Identification of Mycobacterium tuberculosis CTL epitopes restricted by HLA-A*0201 in HHD mice

BACKGROUND: CD8 T cells are thought to play an important role in protective immunity to tuberculosis. The major histocompatibility complex class I subtype HLA-A*0201 is one of the most prevalent class I alleles, with a frequency of over 30% in most populations. HLA-A*0201 transgenic, H-2D(b)/mouse beta2-microglobulin double-knockout mice (HHD) which express human HLA-A*0201 but no mouse class 1, was shown to provide a powerful model for studying induction of HLA-A*0201-restricted immune responses in vivo. METHODS: HHD mice were immunized with plasmid DNA encoding MPB51 by using a gene gun, and IFN-gamma production from the immune spleen cells was analyzed in response to a synthetic overlapping peptide library covering the mature MPB51 sequence. catatonic T lymphocytes (CTL) activity was measured using cytotoxicity assay and the three-color flowcytometry was used to reveal IFN-gamma-producing immune spleen cells. RESULTS: Our findings were shown that only one peptide, p51-70, appeared to stimulate the immune splenocytes to produce IFN-gamma. Flow cytometric analysis with intracellular IFN-gamma and the T-cell phenotype revealed that the p51-70 peptide contains an immunodominant CD8+ T-cell epitope. Further analysis with computer-assisted algorithms permitted identification of a T-cell nona mer epitope, p54-62. Finally, we proved that the p54-62/HLA-A*0201 complex is strongly recognized by HLA class I-restricted CD8+ MPB5 1-specific CTL cells. CONCLUSION: These results suggest that vaccination with MPB51 gene elicited MPB51-specific CTL. In addition, the P54-62 epitope thus represent potential subunit component for the design of vaccines against tuberculosis.     

Molecular characterization of Newcastle disease viruses isolated from rural chicken in northwest Ethiopia reveals the circulation of three distinct genotypes in the country

Newcastle disease (ND) is a highly contagious disease that affects many species of birds and causes significant economic losses to the poultry industry worldwide. Fifteen Newcastle disease virus (NDV) isolates obtained from rural chickens in northwest Ethiopia in 2011 and 2012 were characterized genotypically. The main functional region of the F gene was amplified and sequenced (260 nucleotides). Among the Ethiopian NDV isolates, 2 isolates had the virulent motif ¹¹²R-R-Q-K-R-F¹¹⁷ at the cleavage site of the fusion protein while 13 isolates contained the lentogenic motif ¹¹²G-G/R-Q-G-R-L¹¹⁷. Phylogenetic analysis based on the variable region of the F gene indicated that the two isolates exhibiting the virulent motif belonged to lineage 5 (genotype VII) subgenotype d and the remaining 13 isolates were grouped into lineage 2 (genotype II). The nucleotide sequences of lineage 5 isolates were genetically related to the Sudanese NDV isolates, suggesting potential epidemiological link of ND outbreaks between neighbouring countries. The lentogenic strains shared similarities with La Sota vaccine strain and probably originated from the vaccine strain either through direct exposure of birds to the live vaccine or to infectious La Sota-like strains circulating in rural poultry. This study provides genetic evidence on the existence of different NDV genotypes circulating in the rural poultry in Ethiopia. The virulent NDV continues to be a problem in poultry sector in Ethiopia, and their continuous circulation in rural and commercial poultry calls for improved surveillance and intensified vaccination and other control measures.     

Effect of growth parameters on structural and optical properties of CdS nanowires prepared by polymer controlled solvothermal route

Cadmium sulfide nanowires were successfully obtained via a poly (ethylene glycol)-assisted solvothermal route. In this procedure, cadmium nitrate and thiourea were used as Cd and S sources, respectively, and polyethylene glycol 400 was used as an inducing soft template to control the one-dimensional growth of CdS nanostructures. The effects of different growth parameters in the solvothermal process such as type of the solvents, reaction time, and temperature on the morphology, structural and optical properties of the products were investigated. The provided structures were characterized by X-ray diffractometery, scanning electron microscopy, and UV-visible absorption spectroscopy. The results show that the as-prepared samples have hexagonal phase and grow into long nanowire shape with increasing the reaction time, temperature and volume ratios of ethylenediamine (en) to H₂O. Uniform sized nanowires with the average diameter of 75 nm and the average length of 2.5 µm were obtained using ethylenediamine solvent at 170 °C for 3 days.     

Genetic analysis of flowering and maturity time in high latitude spring wheat

Due to the short growing season in the high northern latitudes, the development of early maturing spring wheat (Triticum aestivum L.) cultivars is important to avoid frost damage which can lower production and quality. We investigated earliness of flowering and maturity, and some associated agronomic traits, using a set of randomly selected high northern latitude adapted spring wheat cultivars (differing in maturity) and their F₁ and F₂ crosses made in a one-way diallel mating design. The parents, and their F₁ and F₂ crosses were evaluated under field conditions over 2 years. Anthesis and maturity times were controlled by both vernalization response and earliness per se genes, mainly acting additively. Non-additive genetic effects were more important in controlling grain fill duration, grain yield and plant height. Additive × additive epistatic effects were detected for all traits studied except time to anthesis. Segregation analyses of the F₂ populations for time to anthesis indicated the presence of different vernalization response genes. Molecular genetic analyses revealed the presence of Vrn-A1 and Vrn-B1 genes in the parental cultivars. Narrow-sense heritability was medium to high (60–86%) for anthesis and maturity times but low to medium (13–55%) for grain fill duration, plant height and grain yield. Selection for early flowering/maturity in early segregating generations would be expected to result in genetic improvement towards earliness in high latitude spring wheats. Incorporation of the vernalization responsive gene Vrn-B1 in combination with vernalization non-responsive gene Vrn-A1 into spring wheats would aid in the development of early maturing cultivars with high grain yield potential for the high latitude wheat growing regions of the northern hemisphere.     

Role of Raf in vascular protection from distinct apoptotic stimuli

Raf kinases have been linked to endothelial cell survival. Here, we show that basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) differentially activate Raf, resulting in protection from distinct pathways of apoptosis in human endothelial cells and chick embryo vasculature. bFGF activated Raf-1 via p21-activated protein kinase-1 (PAK-1) phosphorylation of serines 338 and 339, resulting in Raf-1 mitochondrial translocation and endothelial cell protection from the intrinsic pathway of apoptosis, independent of the mitogen-activated protein kinase kinase-1 (MEK1). In contrast, VEGF activated Raf-1 via Src kinase, leading to phosphorylation of tyrosines 340 and 341 and MEK1-dependent protection from extrinsic-mediated apoptosis. These findings implicate Raf-1 as a pivotal regulator of endothelial cell survival during angiogenesis.     

Nutrition Management and Farm’s Age Affect Saffron Daughter Corms Behavior,Nutrients Uptake and Economic Water and Fertilizer Use Efficiency: A Large Scale On-farm Experiment in Torbat Heydarieh,Iran

ABSTRACT

Evaluation of the relationships between nutritional patterns and farms’ age with the behavior of saffron (Crocus sativus L.) daughter corms based on farmers’ management (on-farm) can be crucial in improving saffron sustainable yield. Furthermore, in commercial saffron production, especially in small farms, the purchased water and fertilizers are the basis of sustainable saffron profitability, hence, recognizing the relationships between economic water use efficiency (EWUE) and economic fertilizer use efficiency (EFUE) can be important. An on-farm experiment was conducted on a large scale based on farmers’ management in Torbat Heydarieh, Iran. The farms’ age (1–6 years old) and fertilizers management approaches (organic, mineral and integrated) were considered as the first and second factors, respectively. Large-sized daughter corms number and weight and corms N and P content increased with increasing farms’ age from 1 to 4 years old. However, these parameters decreased with increasing farms’ age from 4 to 6 years old. Irrespective of saffron farms’ age, the minimum large-sized daughter corms number and weight were observed when mineral fertilizer management was practiced. By contrast, the maximum values were related to integrated fertilizer management. In each type of fertilizer management, EWUE and EFUE (based on large-sized daughter corms monetary value) increased with increasing farms age from 1 to 4 years. However, increase in farms age from 4 to 6 years caused a significant reduction in mentioned indices. Overall, if high-quality saffron corm production is needed, corms should be harvested in the fourth year to gain the maximum yield and profit.     

Monitoring of the fermentation media of citric acid by the trimethylsilyl derivatives of the organic acids formed

In this approach, a derivatization method is described for monitoring of organic acids in fermentation media without any separation step. The aqueous phase of fermentation media was evaporated and heated in a silylation reagent to form trimethylsilyl (TMS) derivatives. The silylated compounds are analyzed by 29Si nuclear magnetic resonance (29Si NMR) and gas chromatography-mass spectrometry (GC-MS). 29Si NMR can qualitatively monitor the components produced in the Krebs cycle. Quantification of these compounds is investigated by using selected ion monitoring mode of mass spectrometry. In this mode, mass to charge (m/z) values of their [M - 15]+ ions, which are 465, 275, 247, 221, 335, 251, and 313 of TMS derivatives of citric, alpha-ketoglutaric, succinic, fumaric, l-malic, oxaloacetic, and palmitic (as an internal standard), acids, respectively, are used. The limit of detection and the linear working range for derivatized citric acid were found to be 0.1 mg L(-1) and 10-3 x 10(4) mg L(-1). The relative standard deviation of the method for five replicates was 2.1%. The average recovery efficiency for citric acid added to culture media was approximately 97.2%. Quantitative results of GC-MS are compared with those obtained by an ultraviolet-visible method.     

Foliar application of iron,zinc, and manganese nano-chelates improves physiological indicators and soybean yield under water deficit stress

Abstract

To investigate the effect of foliar application of nano-chelates of iron, zinc, and manganese subjected to different irrigation conditions on physiological traits, and yield of soybean (cultivar M9), a split plot experiment was conducted in a completely randomized block design with three replications in two crop years (2016–2017). The main plot included four levels of irrigation (I): full irrigation (I ₁), irrigation withhold at flowering stage (I ₂), irrigation withhold at podding stage (I ₃), and irrigation withhold during the grain filling period (I ₄). Also, the subplot included eight levels of foliar application with Fe, Zn, Mn, Fe?+?Zn, Fe?+?Mn, Zn?+?Mn, Fe?+?Zn?+?Mn nano-chelates, and distilled water (control). The results of combined analysis of variance suggested that the effect of irrigation and foliar application of nano-chelate was significant on all traits. Water deficit stress significantly reduced the grain yield. The minimum numbers of pods per plant, number of grains per plant, 100-seed weight per plant, leaf area index, leaf chlorophyll concentration, total dry weight of plant, and the grain yield were obtained by irrigation withhold at podding stage. Foliar application of combined nano-chelates increased the soybean resistance against water shortage more considerably than the separate consumption of these elements. Under drought stress in podding stage, the application of Fe?+?Zn led to the highest yield with a mean of 2613.84?kg ha^?1 where this increase was 61.1% higher than control.     

基于COI基因的入侵物种旧金山卤虫遗传多样性与遗传分化研究

伴随着中国水产养殖业的快速发展,原产美国的旧金山卤虫(Artemia franciscana)被引入至我国渤海湾地区.为研究旧金山卤虫被引入中国后其基因组遗传多样性改变情况,本研究利用COI线粒体分子标记,评估旧金山卤虫在非原生栖息地,即唐山曹妃甸南堡镇(渤海湾,河北省)定殖的旧金山卤虫的遗传多样性现状.研究分析发现:...