SEM represent the spherical structure of silver nanoparticles with a smooth surface. Also, the results of FTIR, XRD, DLS, SEM, and CHN of AgNPs before and after surface coating confirmed the formation of GSH-coated AgNPs. GSH-coated AgNPs and Cip at a concentration of 1/2 and 1/4 MIC had an inhibitory activity on biofilm formation of 87.5% and 83.4% of P. aeruginosa isolates respectively. Conclusion:This study illustrated that the combination of GSH-coated AgNPs and Cip has a synergistic inhibitory activity on P. aeruginosa biofilm formation

The altered expression of long non-coding RNAs: GHET1, BACE1-AS, PANDA, UCA1 associated with non-small cell lung cancer

Objective: Long non-coding RNAs (lncRNAs) are characterized as non-coding transcripts greater than 200 nucleotides. lncRNAs have extensive molecular connections with proteins and microRNAs, which are important in the regulation of gene expression in physiologic and pathologic states including cancer. About 18% of human LncRNAs were recently found to be associated with tumours. Many studies indicated that aberrant expression of LncRNAs play key roles in the progression and metastasis of NSCLC. In this study we evaluated the expression of long non-coding RNAs: GHET1, BACE1-AS, PANDA, UCA1 in non-small cell lung cancer. Material & Methods: In this study, RNA was extracted from tumor tissues of NSCLC and paired adjacent normal lung tissues. After cDNA synthesis, the relative expression level of lncRNA GHET1, BACE1-AS, PANDA, and UCA1 genes was studied by TaqMan Real-Time PCR, and the data were analyzed by 2-∆∆CT. The t-test was used to compare the values and P-value < 0.05 was considered statistically significant. Results: The data of qRT-PCR analysis revealed that the expression level of GHET1 gene in patients with NSCLC is increased (P= 0.0032) and BACE1-AS showed down-regulation (P= 0.0093). There was no significant change in the expression of PANDA and UCA1 genes. Conclusion: Our study sheds lights on the expression signature of several crucial lncRNAs in human lung cancer. This data not only could be further be utilized for different therapeutic approaches but also reveal the changes in biological processes of human lung tumors. &nbsp

Association of kinase insert domain-containing receptor (KDR) gene polymorphism/ haplotypes with recurrent spontaneous abortion and genetic structure

Background: Recurrent spontaneous abortion is one of the diseases that can lead to physical, psychological, and, economical problems for both individuals and society. Recently a few numbers of genetic polymorphisms in kinase insert domain-containing receptor (KDR) gene are examined that can endanger the life of the fetus in pregnant women. Objective: The risk of KDR gene polymorphisms was investigated in Iranian women with idiopathic recurrent spontaneous abortion (RSA). Materials and Methods: A case controlled study was performed. One hundred idiopathic recurrent spontaneous abortion patients with at least two consecutive pregnancy losses before 20 weeks of gestational age with normal karyotypes were included in the study. Also, 100 healthy women with at least one natural pregnancy were studied as control group. Two functional SNPs located in KDR gene; rs1870377 (Q472H), and rs2305948 (V297I) as well as one tag SNP in the intron region (rs6838752) were genotyped by using PCR based restriction fragment length polymorphism (PCR-RFLP) technique. Haplotype frequency was determined for these three SNPs’ genotypes. Analysis of genetic STRUCTURE and K means clustering were performed to study genetic variation. Results: Functional SNP (rs1870377) was highly linked to tag SNP (rs6838752) (D´ value=0. 214; χ2 = 16.44, p<0. 001). K means clustering showed that k = 8 as the best fit for the optimal number of genetic subgroups in our studied materials. This result was in agreement with Neighbor Joining cluster analysis. Conclusion: In our study, the allele and genotype frequencies were not associated with RSA between patient and control individuals. Inconsistent results in different populations with different allele frequencies among RSA patients and controls may be due to ethnic variation and used sample size

Profiling the miRNAs for Early Cancer Detection using DNA-based Logic Gates

Abstract Background: DNA-based computing is an emerging research aspect that enables the in-vivo computation and decision making with significant correctness. Recent papers show that the expression level of miRNAs are related to the progress status of some diseases such as cancers and DNA computing is introduced as a low cost and concise technique for detection of these biomarkers. In this paper, DNA-based logic gates are implemented in the laboratory to detect the level of miR-21 as the biomarker of cancer. Materials and Methods: At the first, required strands for designing DNA gates are synthesized. Then, double stranded gate is generated in laboratory using a temperature gradient that followed by electrophoresis process. This double strand is the computation engine for detecting the miR-21 biomarker. miR-21 is as input in designed gate. At the end, the expression level of miR-21 is identified by measuring the generated fluorescent. Results: at the first stage, the proposed DNA-based logic gate is evaluated by using the synthesized input strands and then it is experimented on a tumor tissue. Experimental results on synthesized strands show that its detection quality/correctness is 2.5x better than conventional methods. Conclusion: Experimental results on the tumor tissues are successful and are matched with those are extracted from real time PCR results. Also, the results show that this method is significantly more suitable than real time PCR in view of time and cost

Comparison of Tissue Culture Plate, Congo red Agar and Tube Methods for Evaluation of Biofilm Formation among Uropathogenic E. coli Isolates

Background and Aims: Microorganisms within biofilms,, formed on medical devices in the body are highly resistant to antimicrobial compounds and host responses and play a major role in nosocomial infections, especially urinary tract infections (UTIs). Uropathogenic Escherichia coli (UPEC) causes 50% of the hospital-acquired urinary tract infections and is capable to form biofilm in the bladder epithelium which plays an important role in its pathogenesisThe identification of biofilm producing UPEC strains by routine laboratory methods is important for the better understanding of the pathogenesis of this bacterium in UTIs. Materials and Methods: A total of 100 UPEC strains were collected from patients with UTIs in a hospital inTehran in 2016 and diagnosed by biochemical tests and the ability of biofilm formation was determined by Tissue Culture Plate (TCP), tube method (TM) and Congo red agar (CRA) methods. Sensitivity and specificity of methods were determined. Results: In tube method, 23% of the isolates formed strong and 59% formed weak biofilm. In Tissue Culture Plate method 40%, 22% and 28% of isolates formed strong, moderate and weak biofilm respectively and 10% were biofilm negative. According to the Congo red agar method only 4% of the isolates formed strong biofilm, 65% and 31% respectively had a weak biofilm and no biofilm. The sensitivity and specificity of Congo red agar and Tube methods were 39.1%, 50.9% and their features were 78.3%, 79.7% respectively. Conclusions: The results showed that Tissue Culture Plate method is important for the determination of UPEC biofilm formation. Tube method and Congo red agar are not reliable methods for this purpose

Evaluation of MiR-20a and MiR-204 Expression Involved in Autophagy in Non-small Cell lung Cancer

Introduction: Non-small cell lung cancer (NSCLC) is the most common type of lung cancer and the most lethal cancer worldwide. It is often diagnosed at advanced stages. One of the mechanisms of the immune system to fight lung cancer cells is autophagy. Autophagy is a conserved catabolic process in which proteins and organelles are deleted by lysosomes. microRNAs are small RNAs containing about 19–22 nucleotides that function as important regulatory elements in the cell and as oncogene or tumor suppressors in lung cancer. The role of miRNAs is important in lung cancer progression by regulating autophagy genes of several proteins. The aim of this study was to evaluate the expression of miR-204-5p and miR-20a expressions involved in the autophagy pathway in non-small cell lung cancer.   Materials & Methods: In this study, miR-204-5p and miR-20a expression levels were studied, using the quantitative Real-Time PCR technique, in 30 patients with non-small cell lung cancer. RNA was extracted from tumor and adjacent normal tissue of NSCLC patients. cDNA was synthesized using specific stem-loop for miR-20a, miR-204-5p, and RNU44 reference gene. Finally, the expression levels of miRNAs were assessed by Real-Time PCR and the data were analyzed using the 2-∆∆CT method. Code of ethics: sbmub.REC.1394.112   Findings: Based on the results of the qRT-PCR analysis it was revealed that miR-20a and miR-204 were upregulated and downregulated in tumor tissues, respectively.   Discussion & Conclusions: These changes in the expression level, suggest that miR-20a and miR-204-5p are oncogenes and tumor suppressors, respectively. So, measuring the expression level of miR-20a could be a biomarker for the diagnosis and progression of non-small cell lung cancer as well as a platform for the targeted treatment of cancer

Investigation of ATG5, ATG12 and LC3 Genes Expression Related to Autophagy in Breast Cancer

Purpose: Breast cancer is caused by uncontrolled growth of the cell in the breast tissue, and the cancer is the most common cancer in women. Among the mechanisms involved in the incidence of cancer, the autophagy mechanism is recognized as one of these important pathways. Therefore, the study of effective genes in this pathway can be of great importance. The genes of LC3, ATG5 and ATG12 are among the important genes in this pathway.Method: The Real Time-PCR method was used to evaluate the expression levels of LC3, ATG5 and ATG12 genes in 30 breast tissues and 30 adjacent normal tissues. Moreover, GenAll and Takara kits were used to extract RNA as well as cDNA synthesis, respectively.Results:  The results of this study showed that there is no significant difference in ATG5, ATG12 and LC3 genes expression in tumor samples compared to normal samples (P> 0.05). The evaluation of expression changes in ATG5, ATG12 and LC3 demonstrate that, out of 30 tumor samples which were considered for each of genes, 18 tumor samples of ATG5 gene, 17 tumor samples of ATG12 gene and 16 tumor samples of LC3 gene illustrate the expression decreasing.Conclusion: Finally, no significant changes were observed for the selected genes in this study. Since this research was designed on a pilot study, more researches and samples are needed to obtain more accurate results

Expression Patterns of miR181a and miR30d in Patients with Breast Cancer

One of the important molecular pathways in breast cancer is the PTEN-PI3K-AKT pathway. Any change in the activity of the PTEN gene can alter the PI3K-AKT pathway. Moreover, there are subsets of genes and pathways their expression changes by post-transcriptional regulations. For instance, gene regulation alters by non-coding RNAs such as micro-RNAs as post-transcriptional regulators that prevent the expression of the target transcript. Therefore, it is essential to assess the related alterations in micro-RNA expression patterns to find out the possible causes of conversions in related transcripts and pathways such as the PTENPI3K-AKT pathway in breast cancer. To determine the expression level of miR-181a and miR-30d in 30 breast tumor samples and 30 adjacent normal samples, the RNA extraction, and cDNA synthesis was performed by RiboEx (GeneAll, Korea). Finally, the Real-Time PCR method was used for quantitative analysis of the expression levels of these miRNAs. all the experimental part of the project in done at Islamic Azad University in 2017. After analyzing comparisons in the expression level of miR-181a and miR-30d in tumor and normal tissues, there was a significant increase in the expression level of miR-181a in tumor samples compared with normal samples. Moreover, the expression level of miR-30d in tumor samples reported a significant decrease in comparison with normal samples (P <0.05). Upregulation of miR-181a may affect the transcription of the PTEN gene resulting in the cell progress to cancer. The Downregulation of miR-30d may also lead to cancer cell growth, due to a reduction in the affecting on the CREB gene transcript

Assessing Methylation of Mir-146b in Patients with Non-Small Cell Lung Cancer

Background: One of the most important epigenetic factors in lung cancer is aberrant DNA methylation. So far, many studies have been performed on the methylation of various genes and microRNAs in various cancers, especially lung cancer. So because of the high importance of microRNAs in cancer. In this study, mir-146b methylation was checked in NSCLC tissues.Method: Analysis methylation of mir146b was investigated in 30 samples NSCLC tissues and 30 adjacent normal tissues using by MS-HRM method.Results: Study on mir146b methylation showed that there was no significant difference between the methylation levels of this microRNA in tumor samples compared with healthy samples (P>0.05). However, this study was designed as a pilot study, and further investigations are required to confirm our findings