Homology Modeling and Functional Characterization of PR-1a Protein of Hordeum vulgare subsp. Vulgare

Pathogenesis-related protein 1a of Hordeum vulgare subsp. Vulgare (HvPR-1a) is induced by various pathogens and stress related factors. It plays important roles in plant defense system. Since the discovery of HvPR-1a a great deal of research has been focused on its isolation and characterization. However, three dimensional structure of HvPR-1a is still unknown. 3D structure can be used for determining protein function, and identifying novel protein folds and potential targets for regulation. The protein model was developed using MODELLER 9v10. Physicochemical characterization and functional annotation of the model carried out with Expasy''s ProtParam server and three different conserved domain finding programs including InterProScan, Proteins Families Database (Pfam), and NCBI Conserved Domains Database (NCBI-CDD). Applying validation programs revealed that the model has good quality and the RMSD value is 0.7. The predicted model submitted in Protein Model Database, PMDB for public use. This model will be used in wide range of studies for functional analysis and improvement activity of the protein.     

Penconazole alleviates salt-induced damage in safflower (Carthamus tinctorius L.) plants

It has been shown that penconazole (PEN) acts as an endogenous signal molecule responsible for inducing stress tolerance in plants. The effect of PEN (15?mg?l^–1) and sodium chloride (0, 100, and 200 mM NaCl) on some biochemical and molecular responses of safflower was studied. Results revealed that chlorophylls and total soluble protein contents decreased under salinity, however total carotenoid, anthocyanin, flavonoid, and carbohydrate contents increased as well as SOS1 and NHX1 genes expression. The exogenous PEN had a positive effect on chlorophylls, carotenoid, anthocyanin, flavonoid, soluble protein and carbohydrate contents. In addition, RT-qPCR analysis showed that the exogenous PEN induced expression of SOS1 and NHX1 genes in both salt-treated and untreated plants. Our data indicate that PEN helps safflower plants to better cope with salt stress. The results can provide new insights to better realizing the responsible mechanisms to regulate salinity resistance in safflower. PEN can be considered in order to ameliorate salinity effects, due to the low price and their availability.     

Nitric oxide production and absorption in trachea, bronchi, bronchioles, and respiratory bronchioles of humans

Different volumesof dead-space gas were collected and analyzed for nitric oxide (NO)content, either immediately after inspiration or after a period ofbreath holding on clean air or NO mixtures. This allowed calculation ofNO equilibrium, NO production, and NO absorption. In seven young,healthy, adult nonsmokers, the mean NO equilibrium values in parts perbillion (ppb) were 56 ± 11 (SE) in the trachea, 37 ± 6 in thebronchi, 21 ± 3 in the bronchioles, and 16 ± 2 in therespiratory bronchioles. At any given NO concentration, the NOabsorption rate (in nl/min) equaled the NO concentration (in ppb) timesA (the absorption coefficient inl/min). A values (in l/min) were 0.11 ± 0.01 in the trachea, 0.17 ± 0.04 in the bronchi, 0.66 ± 0.09 in the bronchioles, and 1.35 ± 0.32 in the respiratorybronchioles. NO equilibrium concentrations and production rates in one74-yr-old subject were three to five times as high as those found inthe young subjects. Mouth equilibrium NO concentrations were 3 and 6 parts per million in two subjects who had oral production rates of 6 and 23 nl/min, respectively. In conclusion, production and absorptionof NO occur throughout the first 450 ml of the airways.

     

A mosquito predator survey in Townsville, Australia, and an assessment of Diplonychus sp. and Anisops sp. predatorial capacity against Culex annulirostris mosquito immatures.

A twelve-month survey for mosquito predators was conducted in Townsville, Queensland, Australia, which is located in the arid tropics. The survey revealed the presence of five predaceous insects but only Anisops sp. (backswimmers) and Diplonychus sp. were common. Predatorial capacity and factors influencing this capacity were then assessed for adult Anisops sp. and adult and nymph stages of Diplonychus sp. against Culex annulirostris mosquito immatures under laboratory conditions. Predatorial capacity bioassays showed that adult Diplonychus sp. preyed upon both larval and pupal stages of Cx. annulirostris quite successfully. Nymphs of Diplonychus sp. proved to be more successful with smaller prey immatures, and Anisops sp adults did not prey successfully on any prey pupae. Increasing the foraging area and introducing aquatic vegetation significantly reduced the predatorial capacity of Diplonychus sp. nymphs, while only vegetation and not foraging area had a significant effect on adult Diplonychus sp. predation capacity. Overall, adult Diplonychus sp. proved to be a more efficient predator than Anisops sp., and field trials are now recommended to further assess the potential of Diplonychus sp. as a biocontrol agent.     

Enhanced chondrogenic differentiation of dental pulp-derived mesenchymal stem cells in 3D pellet culture system: effect of mimicking hypoxia

High incidence of articular cartilage defects resulting from age-related degeneration or trauma injuries is a major problem worldwide. Limited self-regeneration ability of cartilage often leads to inappropriate biochemistry and structure of healed tissue. Considering Impairments of traditional treatments, cell-based therapies are promising. The rapid ex vivo expansion and chondrogenic differentiation capability make dental pulp stem cells (DPSCs) a favorable cell type for therapeutic application, however strategies in order to efficient cartilage tissue-like production are imperative. In the present study the potential role of hypoxia mimicking agent, cobalt chloride (CoCl2), on chondrogenic differentiation of human DPSCs was surveyed. Cell viability assay used to obtain the optimum dose and exposure time of CoCl2. DPSCs were differentiated in pellet culture system after CoCl2 pretreatment. Chondrogenic differentiation efficiency was evaluated by histological and immunohistological analyses. The results showed that CoCl2 led to increased pellet size, integrity and matrix deposition with organizations more resembled typical cartilage lacuna structure. Furthermore, CoCl2 could improve differentiation by elevated chondrogenic markers, glycosaminoglycans (GAGs) and collagen II expression. CoCl2 pretreatment mitigated hypertrophy, as well, which was reflected in decreased collagen X expression. Alkaline phosphatase (ALP) specific activity did not change significantly by CoCl2 preconditioning. Based on current study hypoxia mimicking agent, CoCl2, could be suggested to promote DPSCs chondrogenic differentiation.     

The stochastic nature of biochemical networks

Cell behaviour and the cellular environment are stochastic. Phenotypes vary across isogenic populations and in individual cells over time. Here we will argue that to understand the abilities of cells we need to understand their stochastic nature. New experimental techniques allow gene expression to be followed in single cells over time and reveal stochastic bursts of both mRNA and protein synthesis in many different types of organisms. Stochasticity has been shown to be exploited by bacteria and viruses to decide between different behaviours. In fluctuating environments, cells that respond stochastically can out-compete those that sense environmental changes, and stochasticity may even have contributed to chromosomal gene order. We will focus on advances in modelling stochasticity, in understanding its effects on evolution and cellular design, and on means by which it may be exploited in biotechnology and medicine.     

Mixture of fibroblast, epithelial and endothelial cells conditioned media induce monocyte-derived dendritic cell maturation

Fully matured DCs with large amount cytoplasm and copious dendritic projections were visible at the end of culturing period in the presence of MCM, TNF-α and poly (I:C), with or without FEECM. Thus, DCs generated with these maturation factors are nonadherent and have typical satellite morphology. Flow cytometric analysis using anti-CD14, -CD80, -CD86, -HLA-DR and -CD83 revealed that expression of CD14 is decreased in particular in FEECM treated DCs, on day 5 and expression of CD80, CD86 and HLA-DR was the higher when FEECM are added to maturation factor. Functionally, when DCs matured in the presence of FEECM elicited stronger MLR, reduced phagocytic activity. These results support the use of the FEECM with MCM, TNF-α and poly (I–C) as maturation factor in DC generation that could result in functionally mature monocyte-derived DCs in comparison to either alone.     

An Engineered α1 Integrin-binding Collagenous Sequence

Collagen is an extracellular matrix structural component that can regulate cellular processes through its interaction with the integrins, α1β1, α2β1, α10β1, and α11β1. Collagen-like proteins have been identified in a number of bacterial species. Here, we used Scl2 from Streptococcus pyogenes serotype M28 strain MGAS6274 as a backbone for the introduction of discrete integrin-binding sequences. The introduced sequences GLPGER, GFPGER, or GFPGEN did not affect triple helix stability of the Scl (Streptococcal collagen-like) protein. Using ELISA and surface plasmon resonance, we determined that Scl2_GLPGER and Scl2_GFPGER bound to recombinant human α1 and α2 I-domains in a metal ion-dependent manner and without a requirement for hydroxyproline. We predicted a novel and selective integrin-binding sequence, GFPGEN, through the use of computer modeling and demonstrated that Scl2_GFPGEN shows specificity toward the α1 I-domain and does not bind the α2 I-domain. Using C2C12 cells, we determined that intact integrins interact with the modified Scl2 proteins with the same selectivity as recombinant I-domains. These modified Scl2 proteins also acted as cell attachment substrates for fibroblast, endothelial, and smooth muscle cells. However, the modified Scl2 proteins were unable to aggregate platelets. These results indicate that Scl2 is a suitable backbone for the introduction of mammalian integrin-binding sequences, and these sequences may be manipulated to individually target α1β1 and α2β1.     

PCR-based detection and eradication of mycoplasmal infections from various mammalian cell lines: a local experience

A total of 200 cell lines including different human, monkey, mice, hamster and rat cell types were examined for mycoplasma infection status. PCR assay using generic-specific universal primers showed that 40 (20%) of the cell lines are contaminated with mycoplasma. Employment of species-specific primers within these infected cell lines revealed infection with M. hyorhinis (42.5%), M. fermentas (37.5%), M. arginini (37.5%), M. orale (12.5%) and A. laidlawii (7.5%). A number of the cultures were coinfected with 2 or 3 different species. Contaminated samples were treated with BM-Cyclin, Ciprofloxacin and mycoplasma removal agent (MRA). Mycoplasma eradication was subsequently checked by PCR following 2 weeks continuous culture of treated cells in antibiotic free culture medium. Mycoplasmal infections were eradicated in 100, 70 and 42% of infected cell lines when the samples were treated with BM-Cyclin, MRA and Ciprofloxacin, respectively. However, 12% (BM-Cyclin), 62.5% (MRA) and 82.5% (Ciprofloxacin) of mycoplasma regrowth was observed 4 months after the treatment. Notably, the risk of spontaneous culture death was 17.5, 12.5 and 0% for BM-Cyclin, MRA and Ciprofloxacin, respectively.     

Serological detection of Sharka quarantine disease (Plum pox virus) on stone fruit trees in Golestan province

Sharka disease is one of the most damaging diseases of fruit trees in the world, which is caused by Plum pox virus (PPV) that belongs to the genus Potyvirus in the family Potyviridae. Each year, this virus decreases the yield and causes substantial economic damages to its host plants worldwide. This virus is quarantined in Iran but in recent years, suspicious symptoms of the disease were observed in different grown areas, such as Golestan province. During 2010, 420 samples with mosaic, chlorosis, necrosis, ring pattern, blotches, etc. symptoms were collected from the gardens in Golestan province that included 100 samples from plum, 100 from peach and 240 from nectarine. These samples were evaluated using a double-antibody sandwich-ELISA (DAS-ELISA) method and a polyclonal antibody. The results of this survey indicated that among the total of 420 samples, none of them showed positive reaction in DAS-ELISA test.