A comprehensive multi-directional exploration of phytochemicals and bioactivities of flower extracts from Delonix regia (Bojer ex Hook.) Raf., Cassia fistula L. and Lagerstroemia speciosa L.

Delonix regia (Bojer ex Hook.) Raf., Cassia fistula L. and Lagerstroemia speciosa L. are three ornamental plants that produce colorful flowers. The present study aimed to evaluate the phytochemicals and bioactivities of methanolic extracts of flowers from Delonix regia (DrFME), Cassia fistula (CfFME), and Lagerstroemia speciosa (LsFME). The presence of ten different chemical classes in varying degrees was confirmed while qualitatively screened. During quantitative determination, LsFME possesses the highest amount of total phenolic (418.0 mg/g), flavonoid (50.8 mg/g), and tannin (256.3 mg/g) contents. The extracts showed excellent antioxidant capacity in a concentration-dependent manner with the lowest IC50 value (41.51 μg/mL) displayed by LsFME. LsFME paralyzed the experimental worms at 2.95 min and killed at 3.96 min. DrFME was found to be more effective in thrombolytic (35.5% clot lysis) and anticoagulant activities. Negligible hemolytic activity (IC50 > 200 μg/mL) found for all extracts which suggest their less potential toxicity. The in vivo experiments revealed that the CfFME has the highest analgesic (64.34% pain inhibition) activity while LsFME has the highest antidiarrheal (70.27% inhibition) and antihyperglycemic (46.94% inhibition) activities at 400 mg/kg of body weight doses. This study has shown the presence of phytochemicals and potential bioactivities which indicates the possibility of these flowers to be used as a source of phytochemicals as well as safe and effective natural medicine.     

Recombinant Tandem Repeated Expression of S3 and SΔ3 Antimicrobial Peptides

Background:Antimicrobial peptides (AMPs) are promising candidates for new generations of antibiotics to overcome the threats of multidrug-resistant infections as well as other industrial applications. Recombinant expression of small peptides is challenging due to low expression rates and high sensitivity to proteases. However, recombinant multimeric or fusion expression of AMPs facilitates cost-effective large-scale production of AMPs. In This project, S3 and SΔ3 AMPs were expressed as fusion partners. S3 peptide is a 34 amino acid linear antimicrobial peptide derived from lipopolysaccharide (LPS) binding site of factor C of horseshoe crab hemolymph and SΔ3 is a modified variant of S3 possessing more positive charges.Methods:Two copy tandem repeat of the fusion protein (named as SΔ3S3-2mer-GS using glycine- serine linker was expressed in E. coli. BL21 (DE3). After cell disruption and solubilization of inclusion bodies, the protein was purified by Ni -NTA affinity chromatography. Antimicrobial activity and cytotoxic properties of purified SΔ3S3-2mer-GS were compared with a previously produced tetramer of S3 with the same glycine- serine linker (S3-4mer-GS) and each of monomeric blocks of S3 and SΔ3. Results:SΔ3S3-2mer-GS was successfully expressed with an expression rate of 26%. The geometric average of minimum inhibitory concentration (MIC GM) of SΔ3S3-2mer-GS was 28%, 34%, and 57% lower than SΔ3, S3-4mer-GS, and S3, respectively. SΔ3S3-2mer-GS had no toxic effect on eukaryotes human embryonic kidney cells at its MIC concentration.Conclusion:tandem repeated fusion expression strategy could be employed as an effective technique for recombinant production of AMPs.Key Words: Antimicrobial Peptide, S3, SΔ3 Fusion Expression, Tandem Repeat Expression     

Interactive role of zinc and iron lysine on Spinacia oleracea L. growth,photosynthesis and antioxidant capacity irrigated with tannery wastewater

AbstractUntreated wastewater contains toxic amounts of heavy metals such as chromium (Cr), which poses a serious threat to the growth and physiology of plants when used in irrigation. Though, Cr is among the most widespread toxic trace elements found in agricultural soils due to various anthropogenic activities. To explore the interactive effects of micronutrients with amino acid chelators [iron-lysine (Fe-lys) and zinc-lysine (Zn-lys)], pot experiments were conducted in a controlled environment, using spinach (Spinacia oleracea L.) plant irrigated with tannery wastewater. S. oleracea was treated without Fe and Zn-lys (0 mg/L Zn-lys and 0 mg/L Fe-lys) and also treated with various combinations of (interactive application) Fe and Zn-lys (10 mg/L Zn-lys and 5 mg/L Fe-lys), when cultivated at different levels [0 (control) 33, 66 and 100%) of tannery wastewater in the soil having a toxic level of Cr in it. According to the results, we have found that, high concentration of Cr in the soil significantly (P < 0.05) reduced plant height, fresh biomass of roots and leaves, dry biomass of roots and leaves, root length, number of leaves, leaf area, total chlorophyll contents, carotenoid contents, transpiration rate (E), stomatal conductance (gs), net photosynthesis (PN), and water use efficiency (WUE) and the contents of Zn and Fe in the plant organs without foliar application of Zn and Fe-lys. Moreover, phytotoxicity of Cr increased malondialdehyde (MDA) contents in the plant organs (roots and leaves), which induced oxidative damage in S. oleracea manifested by the contents of hydrogen peroxide (H₂O₂) and membrane leakage. The negative effects of Cr toxicity could be overturned by Zn and Fe-lys application, which significantly (P < 0.05) increase plant growth, biomass, chlorophyll content, and gaseous exchange attributes by reducing oxidative stress (H₂O₂, MDA, EL) and increasing the activities of various antioxidant enzymes such as superoxide dismutase (SOD), peroxidase (POD) catalase (CAT) and ascorbate peroxidase (APX). Furthermore, the supplementation of Zn and Fe-lys increased the contents of essential nutrients (Fe and Zn) and decreased the content of Cr in all plant parts compared to the plants cultivated in tannery wastewater without application of Fe-lys. Taken together, foliar supplementation of Zn and Fe-lys alleviates Cr toxicity in S. oleracea by increased morpho-physiological attributes of the plants, decreased Cr contents and increased micronutrients uptake by the soil, and can be an effective in heavy metal toxicity remedial approach for other crops.Graphic abstract     

Effects of insulin infusion on glucose kinetics in normal and burned guinea pigs

The effects of a constant infusion of insulin (12 mu/kg·min for 90 min) on glucose turnover (determined by means of the primed-constant infusion of 6-³H-glucose) was evaluated in normal and burned (50% BSA) guinea pigs (gp). In burned, untreated gp, the mean plasma glucose level (gl) was increased from 129±8.2 to 205±13.7 mg/dl 90 min after burning, whereas gl was 140±14.5 mg/dl in the burned + insulin-infused animals at 90 min. The insulin infusion reduced gl from 120±5.6 to 69±5.8 mg/dl in unburned gp; the rate of glucose appearance (R_a) was reduced and the metabolic clearance rate (MCR) was increased. In the B+I gp, the insulin effectively minimized the increase in R_a which followed burning in the burned, untreated gp. However, insulin did not increase the MCR of the burned + insulin-infused group above that of the burned, untreated group. On the day following the burn, the insulin infusion decreased gl in the burned gp to the same extent as in the unburned animals and also increased MCR. We concluded that whereas there was a lack of peripheral responsiveness to the insulin infusion in the first 90 min after burning (during the shock phase), no such lack of responsiveness was evident on the second day.     

Sequence-based 5-mers highly correlated to epigenetic modifications in genes interactions

One of the main concerns in biology is extracting sophisticated features from DNA sequence for gene interaction determination, receiving a great deal of researchers’ attention. The epigenetic modifications along with their patterns have been intensely recognized as dominant features affecting on gene expression. However, studying sequenced-based features highly correlated to this key element has remained limited. The main objective in this research was to propose a new feature highly correlated to epigenetic modifications capable of classification of genes. In this paper, classification of 34 genes in PPAR signaling pathway associated with muscle fat tissue in human was performed. Using different statistical outlier detection methods, we proposed that 5-mers highly correlated to epigenetic modifications can correctly categorize the genes involved in the same biological pathway or process. Thirty-four genes in PPAR signaling pathway were classified via applying a proposed feature, 5-mers strongly associated to 17 different epigenetic modifications. For this, diverse statistical outlier detection methods were applied to specify the group of thoroughly correlated genes. The results indicated that these 5-mers can appropriately identify correlated genes. In addition, our results corresponded to GeneMania interaction information, leading to support the suggested method. The appealing findings imply that not only epigenetic modifications but also their highly correlated 5-mers can be applied for reconstructing gene regulatory networks as supplementary data as well as other applications like physical interaction, genes prioritization, indicating some sort of data fusion in this analysis.     

The postnatal methylation of transfer ribonucleic acid in brain. Evidence for the methylation of precursor transfer ribonucleic acid.

Incubation of 3-day-old rat brain with L-[methyl-3H]methionine resulted in the rapid labeling of low-molecular-weight cytoplasmic RNA. Electrophoresis in 15% polyacrylamide gels provided evidence for the methylation of precursor tRNA molecules, and high-performance liquid chromatography demonstrated N2-methylguanine to be the predominant methylated base formed during the first 2 min of labelling.