Designing E1 deleted adenoviral vector by homologous recombination

BACKGROUND: Adenoviruses are used extensively to deliver genes into mammalian cells, particularly where there is a requirement for high-level expression of transgene products in cultured cells, or for use as recombinant viral vaccines or in gene therapy. In spite of their usefulness, the construction of adenoviral vectors (AdV) is a cumbersome and lengthy process that is not readily amenable to the generation of large collection of clones. METHODS: In this project, to delete E1 gene in adenovirus, an adenoviral plasmid containing lateral sites of E1 region of adenovirus was made and recombination in the 293A cells between the homologous region of this linearized plasmid and the adenovirus genome resulted in the formation of the complete adenoviral recombinant. RESULTS: This recombination resulted in loss of E1 region and we constructed a recombinant adenovirus type 5 vector that E1 gene was deleted by homologous recombination. CONCLUSION: Homologous recombination is more easy and fast technique in the production of AdV.     

Herbicide risk assessment during the Wheat Self-sufficiency Project in Iran

BACKGROUND: The present study describes the quantitative changes in herbicide use during the specific observation periods in the Wheat Self-sufficiency Project in Iran from 1994 to 2004 and the associated changes in herbicide risk, area and yield in this context. A risk index, the environmental impact quotient (EIQ), was used to estimate the environmental impacts (EIs) of herbicides applied to wheat agroecosystems. RESULTS: The results suggest that, during this period, the overall risk posed by the herbicides applied to wheat agroecosystems increased substantially, as evidenced by a 71% increase in herbicide usage (weight of pesticide applied) and a 62.2% increase in EI rating, in spite of an 8.2% decrease in overall EIQ rating and an 89.2% decrease in mean application rate. Furthermore, a 0.57% increase in the area and a 23.6% and 22.7% increase in irrigated and rainfed yields, respectively, were observed. The results also indicate that the EIQ(FUR) of two herbicides, dichlorprop-P/mecoprop-P/MCPA and difenzoquat, is much higher than that of other herbicides, and they could be qualified as high-risk herbicides. CONCLUSION: As a whole, an increase in herbicide usage (weight of pesticide applied) was an important factor on herbicide environmental impact boost. However, this increase has not led to a similar increase in wheat yield, which could be attributed in part to the negative impact of high herbicide consumption in wheat fields and subsequent threat to the long-term sustainability of these agroecosystems.     

Chemical composition and in vitro antifungal activity of Sambucus ebulus and Actinidia deliciosa on the fish pathogenic fungus,Saprolegnia parasitica

The aim of the present study was to determine the chemical composition of Sambucus ebulus and Actinidia deliciosa ethanolic extracts as well as their in vitro antifungal activity on the mycelial growth of the water mold, Saprolegnia parasitica. The preliminary minimum inhibition concentration (MIC) and minimum lethal concentration (MLC) were determined by the HeMP method and finally, concentrations of each extract ranging from 1 to 10% were prepared by an agar dilution method to assess the in vitro antifungal activity, quantitatively. Both herbal extracts inhibited growth of Saprolegnia hyphae in vitro. Complete in vitro growth inhibition was found at a concentration of ≥5% for S. ebulus, whereas it was not observed even at 10% concentration of A. deliciosa. Based on gas chromatography coupled with mass spectrometry (GC/MS) analysis, the main constituents of S. ebulus include monophthalate (66.17%), fatty acids (26.47%), phytol (4.25%), and acetic acid (2.11%). Using colorimetric assays, A. deliciosa contained phenolic contents at 162 mg gallic acid (GAE)/g DW and flavonoid contents at 2.31 mg quercetin (QE)/g DW. In conclusion, the results of this study indicated that S. ebulus and A. deliciosa showed some antifungal activities against S. parasitica with formerly exhibiting stronger activity (p < 0.05).

     

Molecular typing of Iranian field isolates Mycoplasma synoviae and their differentiation from the live commercial vaccine strain MS-H using vlhA gene

1. The single-copy domain of the N-terminal region of the vlhA gene of Mycoplasma synoviae was sequenced, analysed and verified and used to type Iranian field isolates of M. synoviae and the MS-H live vaccine strain. In addition, a restriction fragment length polymorphism (RFLP) method was developed to differentiate between field isolates of Iranian and MS-H vaccine strains.

2. All sequences were analysed and aligned; the percentage similarity of the DNA was calculated and dendrograms were constructed. Based on single nucleotide polymorphism (SNP) that existed in all field isolates in Iran, the PCR-RFLP method allowed the differentiation of all M. synoviae field isolates from the vaccine strain.

3. Using phylogenetic analysis, the isolates were assigned to 8 unique genotypes and, within each group, DNA had a high level of similarity.

4. DNA sequence analysis and PCR-RFLP of the amplicon based on percent similarity and evolutionary relationship appeared to be useful tools for strain differentiation whether M. synoviae clinical isolates from infected chickens were derived from the vaccine strain or wild-type strains.

5. This study confirms the potential value of strain typing for epidemiological purposes and suggests that phylogenetic studies are essential to understand the true relationships between strains.     

Immune responses to oral and IM administration of M2e-Hsp70 construct

The eukaryotic expression plasmid of M2e-Hsp70 is a candidate M2e-based DNA vaccine. In order to evaluate the immunization potential of this construct, Specific Pathogen Free chickens were immunized either intramuscularly or orally. Mutant Salmonella typhimurium was used as carrier for oral delivery of the M2e-Hsp70 construct. M2e-specific humoral and cellular responses were tested by ELISA and Lymphocyte Proliferation Assay, respectively. Our results indicate that both humoral and cellular immune responses are conferred against M2e-Hsp70 plasmid in either of the intramuscular or oral routes of administration; however, these responses are significantly higher in intramuscular injection in contrast to oral administration. When it comes to mass vaccination of commercial chicken flocks oral administration is preferred due to the ease of application as well as its capability of eliciting mucosal, humoral and cellular immune responses; so measurements should be taken to improve the immunization potency of our orally delivered DNA vaccine.     

Karyological analysis of endangered Caspian salmon, Salmo trutta caspius (Kessler, 1877)

The karyotype and chromosomal characteristics of endangered Caspian salmon, Salmo trutta caspius, in the southern part of the Caspian Sea were investigated. From the total of 60 mitotic metaphases achieved from 15 individuals, 15, 9 and 36 metaphases were with the mode <76, 76 and 80 representing 25%, 15% and 60% of metaphases respectively. So, the most common pattern of chromosome number was 80 (36 metaphases, 60%) and the number of diploid chromosomes was thus confirmed as (2n=80) of this subspecies. The karyotype consists of seven metacentric, five submetacentric and 28 telocentric pairs. The karyological parameters of the Caspian salmon, the centromic index, arm ratio, relative length and length variation range of chromosomes, were 0–0.5, 1–∞, 0.011–0.045 and 0.507–2.028 μm respectively. The total length of chromosomes in haploid series was 44.776 μm and fundamental number of chromosome arms was 104. This study may provide the first knowledge on chromosome analysis in Caspian salmon and add basic information useful for its chromosomal manipulations.     

Antinociceptive and selected physiological effects of morphine and xylazine on tiletamine‐zolazepam anesthesia in llamas

ObjectiveEvaluate antinociception, anesthesia, and recovery in llamas given tiletamine-zolazepam (TZ) with either morphine, xylazine, morphine and xylazine, or saline.Study designRandomized crossover experimental study.AnimalsSix healthy, adult intact male llamas.MethodsLlamas were given each of four treatments intramuscularly with a 1-week washout: TZ (2 mg kg^?1) combined with either morphine (0.5 mg kg^?1; M), xylazine (0.15 mg kg^?1; X), morphine (0.5 mg kg^?1) and xylazine (0.15mg kg^?1) (MX), or saline (C). Llamas breathed room air during the experiment. Characteristics of anesthesia, recovery, and selected cardiopulmonary variables were recorded. Antinociception was assessed by clamping a claw at 5-minute intervals. Data were analyzed using a mixed-model anova and Tukey-Kramer test, and are expressed as least squares mean ± SEM. Significance was set at p < 0.05.ResultsNo llama in the control group demonstrated antinociception. Antinociception was longest with treatment MX, followed by treatments X and M, respectively. Heart rates in llamas given treatments X and MX were significantly lower than with other treatments. The respiratory rate in llamas given treatment C was greater (p < 0.05) than for all other treatments, however, the respiratory rate was not significantly different among treatments X, M and MX. The PaO₂ for llamas given MX remained <60 mmHg throughout the 20 minute period of blood gas analysis. Mean arterial blood pressure in llamas in treatment MX was less than for treatments M or C.Conclusion and clinical relevanceThe combination of morphine (0.5 mg kg^?1) and xylazine (0.15 mg kg^?1) increased the duration of antinociception compared with xylazine alone, in TZ-anesthetized llamas. Treatments X, M and MX were associated with hypoxemia (PaO₂ < 60 mmHg).     

A bioinformatic insight into the genetic diversity within pomegranate cultivars: from nuclear to chloroplast genes

Genetic Resources and Crop Evolution - The pomegranate (Punica granatum L.) is an ancient fruit plant which has been consumed for its nutritional value and medicinal properties by mankind. Several...     

Quantitative Mapping and Assessment of Environmentally Sensitive Areas to Desertification in Central Iran

Desertification is one of the main environmental and also social and economic problems facing Iran. Seventeen out of 31 Iranian provinces, which are home to approximately 70% of the total population, are affected by desertification. This study aimed to use geographic information system (GIS) and fuzzy logic for mapping environmentally sensitive areas to desertification based on Mediterranean Desertification and Land Use approach in Isfahan province, central Iran. Six desertification indicators including climate, soil, vegetation, soil erosion, groundwater, and management and policy quality were used to determine various types of environmentally sensitive areas to desertification. Seventeen desertification indices affecting the quality of each indicator were spatially mapped and assigned a value between 0 and 1 using a fuzzy logic option of ArcSDM3 software in GIS environment. Results showed that a 21·7% of the study area was classified as critical, 70% as fragile and 5·5% as potential, and 2·9% of the area was not affected by desertification. In the town of Borkhar, 64·2% of the area was classified as critical, followed by the towns of Isfahan and Nayin with 40·2% and 31·8%, respectively. Results at provincial scale indicated that the climate indicator and humidity index with a weighting mean of 0·71 and 0·77 were the most affective factors in the desertification of the study area. The developed model in this study can be used for mapping desertification status in other 16 provinces that contain desert areas. These assessments provide a GIS‐based desertification database that Iran as a member of the United Nation Convention to Combat Desertification can use to report the condition of desertification at national scale. Copyright © 2013 John Wiley & Sons, Ltd.