Molecular cloning and characterization of genes related to the ethylene signal transduction pathway in pomegranate ( Punica granatum L.) under different temperature treatments Abstract Low temperature storage is a common method for storing pomegranates post-harvest; however, unsuitable low temperatures can cause fruit chilling injuries, the molecular mechanism of which is as yet unclear. Ethylene is a major factor affecting the post-harvest storage quality of pomegranates, and functions mainly through the ethylene signal transduction pathway. ERF1, ERF2 and ETR are key genes in the ethylene signal transduction pathway. Here, we used RACE and homologous cloning techniques to obtain PgERF1 (KU058889), PgERF2 (KU058890) and PgETR (KU058891) from Punica granatum cv. Yushizi. Sequence alignment and functional domain analysis revealed that both PgERF1 and PgERF2 contained a DNA-binding-site at the 120th to 177th amino acids of the N-terminus, which is a typical AP2/ERF center structure domain. Analysis of changes in expression of PgERF1, PgERF2 and PgETR following storage for different lengths of time (0, 14, 28, 42 and 56 days) at different temperatures (0°C, 5°C, 10°C and 15°C) revealed that the expression levels of PgERF1 and PgERF2 had a significant positive correlation. At the same time, the expression of both PgERF1 and PgERF2 increased continuously with time when seeds were stored at 0°C. However, there was no obvious linear relationship between time stored and the levels of expression of PgETR. Therefore, we inferred that at 0°C, the ethylene signal transduction pathway might play an important role in fruit chilling injuries during post-harvest storage.

Investigation of the response to salinity of transgenic potato plants overexpressing the transcription factor StERF94 Abstract Salinity is one of the most important constraints threatening the cultivation of potato plants (Solanum tuberosum L.). It affects plant growth and leads to significant yield loss. Consequently, it is important to improve the tolerance of potato plants to salinity. In this context, we investigated the involvement of a potato ethylene responsive factor (StERF94) in plant response to salinity, since our previous genome-wide analysis showed that it may be related to biotic and abiotic stress response. ERF proteins belong to a large family of transcription factors that participate in plant response to abiotic stresses. We have previously identified the StERF94 gene which shows increased expression in potato plants submitted to salt treatment. In this study, transgenic potato plants overexpressing StERF94 were produced and submitted to salt treatment (100 mM NaCl) in vitro and under greenhouse culture conditions. StERF94 transgenic lines showed lower decrease of stem elongation under salt treatment in comparison to non-transgenic wild-type plants. Moreover, these plants showed a low level of H2O2 and Malondialdehyde content, and an increase in catalase and GPX (Gluthation peroxidase) activities compared to non-transgenic plants. In a second step, enhanced expression of some target genes for example CuZn-SOD, DHN25 (Dehydrin) and ERD (Early Responsive to Dehydration) was noted in the StERF94 transgenic plants, submitted to salt treatment. The StERF94 factor was also involved in the activation of osmoprotectant synthesis. Taken together, all these data suggest that overexpression of the StERF94 transcription factor increases the tolerance of potato plants to salinity by improving plant growth, osmoprotectant synthesis and antioxidant activityleading to low oxidative stress damage.

Translin: A multifunctional protein involved in nucleic acid metabolism Abstract Translin, a highly conserved, DNA/RNA binding protein, is abundantly expressed in brain, testis and in certain malignancies. It was discovered initially in the quest to find proteins that bind to alternating polypurines-polypyrimidines repeats. It has been implicated to have a role in RNA metabolism (tRNA processing, RNAi, RNA transport, etc.), transcription, DNA damage response, etc. Studies from human, mice, drosophila and yeast have revealed that it forms an octameric ring, which is important for its function. Translin is a cytoplasmic protein, but under genotoxic stress, it migrates into the nucleus, binds to the break point hot spots and therefore, thought to be involved in chromosomal translocation events as well as DNA damage related response. Its structure is known and DNA binding regions, GTP binding region and regions responsible for homotypic and heterotypic interaction are known. It forms a ball like structure with open central channel for accommodating the substrate nucleic acids. Besides this, translin protein binds to 3′ and 5′ UTR of certain mRNAs and probably regulates their availability for translation. It is also involved in mRNA transport and cell cycle progression. It forms a heteromeric complex with translin associated factor-X (TRAX) to form C3PO complex which is involved in RNA silencing process. Recently, it has been shown that translin is upregulated under starvation conditions in Drosophila and is involved in the integration of sleep and metabolic rate of the flies. Earlier studies classified translin as a DNA repair protein; however subsequent studies showed that it is a multifunctional protein. With this background, in this review we have summarized the translin biochemical activities, cellular function as well as structural properties of this important protein.

Biophysical methods for quality evaluation of decellularized and recellularized tissue-engineered constructs of organs and tissues Abstract Tissue engineering is rapidly growing now and can become a promising alternative to transplantation of organs and tissues, as it is devoid of major shortcomings of transplantology, such as acute shortage, complexity of selection, delivery and storage of donor material, lifelong immunosuppressive therapy. One of the most widely known methods of obtaining biological scaffolds for the subsequent creation of tissue-engineered constructs of organs and tissues is decellularization. The evaluation of the quality of the obtained scaffolds, based on the study of the viability of cell structures in decellularized and recellularized matrices, is one of the priorities of modern regenerative medicine worldwide. In this investigation, the biophysical criteria of decellularization and recellularization of tissue-engineered constructs based on the evaluation of the generation of free radicals in native, decellularized and recellularized tissues by EPR spectroscopy and chemoluminescence in a complex assessment of the quality of biological matrixes obtained are considered using intrathoracic organs and tissues of rats. It has been established that the intensity indices of free radical generation in native and recellularized tissues of animal organs, as well as in decellularized matrices, can serve as one of the express criteria for quantitative assessment of cell structures viability.

Iris yellow spot virus –induced chloroplast malformation results in male sterility Abstract Iris yellow spot virus (IYSV) is one of the most devastating viral pathogens, which causes high economic losses in the onion yield. Physiological and genetic changes are associated with the appearance of chlorotic symptom in the infected plants. IYSV-N gene sequence analysis revealed that it shared sequence identity of 99% with other Egyptian isolates, at both genomic and proteomic levels. In addition, N protein sequence with computational examination indicated many motifs involved and played different roles in the virus activity and its regulation and stability were detected. In the Differential Display-Polymerase Chain Reaction (DD-PCR) study, a highly up-regulated gene at 15 days post-biological IYSV inoculation (dpi) was selected for sequencing. Based on the sequencing results that showed the identified gene was coding for a chloroplast-related gene, degenerate specific primers were designed for Real-Time PCR analysis. A significant change in the transcription level of the chloroplast-related gene after 15 dpi suggested that some IYSV proteins interact and/or regulate with chloroplast proteins and this finding supports the DD-PCR results. At 20 dpi, the ultrathin sections showed that IYSV infection caused many dramatic chloroplasts malformations. The malformation appeared as chloroplast broken envelope with the presence of numerous spherical particles inside it and chloroplasts with long stromule. Our findings indicated that IYSV interrupts normal chloroplast functions, as a part of the onion defence response, however many crucial factors remain to be elucidated and further studies are needed at both biological and molecular levels.

Presence of carbohydrate binding modules in extracellular region of class C G-protein coupled receptors (C GPCR): An in silico investigation on sweet taste receptor Abstract Sweet taste receptor (STR) is a C GPCR family member and a suggested drug target for metabolic disorders such as diabetes. Detailed characteristics of the molecule as well as its ligand interactions mode are yet considerably unclear due to experimental study limitations of transmembrane proteins. An in silico study was designed to find the putative carbohydrate binding sites on STR. To this end, α-D-glucose and its α-1,4-oligomers (degree of polymerization up to 14) were chosen as probes and docked into an ensemble of different conformations of the extracellular region of STR monomers (T1R2 and T1R3), using AutoDock Vina. Ensembles had been sampled from an MD simulation experiment. Best poses were further energy-minimized in the presence of water molecules with Amber14 forcefield. For each monomer, four distinct binding regions consisting of one or two binding pockets could be distinguished. These regions were further investigated with regard to hydrophobicity and hydrophilicity of the residues, as well as residue compositions and non-covalent interactions with ligands. Popular binding regions showed similar characteristics to carbohydrate binding modules (CBM). Observation of several conserved or semi-conserved residues in these binding regions suggests a possibility to extrapolate the results to other C GPCR family members. In conclusion, presence of CBM in STR and, by extrapolation, in other C GPCR family members is suggested, similar to previously proposed sites in gut fungal C GPCRs, through transcriptome analyses. STR modes of interaction with carbohydrates are also discussed and characteristics of non-covalent interactions in C GPCR family are highlighted.

Correction to: Protein-rich extract of Musca domestica larvae alleviated metabolic disorder in STZ-induced type 2 diabetic rat model via hepatoprotective and pancreatic β-cell protective activities In the December 2018 issue of the Journal of Biosciences, in the article titled ‘‘Protein-rich extract of Musca domestica larvae alleviated metabolic disorder in STZ-induced type 2 diabetic rat model via hepatoprotective and pancreatic β -cell protective activities’’ by Hanfang Mei et al. (DOI: 10.1007/s12038-018-9804-z; Vol. 43, No. 5, pp. 969–983), the affiliations of Hanfang Mei have been incompletely mentioned as:

A cross-eyed geneticist’s view V. How Sydney Brenner, Leslie Barnett, Eugene Katz, and Francis Crick inferred that UGA is a nonsense codon

Involvement of putrescine in osmotic stress-induced ABA signaling in leaves of wheat seedlings Abstract To elucidate one mechanism by which putrescine (Put) functions in plant signaling under osmotic stress, Put and ABA contents, and plasma membrane-NADPH oxidase (PM-NOX) activity were detected in wheat seedling leaves. Under osmotic stress, ABA and Put contents, PM-NOX activity, and PM-NOX-dependent O 2 .− production all increased. The inhibitor tungstate (T) of ABA bio-synthesis reduced the increases in ABA and Put contents under osmotic stress. The inhibitor D-arginine (D-Arg) of Put bio-synthesis didn’t reduce osmotic-induced increase of ABA, but it inhibited the increases of PM-NOX activity and O 2 . − production, and the inhibitory effects were reversed by exogenous Put. These findings suggested that ABA might regulate Put biosynthesis, and Put might regulate PM-NOX activity. Treatments with three inhibitors imidazole (I), diphenylene iodonium (DPI) and pyridine (P) of PM-NOX reduced significantly not only O 2 . − production, but also the stress-induced increase of Put content, which indicated that O 2 . − production might regulate Put biosynthesis. Treatments with EGTA (Ca2+ chelator), La3+ and verapamil (V) (Ca2+ channel blockers) reduced significantly the stress-induced increase of Put content, which suggested that Ca2+ might regulate Put biosynthesis. With these findings, it could be concluded that Put was involved in ABA signaling induced by osmotic stress via regulating PM-NOX activity in wheat seedling leaves.

Laser-ablation-synthesized nanoparticles and animal cell lines studies Abstract Nanoparticles (NPs) synthesized by laser ablation in distilled water were used to study their biological effect on normal and cancer cells. Parameters such as cell morphology, cell proliferation and viability were examined for treated cell lines, and the effect was represented in terms of cells cytotoxicity using standard procedures. The study reveals the higher cytotoxic effect of nanoparticles on cancerous cells of breast, melanoma and colon origin compared to normal fibroblast cells NIH-3T3. Furthermore, DNA fragmentation assay results demonstrated the apoptosis mediated cell death in nanoparticle-treated cancer cells. The distinct role of nanoparticles in normal and cancer cells of different origin showed that nanoparticles were specific to cause cytotoxicity in particular cancer cells type. NPs exhibit cytotoxic effects in cancer cells by inducing apoptosis. These studies provide fundamental evidence for the easy, simple and safe mode of nanoparticles synthesis and their application in cancer cells death.