Standardization of the outbred BALB/c mice as a suitable animal model for <Emphasis Type="BoldItalic">Besnoitia caprae</Emphasis> studies

The Apicomplexan parasite Besnoitia caprae infects wild and domestic goats. Knowledge on Besnoitia caprae specially an optimized animal model is sparse. Experimental infections with tachyzoites of BC-Pars obtained from BALB/c mice were conducted in outbred mice to determine the infectivity and LD50 of Besnoitia caprae. Six groups of five mice were intraperitoneally infected with 12.5 × 10³, 25 × 10³, 5 × 10⁴, 1 × 10⁵ and 2 × 10⁵ tachyzoites and a control inoculum of DMEM, respectively. Although morbidity and mortality were observed in all groups, two mice in the 12.5 × 10³ group showed alopecia and skin lesions on 60 days post-infection (DPI). Histopathological and molecular examination of skins confirmed B. caprae infection. The LD₅₀ was calculated as 25 × 10^3.2 tachyzoites per mouse. The results indicate that outbred BALB/c mice can be used as a suitable model of besnoitiosis and to screen candidate treatments and to test the efficacy of vaccines for besnoitiosis.     

Full sequence analysis of hemagglutinin and neuraminidase genes and proteins of highly pathogenic avian influenza H5N1 virus detected in Iran, 2015

Over the last two decades, the highly pathogenic avian influenza H5N1 virus has gained a lot of attention due to its zoonotic and mutative nature. Iran is among the countries significantly affected by the virus as it hosts migratory birds during seasonal migration. In this study, the molecular characterizations of hemagglutinin (HA) and neuraminidase (NA) genes and proteins of H5N1 strain A/chicken/Iran/8/2015 detected in backyard poultry, Mazandaran province, were investigated. Phylogenetic analysis classified this virus as a member of subclade 2.3.2.1c, with the cleavage site motif of “PQRERRRK-R/GLF”. HA carried a few mutations altering affinity to mammalian cells; however, the virus was categorized as avian. NA protein had the 20-amino acid deletion at aa position 49–69 similar to those isolated since 2000. Mutations of H253Y and H274Y contributing to antiviral resistance were present in NA. From this analysis, it can be concluded that the wild migratory birds flying from Western Asia to Eastern Africa are probably the main carriers of seasonal H5N1 in the country.

     

Effect of grape seed extract on lead induced hypertension and heart rate in rat

The main objective of this study was to evaluate the potential protective effect of red Grape Seed Extract (GSE) on lead induced hypertension (HTN) and Heart Rate (HR) in male Wistar rats. The rats were randomly assigned to one of 4 groups: Each group received lead acetate (100 ppm in drinking water), GSE (100 mg kg(-1), orally) or Lead + GSE for 45 days. Another group assigned as control group provided with tap water and regular pellet food. The Systolic Blood Pressure (SBP) and heart rate were determined by tail plethysmography coupled to a computer system. There was a sustained elevation of SBP in lead exposed rats that significantly increased at day 18 (lead treated, 112.7 +/- 2.7 mmHg, vs. control, 105.6 +/- 2.6 mmHg, n = 10, p < 0.05) and reached a maximum level at day 36 (lead treated, 124.9 +/- 2.3 mmHg, vs. control, 103.6 +/- 3.1 mmHg, n = 10, p < 0.001). However, the other three groups; showed no significant changes in SBP. Furthermore, the heart rate was increased sustainly in lead exposed animals that was statistically significant at days 36 and 45 (lead treated group, 404.5 +/- 9.4 vs. control group, 381.7 +/- 6.7, n = 10, p < 0.05). The blood lead level in both lead and lead + GSE treated groups was increased significantly compared with control and GSE treated groups (p < 0.001). However, GSE administration had no effect on the blood lead level in lead treated group. According to the result of this study, it may be concluded that GSE could have beneficial effect in protecting the cardiovascular system through its antioxidant activity against oxidative stress.     

Cytotoxic Effects of Serum from Equine Grass Sickness Cases on Neuro-2a and PC12 Tet-Off Cell Lines: Implication for Using In Vitro Methods as Antemortem Diagnostic Tools

A wide variety of clinical and paraclinical methods have been used for diagnosis of equine grass sickness (EGS), but none of them could absolutely confirm the diagnosis, and postmortem pathologic examination is still considered the final step in precise diagnosis of EGS. Use of in vitro cell toxicity caused by EGS serum on neuronal cell lines was investigated. Three well-known cytotoxic methods were used to investigate the cytotoxicity of EGS serum on neuro-2a and genetically engineered PC12 Tet-Off P53 cells. The results of alamar blue reduction as an index for mitochondrial activities and intracellular adenosine triphosphate content assays, but not neutral red uptake, indicate that the EGS serum may affect the mitochondrial function and cellular metabolism at up to 60% of the cases. The results of present study might be used for diagnosis of EGS cases. Further studies with high sample size may lead us to uncover the pathogenesis of EGS and to increase the sensitivity and applicability of in vitro techniques as diagnostic tools.     

Long-term charcoal-induced changes to soil properties in temperate regions of northern Iran

The long-term performance and benefits of charcoal application on the carbon sequestration and properties of forest soils in temperate or non-tropical regions has not been studied in detail in spite of its important role in global warming. This study was conducted to describe the long-term charcoal-induced changes in organic carbon (OC) content and other soil properties of temperate deciduous forests in Mazandaran province, northern Iran. Three sites were sampled to collect composite soil samples from two depths (0–20 and 20–40 cm) inside and outside of a plot of charcoal-enriched soils surrounding a historical charcoal production site (abandoned for more than 120 years). The presence of charcoal in soils for about 120 years elevated significantly the black carbon, total OC, natural soil OC, total nitrogen, dissolved organic matter, soil OC density, exchangeable bases, saturated hydraulic conductivity, available water capacity and available Fe, Mn and Zn compared to the adjacent reference soils. Cation exchange capacity (CEC) and pH were 15.5 cmolc kg^-1 and 0.5 units, respectively, higher than the adjacent reference soils at 0–20 cm soil depth. However, electrical conductivity (EC), bulk density and available Cu were higher in the adjacent reference soil. The aged charcoal had no significant effect on the microbial respiration rate of studied soils. The results of this study provide new insights and strong support for the long-term benefits of biochar application as a management strategy for improving soil productivity as well as sequestering large quantities of durable carbon in soils of the region and mitigating global warming.     

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.     

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.

     

Rabies Infection: An Overview of Lyssavirus-Host Protein Interactions

Viruses are obligatory intracellular parasites that use cell proteins to take the control of the cell functions in order to accomplish their life cycle. Studying the viral-host interactions would increase our knowledge of the viral biology and mechanisms of pathogenesis. Studies on pathogenesis mechanisms of lyssaviruses, which are the causative agents of rabies, have revealed some important host protein partners for viral proteins, especially for most studied species, i.e. RABV. In this review article, the key physical lyssavirus-host protein interactions, their contributions to rabies infection, and their exploitation are discussed to improve the knowledge about rabies pathogenesis. Key Words: Host-Pathogen Interactions, Lyssavirus, Rabies     

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     

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...