Production and introduction of a novel immunotoxin based on engineered RNase A for inducing death to Her1-positive cell lines

The present study was performed to design an immunotoxin consisting of engineered RNase A and scFv of Cetuximab. To accomplish this study goal, at first to evade RNase A from its inhibitors in the cytoplasm, six amino acids of RNase A were substituted, then the physicochemical features of engineered RNase A were assessed. To investigate the interaction between the engineered RNase A and the ribonuclease inhibitor, protein–protein docking was performed. After engineering the RNase A, it was theoretically conjugated with scFv of Cetuximab using a cleavable linker to produce scFv-engineered RNase A. Then, wild-RNase A (14 kD), engineered RNase A (14 kD) and scFv-engineered RNase A (42 kDa) were expressed in the BL21 (DE3) strain of Escherichia coli and purified by Ni-NTA columns. To confirm the expressed proteins, western blot analysis was performed. The functioning of wild-RNase A and engineered RNase A were investigated by RNA fragmentation assay. Finally, to evaluate the cytotoxicity of scFv-engineered RNase A, a dose–response cytotoxicity assay was performed on Her1-positive and Her1-negative cell lines. The results showed that engineered RNase A could maintain its structure and disulfide bonds and evade its inhibitor. Expression and purification were successfully conducted and both enzymes could degrade yeast RNA. The result of cytotoxicity showed that the engineered immunotoxin could induce cell death to Her1-positive cell lines with an IC₅₀ of 50 nM. It appears that scFv-engineered RNase A can be a promising molecule for use.     

Synergy between parasitoids and pathogens for biological control of Helicoverpa armigera in chickpea

The chickpea pod borer, Helicoverpa armigera Hübner (Lepidoptera: Noctuidae), is an important pest of many legume crops, including chickpea, Cicer arietinum L. (Fabaceae). We field-tested the efficacy of two pathogens, Helicoverpa armigera nucleopolyhedrosis virus (HaNPV) and Bacillus thuringiensis Berliner subsp. kurstaki (Btk), both alone and in combination with the parasitoid Habrobracon hebetor Say (Hh) (Hymenoptera: Braconidae), for control of H. armigera larvae in chickpea fields. A randomized complete block design was used at two sites, so that wasp release treatments at one site would not contaminate pathogen-only treatments at the other. Pathogens were applied when most pest larvae were in the second instar, and wasp releases were made 2 days later. Number of H. armigera larvae per plant, number of damaged pods, yield (kg ha⁻¹), treatment cost (US$ ha⁻¹), and benefit:cost ratio were all determined. Treatments separated significantly at 14 days after treatment, with the HaNPV+Hh and Btk+Hh treatments providing the best control, suggesting a synergistic impact of the wasp and the pathogens when applied together. These treatments also produced the fewest damaged pods, the highest yields, and the greatest yield increments in comparison to untreated controls. However, costs were also highest for these combined treatments, which reduced their net benefit in comparison to use of Hh or Btk alone, although more efficient mass production of the pathogens might make combined applications feasible. We conclude that both HaNPV and Btk are suitable for integration with releases of H. hebetor against H. armigera in chickpea fields, provided parasitoid releases are delayed until 2 days after pathogen application.     

The Effect of Cerium Oxide During Pregnancy on the Development of the Testicular Tissue of Newborn NMRI Mice

Biological Trace Element Research - Cerium(IV) oxide is widely used as a catalyst in all aspects of human life and human beings are exposed to these materials. The purpose of this experimental...     

Opportunistic Fungal Infections in the Epidemic Area of COVID-19: A Clinical and Diagnostic Perspective from Iran

Mycopathologia - The coronavirus disease 2019 (COVID-19) pandemic emerged in Wuhan, China, in late 2109, and has rapidly spread around the world. Until May 25, 2020, there were 133,521 confirmed...     

Geographic Wormhole Detection in Wireless Sensor Networks

Wireless sensor networks (WSNs) are ubiquitous and pervasive, and therefore; highly susceptible to a number of security attacks. Denial of Service (DoS) attack is considered the most dominant and a major threat to WSNs. Moreover, the wormhole attack represents one of the potential forms of the Denial of Service (DoS) attack. Besides, crafting the wormhole attack is comparatively simple; though, its detection is nontrivial. On the contrary, the extant wormhole defense methods need both specialized hardware and strong assumptions to defend against static and dynamic wormhole attack. The ensuing paper introduces a novel scheme to detect wormhole attacks in a geographic routing protocol (DWGRP). The main contribution of this paper is to detect malicious nodes and select the best and the most reliable neighbors based on pairwise key pre-distribution technique and the beacon packet. Moreover, this novel technique is not subject to any specific assumption, requirement, or specialized hardware, such as a precise synchronized clock. The proposed detection method is validated by comparisons with several related techniques in the literature, such as Received Signal Strength (RSS), Authentication of Nodes Scheme (ANS), Wormhole Detection uses Hound Packet (WHOP), and Wormhole Detection with Neighborhood Information (WDI) using the NS-2 simulator. The analysis of the simulations shows promising results with low False Detection Rate (FDR) in the geographic routing protocols.     

Migrasomes and exosomes; different types of messaging vesicles in podocytes

Podocytes, highly specified kidney epithelial cells, live under several pathological stimuli and stresses during which they adapt themselves to keep homeostasis. Nevertheless, under extreme stress, a complex scenario of podocyte damage and its consequences occur. Podocyte damage causes foot process effacement and their detachment from the glomerular basement membrane, leading to proteinuria. Podocyte-derived extracellular vesicles (pEVs), mainly microparticles and exosomes are considered as signaling mediators of intercellular communication. Recently, it has been shown that throughout the injury-related migration procedure, podocytes are capable of releasing the injury-related migrasomes. Evidence indicates that at the early stages of glomerular disorders, increased levels of pEVs are observed in urine. At the early stage of nephropathy, pEVs especially migrasomes seem to be more sensitive and reliable indicators of podocyte stress and/or damage than proteinuria. This review highlights the current knowledge of pEVs and their values for the diagnosis of different kidney diseases.     

Designing of a Functional Chimeric Protein for Production of Nanobodies Against Human CD20: Molecular Dynamics Simulation and In Vitro Verification

International Journal of Peptide Research and Therapeutics - One major issue in cancer immunotherapy is the large size of mAbs (150 kDa) which usually causes limitation in tumor...     

Correlation of micro vessel density and c-Myc expression in breast tumor of mice following mesenchymal stem cell therapy

Stem cell therapy for degenerative diseases has been established; however there are controversies over the treatment of solid tumors with stem cell transplantation. In the present study, the anti-tumor action of mesenchymal stem cells (MSCs) has been examined in a mouse model of breast cancer with emphasize on tumor growth, angiogenesis and c-Myc expression in breast tumors. For this purpose, MSCs were isolated from bone marrow of Balb/c mice and characterized. A Balb/c mouse model of breast cancer was developed and subjected to cell therapy intra venous (I.V) or intra tumor (I.T) with MSCs. Tumor growth was measured using a digital caliber for until the end of experiment (30 days). Then the mice were sacrificed and their tumors were removed and processed for histopathological examination, immunohistochemical assay of CD31 and measuring of c-Myc expression using quantitative PCR. Detection of the labeled-MSCs in tumors following injection of the cells (I.V or I.T) clearly showed the homing of MSCs into tumors. Tumor growth in case of MSC-treated mice by I.V and I.T routes was inhibited by approximately 28% and 34% respectively compared to controls. The suppression of angiogenesis was reflected in Micro Vessel Density (MVD) following I.V or I.T delivery of the MSCs. c-Myc gene expression in tumor tissues of mice treated I.V or IT with MSCs was down-regulated to 28.0% and 16.0% respectively compare to control groups. In conclusion, growth inhibition of breast tumors in mice due to MSC therapy is associated with modulation of c-Myc activation and angiogenesis markers.     

A Clean Kill: Tracking the Socio-technological Aspects of Slaughtering Animals in Iran

The slaughterhouse, whether it is seen as an institution, as an industry, or as a unique technological system, is arguably right at the heart of modernism. Human–animal relationships and more generally human–nature relationships bring about issues of rationalization, alienation, and commodity fetishism, while raising philosophical concerns of the human worldview. Current practices of animal slaughter in Iran challenge the core values of Islamic teachings. Slaughterhouses are hidden from the public, segregated into various sections, and follow the exact same models of industrial production as in other parts of the world. Since they must abide by Islamic ways of slaughter and therefore become Halal, these slaughterhouses are slightly modified accordingly. When meat, or what is recently referred to as protein, shows up in the market, it conceals any sign of its true origin—it is neatly packaged with layers of plastic and modern colorful labels showing animals happily grazing on open, green pasture. The findings of this research point to the subtle imbalance and alienation created between humans and animals in Iran.     

Co-transformation of canola by chimeric chitinase and <Emphasis Type="Italic">tlp</Emphasis> genes towards improving resistance to <Emphasis Type="Italic">Sclerotinia sclerotiorum</Emphasis>

Canola (Brassica napus) plants were co-transformed with two pathogenesis-related protein genes expressing a Trichoderma atroviride chitinase with a chitin-binding domain (chimeric chitinase) and a thaumatin-like protein (tlp) from Oryza sativa conferring resistance to phytopatogenic fungi by Agrobacterium-mediated transformation. The putative transgenic plants were confirmed by PCR. After measuring the specific activity of the chimeric chitinase and glucanase activity for tlp genes, transgenic plants with high specific activity were selected for southern blot analysis to confirm the copy number of the genes. In vitro assays, the antifungal activity of crude extracted protein against Sclerotinia sclerotiorum showed that the inhibition percentage in double transgenic plants was between 55 and 62, whereas the inhibition percentage in single-gene transformants (chimeric chitinase) ranged from 35 to 45 percent. Importantly, in greenhouse conditions, the double transgenic plants showed significant resistance than the single-gene transformant and wild type plants. The results in T₂ generation using the intact leaf inoculation method showed that the average lesion diameters were 10, 14.7 and 29 mm for the double transformant, single-gene transformant and non-transgenic plants, respectively. Combined expression of chimeric chitinase and tlp in transgenic plants showed significantly enhanced resistance against S. sclerotiorum than the one that express single-gene transformant plants. These results suggest that the co-expression of chimeric chitinase and tlp can confer enhanced disease resistance in canola plant.