Simultaneous regulation of miR-451 and miR-191 led to erythroid fate decision of mouse embryonic stem cell

Objective(s): Various microRNAs (miRNAs) are expressed during development of mammalian cells, when they aid in modulating gene expression by mediating mRNA transcript cleavage and/or regulation of translation rate. miR-191 and miR-451 have been shown to be critical regulators of hematopoiesis and have important roles in the induction of erythroid fate decision. So, the aim of this study is investigation of the miR-191 and miR-451 roles in the controlling mouse embryonic stem cell (mESC) differentiation toward the erythroid lineage.Materials and Methods: mESCs were infected with either pCDH-miR-Off-191 viruses in pCDH-miR-Off-191 group or simultaneously with pCDH-miR-Off-191 and pCDH-miR-451 lentiviruses in simultaneous  group. Then, the expression profiles of erythroid specific transcription factors and globin genes were analyzed using QRT-PCR on day 14 and 21 of differentiation. Flow cytometry analysis was used to evaluate of TER119 and CD235a erythroid specific surface markers. Results: Gata-1, Klf-1, Epor and globin chains were found to be expressed in pCDH-miR-Off-191 and in simultaneous groups. The majority of globin chains showed changes in their expression levels with progression of differentiation from day 14 to day 21. Flow cytometry results showed that miR-451 up- regulation and miR-191 down-regulation is associated with the expression of TER119 and CD235a. Of these two groups analyzed, simultaneous group was most significantly potent in stimulation of erythroid fate decision of mESCs. Conclusion: Together, present data demonstrate that down-regulation of miR-191 alone can enhance the differentiation of mESCs. However, the simultaneous effect of miR-451up-regulation and miR-191 down-regulation is much stronger and can have more practical use in artificial blood production

In vitro antibacterial effect of Pimpinella anisum essential oil on Enterococcus faecalis, Lactobacillus casei, Actinomyces naeslundii, and Aggregatibacter actinomycetemcomitans

Introduction: Pimpinella anisum is a medicinal plant with antimicrobial, antifungal, and anti-oxidative properties. Limited studies have assessed the antibacterial properties of Pimpinella anisum on oral and dental pathogens. Aim: This in vitro study aimed to assess the antibacterial effect of Pimpinella anisum essential oil on Enterococcus faecalis, Lactobacillus casei, Actinomyces naeslundii, and Aggregatibacter actinomycetemcomitans. Materials and methods: After obtaining the essential oil of Pimpinella anisum, its antimicrobial activity was evaluated using the agar disc diffusion test. The minimum inhibitory concentration and minimum bactericidal concentration of the essential oil were also determined; 0.2% chlorhexidine was used as the positive control. Results: The mean diameter of growth inhibition zone was 39 mm for Enterococcus faecalis, 40 mm for Lactobacillus casei, 42 mm for Actinomyces naeslundii, and 18.5 mm for Aggregatibacter actinomycetemcomitans. The mean diameter of the growth inhibition zones for Enterococcus faecalis, Lactobacillus casei, and Actinomyces naeslundii was significantly greater than that of Aggregatibacter actinomycetemcomitans (p=0.001). Also, the mean diameter of the growth inhibition zone of Actinomyces naeslundii was significantly larger than that of Enterococcus faecalis (p=0.05). The minimum inhibitory concentration and minimum bactericidal concentration of the essential oil for Enterococcus faecalis were 4.88% and 4.88%, respectively. These values were 9.76% and 9.76% for Lactobacillus casei, 9.76% and 4.88% for Actinomyces naeslundii, and 9.76% and 9.76% for Aggregatibacter actinomycetemcomitans, respectively. Conclusions: Pimpinella anisum essential oil was effective against all four microorganisms evaluated in this study. Since the lowest minimum inhibitory concentration and minimum bactericidal concentration were recorded for Enterococcus faecalis, this essential oil has maximum effects on Enterococcus faecalis. Future clinical studies are required to assess the antimicrobial efficacy of Pimpinella anisum essential oil in clinical samples