The effect of Bacillus subtilis producing Surfactin in ROS production and transformation efficiency of tobacco cells

Surfactin secreted by bacilli has biological functions in plant. Surfactin C14 and C15 have the highest effect on inducing hydrogen peroxide species release in the plant. Surfactin production in the two Bacillus strains ACCT21332 and FKR3 were analysed by HPLC and the phytotoxicity of the Bacilli-derived surfactins was determined in Tobacco cell culture. Surfactin C14 and C15 were detected in ACCT21332 but not in FKR3 strain. Extracellular hydrogen peroxide produced by tobacco cell culture cells exposed to ATCC21332 and FKR3 strains increased compared to untreated ones. The Agrobacterium mediated transformation rate of tobacco cells drops from 4% transformed cells to 0.8 and 1.2% when pretreated with ATCC21332 or FKR3 strain, respectively. The strong drop in transformation rate of plant cell culture after FKR3 strain pre-treatment indicates that Surfactin C14 and C15 are not the major or the only cause in protecting plant cells from Agrobacterial infection and transformation.     

Cortactin, an actin binding protein, regulates GLUT4 translocation via actin filament remodeling

Insulin regulates glucose uptake into fat and skeletal muscle cells by modulating the translocation of GLUT4 between the cell surface and interior. We investigated a role for cortactin, a cortical actin binding protein, in the actin filament organization and translocation of GLUT4 in Chinese hamster ovary (CHO-GLUT4myc) and L6-GLUT4myc myotube cells. Overexpression of wild-type cortactin enhanced insulin-stimulated GLUT4myc translocation but did not alter actin fiber formation. Conversely, cortactin mutants lacking the Src homology 3 (SH3) domain inhibited insulin-stimulated formation of actin stress fibers and GLUT4 translocation similar to the actin depolymerizing agent cytochalasin D. Wortmannin, genistein, and a PP1 analog completely blocked insulin-induced Akt phosphorylation, formation of actin stress fibers, and GLUT4 translocation indicating the involvement of both PI3-K/Akt and the Src family of kinases. The effect of these inhibitors was even more pronounced in the presence of overexpressed cortactin suggesting that the same pathways are involved. Knockdown of cortactin by siRNA did not inhibit insulin-induced Akt phosphorylation but completely inhibited actin stress fiber formation and glucose uptake. These results suggest that the actin binding protein cortactin is required for actin stress fiber formation in muscle cells and that this process is absolutely required for translocation of GLUT4-containing vesicles to the plasma membrane.     

The effect of cereal seed extracts on amylase activity of the rose sawfly,Arge rosae Linnaeus (Hymenoptera: Argidae)

The sawfly, Arge rosae Linnaeus (Hymenoptera: Argidae) is a serious pest of rose plants in a vast area of the world including Iran. Pesticides are applied in order to control in areas where infestation is high. So, there is a need for more environmentally benign alternatives to control this insect pest in urban areas. Therefore, the aim of the current study was to study the effect of cereal seed proteinaceous extracts including wheat and triticale on the insect α-amylase. Wheat and triticale seed proteins as well as fourth instars larvae α-amylase were extracted. The results showed that there are two different α-amylase isoenzymes in the insect gut, one major band and the other minor band, which had optimal activity at alkaline pH (pH 8.0). The effect of five concentrations including 0.42, 0.21, 0.105, 0.05 and 0.025?mg?ml^?1 of each seed extract was tested on α-amylase activity of the larvae gut. At the highest concentration (0.42?mg?ml^?1), wheat seed extract caused 74% inhibition of the enzyme activity while triticale seed extract inhibited 62% enzyme activity. Experiments proved that seed proteinaceous extract affected the enzyme activity in a pH-dependant manner.     

A Genetic Framework for Grain Size and Shape Variation in Wheat

Grain morphology in wheat (Triticum aestivum) has been selected and manipulated even in very early agrarian societies and remains a major breeding target. We undertook a large-scale quantitative analysis to determine the genetic basis of the phenotypic diversity in wheat grain morphology. A high-throughput method was used to capture grain size and shape variation in multiple mapping populations, elite varieties, and a broad collection of ancestral wheat species. This analysis reveals that grain size and shape are largely independent traits in both primitive wheat and in modern varieties. This phenotypic structure was retained across the mapping populations studied, suggesting that these traits are under the control of a limited number of discrete genetic components. We identified the underlying genes as quantitative trait loci that are distinct for grain size and shape and are largely shared between the different mapping populations. Moreover, our results show a significant reduction of phenotypic variation in grain shape in the modern germplasm pool compared with the ancestral wheat species, probably as a result of a relatively recent bottleneck. Therefore, this study provides the genetic underpinnings of an emerging phenotypic model where wheat domestication has transformed a long thin primitive grain to a wider and shorter modern grain.     

Mosaicism in CRISPR/Cas9-mediated genome editing

The CRISPR/Cas9 system is a rapid, simple, and often extremely efficient gene editing method. This method has been used in a variety of organisms and cell types over the past several years. However, using this technology for generating gene-edited animals involves a number of obstacles. One such obstacle is mosaicism, which is common in founder animals. This is especially the case when the CRISPR/Cas9 system is used in embryos. Here we review the pros and cons of mosaic mutations of gene-edited animals caused by using the CRISPR/Cas9 system in embryos. Furthermore, we will discuss the mechanisms underlying mosaic mutations resulting from the CRISPR/Cas9 system, as well as the possible strategies for reducing mosaicism. By developing ways to overcome mosaic mutations when using CRISPR/Cas9, genotyping for germline gene disruptions should become more reliable. This achievement will pave the way for using the CRISPR technology in the research and clinical applications where mosaicism is an issue.     

Gonadal histology and plasma sex steroid concentrations in maturing and mature spring migrants of Caspian lamprey Caspiomyzon wagneri in the Shirud River,southern Caspian Sea,Iran

Maturing degree-days, gonadal histology, and changes in serum sex steroids (progesterone, P; testosterone, T; and 17β-estradiol, E2) were examined in maturing and mature spring migrant Caspian lamprey Caspiomyzon wagneri (Kessler, 1870) in the Shirud River (southern Caspian Sea). Blood and gonad samples were collected from ten fish when they first entered the river (maturing stage) and from ten fish that showed spawning readiness after being held in cages in the river (mature stage). The maturing degree-days of Caspian lamprey from the start of upstream migration to maturation was 208–470°C.day. Serum P and E2 concentrations in maturing females were significantly higher than in maturing males, but in the mature stage, serum P and E2 concentrations of females were lower than males. In both stages, there were no differences in serum concentration of T between females and males. In both males and females, P increased significantly with maturation; T levels likewise appeared to increase, but the difference was not significant. E2 increased significantly with maturation in males, but females showed a significant decrease. Maturing females had similar stage gonads with the germinal vesicle in the polar position. Maturing males had testes that primarily contained secondary spermatocytes with an occasional occurrence of spermatozoa. These results suggest that males mature earlier than females, which is a pattern similar to that found in the sea lamprey.     

Betaine Down Regulates Apelin Gene Expression in Cardiac and Adipose Tissues of Insulin Resistant Diabetic Rats Fed by High-Calorie Diet

Apelin is a newly discovered peptide that its serum level increases in diabetic patients with cardiovascular dysfunction. Recent studies indicate the beneficial actions of betaine in reducing the cardiovascular and metabolic complications, however data related to its effect on adipocytokine expression is limited. The aim of this study was to evaluate the effect of betaine supplementation on Apelin gene expression in cardiac muscle and adipose tissue of insulin resistance, diabetic rats fed by a high calorie diet. To induce insulin resistance rats were fed with high fat/high carbohydrate diet for five weeks and then 30 mg/kg STZ was injected intraperitoneally. After confirming of diabetes incidence (serum glucose above 7.5 mmol/l) the animals were treated with 1 % betaine in drinking water for 28 days. At days 14 and 28 after treatment, animals were euthanized and Apelin gene expression was evaluated by real time PCR and western blot in heart and adipose tissues. Serum levels of insulin, Apelin and glucose and HOMA–IR were also measured. Our results showed that feeding of rats by a high calorie diets caused insulin resistance, which was manifested by elevated plasma insulin, glucose and Apelin levels and also HOMA–IR. Apelin gene expression in heart and adipose tissues were significantly increased simultaneously with the progression of diabetes. Betaine supplementation decreased serum Apelin and down regulated Apelin expression in adipose tissue and cardiac muscle, particularly at day 28 of treatment. We concluded that betaine might improve metabolic and cardiovascular complications in diabetic patients by regulation of Apelin expression and secretion.     

A glimpse into molecular mechanisms of embryonic stem cells pluripotency: Current status and future perspective

Embryonic stem cells have potential differentiation ability into a large variety of cell lineages and proved to be an effective therapeutic modality. However, prolonged in vitro and ex-vivo expansions impair embryonic stem cells multipotentiality, and thereby limit their clinical application. In the past few years, research collected attempts to explore new insights into the molecular mechanisms participate in the stemness capacity of embryonic stem cells. Along with these comments, modalities and strategies with the potential to maintain embryonic stem cells multipotentiality are of great interest. In this review, the authors attempted to discuss the pathways participating in the preservation of embryonic stem cells multipotentiality and emphasized the novel strategies that help to harness regenerative potential.     

Effect of culture system on survival rate of vitrified bovine embryos produced in vitro

This study was designed to evaluate the effect of in vitro culture system on bovine blastocyst yield and quality after vitrification. In Experiment 1, IVM/IVF zygotes were allocated to three culture conditions: (I) Oviductal cells-SOF (OCM-SOF); (II) Oviductal cells-TCM (OCM-TCM); and (III) SOF for 8 days. There was no significant difference between blastocyst rates among groups.In Experiment 2, the IVP-blastocysts in three above culture conditions were vitrified within groups segregated according to age (Day 7 and 8) and blastocoelic cavity size (early and expanded blastocysts). A trend of higher survival rate was obtained in vitrified/warmed early blastocysts compared with expanded ones, so that the difference in OCM-TCM group was significant (P < 0.001). Higher survival and hatching rates (P < 0.001) were obtained in OCM-SOF and OCM-TCM groups (co-culture) compared with SOF group and the age of blastocyst had no effect on post-thaw survival and hatching rates. In Experiment 3, after staining of blastocysts, in fresh blastocysts the highest number of trophectoderm cells was observed in OCM-TCM group and the number of inner cell mass (ICM) was higher in co-culture groups than SOF group (P < 0.001). In vitrified/warmed blastocysts the number of ICM and trophectoderm cells in co-culture groups was higher than SOF group (P < 0.001) except for the ICM of expanded blastocysts. In conclusion, in our culture conditions, the blastocyst yield is not influenced by culture system, while the cryotolerance of IVP-blastocysts is positively influenced by the presence of somatic cells. Moreover, the expanded blastocysts are more susceptible to cryoinjury than early blastocysts.     

Experimental and theoretical investigations of Dy(III) complex with 2,2′-bipyridine ligand: DNA and BSA interactions and antimicrobial activity study

Abstract

In this study, the interactions of a novel metal complex [Dy(bpy)₂Cl₃.OH₂] (bpy is 2,2'-bipyridine) with fish salmon DNA (FS-DNA) and bovine serum albumin (BSA) were investigated by experimental and theoretical methods. All results suggested significant binding between the Dy(III) complex with FS-DNA and BSA. The binding constants (K_b), Stern-Volmer quenching constants (K_SV) of Dy(III)-complex with FS-DNA and BSA at various temperatures as well as thermodynamic parameters using Van’t Hoff equation were obtained. The experimental results from absorption, ionic strength, iodide ion quenching, ethidium bromide (EtBr) quenching studies and positive ΔH? and ΔS? suggested that hydrophobic groove-binding mode played a predominant role in the binding of Dy(III)-complex with FS-DNA. Indeed, the molecular docking results for DNA-binding were in agreement with experimental data. Besides, the results found from experimental and molecular modeling indicated that the Dy(III)-complex bound to BSA via Van der Waals interactions. Moreover, the results of competitive tests by phenylbutazone, ibuprofen, and hemin (as a site-I, site-II and site-III markers, respectively) considered that the site-III of BSA is the most possible binding site for Dy(III)-complex. In addition, Dy(III) complex was concurrently screened for its antimicrobial activities. The presented data provide a promising platform for the development of novel metal complexes that target nucleic acids and proteins with antimicrobial activity.

Communicated by Ramaswamy H. Sarma