Woodchuck Hepatitis Virus Post-Transcriptional Regulation Element (WPRE) Promotes Anti-CD19 BiTE Expression in Expi293 Cells

Background:Bispecific antibodies represent an important class of mAbs, with great therapeutic potentials due to their ability to target simultaneously two distinct epitopes. The generation of functional bispecific antibodies with the highest possible yields is particularly critical for the production of these compounds on industrial scales. Anti- CD3 × CD19 bsAb is a bispecific T-cell engager (BiTE) currently used for treating ALL. Herein, we have tried to optimize the expression level of this antibody in mammalian hosts. Methods:WPRE sequence was incorporated at the 3’ end of the expression cassette. This modification resulted in a notable about two-fold increase in the expression of the bsAb in the Expi293 cell line. Results & Conclusion:Follow-up flow cytometry analysis demonstrated the binding properties of the produced antibody at acceptable levels, and in vitro bioactivity assays showed that this product is potent enough for targeting and destroying CD19-positive cells. Our findings show that WPRE enhances the expression of this type of bispecific mAbs in HEK-293 family cell lines. This approach can be used in biopharma industry for the mass production of anti-CD3 × CD19 bispecific antibody. Key Words: Acute lymphoblastic leukemia, Bispecific antibodies, Monoclonal antibody     

The potential of rice to offer solutions for malnutrition and chronic diseases

Elevated Na⁺ levels in agricultural lands are increasingly becoming a serious threat to the world agriculture. Plants suffer osmotic and ionic stress under high salinity due to the salts accumulated at the outside of roots and those accumulated at the inside of the plant cells, respectively. Mechanisms of salinity tolerance in plants have been extensively studied and in the recent years these studies focus on the function of key enzymes and plant morphological traits. Here, we provide an updated overview of salt tolerant mechanisms in glycophytes with a particular interest in rice (Oryza sativa) plants. Protective mechanisms that prevent water loss due to the increased osmotic pressure, the development of Na⁺ toxicity on essential cellular metabolisms, and the movement of ions via the apoplastic pathway (i.e. apoplastic barriers) are described here in detail.     

Nickel supplementation effect on the growth,urease activity and urea and nitrate concentrations in lettuce supplied with different nitrogen sources

Effect of nickel (Ni) in the nutrient solution on yield, N metabolism, and nitrate content of leafy vegetables is poorly understood. The aim of this nutrient solution culture experiment was to investigate the effects of Ni supplementation on the nitrogen (N) metabolism and growth of lettuce (Lactuca sativa L. cv. Baker) with either urea or nitrate as the N source. Nickel supplement at 0.04 μM reduced urea toxicity to the urea-fed plants. Addition of Ni to the nutrient solution significantly increased the leaves and root growth of the urea-fed lettuce plants while it increased the growth of lettuce plants fed with nitrate only at N level of 20 mM. At N level of 20 mM, the leaves fresh weight of the urea-fed plants promoted by Ni supplement was comparable with the nitrate-fed plants untreated with Ni. Nickel supplementation increased the leaf total N concentrations in the urea-fed plants, although the nitrate-fed plants accumulated greater N in their leaves compared with urea treated plants at without Ni treatment. Nickel addition decreased the concentrations of leaf urea-N in the urea-fed plants and NO₃-N in the nitrate-fed plants. Nickel addition enhanced urease activity in the leaves of urea-fed plants. The results indicated Ni supplementation enhances the growth of the urea-fed lettuce plants while it has role in decreasing leaf nitrate concentration and thus, improving the health quality of the nitrate-fed plants.     

Association of allelic polymorphisms of IGFALS gene with growth traits in Makouei and Ghezel sheep breeds

Insulin-like growth factor-binding protein acid-labile subunit (IGFALS) encodes a protein which binds to IGF1 and IGFBP-3 to regulate the growth, differentiation, and other physiological processes. The aim of this study was the identification of allelic polymorphisms of the IGFALS gene using the PCR-RFLP technique and evaluation of their association with growth traits. For this end, 120 blood samples were randomly collected from each breed. Following amplification of an 1113-bp fragment of exon 1 and a part of intron 1 of the IGFLAS gene, genotyping was conducted by three restriction enzymes including HinfI, MscI, and PvuII. The results showed that only one allele was observed in IGFALS-PvuII site, while in IGFLAS-MscI site, three AA, AB, and BB genotypes with the frequencies of 17.5%, 32%, and 50.5% and 11%, 37.5%, and 51.5% were observed in Makouei and Ghezel sheep breeds, respectively. Additionally, in the IGFLAS-HinfI site, two AB and BB genotypes with the frequency of 34.2% and 65.8% were observed in Makouei sheep and AA, AB, and BB genotypes with the frequency of 9%, 21%, and 70% were observed in Ghezel sheep. So that, Makouei sheep with AB genotype had more chest girth (CG) compared with other genotypes. Furthermore, a significant association was observed between the genotypes of IGFLAS-HinfI with birth weight (BW) in Ghezel and BW, weaning weight (BW3), and CG in Makouei sheep. Haplotype analysis revealed an association between paternal haplotypes and BW in both Ghezel and Makouei breeds. So that, AAB and ABB haplotypes showed more BW than others in Makouei and Ghezel sheep, respectively.

     

Genetic variation,heritability and correlations of agro-morphological traits in tall fescue (<Emphasis Type="Italic">Festuca arundinacea</Emphasis> Schreb.)

To study the genetic variation, heritability and genetic correlations of some agro-morphological traits in tall fescue, 25 parents from a genetically broad-base germplasm were polycrossed and their respective half-sib families were generated. Clonally propagated parents and their half-sib families were grown as spaced single plants using a randomized complete-block design with three replications, and observed for seven traits in 2006 and 2007. The estimates of broad-sense heritability were moderate to high for the traits studied. Narrow-sense heritability estimates from analyses of progenies and from regression of half-sib (HS) progenies on parents suggested that genetic variation for these traits was largely controlled by additive gene action. Association of dry matter yield (DMY) with plant height, number of fertile shoot, curbs width and spring growth was positive and significant. With the exception of number of days to pollination, correlation coefficients of the traits between the parents and offspring were not significant. Based on parent-offspring regression, genetic gain from selection for DMY was high, demonstrating genetic potential for improving this trait. Overall, there was high genetic variation and moderate heritability for most traits in the tall fescue populations evaluated. In conclusion, to improve herbage yield, selection would be more effective based on forage yield components.     

Gibbsiella quercinecans and Brenneria roseae subsp. roseae associated to the canker disease of walnut trees in northwestern Iran

European Journal of Plant Pathology - A bacterial disease of walnut (Juglans regia) has been observed in northwestern Iran during the summer of 2018 with symptoms similar to the shallow bark canker...     

Functional analysis of walnut polyphenol oxidase gene (JrPPO1) in transgenic tobacco plants and PPO induction in response to walnut bacterial blight

Walnut (Juglans regia) is economically important for both its wood and nut nutritional value, but it is susceptible to diseases such as walnut bacterial blight, caused by Xanthomonas arboricola pv. juglandis (Xaj). Walnuts contain many phenolic compounds, providing a good model on which to study polyphenol oxidase (PPO). We inoculated the detached walnut fruits of cultivars Ford, Chandler, Franquette, Robert Livermore, and Payne with Xaj and measured the induction of PPO activity in infected sites and adjacent to infected sites. Compared to infected and uninfected sites, PPO activity was induced significantly in areas adjacent to infected sites in all cultivars except Ford. Ford and Franquette, presenting the lowest and highest PPO activity, showed the largest and smallest mean diameter spots in response to Xaj, respectively. Polyacrylamide gel electrophoresis confirmed monophenol oxidase activity of walnut PPO in the assessed tissues. Then, we revealed the antipathogenic potential of walnut PPO through Agrobacterium tumefaciens-mediated walnut JrPPO1 gene transfer into tobacco (Nicotiana tabacum). Two transformed tobacco lines overexpressing the JrPPO1 gene were regenerated successfully and challenged with Pseudomonas syringae pv. tabaci. Transgenic lines showed significantly higher PPO activity and lower disease severity to the pathogen compared to the control. However, a significant difference in disease severity and PPO activity level was observed between the two transgenic lines. Our results demonstrate a potential defence-related role of PPO in transgenic tobacco and its induction in areas adjacent to infection sites in walnut cultivars treated with Xaj.     

Owner complaints of canine behavior in Iran–A preliminary survey

The current survey assessed 17 common behavioral complaints reported by 234 dog owners attending the Small Animal Hospital, University of Tehran using a questionnaire. According to the results, nearly 95% of dog owners reported at least 1 undesirable behavior in their dogs. Aggression toward other dogs and people, jumping up, and excessive barking were the most common behavioral complaints. Owners of large-breed dogs were more likely to complain about their dogs’ aggressive behaviors toward people/dogs and excessive barking compared with owners of small-breed dogs. Dogs without access to a yard were more likely to show phobia-associated behaviors, fearfulness, and separation-related behaviors, whereas those having access to a yard were more likely to show aggressive behaviors toward people/dogs and excessive barking. Interaction of dogs with people and type of reaction used by owners in response to the behavior were also related with some other behavioral characteristics of the animals.