Modares Journal of Biotechnology

Abstract In the production of fuel briquettes, different additives are used in order to improve the technical parameters. In the present study, two types of lignocellulosic binders including nanocellulose and lignin have been used. Due to the different chemical structure and difference in calorific value of each of these two materials (lignin and nanocellulose) and the difference in their mechanism of action on improving the thermal properties of fuel briquettes, in order to evaluate the final product of thermal analysis using calorific value and TGA and DTA diagrams were used. The results show the positive effects of the use of cellulosic binders in improving the thermal behavior of biofuel briquettes. Thermal analysis showed that 9% nanocellulose and 9% lignin treatments with 19.85 MJ / Kg and 25.75 MJ / Kg had the highest calorific value compared to the control sample, respectively. The diagram obtained from thermal analysis (TGA) of the control sample and the samples treated with lignin and nanocellulose show that the treated samples have lower weight loss rate, higher burning rate and higher burning temperature.

Abstract Nowadays, the peptides and proteins possessing anti-cancer, anti-allergic and anti-inflammatory properties are used for disease treatment. Brazzein is a sweet protein containing 54 amino acids and according to reports, it has anti-cancer properties based on sequence and structurehas sequence. In this study, the role of position 40 aspartate in the structure and function of wild brazzein protein and mutants as well as the anti-cancer properties of the peptides obtained on the TLR5 receptor were investigated. For this, several models of mutated forms were designed and constructed using Modeller.v.9.20 software. Then, the accuracy of the models and the physico-chemical properties of wild type (WT) and mutants of D40N, D40R and D40Deletion were evaluated using various bioinformatics servers and softwares including ProtParam, ProtScale, SAVES, PIC, ModEval, and PredyFlexy. For predicting anticancer properties, the sequence of WT protein and mutants was examined and compared using ACPred and iACP servers. The quality and analysis of WT protein and mutants binding as a ligand with TLR5 receptor, triggering an anti-cancer signaling pathway, were investigated through molecular docking using HADDOCK software.The results of bioinformatics parameters analysis indicated the possibility of improving the stability of brazzein structure and function, and the probability of increasing the available surface to bind to the receptor. Moreover, based on the results of molecular docking analyses, the ability binding TLR5 receptor was higher in D40R than the other proteins indicating an increased probability in anti-cancer properties of the mutant.

Abstract ntroduction: Colorectal cancer (CRC) is one of the most important cancers and the second leading cause of cancer mortality in Iran. CRC is spread through genetic and epigenetic changes. Understanding the molecular pathways in these genetic and epigenetic changes can provide a clear perspective on cancer treatment.
Materials and Methods: This study was performed on 40 samples of intestinal tumor growths (polyps), 30 tumor samples and 40 normal tissues adjacent to the tumor. RNA and tissue DNA extraction, study of gene expression, miR-22, miR-194 LncRNA MINCR using Real time PCR was performed.
Results: In this study, the expression of MINCR LncRNA in tumor and polyp samples increased compared to the adjacent normal tissue and the expression of miR-194 miR-22 was decreased. Negative correlation coefficient was observed between miR-22, miR-194 and MINCR expression levels and also a significant relationship was observed between the expression of these lncRNAs and microRNAs.

Abstract Leguminosae is one of the most important plant families. In this study, we searched Arachis hypogaea, Glycine max, G. soja, Lotus japonicas, Medicago truncatula, Phaseolus vulgaris, and Vigna unguiculata conserved miRNAs through Expressed Sequence Tag (EST) based-homology method. All candidate sequences with appropriate fold-back structures were screened and characterized according to several filtering criteria. Chromosomal MIR locus and their distributions determined. The Dollo maximum parsimony was employed to construct the species relationship based on the MIR birth and death. Also, the number of MIRs gains and losses in the evolutionary process. In addition, we estimated the numbers of MIRs in their ancestral species using Dollo maximum parsimony in their phylogenetic tree. We found 414 novel miRNAs from 130 MIR families meeting the restricted filtering criteria. Either evolutionary time or the number of miRNAs gains and losses are estimated and characterized in the ancestral species based on the taxon-based phylogenetic tree. It speculates that gains of miRNA gene families within Leguminosae have accelerated during its evolutionary time. In addition, several taxon-specific MIR families find to assign diverse taxon and their species. Our thorough analyses resulted in the definition of some miRNA families as being lineage-specific. Therefore, they can use as markers in future systematic studies.

Abstract Enriching human food using new technology such as lipid nanocarriers is a simple and accessible tool. Accordingly, the present study aimed to evaluate the sensory and production of healthy and useful food products to evaluate the enrichment of milk with zeaxanthin lipid nanocapsules and to evaluate its cryoprotectants. During experimental-laboratory research, zeaxanthin extraction from Spirulina platensis, and nanocarriers produced for milk enrichment were used as a food model system. Three samples of milk, milk enriched with lipid nanocarriers containing zeaxanthin, and milk enriched with lipid nanocarriers were examined (at similar concentrations of nanocarriers). In order to check the efficiency of produced nanocarriers, cold protective compounds (glucose, sorbitol, glycerin, lactose, and sucrose) were added to milk. Sucrose was recognized as the best cryoprotectants. Sensory evaluation of enriched milk was performed on a five-degree hedonic scale and different sensory parameters were examined. Data were analyzed using Minitab (v. 2016). Results No significant difference was observed between the sensory characteristics of control milk and milk enriched with nanocarriers (P<0.05). The lowest particle size and dispersion index were obtained in the coating of nanocarriers with cold protective compounds, respectively, 320.82 and 0.26 to 0.31. Zeta potential was reported as -6.03. By enriching milk with zeaxanthin-containing nanocarriers, in addition to visual and skin health, problems related to the lack of useful natural additives and insolubility of food products can be eliminated.

Abstract In this study, the wheat germ was fermented with industrial bakery yeast powder to produce FWGE with high 2,6-DMBQ content in a Bench-scale bioreactor by scale-up approach. The 2,6-DMBQ content of FWGE was increased by optimizing the three initial variables of pH, fermentation temperature, and agitation rate at two levels using the Taguchi method. The 2,6-DMBQ content of the samples was determined at 14, 16, and 18 hours of the fermentation process. Then, the results were analyzed by Qualitek software. The effect of centrifugation speed on turbidity and the yeast's number in the final supernatant was then investigated. Finally, the supernatant was dried by spray dryer with an inlet temperature of 120 °C and outlet temperature of 70°C, and the amount of active 2,6-DMBQ, pH, moisture, and ash was determined. Under optimal conditions: initial pH of 6, fermentation temperature of 32 °C, and agitation rate of 80 rpm, maximum 1.527 mg of 2,6-DMBQ per gram of FWGE obtained. The separation results showed that the centrifugation rate doesn't have a significant effect on the final turbidity and the number of yeasts left, and thus 3000 g was selected as the optimal speed. However, because of the high content of yeast in the supernatant, filtration was required after centrifugation. Due to the high speed of sample drying, the low moisture of the final product, and high efficiency on an industrial scale, the samples were dried using a spray dryer. Finally, the moisture, protein, ash, and pH of the final product were measured.

Abstract Given the global epidemic of COVID-19, it is important to design a vaccine for prevention. The virus belongs to the beta-coronavirus family and forms appendages on the surface of the glycoprotein spike virus membrane. Studies on SARS-CoV-1 and related MERS-CoV vaccines have shown that the spike protein on the virus surface is a suitable target for the vaccine. In this experimental study, we compare the recombinant fragment of spike protein (rfsp) expressed in the eukaryotic host CHO-K1 cell and prokaryotic E. coli in terms of immunogenicity, neutralizing activity, and epitopes recognition Similar to the virus strain and the ability to bind to the serum of improved patients, in two types of alpha and delta variants. The results showed that both rfSP proteins are a potential new antigen candidate for the development of the Covid 19 vaccine, but the CHO cell maintains the biological activity of the protein by performing post-translational modification processes such as glycolysis. This increases the present likelihood of developing virus-like epitopes and increases the titer of rfsp-specific antibodies in the serum of immunized mice. Therefore, priority is given to rfsp expressed in CHO cells to evaluate vaccine efficacy.

Abstract Flowering transition is one of the most important developmental processes of higher plants, which is controlled by endogenous and external environmental signals. These signaling cues are perceived in leaves and shoot apical meristem (SAM) to induce flower formation. APETALA1 (AP1) is one of floral meristem identity genes that regulate the specification and formation of floral meristems and is required for sepals and petals formation. In this study, the expression of this gene in different organs of Eruca sativa as well as the effect of brassinosteroids (BRs) on flowering and the gene expression was investigated. RNA was extracted from different organs and first-strand cDNA was synthesized. Specific primers were designed based on the sequence alignment of AP1 isoform genes from other plants. In the vegetative stage, no expression was observed in different organs. Brassinosteroid treatment from 28 days (vegetative stage) to flower buds formation caused early flowering, so that the treated plants flower about 10 days earlier than the control. In addition, plant size and its organs were larger in plants treated with brassinosteroids. Evaluation of EvsAP1 gene expression in reproductive phase showed its expression in flower buds, sepals and petals but no was seen in roots, stems, leaves, stamen and gynocium. Also, the onset of expression of this gene was observed earlier, indicating that the transition to flowering and flower bud formation occurs faster in treated plants; therefore, expression occurs earlier. However, expression levels did not affected by brassinosteroids and no significant difference was observed between treated and control samples.

Abstract Production of drought tolerant crop is an important strategy for avoiding water scarce crisis. Improvement of the root structure leading to the higher yield and seed quality. In this study, three genes affecting root structure, drought tolerance and phosphorous absorbance are used in producing hybrid constructs used for the rice transformation. Three genes: a serine/threonine protein kinase (PSTOL1), a gene from the cytokinin oxidase/dehydrogenase family (OsCKX4) and a transcription factor induced under stress from the NAM-ATAF-CUC family (OsNAC5) isolated from the rice wild cultivars are cloned under separate regulatory elements in the T-DNA region of the Agrobacterium binary vector. OsNAC5 gene was cloned under RCc3 root specific promoter and PSTOL1 gene under ubiquitin promoter. Also, OsCKX4 gene was cloned once under ubiquitin promoter and once under RCc3 promoter. Two hybrid multi-gene constructs named pUhrN5CkPstol and pUhrCkPstol harboring multiple genes are synthetized and used for the gene transformation into the Hashemi cultivar. Gene transfer was done to callus obtained from mature rice seeds. Transgenic plants were confirmed using PCR analysis. From the number of 107 regenerated plants in which the presence of transgenes was proved, 14 transgenic events were finally obtained. Root structure of the T0 plants showed drastic phenotypic difference in comparison to the non-transgenic ones. By now, one transgenic event harboring CKX4 and PSTOL1 is confirmed to have a homozygous line in T2 generation. It is hoped that genetic engineering of rice for enhanced root structure lead to drought tolerance, reduce water consumption and improve yield under stress conditions.

Abstract Phenols are organic and highly toxic compounds commonly found in the effluents of various industries due to their wide range of applications. The inhibitory effect of phenol at high concentrations, as well as the high salinity of industrial effluents, poses a serious challenge for treatment by microorganisms. One of the most common approaches to overcome this problem is the immobilization of phenol-degrading microorganisms. The aim of this study was to study the immobilization effect on the phenol removal efficiency of native Janibacter halotolerant bacterium. For this purpose, mica was used as a carrier for bacterial immobilization and the protein concentration assay was applied to determine the immobilization efficiency. The phenol removal by free and immobilized cells was studied as well as the effect of different parameters on phenol removal efficiency. The immobilization efficacy on mica was %68.75, based on protein concentration measurements. The removal time of 100 mg/L phenol by suspended cells was 88 h, while the immobilized cells degraded it in 40 h. Immobilized cells, unlike free cells, were able to remove phenol at lowered temperatures up to 16℃ , salt concentrations greater than 7/5%, and pH levels below 7/5 and above 8/5. Similar results regarding the superior performance of immobilized cells have been obtained in other studies. As a result, the immobilization process considerably improves the efficiency of phenol removal and makes the cells resistant to harsh environmental conditions by protecting the cells from the toxic effects of phenol.

Abstract Resistance to chemotherapy drugs always has been an obstacle in the definitive treatment of cancers. Therefore, the discovery of molecular events leading to drug resistance improves therapeutic methods. Non-coding RNAs (ncRNAs) are a group of molecules that regulate intracellular events, including carcinogenesis and drug resistance pathways. For example, the competitive network of endogenous ncRNAs (ceRNA) regulates the mRNA expression of target genes by binding to miRNAs and limiting their regulatory effect. So far, limited studies have been reported on the role of ceRNA in drug resistance in ovarian cancer. In this study, large-scale RNAseq sequencing data obtained from cisplatin-resistant and sensitive cells were used to search for ceRNAs that are possible regulators of drug resistance in ovarian cancer. For this purpose, the A2780 sensitive and resistant cisplatin ovarian cancer cell line was selected, and the SRA data prepared by RNAseq method was screened. During this process, lncRNAs, microRNAs and mRNAs with expression changes were separated and classified. In the bioinformatic analysis of resistant and sensitive cells, 16 mRNAs, 10 lncRNAs, and 149 miRNAs were overexpressed, and 622 mRNAs, 263 lncRNAs, and 177 miRNAs were underexpressed. These genes were involved in 57 cellular pathways, and by mapping the regulatory ceRNA network, ZNRF3-AS1-miR-33-DUSP1 and ZNRF3-AS1-miR33-HSPA2 axes were identified as potential ceRNA networks involved in cisplatin-resistant ovarian cancer.

Abstract Aptamers or chemical antibodies are mostly nucleic acid sequences that can bind to various diverse targets including small molecules, large molecules, and cells. Among the advantages of aptamers compared to antibodies, we can mention the in vitro production process, the possibility of choosing aptamers against toxic and non-immunogenic molecules, long-term storage, and much lower production costs. Aptamers can also be easily modified or stabilized, and their synthesis is associated with high purity and reproducibility, they are chemically stable, and due to their nucleic acid structure, they are much more flexible than antibodies, and they can be used in the form of molecular beacons probes, a combination of use aptamer-target interaction and nucleic acid amplification to achieve highly sensitive detection ranges. These interesting features have made aptamers ideal diagnostic elements for analytical tools such as biosensors, colorimetric methods, surface plasmon resonance, and lab-on-a-chip. However, all these methods require skilled workers and laboratory-based instruments, thus limiting their application. In this regard, lateral flow assay or paper strip kits provide fast and reliable results and only require user intervention in the sample addition phase. Due to their simplicity is widely used in various fields including medicine, food product quality inspection, and environmental safety. In this work, while introducing aptamers, , an overview of its unique application in lateral flow assay and the future of this technology should be studies.