Modares Journal of Biotechnology

Abstract Fruite juices were already under intensive analytical authenticity control for decades. Adulteration is releativly easy to perform because of the chemical composition and the wide natural variation seen in juice. Addition of water and/or sugar, acidification, cheaper fruit, colorants, flavor, and vitamins are typically frauds that are performed with criminal intent and sometimes with high level sophistication to make the adulterated products as difficult as possible to detect. In this sense, effective, reliable and rapid food authentication methods represent a valuable and irreplaceable tool for the authorities to set up control systems to ensure juice quality and safety and to promote mitigation of fraud. D-Isocitric acid and citric acid are two organic acid found in most fruit juices.  They are an important marker in multicomponent procedures for the evaluation of authenticity and quality of fruit products; high citric/isocitric acid ratios can be used as an indicator of citric acid addition in some juices especially in lime juice. The measurement of D-Isocitric acid by HPLC method is very difficult and time consuming in addition in this time, enzymatic method is one of the main international standard method for this component. One of the main aim in this research set up enzymatic method for measurement of these organic acids as one of the most commonly used analytical techniques (enzymatic method) and it represents a very attractive choice because this method is simple, rapid and cost effective.
 

Abstract A critical aspect of laboratory techniques in plant in vitro culture involves the disinfection of explants and the maintenance of sterile environments. Ideally, disinfectants should demonstrate efficacy against a broad spectrum of microorganisms at minimal concentrations. However, the rising application of antibiotics has led to the development of microbial resistance to these agents. Currently, compounds such as essential oils and nanoparticles are being explored in microbiological research. This study aimed to evaluate the effectiveness of three antibiotic groups (cefazolin, penicillin, and chloramphenicol) alongside cumin essential oil and silver nanoparticles in managing in vitro contamination in potato plants (Solanum tuberosum L.). The explants underwent 1 min disinfection with 70% ethanol, followed by a 10 min treatment with a commercial bleach solution. Additional disinfection treatments, including cefazolin, chloramphenicol, penicillin, silver nanoparticles, and cumin essential oil, were applied at concentrations of 10, 20, 40, and 80 mg l-1 for nanoparticles, and 100 and 200 mg l-1 for the other agents in the cultivation medium. The findings indicated that cumin essential oil exhibited the highest level of microbial inhibition, while cefazolin showed the least. Notably, the application of nanosilver at 40 mg L-1 resulted in a 73% reduction in bacterial contamination, whereas penicillin at 100 mg L-1 achieved a 67% inhibition rate. Ultimately, essential oils and nanoparticles present viable alternatives to conventional chemical treatments for the elimination and management of in vitro contamination in plant explants under laboratory conditions.

Abstract Iran stands out as a significant center of genetic diversity for alfalfa (Medicago sativa) worldwide, harboring diverse types of this plant. Ensuring the authenticity of alfalfa populations and varieties is crucial for farmers and seed producers, as the genetic makeup of this species directly influences forage and seed yield quality. In this study, we developed a method to identify and differentiate key Iranian cultivated alfalfa populations using microsatellite markers. We collected random samples, each containing 100 seeds, from various alfalfa accessions. Nine microsatellite loci were screened and employed to differentiate these populations based on specific allelic genotypes. Notably, the MTIC233, BI90, ACT009, TC7, MTIC183, MS30, MTIC238, and AFCA11 markers exhibited the highest differentiation ability. The genetic distance analysis revealed that 5-B and foreign accessions, as well as 29-N and foreign accessions, were the most distant from each other. Conversely, 27-G, 9-H, and 21-R exhibited the closest genetic similarity. The results revealed that, accessions 9-H, 21-R, 27-G, 25-B, 5-B, and 2-G shared a common genetic background, suggesting their close relatedness. Our proposed method allows straightforward identification of target alfalfa accessions within a short timeframe (one to two days) without the need for DNA extraction from leaves.

Abstract The production of secondary metabolites of medicinal plant

Abstract Pathogen growth in vitro is one of the major problems in plant micropropagation, so the most important stage of in vitro culture of plants is disinfection of the cultures. In the common methods of disinfection, the media and plant materials, apparatus such as autoclaves and chemicals disinfectents are used, which causes the time of this process and the costs to increase. This research objective to improve the disinfection of Iris hollandica cv. Apollo scales using different plant essential oils (thyme, cumin and savory) and the methods of using essential oils as disinfectants, the use of essential oils in the medium and the use of essential oils fumigation was done in four independent experiments. The use of essential oils of thyme, cumin, and savory completely prevented both contamination of the culture medium and contamination of the explant. The best disinfection method was when the essential oils were used in combination with the culture medium. Bacterial contamination was better controlled at concentrations of 0.125 to 0.25%, but the concentration of 0.25% of essential oils resulted in better control of fungal infection. The least browning of iris scale explants was observed at a concentration of 0.125. The technique presented in this study can significantly reduce the cost of electricity and lighting, as well as personnel costs. Therefore, this method can introduce a practical and cost-effective technique for plant micropropagation.

Abstract Every year, more than 600,000 tons of rice husk are produced in rice mills in Iran, which are left in nature without proper use or burned. But, with biotechnological methods, it is possible to convert these agricultural wastes into compost with high content of nitrogen and minerals. Accelerating the production of high quality organic fertilizer and reducing the process time requires determining and studying the factors affecting the composting process. In this research, rotten wood waste in the presence of whey was used as an inoculum and the process was studied for one hundred days. The results showed that the process of rice husk composting decreased the ratio of carbon to nitrogen from 161.2 to 21.1 and also increased the nitrogen content from 0.28 to 1.33 percent. By studying temperature changes during the process, it was determined that the degradation of easily biodegradable compounds at the beginning of the process caused an increase in the population of microorganisms and an increase in temperature, and as the process continued, the temperature of the composting environment gradually decreased. On the other hand, the results indicated that the addition of tree bark fertilizer does not affect the quality of the compost content, but it changed the color of the product. The results of the evaluation of the produced compost showed that adding 10 percent by weight of compost to the soil had a significant effect on the length and weight of cowpea.

Abstract Over the past few years, due to the shortage of forest resources as well as the increasing consumption of paper and paperboard, many pulp and paper manufacturers have developed the use of different sources of recycled (secondary) fibers. Therefore, recycling of paper is an effective and eco-friendly way to preserve forest resources, which eventually save the natural diversity and energy. The use of biotechnology in various sectors of the cellulosic products industries such as bio-pulping, bio-bleaching, bio-deinking, bio-wastewater treatment, etc. has been considered and good achievements have been obtained in this field. One of the most important applications of biotechnology in the cellulosic industries is the use of enzymes in the processing of recycled fibers. The use of enzymatic technologies, as environmental friendly process, has led to changes in industrial processes as much as possible and indicated great potential in solving many problems of recycled fibers, especially problems related to waste paper deinking, pulp drainage rate, fiber hornification, refining and stickies materials. In general, deinking with enzymes under acidic or neutral conditions declines the chemicals usage and reduces the yellowing of recycled paper under conventional alkaline deinking conditions. Today, the use of cellulosic enzymes (cellulase and hemicellulase) and oxidative enzymes (such as laccase) as well as amylase and pectinase have shown acceptable results for deinking different types of waste paper and most experiments in semi-industrial as well as industrial units have shown that deinking with them can reduce the cost of chemicals, increase the separation of ink particles

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 In this study, the genetic diversity of 10 different accessions of Andrographis paniculata was investigated using protein and SRAP markers. In the vegetative stage, protein and DNA were extracted from the leaves. The results of protein profile indicated a total of 20 bands with 64.15% polymorphism. To evaluate genetic diversity at the DNA level, 6 SRAP primers were used and a total of 583 scalable bands were observed. A total of 549 bands had polymorphism with an average of 91.5 for the studied primers. The highest polymorphism (99.12%) and the lowest polymorphism (84.21%) were observed in E1/M1 E2/M2 primers, respectively. Cluster analysis produced four main clusters. Genetic diversity indices were calculated for all gene loci, including the average genetic diversity of Nei’s (0.27) and the mean of Shannon’s index with a value of 0.41. High level of population differentiation (Gst = 0.79) and good level of gene flow (Nm = 1.3) were estimated between the grouped populations. Molecular analysis of variance showed that intra-population variance (58%) was higher than inter-population variance (42%). Overall, the results of study showed a high genetic diversity in both protein electrophoresis pattern and in polymorphic bands separated using SRAP markers with emphasis on the greater efficiency of SRAP markers than protein markers, which can be selected in parents with genetic distance. It is widely used to produce dispersing and mapping populations in hybridization programs and to breed or improve desirable traits, as well as to protect and manage the germplasm of this plant.

Abstract Transforming growth factor beta (TGF-β), is a small homodimeric signaling protein. The TGF-β isoforms (TGFβ1, β2 and β3) are involved in many cellular processes including growth inhibition, extracellular matrix remodeling, tissue development, cell migration, invasion and immune regulation. For research aims, TGFβs are overexpressed using recombinant eukaryotic cell or bacterial expression systems. For achieving an efficient purification of TGF-β by immobilized metal ion affinity chromatography (IMAC), a histidine tag was placed either at the C-terminal (C-TGFβ) or N-terminal (N-TGFβ) region of the sequence and the effect of His-tag on TGF-β structure has been studied by computational tools. Proteins 3D structures were modeled using MODELLER software and molecular dynamics simulation of native TGF-β and modelled proteins, N-TGFβ and C-TGFβ were studied in water by GROMACS package. Protein dynamics modeling indicated that the His-tag attached at the C-terminus but not at the N-terminus of the TGF-β can affect the fluctuations of amino acids and protein structure. It is concluded that the C-terminal tagging may cause distortion and misfolding in the structure.

Abstract Different defense pathways in plants evolved in reaction to pathogens. The main aim of this study was to investigate the mechanism of action of glyphosate in resistance induction to bacterial phytopathogens. To do so, glyphosate at an optimal concentration of 1.8 mg / l was used on transgenic potato, to induce resistance to two strains of pathogenic bacteria (21A of Pectobacterium atrosepticum and ENA49 of Dickeya dadantii). It was been shown that plant defense responses to pathogens can be stimulated by treatment plants at an optimal concentration of glyphosate. Transgenic potato leaves infected with potato pathogenic bacteria, and then treated with glyphosate showed a high level of expression of pathogenesis-related genes (PR-2, PR-3, PR-5), especially PR-2 gene and defense response genes (HSR-203j, HIN1), especially HSR-203j gene. The expression of PR-2 gene in leaves infected with these two bacteria were 1.5 and 2.9 times, for PR-3 gene 1.7 and 1.7 times, for PR-5 gene to 1.3 and 1.5 times and expression of HSR-203J gene to 2.5 and 2.4 times and - HIN1 gene to 1.7 and 1.7 times, with Dickeya dadantii and Pectobacterium atrosepticum infection, respectively. The expression of these genes in control samples didn’t significantly change. The results showed that there was a significant difference between the expression of genes in the experimental and control samples (plants treated by glyphosate compared to untreated plants). The results showed that the treatment of plants by glyphosate can induce a systemic acquired resistance to phytopathogens by inducing proteins and defense response genes.

Abstract Rye is one of Iran's most important crops, known as Secale, belonging to the Poaceae. In this research, genetic diversity of 39 families of rye populations from different regions of Iran, the USA, and the Soviet :union: was evaluated with the ISSR marker. The results showed that 8 ISSR initiators produced 48 bands which included 18 polymorphic bands (37.5% polymorphism). The mean of polymorphic information content (PIC) and marker index value (MI) for ISSR primers was 0.15 and 7.2 respectively. The highest PIC (3.0) was related to primer 5+6 and the highest MI (0.96) reached for primer 1+6. After observing polymerase chain reaction products on an agarose gel and scoring DNA bands, the analysis was performed with NTSYS software. The cluster analysis dendrogram of UPGMA and Jaccard's similarity coefficient divided the rye populations into 9 groups, the results were compiled with grouping by principal component analysis. The results of analysis of Molecular Variance indicated an in-species variation more than inter-species variation. The mean Nei genetic variation (h) was 350 and the mean of Shannon index (I) in rye species was 523, which indicates a relatively good variety within species. The results showed that the ISSR marker was a useful tool in determining genetic variation of inter and intra specific of rye.

Abstract Aims The secondary metabolites of plants have been considered due to their diversity and roles in plants and human health. Grapevine is one of the plants that have secondary compounds with medicinal properties. such compounds include resveratrol which is a phenolic compound from the stilbenoid group. In order to investigate the resveratrol production under the effect of an elicitor, a CRD design with four replications using Soltani cultivar was done.
Materials & Methods. The salicylic acid (SA) was used as an elicitor with variable concentrations including 0, 10^-4, 10^-5, 10^-6 and was introduced into MS medium without hormones to examine it`s stress effects. At the molecular level, the effect of SA on the expression of stilbene-synthase gene was evaluated by semi-quantitative RT-PCR. In the biochemical experiment, the rate of resveratrol production was measured by HPLC.
Findings Stilbene-synthase expression analysis showed that SA with a concentration of 10^-4 M had a positive and incremental effect on gene expression and showed a 35.48% of increase compared to the control state, and also the concentration of 10^-5 M increased the gene expression 5.65% in relation with control state. In the biochemical experiment, increasing in production of resveratrol was observed in 10^-4 M treatment compared to the control treatment (6.1 µg) and 10^-5 M treatment (3.25 µg) did not show a significant difference with the control sample.
Conclusion SA as an elicitor and stimulant of resveratrol production could enhance the expression of stilbene-synthase gene followed by enhancing the medicinal properties of the Vitis plant.

Abstract Although biosurfactants have great advantages over chemical surfactants, their wider industrial applications have been constrained by their relatively high production cost. Using renewable, sustainable and cheap substrates such as different industrial by-products and wastes maybe decrease biosurfactant production costs. Since in different countries, there are a variety of by-products and wastes so use of these substrates rely on their types and concentrations in countries. In addition to hydrocarbon compounds, molasses has been considered as a dominant by-product in Iran. In this study, among 16 crude oil degrading isolates, strain Pseudomonas aeruginosa ZN was selected as an efficient biosurfactant producer by screening methods for detection of biosurfactant producing bacteria. For investigation of molasses concentrations effect on bacterial growth and biosurfactant production, a wide range of molasses concentrations from 2-12% (v/v) were used. This strain was able to grow and produce biosurfactant in all range of molasses concentrations while the best concentrations were 4-6%. Also, at the optimum molasses concentration, reduction of surface tension from 70 to 32-34 mN/m was observed. The concnetrations more than these values decreased the growth and production process. Acid precipitation and solvent extract (ethyl acetate: hexane) methods were carried out for recovery of biosurfactant from the culture broth, then results of spraying on developed TLC and staining fermentation broth without bacterial cells showed the two produced biosurfactants were glycolipid.

Abstract Diabetes mellitus is a major health problem in the worldwide. Inhibition of DPP-IV is one of the methods to control diabetes type 2. Inhibition of this enzyme may improve glycemic control in diabetics by preventing the rapid breakdown and there by prolonging the physiological action of incretin hormones. Furthermore, improving the antioxidant system in diabetic patients can prevent the occurrence of secondary disease caused by oxidative stress. Therefore, the head of skipjack tuna was hydrolyzed with alcalase enzyme (1/5% of raw material weight) for 4 hours, in order to produce a product with antidiabetic and antioxidant activities. The DPP-IV inhibition activity, DPPH radical scavenging activity and reducing power of hydrolysate were measured. The results showed that the skipjack tuna head protein hydrolysate possess bioactive properties in a concentration dependent manner and increasing the protein concentration leads to a significant increase in bioactive properties of hydrolyzed product (p≤0.05). The IC50 of protein hydrolysate in DPP-IV inhibition and DPPH radical scavenging activities were obtained 1.016±0.02 mg/ml and 0.297±0.015 mg/ml, respectively. Also the reducing power of hydrolysate was 0.176±0.002 in 2.5 mg/ml protein concentration. Overall, according to the obtained results, it can be concluded that protein hydrolysate of skipjack tuna head possess high antioxidant and antidiabetic activities in vitro, and can be used as a food additive to enhance health level if additional research be conducted.

Abstract Abstract. Phycocyanine (PC) belongs to a group of protein receptor proteins called phycobiliprotein. All of the phycobiliprotein are multi-chain proteins made up of apoproteins. Which are covalently attached to the phyclobilins. This experimental study was carried out on strain of native Anabaena doliolum, Isolated from soils and waters of south Iran were Masjed Soleyman area. The cyanobacteria were grown and stored in BG11 medium. Then, the amount of phycocyanin produced under different light treatment and the amount of phycocyanin extracted using different ratios of multi-buffer and at two different temperatures were evaluated. The results of this study showed that the highest growth rate is when the sample is exposed to green light for three to five days. The best amount of extraction for distilled water and at a refrigerator temperature (0^◦C) with a ratio of 3:1 biomass/solvent is equal to 0.03 ± 15 µg/ml. Also, at the environment temperature, phosphate buffer is a more suitable solvent for extracting phycocyanine at a ratio of one to two with a value of 0.05 ± 8 µg/ml. In general, it can be said that the growth rate, pigment production and optimum extraction conditions for each species are quite different, and the optimal extraction of phycocyanin in a species is also dependent on various factors such as time, temperature, solvent and the ratio of biomass to solvent.

Abstract Today, nanosilver is one of the most commercialized nanomaterials. The demand for synthesis of Nanosilver through biocompatible routs due to wide biomedical application has increased. Use of plants and plant products as sustainable and renewable resources in the synthesis of nanoparticles is more advantageous over other biological routes. In this study, biosynthesis of silver nanoparticles (AgNPs) using aqueous extract of Withania somnifera as reducing agent is reported. Effect of parameters such as AgNO3 concentration, aqueous extract, pH and formation time were investigated and optimized by UV-visible spectroscopy in the synthesis of nanoparticles. At room temperature, the solution color started to change from pale yellow to dark brown due to the reduction of silver ion. The transmission electron microscopy (TEM) was applied for size and morphological analysis of nanoparticles. TEM result shows a spherical structure with an average size ranging from 24-35 nm for silver nanoparticles.

Abstract Aims: Breast cancer is the most common malignancy in women throughout the world. Among the various methods for preventing and treating cancer, plant natural compounds have more benefits than chemical drugs and have less side effects. Recently, many studies have been carried out on the antioxidant, anticancer, antiproliferation, and antiinflammatory properties of plant lignans, indicating the importance of these compounds in the prevention and treatment of various diseases. In the present study, the cytotoxic effects and inducing apoptosis of pinoresinol and lariciresinol lignans were investigated on SKBr3 breast cancer cell line.
Materials & Methods: SKBr3 cells were treated with different concentrations of both pinoresinol and lariciresinol, separately for 72 hours. Then, cell viability and cells morphological changes were determined using MTT assay and inverted light microscope, respectively. Also, apoptosis induction was analyzed by flow cytometry using an Annexin V-FITC apoptosis detection kit.
Findings: Both pinoresinol and lariciresinol treatments induced morphological changes, decreased cell growth, survival, proliferation and a significant increase in apoptosis induction in SKBr3 cell line compared to control cells in a concentration-dependent manner.
Conclusion: Inducing apoptosis and preventing the growth and proliferation of cancer cells are important mechanisms in the treatment of cancer. Pinoresinol and lariciresinol can be used to reduce cell proliferation and increase apoptosis induction in prevention and treatment of breast cancer.

Abstract This study was conducted to assess genetic diversity among the 32 tall fescue half-sib families using a randomized complete block design (RCBD) experiment in the four replicates. Based on analysis of variance, significant differences were observed among studied genotypes at the probability of 1% for plant height, canopy diameter, days to heading, days to pollination, crown diameter, fresh forage yield, dry forage yield, number of stem and seed yield in first harvest and in canopy diameter, crown diameter, fresh forage yield and dry forage yield in second harvest. Based on the results of mean comparisons, highest dry forage yield in the first harvest was obtained in genotype 32 by 758.5 grams. Principal component analysis by considering eigenvalues greater than one, caused to introduction of three components which determined 80.5% of the variation among the samples. In cluster analysis, the greatest of distinction between the groups was achieved with three clusters, and by cutting the dendrogram genotypes in three groups. According to the results, the third cluster was superior to other two clusters in terms of most traits. The genotypes of third cluster, according to the value of this cluster in terms of forage yield and seed yield will be of particular importance in breeding programs. In the breeding of cross-pollinated forage crops, success in selection depends on creating diversity by genetic recombination and achievement of heterosis. Due to the distance between groups 1 and 3, probably the most successful crosses will be achieved among genotypes of these two groups.

Abstract Aims: Significant amounts of waste, including feathers, bones, blood, etc. are yearly produced by the poultry industry. Feathers are composed of 90% keratin protein, and the rest is composed of lipids and water. Keratinases are one of the most diverse and usable enzymes, which can be produced by bacterial and fungal microorganisms. These enzymes show a wide range of application in various fields.
Materials and Methods: In this study, the keratinolytic activity of the isolated strain from a poultry farm in Mashhad was evaluated and then the medium conditions for keratinase production were optimized. The strains were identified based on the morphological and biochemical methods. 16SrRNA gene of the strain was amplified by PCR and then sequenced. The strain proteolytic activity was examined and compared with its keratinolytic activity. Finally, strain growth ability tested in variety substrate.
Findings: Using 16SrRNA gene sequencing, morphological and biochemical identification, the strain shared 99.9% similarity with Bacillus mojavensis. Optimization of various factors, including temperature, pH, incubation time, carbon and nitrogen sources, aeration and inoculum size showed that the isolated strain has the highest keratinolytic activity at 37°C, 48 hour incubation period, pH=9.5, sucrose 1%, 3% substrate, aeration 75% and 6% (v/v) inoculum amount. None of the nitrogen sources had a positive effect.
Conclusion: The FUM-1 keratinolytic activity was increased approximately 3.38 fold by condition optimization of the medium, indicating the importance of environmental conditions. In the study, the strain with high keratinolytic activity was suggesting its potential use in biotechnological.

Abstract Revealing DNA sequences is vital for all branches of biological sciences. Next-Generation Sequencing (NGS) is a different approach in this area so that it has created a great evolution in biology science and covers various aspects of genome, transcriptome, epigenome and metagenome-level studies. NGS is considered as a high-performance method for genomic and transcriptomic information analysis in comparison with traditional methods due to providing good genomic coverage, determining each single pairs of bases and eliminating the first generation sequencing disadvantages (Sanger sequencing). Use of NGS has begun since 2005 and 2006, after the commercialization of various apparatus companies such as ABI/SOLiD Illumina, Science Roch/454Life, and Solexa to study the transcriptome of the model and non-model organisms. Recently, RNA sequencing is used widely to identify genes associated with growth and development processes and their expression patterns in response to a variety of biological and non-biological stresses, in various organs and growth stages in different organisms. It helps scientists to determine the amounts of gene expression, differentiation of different isoforms of genes, detection of gene fusions and characterization of small RNA as well as alternative splicing events, duplicate elements, exon of genes, new transcripts, UTRs, SNPs, and somatic mutations. The RNA-seq method typically consists of providing suitable biological samples, isolation of total RNA, enrichment of non-ribosomal RNAs, conversion of RNA to cDNA, construction of a fragment library, selecting size and adding linkers and sequencing on high-throughput sequencing platform, alignment, and assembly of the reads and downstream analysis.

Abstract The stimulants are materials that increase alertness and reduce physical and mental fatigue. These drugs increase the activity of excitatory receptors and reduce the activity of inhibitory receptors in the central nervous system. Methamphetamine, also known as crystal, is a psychoactive substance. This drug is stimulating nerves and by a direct effect on the brain, mechanisms cause joy and excitement in people. Methamphetamine in low to moderate doses (5 to 30 mg) causes euphoria; excitement, increased heart rate, and blood pressure, mydriasis, increased body temperature and decrease appetite. High but non-lethal doses of methamphetamine cause mental disorders and psychotic symptoms, seizures, and rhabdomyolysis. Cardiovascular toxicity of methamphetamine-induced hypertension, arrhythmia, acute coronary syndrome and ischemic ventricular. The most important cellular mechanisms involved in the damage caused by Methamphetamine are oxidative stress, excitotoxicity, and mitochondrial damage. The synthesis of methamphetamine in illegal workshops mainly of six methods which are based on the raw materials are divided into two groups. Raw materials in the synthesis of methamphetamine are ephedrine and phenyl propanol. In reduction, methods involve Birch reduction, Nagai and hydrogenation Rosenmund ephedrine and pseudoephedrine are used as raw material, in Lockhart methods and amination reduction methods based on phenyl propanol as raw material.

Abstract Burns are one of the most important accidents related to human health. Receiving proper treatment is very important due to the intense complications associated with them. The improvement and elimination of ulcer effect can be achieved by controlling the wound infection. According to this, wound dressing containing antibiotics is one of the effective methods in wound's infection treatment. The use of silver in burns caring has been considered a long time ago, but silver deposits on the liver that causes some problems which can be overcome with the help of nanotechnology. In this study, Silver-Oak nanoparticles were green synthesized by using the ethanol extraction of Iranian Oak with the help of reducing agent. Nanoparticles formation were followed by UV-Visible spectrum and they characterize with SEM images and XRD spectrum. Then Gelatin-Alginate Hydrogel was prepared as a wound dressing and their properties were investigated by the presence of nanoparticles, oak extraction and without any additives. Nanoparticles' diameter is about 30-65nm which are dispersed in the hydrogel with regular pore size about 30-100μm. Oak extraction increase the hydrogel water uptake that improve the wound hilling. Antibacterial properties of wound dressing against Staphylococcus aurous and pseudomonas are investigated by inhibiting zone.

Abstract Ibuprofen, 4-isobutyl-propionic acid, is an important well-known member of NSAIDs which is widely utilized in inflammatory therapy like treatment of rheumatoid arthritis and various degrees of analgesic. Despite the high medical activity and low toxicity of ibuprofen, it is supplied as a racemic mixture. In this research enantioselective resolution of (R, S)-ibuprofen by immobilized preparations of Rhizomucor miehei lipase (RML) on silica and silica nanoparticles was investigated. For this, chemical modification of silica and silica mesoporous nanoparticles was performed by the simultaneous use of two coupling linkers; Octyltriethoxysilane (OTES) for hydrophobic interaction and glycidoxypropyltrimethoxylsilane (GPTMS) for covalent linkage of RML. The results showed that immobilization of RML on octyl-functionalized supports produces specific activity almost 1.5-2 folds greater than the specific activity of the free enzyme. The observed hyper-activation decreased with increasing epoxy groups on the supports confirming the enhancement of covalent nature of the attachment. Regarding the specific activity of the immobilized preparations and desorption percentages of RML from each support, the most suitable carrier obtained from the functionalization of the supports in the presence of GPTMS and OTES in the ratio of 1:1. The selected biocatalysts were then used for enantioselective resolution of (R, S)-ibuprofen by esterification reaction at different conditions. The results revealed that the most suitable biocatalysts are those prepared by immobilization of RML on SBA-15 and silica modified with GPTMS and OTES in the ratio of 1:1 which produced high E values at ambient temperature.