Determine exogenous human DDAH2 gene function in rabbit bone marrow–derived endothelial progenitor cells in vitro

The in vitro amplification of endothelial progenitor cells (EPCs) is an important method because of its role in gene transferring and regenerative medicine. In this study, we isolated rabbit bone marrow–derived EPCs to further manipulation and overexpression of dimethylarginine dimethylaminohydrolase (DDAH) in EPCs. Isolated EPCs were cultured, expanded in endothelial basal medium. Morphology of EPCs and expression levels of surface markers detected using immunocytochemistry staining and through the use of flow cytometery. Endothelial progenitor cells were transfected with plasmid vectors expressing human DDAH2 (DDAH2‐EPCs). Three days after gene transfer, positive transfected‐EPCs proliferation and DDAH activity were assayed. We observed colonies conformation and endothelium‐like morphology gradually in the third week of culture. Characterization results revealed positive expression of EPC surface markers CD106, Flk‐1, vWF, and CD34 using few identification techniques. Overexpression of DDAH2 increased citrulline production after 96 hours of transfection, 235.34 ± 0.69 vs 95.26 ± 5.76 ng/mL; P = .023. These results suggest that cell population with EPC characteristics can be simply isolated from rabbit bone marrow and successfully engineered to overexpress exogenous gene. In this study, we offer a feasible method to isolate and identify EPCs from bone marrow. In addition, an efficient transfection with a plasmid vector (without risk of interference) can be constructed a hybrid structure with EPC and DDAH2 gene to examine their function in vitro.     

Leaf gas exchange and grain yield of common bean exposed to spermidine under water stress

Three prevalent aliphatic polyamines (PAs) include putrescine, spermidine, and spermine; they are low-molecular-mass polycations involved in many physiological processes in plants, especially, under stressful conditions. In this experiment, three bean (Phaseolus vulgaris L.) genotypes were subjected to well-watered conditions and two moderate and severe water-stressed conditions with and without spermidine foliar application. Water stress reduced leaf relative water content (RWC), chlorophyll contents, stomatal conductance (gs), intercellular CO₂ concentration (C_i), transpiration rate, maximal quantum yield of PSII (F_v/F_m), net photosynthetic rate (P_N), and finally grain yield of bean plants. However, spermidine application elevated RWC, gs, C_i, F_v/F_m, and P_N, which caused an increase in the grain yield and harvest index of bean plants under water stress. Overall, exogenous spermidine could be utilized to alleviate water stress through protection of photosynthetic pigments, increase of proline and carotenoid contents, and reduction of malondialdehyde content.

     

Germination and emergence of Astrodaucus orientalis (L.) Drude populations influenced by environmental factors and seed burial depth

Astrodaucus orientalis is a weed species in cropping systems and rangelands in Iran. The effects of temperature, light, NaCl concentration, water potential, seed burial depth and crop residue cover were assessed on seed germination and seedling emergence of two populations of A. orientalis from Ardabil (Meshginshahr population) and East Azarbayjan (Tabriz population) provinces of Iran. The A. orientalis populations indicated different responses to environmental factors and burial depth. In the Tabriz population the greatest germination (88.5%) was observed in 20/12°C day/night temperature but in the Meshginshahr population (83.2%) it was obtained in 24/16°C day/night temperature. Over a broad range of light period (10–24 hr light) germination was 74–83%, but it decreased (less than 37%) under 24 hr dark in both A. orientalis populations. With respect to water potential, the C₅₀ parameters were −0.62 and − 0.49 MPa for Tabriz and Meshginshahr populations, respectively. The D₅₀ parameters (the burial depth that caused 50% decrease in emergence) for Tabriz and Meshginshahr populations were 2.42 and 3.13 cm, respectively. Generally, the results showed that emergence of both populations of A. orientalis was delayed as depth of burial increased up to 4 cm and in cropping systems a shallow tillage that locates the seeds to >4 cm of depth in soil could be used in order to suppress seedling emergence. Our findings also could be useful in integrated management of A. orientalis in winter annual crops and rangelands.     

The biological effects of vanadyl curcumin and vanadyl diacetylcurcumin complexes: the effect on structure,function and oxidative stability of the peroxidase enzyme,antibacterial activity and cytotoxic effect

Curcumin has multiple pharmacological effects, but it has poor stability. Complexation of curcumin with metals improves its stability. Here, the effects of vanadyl curcumin and vanadyl diacetylcurcumin on the function and structure of horseradish peroxidase enzyme were evaluated by spectroscopic techniques. Cytotoxic effect of the complexes was also assessed on MCF-7 breast cancer, bladder and LNCaP prostate carcinoma cell line. The results showed that the complexes improve catalytic activity of HRP, and also increase its tolerance against the oxidative condition. The result also indicated that the affinity of HRP for hydrogen peroxide substrate decreases, while the affinity increases for phenol substrate. Circular dichroism and fluorescence spectroscopies showed that compactness of the enzyme structure around the catalytic heme group and the distance between the heme group and tryptophan residue decreases after the binding. The antibacterial and cytotoxic results indicated that the complexes have anticancer potential, but they have no considerable antibacterial activity.     

Extracellular vesicles: Regenerative medicine prospect in hematological malignancies

Extracellular vesicles (EVs) either as endocytic or plasma membrane-emerged vesicles play pivotal role in cell-to-cell communication. Due to the bioactive molecules transformation, lymphoma cell-derived vesicles can alter a recipient cell's function and contribute to signal transduction and drug resistance. These vesicles by acting not only in tumor cells but also in tumor-associated cells have important roles in tumor growth and invasion. On the other hand, the total protein level of circulating exosomes reveals the disease stage, tumor burden, response to therapy, and survival. In residual disease, leukemic blasts are undetectable in the bone marrow by conventional methods but exosomal proteins are elevated significantly. In this manner, new methods for measuring exosomes and exosomal components are required. In this review, we try to reveal the concealed role of EVs in hematological malignancies besides therapeutic potentials.     

HIF-1α in the Crosstalk Between Reactive Oxygen Species and Autophagy Process: A Review in Multiple Sclerosis

Cellular and Molecular Neurobiology - Cellular stress can lead to the production of reactive oxygen species (ROS) while autophagy, as a catabolic pathway, protects the cells against stress....     

Effect of Piriformospora indica and Sebacina vermifera on plant growth and essential oil yield in Thymus vulgaris in vitro and in vivo experiments

Thymus vulgaris of family Lamiaceae is one of the most plants in pharmacy industries. In this study effect of Piriformospora indica and Sebacina vermifera on growth and development plant, yield and composition of the essential oil in a completely randomized design were evaluated in vitro and in pot culture experiments. Plants were studied by means of plant height, shoot fresh and dry weights, number of shoots, root length, root fresh and dry weights and essential oil analyses. The oil was extracted from the dry matter of shoots by hydro distillation, and their composition was determined by GC/MS. In vitro and in vivo cultures showed that plant height and root length increased in pots inoculated with S. vermifera and P. indica. Maximum fresh and dry weight (shoot and root), number of shoots were observed in pots inoculated with P. indica. In thyme inoculated with S. vermifera and P. indica oil yield increased as compared to non-inoculated control plants. GC and GC/MS revealed that the level of thymol was enhanced as the effect of S. vermifera and P. indica.     

Genetic diversity in a collection of old and new bread wheat cultivars from Iran as revealed by simple sequence repeat-based analysis

Genetic diversity in a collection of 70 bread wheat ( Triticum aestivum ) genotypes was studied using 73 microsatellite [simple sequence repeat (SSR)] loci evenly spaced on wheat chromosomes. A total of 592 alleles with an average of 8.53 allele/locus were detected, of which 185 (31.25%) occurred only in a specific group of genotypes. A set of SSR markers consisted of 22 loci with polymorphic information content values of 0.80 or higher were selected for rapid fingerprinting of many genotypes. Average of gene diversity was 0.74 ± 0.017, and significant difference between observed and maximum theoretical values of gene diversity in the analysed SSR loci was obtained using a paired t -test. Genetic distance-based clustering methods including unweighted pair group method with arithmetic average and neighbour joining (NJ) were used for grouping of genotypes. The resulted dendrogram based on NJ and number of differences coefficient hinted of the existence of three groups. This grouping was in agreement with the pedigree information and confirmed by high within-group bootstrap value. A comparatively higher genetic diversity in the studied wheat collection as revealed by presence of high allelic diversity and large number of specific alleles could be utilised in development of new cultivars with desired characteristics.