Hessellundbartlett0517
Cichorium intybus is rich in inulin and has several pharmacological applications. Hairy roots culture is a valuable biotechnological tool used to produce plant secondary metabolites. Agrobacterium rhizogenes-mediated genetic transformation of chicory to hairy roots was investigated using Agrobacterium Strains A4, A13, A7, and ATCC15834. Several hairy roots were tested, from which 17 lines were selected based on their fast-growing characteristics. Results of PCR analysis revealed foreign DNA integration into the selected transgenic hairy root lines. Finally, four Adventitious roots that contained the highest ratio of total sugar to total weight (µg/gr DW), were selected. This study investigated the effects of various levels of minerals and sucrose on the production of inulin in Cichorium hairy root culture. Different levels of sucrose, phosphate (Pi) and Iron (Fe) were evaluated, separately. It was found that an increase in sucrose levels from 3 to 5% could decrease the root growth; however, 60 g/l sucrose remarkably enhanced the inulin production rate in all the examined lines. The highest biomass was achieved by applying 3.75 mM Pi but it ended in the decreasing the inulin content per unit weight. Selleck Brr2 Inhibitor C9 In contrast, the highest inulin accumulation and the lowest amount of biomass were observed in 0.5 mM Pi. Fe starvation caused the biomass decrease and a significant increase in inulin accumulation. Results of this study suggest a successfully optimized culture medium to initiate the induction of Cichorium intybus hairy root cells to produce inulin as a valuable medicinal secondary metabolite.Generally, the high widespread presence of antimicrobial resistance, and the next freeing into aquatic environments which provide a situation for transmission of these genes in water is because of the abuse of the antimicrobial drugs in both medicine and veterinary medicine. In aquatic environment, bacteriophages could have an important role in sharing antimicrobial resistance genes. The purpose of this study was to assess three important antibiotic resistance genes including two β-lactamases (blaTEM, blaSHV) and sul1 gene, referring to resistance to sulfonamides, in both bacteria and phage DNA fractions of wastewater samples, Shiraz, Iran, using polymerase chain reaction. The prevalence of those genes was extremely high and equal to 100% in bacterial DNA, while these rates were lower in phage DNA fractions as 66.66%, 66.66% and 58.33% for blaTEM, blaSHV and sul1, respectively. In conclusion, detection of mentioned genes in bacterial and phage DNA fractions from ambient water is considerable, so the possibility of harbouring and transferring of antibiotic resistance genes by phages needs to be explored in the future. Also, available data is a reputable endorsement that wastewater is a hotspot for these kinds of genes to spread in the environment. Based on our knowledge, this is the first report of blaTEM and bla SHV and sul1 genes in bacterial and phage DNA fractions insulated from urban wastewater and environment in Iran.The application of mesenchymal stem cells (MSCs) is rapidly expanding due to their unique properties in cell therapy, especially as the feeder layer in the ex-vivo expansion of immune cells. Also, Interleukin 2 (IL-2) is an essential human cytokine in the expansion of hematopoietic precursors and progenitors, i.e., NK cells and T cells, while there is no endogenous expression of IL-2 in MSCs. This study aimed to examine the potency of amniotic membrane (AM)-MSCs as the IL-2 secretory cells. IL-2-containing pCMV3-C-GFPspark shuttle vector was transformed in E.coli DH5-alpha. After cloning, the plasmid DNA was extracted and transfected in isolated AM-MSCs, by lipofectamine-2000. Then, the RNA and protein expression levels of exogenous IL-2 were evaluated 3 to 15 days after transfection, using ELISA and qRT-PCR. Fluorescent microscopy and flowcytometry assays were used for evaluating the GFP-positivity of transfected AM-MSCs, as IL-2 expression control. There was a significant increase in RNA expression of exogenous IL-2 in transfected AM-MSCs in 3 to 15 days after transfection. (p less then 0.001) Also, IL-2 concentration released in the medium was increased in 3rd day after transfection (611 pg/ml). However, the RNA and protein expression of IL-2 was reduced through passing the time. The results show AM-MSC is a suitable host for the expression and secretion of IL-2 as a critical cytokine in the ex-vivo expansion of hematopoietic precursors and progenitors, i.e., NK cells and T cells. Also, the survival time of IL-2 expression in AM-MSCs was long enough for use as a feeder layer.Hemophilia A is an X-linked bleeding disorder that occurs due to the deficiency of Factor VIII (FVIII) protein clotting activity. The mutations in the F8 gene, which encodes FVIII coagulating protein have been widely reviewed. However, there is a wide range of criteria that in addition to F8 gene mutations, different molecular mechanisms may be associated with hemophilia A. Various functions of FVIII could be related to the hypothetical small non-coding RNAs, located within the F8 gene sequence. Therefore, miRNAs that can post-transcriptionally regulate gene expression might confer susceptibility to developing hemophilia A. Here, we have selected a bioinformatically predicted hairpin structure sequence in the first intron of the F8 gene that has the potential to produce a real miRNA (named put-miR1). We tried to experimentally detect the predicted miRNA via RT-PCR following its precursor overexpression in HEK 293 cell lines. Despite the accuracy of miRNA prediction, according to the reliable bioinformatics studies, we couldn't confirm the existence of considered mature miRNA in transfected cells. We hope that through changing experimental conditions, designing new primers, or altering cell lines and expression vectors, the exogenous and endogenous expression of the predicted miRNA will be confirmed.Given the significant physical, mental, and economic problems of coronary artery disease (CAD), it is important for communities to help reduce these costs. The Cytochrome P450 Family 1 Subfamily A Member 1) CYP1A1 (enzyme is known to cause coronary artery disease through various mechanisms. Therefore, it is important to investigate the polymorphisms that affect the activity of this enzyme. After collecting samples from 191 patients with angiographically verified CAD and 191 healthy individuals, genotyping for CYP1A1 rs4646903 polymorphism was carried out. Lipid profile was assessed by conventional colorimetric method. The results showed that the frequency of heterozygous and homozygous mutant genotypes of rs4646903 polymorphism was 36.6% and 5.2% in patients and 20.9% and 2.1% in controls, respectively. The heterozygous genotype (OR=2.24; 95% CI=1.30-3.84, P=0.003), homozygous mutant genotype (OR=3.97; 95% CI=1.05-14.98, P=0.042) and mutant C allele (OR=2.15; 95% CI=1.46-3.15, P0.05). In conclusion, the rs4646903 polymorphism contributed to the susceptibleness of people to CAD.A week-long, city-wide science festival called Everything is Science (EiS) was developed to educate the community in an informal manner. The festival serves as a platform for presenters from diverse professions to give engaging talks (without PowerPoint slides) to the public, free of charge, in restaurants and bars around town. Over 350 people attended the events over 5 days with 33 presenters. Surveys completed by attendees and session coordinators indicate strong support for this festival. Altogether, the EiS festival serves as a no-cost method to engage with the community and improve science literacy with potential for adoption in other cities.Folates are essential micronutrients, and folate deficiency still occurs in many countries. Lactic acid bacteria (LAB) are known to be able to synthesize folates during fermentation, but the folate production is strain-dependent and influenced by the fermentation medium, presence of a folate precursor, and fermentation time. This study aimed to screen extracellular folate-producing LAB from local food sources and evaluate the factors influencing their folate biosynthesis during milk fermentation. The selection of folate-producing LAB was based on their ability to grow in folate-free medium (FACM), with folate concentrations quantified by microbiological assay. Growth of the 18 LAB in FACM varied between isolates, with only 8 isolates growing well and able to synthesize extracellular folate at relatively high concentrations (up to 24.27 ng/mL). The isolates with highest extracellular folate levels, Lactobacillus fermentum JK13 from kefir granules, Lactobacillus plantarum 4C261 from salted mustard, and Lactobacillus rhamnosus R23 from breast milk, were applied to milk fermentation. The last two isolates were probiotic candidates. The three isolates consumed folate when it was present in the milk, and its consumption was in line with their growth. The availability of folate precursors affected the amount of folate consumed, but did not lead to increased folate concentrations in the medium after 72 h fermentation. The results of this study indicate that these isolates cannot be utilized for producing folate in folate-containing milk, as it shows feedback inhibition on folate biosynthesis.This study evaluated the relative abundance of proximate, mineral, and phytochemical constituents of the anatomical parts of Xylopia aethiopica (XA) fruit using standard analytical procedures. The results showed that whole fruits (WF) have higher contents of crude protein, crude fiber, fat, ash, and moisture than the seeds (S) and pericarps (P). However, highest contents of crude carbohydrate and nitrogen free extracts were found in the P, followed by the S. The content of minerals (sodium, potassium, calcium, phosphorus, iron, zinc, magnesium, and copper) and phytochemicals were present in the following order of abundance WF>S>P. Furthermore, the phytochemical constituents in each XA parts were present in the following order of relative abundance total flavonoids>tannins>total phenolics>cardiac glycoside>alkaloids>steroids. Vitamin A was abundant in all three parts, with the abundance highest in WF [4.83±0.06 g vitamin A equivalent (vit A eq)/100 g] and lowest in P (1.64±0.02 g vit A eq/100 g). This preliminary study indicates XA fruits are rich in minerals, anti-nutrients, and phytochemicals. Therefore, these data could represent a biochemical rationale for inclusion of XA as a spice or functional ingredient in many Nigerian local soups to help prevent ailments.Melanosis in shrimp usually leads to reduction in its shelf life and quality, which causes a significant loss in economic value of shrimp products. This study reports potential applications of nine ethanolic extracts of by-products, i.e., peel and/or seed from three Vietnamese avocado varieties as effective inhibitors of melanosis in whiteleg shrimp. Six out of nine shrimp samples treated with the prepared extracts (0.025%, w/v) reduced melanosis and lipid oxidation more significantly as compared to those treated with sodium metabisulfite (SMS, 1.25%, w/v) and control groups (treated with water) during 8-day storage at 4°C (P0.50). The findings obtained from this study suggest potential utilization of avocado by-product extracts as safe and cheap natural alternatives to traditional sulfites for anti-melanosis and shelf life extension of whiteleg shrimp.