Baqiyatallah University of Medical SciencesJournal of Applied Biotechnology Reports2322-11868420211201The Effect of Probiotics Supplements on Oxidative Stress in Various Diseases: A Systematic Review Study32633111786310.30491/jabr.2020.117863ENSadrollah MahmoudiTrauma Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran0000-0001-5251-3949Mohammad Reza GhaneTrauma Research Center, Baqiyatallah University of Medical Sciences, Tehran, IranMehrdad FarajiTrauma Research Center, Baqiyatallah University of Medical Sciences, Tehran, IranHassan GoodarziTrauma Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran0000-0002-8640-6639Fahimeh ShahjooieTrauma Research Center, Baqiyatallah University of Medical Sciences, Tehran, IranHamid Reza JavadzadehTrauma Research Center, Baqiyatallah University of Medical Sciences, Tehran, IranJournal Article20191023<span class="fontstyle0"><strong>Introduction:</strong> </span><span class="fontstyle2">Recently, the consumption of probiotics to decrease Oxidative Stress (OS) has been recommended. The present study aimed to assess the effect of probiotics on OS in various diseases as well as in different models in a systematic review study.<br /></span><span class="fontstyle0"><strong>Materials and Methods:</strong> </span><span class="fontstyle2">Articles were searched through scientific sources such as PubMed, MEDLINE, Wiley, EMBASE, ISI Web of Knowledge, and Scopus by two review authors independently. The keywords that applied to search these articles included: Probiotics, OS, Stresses, Oxidative, and Stress. Results were then combined and reviewed regarding the type of study, type of disease/participants, and outcomes.<br /></span><span class="fontstyle0"><strong>Results:</strong> </span><span class="fontstyle2">In order to conduct this study, 19 eligible studies were investigated. The results showed that various probiotics supplements could improve OS parameters in pregnant women, patients with type 2 diabetics, Diabetic Kidney Disease (DKD) type 2, gestational diabetes mellitus, and the aging process, inflammatory factors in petrochemical workers, acute necrotizing pancreatitis, and polycystic ovary syndrome. However, no significant effect was reported on the oxidative status in patients with Rheumatoid Arthritis.<br /></span><span class="fontstyle0"><strong>Conclusions:</strong> </span><span class="fontstyle2">Probiotics supplements could improve OS biomarkers in various diseases. There are still some controversies about these methods and types of probiotics supplementations. It is recommended that more studies should be conducted for a better decision.</span> Baqiyatallah University of Medical SciencesJournal of Applied Biotechnology Reports2322-11868420211201A Review on the Applications of Listex™ P100 Bacteriophage33233614252910.30491/jabr.2020.248851.1286ENBehzad HeshmatiApplied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran0000-0002-7673-9376Hadi Esmaeili Gouvarchin GalehApplied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran0000-0001-8562-2295Ruhollah DorostkarApplied Virology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran0000-0002-2574-1150Journal Article20200919<span class="fontstyle0"><em>Listeria monocytogenes</em> </span><span class="fontstyle0">is a Gram-positive, small rod-shaped bacterium that causes listeriosis in animals and humans from eating contaminated food. It can mostly affect susceptible individuals with defective immune systems. Also, the mortality rates, as a result of listeriosis, can be varied between 30 to 75%. <em>Listeria</em> had been reported to be more allergic to antibiotics than Gram-positive bacteria. However, according to reports, <em>Listeria</em> has recently been allergic to the resistance to these antibiotics. Such an increase in antibiotic resistance in <em>Listeria</em> is in line with the worldwide pattern of the increasing prevalence of antibiotic resistance, including multiple antibiotic resistance in many bacterial groups.<br />Therefore, one of the ways to reduce the incidence of this disease in animals and humans is the biological control of this bacterium by bacteriophage. Bacteriophages have been shown to be effective in controlling </span><span class="fontstyle0"><em>L. monocytogenes</em> </span><span class="fontstyle0">in food</span><span class="fontstyle2">. </span><span class="fontstyle0">In 2006, the US Food and Drug Administration approved the preparation of two bacteriophages (Listex™ P100 and LMP-102) for use in certain foods to control </span><em><span class="fontstyle0">L. monocytogenes</span></em><span class="fontstyle0">. The prevalence of foodborne pathogens is the use of phages as biocontrol agents in food. Also, the activation of Phage Listex™</span><span class="fontstyle0"> </span><span class="fontstyle0">P100 against multiple </span><span class="fontstyle0"><em>L. monocytogenes</em> </span><span class="fontstyle0">serovars allows it to kill </span><span class="fontstyle0"><em>L. monocytogenes</em> </span><span class="fontstyle0">and its immunogenicity in food and clinical products</span><span class="fontstyle2">. </span><span class="fontstyle0">The prevalence of foodborne pathogens is the use of phages as biocontrol agents in food. Also, the activation of Phage Listex™</span><span class="fontstyle0"> </span><span class="fontstyle0">P100 against multiple </span><span class="fontstyle0"><em>L. monocytogenes</em> </span><span class="fontstyle0">serovars allows it to kill </span><span class="fontstyle0"><em>L. monocytogenes</em> </span><span class="fontstyle0">and its immunogenicity in food and clinical products.</span> Baqiyatallah University of Medical SciencesJournal of Applied Biotechnology Reports2322-11868420211201In Vitro Investigation of the Anti-Diabetic Effects of Imperialine on Beta-TC6 Pancreatic and C2C12 Skeletal Muscle Cell Lines33734512532310.30491/jabr.2021.125323ENMassoud Mashhadi Akbar BoojarDepartment of Cell and Molecular Biology, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran0000-0003-0145-5472Mahdi Mashhadi Akbar BoojarDepartment of Pharmacology and Toxicology, Faculty of Pharmacy, Baqiyatallah University of Medical Sciences, Tehran, Iran0000-0002-2002-9332Yaghuob FiroozivandDepartment of Pathobiology, Malekan Branch, Islamic Azad University, Malekan, IranJournal Article20200112<span class="fontstyle0"><strong>Introduction:</strong> </span><span class="fontstyle2">Imperialine (Imp) is a steroidal alkaloid present as the main active constituent of medicinal herb, </span><span class="fontstyle2"><em>Fritillaria imperialis</em> </span><span class="fontstyle2">with many biological and therapeutic effects. However, it has not been investigated in vitro for hypoglycemic effects. Herein, the effects of Imp on cell survival, carbohydrate-hydrolyzing enzymes (alpha-amylase and alpha-glucosidase), glucose uptake ability, insulin secretion levels, Advanced Glycation End products (AGEs) including pentosidine, methylglyoxal, and 3-deoxyglucosone levels and the activity of glyoxalase I as the main factor for degradation of AGEs were examined.<br /></span><span class="fontstyle0"><strong>Materials and Methods:</strong> </span><span class="fontstyle2">C2C12 skeletal muscle and beta-TC6 pancreatic cells were incubated with Imp at concentrations of 0, 25, 50, 75, and 10 μg/ml and the cells were evaluated separately. The biological assays were based on ultraviolet-visible (UV/VIS) spectrophotometric and/or high-performance liquid chromatography (HPLC) methods.<br /></span><span class="fontstyle0"><strong>Results:</strong> </span><span class="fontstyle2">Imp had considerable and dose-dependent effects on glucose uptake and insulin secretion (<em>P</em><0.05). The highest levels of glucose uptake were achieved at a concentration of 100 μg/ml of Imp. Increased glycation index, cytotoxicity, and decreased glyoxalase I activity appeared mostly at concentrations of 75 μg/ml and higher. The studied alkaloid demonstrated remarkable hypoglycemic effect by inhibition of alpha-amylase and alpha-glucosidase.<br /></span><span class="fontstyle0"><strong>Conclusions:</strong> </span><span class="fontstyle2">Consequently, the results of the present study revealed possible hypoglycemic effects of Imp and it could be suggested for future studies in the treatment of diabetes mellitus.</span> Baqiyatallah University of Medical SciencesJournal of Applied Biotechnology Reports2322-11868420211201Production and Optimization of Polyhydroxyalkaonoate Obtained from Bacillus megaterium JHA34636014201610.30491/jabr.2020.242493.1263ENJoyline MascarenhasDepartment of Microbiology, Wilson College, Mumbai 400007, Maharashtra, India0000-0002-8304-4011Aruna KDepartment of Microbiology, Wilson College, Mumbai 400007, Maharashtra, India0000-0002-7776-2690Journal Article20200804<span class="fontstyle0"><strong>Introduction:</strong> </span><span class="fontstyle2">The last four decades have recognized the critical environmental issues pertaining to the use of non-degradable synthetic plastics. Currently, the synthesis of biodegradable polymers, like polyhydroxyalkaonoates (PHA) has been gaining considerable interest as a sustainable approach to overcome these issues. The current study was carried out with an objective to optimize the PHA production from </span><span class="fontstyle2"><em>Bacillus megaterium</em> </span><span class="fontstyle2">JHA.<br /></span><strong><span class="fontstyle0">Materials and Methods: </span></strong><span class="fontstyle2">Various nutritional and physico-chemical parameters were optimized using the ‘one factor at a time’ approach. The findings were analysed statistically by one-way ANNOVA and linear regression using the open-source R software.<br /></span><span class="fontstyle0"><strong>Results:</strong> </span><span class="fontstyle2">The optimum physico-chemical parameters for maximum PHA production were attained when 5% inoculum size of </span><span class="fontstyle2"><em>B. megaterium</em> </span><span class="fontstyle2">JHA was added to the JHA medium (pH 8) and incubated at 28 °C for 96 h under shaker conditions (120 rpm). The nutritional parameters were further optimized by addition of 0.6 g% K</span><sub><span class="fontstyle2">2</span></sub><span class="fontstyle2">HPO</span><sub><span class="fontstyle2">4</span></sub><span class="fontstyle2">, 0.4 g% KH</span><sub><span class="fontstyle2">2</span></sub><span class="fontstyle2">PO</span><sub><span class="fontstyle2">4</span></sub><span class="fontstyle2">, 21 g% glucose, 12.5 mM microcosmic salt, 0.04 g% KNO</span><sub><span class="fontstyle2">3</span></sub><span class="fontstyle2">, 1 mM MgSO</span><span class="fontstyle2"><sub>4</sub>, </span><span class="fontstyle2">and 2 ml trace elements to the JHA medium. The C:N and C:P ratio was adjusted to 70:1 and 20:1 respectively. The growth of bacteria under these optimized parameters allowed 13.71 g/L accumulation of PHA leading to a yield of 54.51%. This resulted in 66.88% increase in yield as compared to the original medium. The scale-up study using a 2 L fermenter further increased PHA accumulation by 4.39% under optimized conditions.<br /></span><span class="fontstyle0"><strong>Conclusions:</strong> </span><span class="fontstyle2">The above results clearly indicate that </span><span class="fontstyle2"><em>B. megaterium</em> </span><span class="fontstyle2">JHA is a promising isolate that can be exploited for the industrial production of PHA.</span>Baqiyatallah University of Medical SciencesJournal of Applied Biotechnology Reports2322-11868420211201Adipose-Derived Stem Cells Growth and Proliferation Enhancement Using Poly(Lactic-co-Glycolic Acid) (PLGA)/Fibrin Nanofiber Mats36136912549110.30491/jabr.2020.223551.1199ENMohsen NorouziDepartment of Biomedical Engineering, Russ College of Engineering and Technology, Ohio University, Athens, Ohio, USADepartment of Tissue Engineering, Faculty of Materials Engineering, Najafabad Branch, Islamic Azad University, Najafabad, Isfahan, Iran0000-0002-7150-2716Mohammad RafieniaDepartment of Biomaterials, Tissue Engineering and Nanotechnology, School of Advanced Technologies in Medicine, Isfahan University of Medical Sciences, Isfahan, Iran0000-0001-6030-5082Elahe PooraziziDepartment of Biochemistry, Faculty of Medicine, Najafabad Branch, Islamic Azad University, Najafabad, Isfahan, Iran0000-0003-4509-1885Mohsen SetayeshmehrDepartment of Biomaterials, Tissue Engineering and Nanotechnology, School of Advanced Technologies in Medicine, Isfahan University of Medical Sciences, Isfahan, Iran0000-0001-6044-1279Journal Article20200314<strong>Introduction: </strong>A Synthetic biomaterial, such as Poly Lactic-co-Glycolic Acid (PLGA) with superior mechanical properties, along with a natural polymer such as fibrin, which facilitates cell attachment and enhances biocompatibility, can be used in the production of novel composite tissue engineering scaffolds.<br /><strong>Materials and Methods: </strong>To carry out this study, 10% polymer solutions with different ratios of PLGA: Fibrin, including 10:0, 9:1, 8:2, and 7:3, were prepared and used in the production of aligned and unaligned electrospun nanofiber scaffolds. Human Adipose-Derived Stem Cells (h-ADSCs) were cultured on the scaffolds, and they were characterized using Fourier Transform Infrared Spectroscopy (FTIR), Energy-dispersive X-ray spectroscopy (EDX), Scanning Electron Microscopy (SEM), mechanical, hydrophilic, degradation, water absorption, and biocompatibility tests.<br /><strong>Results: </strong>The obtained scaffolds consisted of homogeneous fibers, without any beads and water droplets. The percentage of porosities and internal correlation of the cavities were not significantly different between aligned and unaligned electrospun scaffolds <span class="fontstyle0">(</span><span class="fontstyle1"><em>P</em></span>><span class="fontstyle0">0.05)</span> and by adding fibrin, these properties improved, while tensile strength and elasticity decreased. All the scaffolds were hydrophobic and the highest and lowest swelling rates belonged to PLGA/30% Fibrin scaffolds and pure PLGA scaffolds (more than 90% and less than 45%, respectively). There is a significant difference in degradation rates between fibrin-contained scaffolds and pure PLGA scaffolds. Moreover, compared to the aligned electrospun scaffolds, the highest degradation rate of unaligned ones was observed.<br /><strong>Conclusion: </strong>Considering the results of SEM and bio-compatibility experiments, the aligned electrospun PLGA/10% Fibrin scaffold with numerous spindle shape h-ADSCs and unaligned electrospun PLGA/20% Fibrin scaffold with many spindle shape cells together with round shape cells are introduced as optimal options.Baqiyatallah University of Medical SciencesJournal of Applied Biotechnology Reports2322-11868420211201Role of Intracellular Divalent Cations on the Adenylate Cyclase Activation by Human LH in mLTC-1 Leydig Cells37037414202210.30491/jabr.2021.286454.1382ENThi Mong Diep NguyenDepartment of Applied Biology & Agriculture, Faculty of Natural Sciences, Quy Nhon University, Binh Dinh 820000, Vietnam0000-0001-6534-9041Van Giang TranFaculty of Biology, Hue University of Education, Hue University, Hue 530000, VietnamThi Chi Hieu NguyenFaculty of Applied Biology, Quang Trung University, Quy Nhon, VietnamBa Nghi NguyenFaculty of Applied Biology, Quang Trung University, Quy Nhon, VietnamJournal Article20210516<span class="fontstyle0"><strong>Introduction:</strong> </span><span class="fontstyle2">The Luteinizing Hormone (LH) regulates Leydig cell activities through LHR occupation, promoting Gs protein and/or </span><span class="fontstyle3">β</span><span class="fontstyle2">-arrestin activation. The activated Gs</span><span class="fontstyle3">α</span><span class="fontstyle2">GTP </span><span class="fontstyle2">subunit stimulates Adenylyl Cyclase (ACs) and, therefore, intracellular cyclic Adenosine Monophosphate (cAMP) accumulation from the AC substrate Adenosine Triphosphate (ATP). The divalent cations, magnesium (Mg</span><sup><span class="fontstyle2">2+</span></sup><span class="fontstyle2">) or manganese (Mn</span><span class="fontstyle2"><sup>2+</sup></span><span class="fontstyle2">) associate with ATP to form the real substrates of ACs. In addition, ACs are sensitive to calcium (Ca</span><sup><span class="fontstyle2">2+</span></sup><span class="fontstyle2">) but in a different way than Mg</span><span class="fontstyle2"><sup>2+</sup> </span><span class="fontstyle2">or Mn</span><sup><span class="fontstyle2">2+</span></sup><span class="fontstyle2">. Indeed, LH increases the cytoplasmic calcium ion concentration ([Ca</span><sup><span class="fontstyle2">2+</span></sup><span class="fontstyle2">]</span><span class="fontstyle2">cyt</span><span class="fontstyle2">) but only when Ca</span><span class="fontstyle2"><sup>2+</sup> </span><span class="fontstyle2">is present in the extracellular medium.<br /></span><span class="fontstyle0"><strong>Materials and Methods:</strong> </span><span class="fontstyle2">In the present study, the effects of intracytoplasmic Ca</span><sup><span class="fontstyle2">2+</span></sup><span class="fontstyle2">, Mg</span><sup><span class="fontstyle2">2+</span></sup><span class="fontstyle2">, and Mn</span><span class="fontstyle2"><sup>2+</sup> </span><span class="fontstyle2">on the cyclic AMP response to human LH in mLTC-1 Leydig tumor cells were investigated. The mLTC-1 cells were incubated at 37 °C in media supplemented with and without 5 µM Ca</span><sup><span class="fontstyle2">2+</span></sup><span class="fontstyle2">, 5 µM Mg</span><sup><span class="fontstyle2">2+</span></sup><span class="fontstyle2">, or 5 µM Mn</span><sup><span class="fontstyle2">2+</span></sup><span class="fontstyle2">. The intracellular cyclic AMP accumulation was then monitored under LH stimulation.<br /></span><span class="fontstyle0"><strong>Results:</strong> </span><span class="fontstyle2">Our findings revealed that only Mg</span><span class="fontstyle2"><sup>2+</sup> </span><span class="fontstyle2">and Mn</span><span class="fontstyle2"><sup>2+</sup> </span><span class="fontstyle2">in the extracellular medium potentiate the cAMP response to hLH, in contrast to Ca</span><sup><span class="fontstyle2">2+</span></sup><span class="fontstyle2">. In addition, we also showed that HCO</span><span class="fontstyle2">3- </span><span class="fontstyle2">increased the stimulation of the adenylyl cyclase enzyme by Ca</span><sup><span class="fontstyle2">2+</span></sup><span class="fontstyle2">, Mg</span><span class="fontstyle2"><sup>2+</sup> </span><span class="fontstyle2">or Mn</span><sup><span class="fontstyle2">2+</span></sup><span class="fontstyle2">.<br /></span><strong><span class="fontstyle0">Conclusions: </span></strong><span class="fontstyle2">In mLTC-1 cells, extracellular Mg</span><span class="fontstyle2"><sup>2+</sup> </span><span class="fontstyle2">and Mn</span><span class="fontstyle2"><sup>2+</sup> </span><span class="fontstyle2">might potentiate LH-stimulated ACs activity by favoring LH interaction with its receptor, whereas Ca</span><span class="fontstyle2"><sup>2+</sup> </span><span class="fontstyle2">from internal stores might be mobilized towards the cytoplasm to increase ACs activity, possibly through the soluble isoform.</span> Baqiyatallah University of Medical SciencesJournal of Applied Biotechnology Reports2322-11868420211201Genetic Diversity among Economically Important Zataria multiflora Accessions through ISSR Markers: The Main Step for Breeding and Exploitation Programs37538414240310.30491/jabr.2021.254653.1305ENSoolmaz MeamariDepartment of Horticultural Sciences, Faculty of Agriculture and Natural Resources, University of Hormozgan, Bandar Abbas, IranAlireza YavariDepartment of Horticultural Sciences, Faculty of Agriculture and Natural Resources, University of Hormozgan, Bandar Abbas, Iran0000-0002-4786-0759Mahdi BikdelooDepartment of Horticultural Sciences, Faculty of Agriculture & Natural Resources, Arak University, Arak, IranTahereh SadatHashemiDepartment of Horticultural Sciences, Faculty of Agriculture and Natural Resources, University of Hormozgan, Bandar Abbas, IranJournal Article20201028<strong>Introduction:</strong><em> Zataria multiflora</em> is an important medicinal plant of the Lamiaceae family in Central and Southern Iran. This plant is at the risk of extinction as a result of wasteful harvesting due to growing demand and high economic value.<br /><strong>Materials and Methods:</strong> In this study, the genetic diversity of 25 different accessions of <em>Z. multiflora, </em>collected from provinces including Hormozgan, Fars, Sistan and Balouchestan, Bushehr, Yazd, Kerman, and Isfahan were examined using Inter Simple Sequence Repeat (ISSR) markers. To extract the DNA, five samples taken from the leaves of each accession were transformed into bulks; then, their concentrations were homogenized and 15 primers were used for the remainder of the experiment.<br /><strong>Results:</strong> The primers produced 83 polymorphic strands altogether, with an average polymorphism percentage of 77.30%; meanwhile, the highest polymorphism percentage (100%) was achieved via primers including 816D, 824H, 836P, and 844S. The lowest polymorphism percentage was obtained from 811C and 834N primers. The results obtained from the Jaccard similarity coefficient in the NTYSIS software showed that the genetic similarity of <em>Z. multiflora</em> accessions varies between 0.32-0.82. The lowest similarities were observed in accessions taken from Fanuj and Ashar, Mehriz, and NasrAbad, and two accessions of Mehriz and Khafr. However, the highest similarities were seen among accessions of NalShah GhandAab and Kerman. In principal component analysis, three of the first components explained 40.44% of changes in the entire data. Following a cluster analysis based on the UPGMA algorithm, accessions were classified into six groups.<br /><strong>Conclusions:</strong> It can be concluded that the ISSR markers are suitable for examining the genetic diversity of <em>Z. multiflora</em> accessions.Baqiyatallah University of Medical SciencesJournal of Applied Biotechnology Reports2322-11868420211201Elucidating the Role of Plant Growth Promoting Bacteria for Nitrate and Phosphate Bioremediation: A Sustainable Approach Towards Crop Productivity and Environmental Protection38539414205110.30491/jabr.2020.240413.1260ENSiddiqua KhotSchool of Biotechnology & Bioinformatics, DY Patil Deemed to be University, Navi Mumbai, Maharashtra-400614, IndiaSourav GhoshSchool of Biotechnology & Bioinformatics, DY Patil Deemed to be University, Navi Mumbai, Maharashtra-400614, IndiaJanvi ShahSchool of Biotechnology & Bioinformatics, DY Patil Deemed to be University, Navi Mumbai, Maharashtra-400614, IndiaJournal Article20200728<span class="fontstyle0"><strong>Introduction:</strong> </span><span class="fontstyle2">The production of healthy food along with environmental sustenance is an everlasting mission for the coming ages. The utilization of chemical fertilizers may fulfil the requirement but it could also compromise the environment. The present study aimed to tap beneficial microbes that could not only harbour plant growth-promoting traits but could also remove environmental pollutants, nitrate and phosphate.<br /></span><span class="fontstyle0"><strong>Materials and Methods:</strong> </span><span class="fontstyle2">The culture-dependent approach was taken into account to isolate and characterize the bacterial population from dumping ground and mangrove soil. The selected isolates were further tested for remediating nitrate and phosphate by standard biochemical tests.<br /></span><span class="fontstyle0"><strong>Results:</strong> </span><span class="fontstyle2">Three isolates from dumping ground and four bacterial strains were proved to contain three out of four Plant Growth-Promoting (PGP) traits (HCN, IAA, Phosphate-solubilisation, and Nitrogen fixation). Two out of three selected bacterial strains were found to have the ability to remove nitrate and phosphate up to 74% and 62% (</span><span class="fontstyle2"><em>p</em>< 0.05</span><span class="fontstyle2">) respectively. This is while all the other selected bacterial strains from mangrove soil could effectively remove nitrate and phosphate in the range of 62% to 79% and 24% to 100% (</span><span class="fontstyle2"><em>p</em><0.05</span><span class="fontstyle2">) respectively. Two novel strains of </span><span class="fontstyle2"><em>Streanomonas</em> sp., <em>Alkaligens</em> sp </span><span class="fontstyle2">and one each </span><span class="fontstyle2"><em>Bacillus</em> sp., <em>Corynebacterium, Pseudomonas, </em></span><span class="fontstyle2">and </span><span class="fontstyle2"><em>Citrobacter</em> </span><span class="fontstyle2">species isolates were found in the present case.<br /></span><span class="fontstyle0"><strong>Conclusions:</strong> </span><span class="fontstyle2">The study represents the dual ability of the novel bacterial strains in plant growth promotion as well as remediation of environmental pollutants. Thus, the study might aid in designing a microbe-based bio-fertilizer for plant growth promotion along with maintenance of soil and environmental health.</span> Baqiyatallah University of Medical SciencesJournal of Applied Biotechnology Reports2322-11868420211201Cloning, Expression, and Purification Strategies for Enhanced Production of Enterokinase using TrpE fusion tag in Bench Scale Bioreactor39540514258510.30491/jabr.2021.250014.1293ENSanthosh Nagaraj NanjundaiahDiscovery Biology, Anthem Biosciences Pvt Ltd, Bommasandra, Bangalore, India School of Biosciences and Technology, Vellore Institute of Technology, Vellore, India0000-0003-3894-3904Jayasri MASchool of Biosciences and Technology, Vellore Institute of Technology, Vellore, IndiaSunilkumar SukumaranDiscovery Biology, Anthem Biosciences Pvt Ltd, Bommasandra, Bangalore, IndiaGanesh SambasivamDiscovery Biology, Anthem Biosciences Pvt Ltd, Bommasandra, Bangalore, IndiaJournal Article20200928<strong>Introduction:</strong> Enterokinase (EK) is an enzyme of the serine protease family which is widely used in protein purification. EK acts at the C terminus of the DDDDK site in a protein chain. The enzyme has gained commercial importance in recent times owing to its specificity in biosimilar processing and also while removing the fusion tags in the course of protein purification.<br /><strong>Materials and Methods:</strong> The commercial production of EK has faced several challenges and demands the usage of novel strategies. This research shows the construction of a vector using TrpLE1413 (TrpE) as a fusion tag that pushes the produced EK inside the cell towards the inclusion body fraction and produced more of the desired protein in the BL21 (DE3) strain of <em>Escherichia coli</em>. The inclusion bodies produced by fed-batch fermentation were solubilized, refolded, activated, and purified by a single step of anion exchange chromatography.<br /><strong>Results:</strong> We purified 241 mg/L of recombinant EK, and its purity confirmed by RP-HPLC was greater than 97%. However, the maximum EK yield reported by other researchers is only 106 mg/L.<br /><strong>Conclusions:</strong> Overall, our results demonstrate the potential of the TrpE fusion tag along with novel expression and purification strategies to increase the enzyme yield by 2-2.5 times when compared to the yield achieved using traditional methods. Hence, this study has paved the way for the industrial production of EK in an economically viable manner. The same strategy could possibly be implemented on the expression of other industrially important recombinant enzymes depending on the protein characteristics.Baqiyatallah University of Medical SciencesJournal of Applied Biotechnology Reports2322-11868420211201Computational Analysis of Responsive Transcription Factors Involved in Drought and Salt Stress in Rice40641314252810.30491/jabr.2020.243913.1272ENAbbas SaidiDepartment of Plant Sciences and Biotechnology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran5389-6721-0001-0000Zahra HajibaratDepartment of Plant Sciences and Biotechnology, Faculty of Life Sciences and Biotechnology,
Shahid Beheshti University, Tehran, IranAsadollah AhmadikhahDepartment of Plant Sciences and Biotechnology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran0000-0001-5100-9740Journal Article20200815<span class="fontstyle0"><strong>Introduction:</strong> <span class="fontstyle2">Rice is one of the most important crops to more than half the world's population. The reduction of rice yield has been severely influenced by drought and salt stress. The main purpose of this survey was to detect Transcription Factors (TFs) involved in drought and salt stress in rice.<br /></span><strong>Materials and Methods:</strong> <span class="fontstyle2">In this study, microarray data in PlnTFDB and DRTF were taken to evaluate the expression of responsive TFs to drought and salt stress in the growth stages. A comprehensive analysis of responsive TFs were performed containing gene network, expression analysis in different tissues, and detection of Transcription Factor Binding (TFBs) sites.<br /></span><strong>Results:</strong> <span class="fontstyle2">A total of 80 TFs were found differentially expressed (DEGs) under drought and salt stress in rice. Gene Ontology (GO) revealed that biological processes included transcription, regulation of transcription, regulation of RNA metabolic, and RNA metabolic. In addition, some molecular functions such as organic cyclic compound binding, heterocyclic compound binding, DNA binding, and cellular component are enriched in intracellular and nucleus. To survey selection pressure in responsive TFs under drought and salt conditions, Tajima’s and Fu test analysis revealed balancing selection. Analysis of TFBs illustrated that several TFBs namely AP2, bZIP, and MYB/SANT act as basic TFBs linked to abiotic stress responses as well as different growth stages in rice. The current study revealed that most TFs related to histone modification were up-regulated whereas, TFs associated with the regulation of repression/ activation transcription were down-regulated.<br /></span><strong>Conclusions:</strong> <span class="fontstyle2">Our results can provide an insight into the regulatory mechanisms involved in response to drought and salt stress which can aid to improve rice varieties.</span> </span>Baqiyatallah University of Medical SciencesJournal of Applied Biotechnology Reports2322-11868420211201Purification and Characterization of Arginase from the Stomach of Cane rat (Thryonomys swinderianus, Temminck 1827)41442014215410.30491/jabr.2021.268720.1343ENOluwaseun EmmanuelAgboolaDepartment of Biochemistry and Molecular Biology, Obafemi Awolowo University, Ile-Ife, Osun State, Nigeria0000-0002-6794-4749Scholes TaiwoAdewoleDepartment of Chemical Sciences, Kings University, Odeomu, Osun State, NigeriaOlutosin SamuelIlesanmiDepartment of Chemical Sciences, Achievers University, Owo, Nigeria0000-0002-8819-5935Journal Article20210116<span class="fontstyle0"><strong>Introduction:</strong> </span><span class="fontstyle2">Rodent pests are unique threats to agriculture and food security in many developing countries. Hence, this study investigated some physicochemical properties of partially purified arginase from the stomach of cane rats (</span><em><span class="fontstyle2">Thryonomys swinderianus</span></em><span class="fontstyle0">)</span><span class="fontstyle2">. This was with a view to understanding the significance of the enzyme in nitrogen metabolism, which could be exploited in the control of rodent pests.<br /></span><span class="fontstyle0"><strong>Materials and Methods:</strong> </span><span class="fontstyle2">Arginase was isolated and subjected to 70% ammonium sulfate (NH</span><sub><span class="fontstyle2">4</span></sub><span class="fontstyle2">)</span><sub><span class="fontstyle2">2</span></sub><span class="fontstyle2">SO</span><span class="fontstyle2"><sub>4</sub> </span><span class="fontstyle2">precipitation and dialyzed. The dialysate was further purified using ion-exchange chromatography on CM-Sephadex C-50 matrix.<br /></span><span class="fontstyle0"><strong>Results:</strong> </span><span class="fontstyle2">Apparent </span><span class="fontstyle2">K</span><span class="fontstyle2">m </span><span class="fontstyle2">of the enzyme for L-arginine and V</span><span class="fontstyle2">max </span><span class="fontstyle2">were 54 ± 0.7 mM and 0.057 ± 0.1 </span><span class="fontstyle3">μ</span><span class="fontstyle2">mol/minute/ml, while the optimum pH and temperature obtained for the enzyme were 7 and 70 °C, respectively. NaCl</span><span class="fontstyle2">, </span><span class="fontstyle2">MnCl</span><span class="fontstyle2"><sub>2</sub>, </span><span class="fontstyle2">and HgCl</span><span class="fontstyle2"><sub>2 </sub></span><span class="fontstyle2">activated the enzyme activity, while FeCl</span><span class="fontstyle2"><sub>3</sub> </span><span class="fontstyle2">totally inactivated the enzyme at tested concentrations. </span><span class="fontstyle3">β</span><span class="fontstyle2">-mercaptoethanol, urea, and EDTA profoundly potentiated the activity of the enzyme. Enzyme activity (81%) was retained when arginine was substituted with cysteine, while histidine, proline, alanine, tyrosine, and tryptophan had potent inhibitory effects on the enzyme activity.<br /></span><span class="fontstyle0"><strong>Conclusions:</strong> </span><span class="fontstyle2">The study established the presence of arginase in the stomach of cane rat and illuminated some physicochemical properties of the enzyme, which could be exploited in its deployment as a viable strategy to control </span><em><span class="fontstyle2">T. swinderianus</span></em><span class="fontstyle2">. However, further studies including structure-function investigations of the enzyme are recommended to fully exploit its potential in the control of rodent pests.</span> Baqiyatallah University of Medical SciencesJournal of Applied Biotechnology Reports2322-11868420211201Development of an IgG/IgY Sandwich-ELISA for the Detection of Cholera Toxin Subunit B42142714200210.30491/jabr.2020.244928.1276ENAmir Namvar VansoflaBiology Research Center, Faculty of Basic Sciences, Imam Hossein University, Tehran, IranShahram NazarianBiology Research Center, Faculty of Basic Sciences, Imam Hossein University, Tehran, Iran0000-0002-4693-877XDavoud SadeghiBiology Research Center, Faculty of Basic Sciences, Imam Hossein University, Tehran, IranMohamad Ebrahim MinaeBiology Research Center, Faculty of Basic Sciences, Imam Hossein University, Tehran, IranAbbas HajizadeBiology Research Center, Faculty of Basic Sciences, Imam Hossein University, Tehran, Iran0000-0003-3662-6681Journal Article20200822<span class="fontstyle0"><strong>Introduction:</strong> </span><span class="fontstyle2">Cholera is a lethal diarrheal disease caused by </span><em><span class="fontstyle2">Vibrio cholerae</span></em><span class="fontstyle2">. Cholera toxin (CTX) is one of the major virulence factors in </span><span class="fontstyle2">V. cholerae </span><span class="fontstyle2">pathogenesis. One of the major strategies in dealing with the poisoning of the bacteria is their rapid detection. The aim of this study was to design and set up a double-sandwich ELISA diagnostic method for the direct detection of cholera toxin B (CtxB) based on chicken immunoglobulin Y (IgY) and rabbit immunoglobulin G (IgG).<br /></span><span class="fontstyle0"><strong>Materials and Methods:</strong> </span><span class="fontstyle2">Recombinant CtxB protein was expressed in </span><span class="fontstyle2"><em>E. coli</em> </span><span class="fontstyle2">BL21 (DE3) cells by addition of IPTG and was purified using an NiNTA column. Chickens and rabbits were immunized subcutaneously and the generated antibodies were puri</span><span class="fontstyle3">fi</span><span class="fontstyle2">ed from egg yolks by polyethyleneglycol (PEG) precipitation and from the rabbits' sera by protein G column. These antibodies were used to set up the ELISA method. The sensitivity of the designed ELISA method was evaluated using serial dilutions of the protein and the specificity of this method was evaluated.<br /></span><span class="fontstyle0"><strong>Results:</strong> </span><span class="fontstyle2">Recombinant protein expression analysis showed an appropriate expression of the protein (300 µg/ml). ELISA assay results showed an increased serum antibody levels against the protein in rabbits’ and chickens’ sera after each injection. The yield of the purified IgY and IgG was 10 and 2.5 mg/ml, respectively. The sensitivity of the ELISA method was about 39 ng for recombinant CtxB. The results showed the high specificity of this technique.<br /></span><span class="fontstyle0"><strong>Conclusions:</strong> </span><span class="fontstyle2">Results suggest that IgY/IgG-based sandwich ELISA in this study provides a convenient preparation for the development of an immune-based method for the detection of CtxB.</span> Baqiyatallah University of Medical SciencesJournal of Applied Biotechnology Reports2322-11868420211201Evaluation of Bacterial Consortium and Optimization of Growth Parameters for Effective Decolorization of Azo Dye Reactive Red 12042843914215710.30491/jabr.2020.243280.1269ENRadhika BirmoleDepartment of Microbiology, Wilson College, Mumbai 400007, Maharashtra, India0000-0002-7212-7575Aruna K.Department of Microbiology, Wilson College, Mumbai 400007, Maharashtra, India0000-0002-7776-2690Journal Article20200810<span class="fontstyle0"><strong>Introduction:</strong> </span><span class="fontstyle2">The effluents produced as a result of the dyeing process, especially by textile industries are a major threat to sustainable environmental development. Several challenges are observed in the treatment and disposal of complex azo dyes like Reactive Red 120 (RR120). The aim of the present study was to optimize the dye decolorization/degradation process by bacterial consortium.<br /></span><span class="fontstyle0"><strong>Materials and Methods:</strong> </span><span class="fontstyle2">The consortium consisted of three potential azo dye degraders i.e., </span><span class="fontstyle2"><em>Shewanella haliotis</em> </span><span class="fontstyle2">RDB_1, </span><span class="fontstyle2"><em>Shewanella putrefaciens</em> </span><span class="fontstyle2">RDB_2, and </span><span class="fontstyle2"><em>Aeromonas hydrophila</em> </span><span class="fontstyle2">RDB_3. It was prepared in 1:1:1 ratio and was named as RAR. This consortium was optimized under several nutritional and physicochemical parameters for effective decolorization of RR120.<br /></span><span class="fontstyle0"><strong>Results:</strong> </span><span class="fontstyle2">Complete decolorization of 50 ppm RR120 was achieved with 10% inoculum of 1.0 OD</span><span class="fontstyle2"><sub>540 nm</sub> </span><span class="fontstyle2">in 3% Yeast Extract (YE) medium under static conditions in 4 h. The optimum decolorization was observed between pH 7-8 and temperature 30°C-35°C. However, the consortium RAR showed significant activity between a pH range of 6-10, temperature 25°C-45°C and NaCl concentration up to 10%. The electron acceptors like nitrate and nitrite salts, and electron donors like urea and casamino acids negatively affected the decolorization rate of RR120. The sugars and organic acids failed to support decolorization in M9 medium. However, a varying effect was observed in the 3% YE medium. Soymeal peptone (prepared in distilled water) supported considerable decolorization but was not as effective as 3% YE medium.<br /></span><span class="fontstyle0"><strong>Conclusions:</strong> </span><span class="fontstyle2">Considering the above features and tolerance of consortium RAR to varied range of pH, temperature and NaCl concentration, it may be a suitable candidate for biodecolorization of textile effluents.</span> Baqiyatallah University of Medical SciencesJournal of Applied Biotechnology Reports2322-11868420211201Molecular Characterization and Phenotype Analysis of a Novel Psychrotrophic Exiguobacterium Species from the Ilam Mountains44045214305710.30491/jabr.2022.308621.1453ENMehrdad Nasrollahzadeh SabetSchool of Medicine, AJA University of Medical Sciences, Tehran, IranCancer Epidemiology Research and Treatment Center, AJA University of Medical Sciences, Tehran, IranMostafa AkbariqomiApplied Biotechnology Research Centre, Baqiyatallah University of Medical Sciences, Tehran, Iran0000-0002-3533-2393Garshasb RigiDepartment of Genetics, Faculty of Basic Science, Shahrekord University, Rahbar Blvd, Shahrekord, Iran0000-0003-2450-912XAli SaberianDepartment of Food Science and Technology, Shahrekord Branch, Islamic Azad University, Shahrekord 88137-33395, IranJavad BehrooziCancer Epidemiology Research and Treatment Center, AJA University of Medical Sciences, Tehran, IranMohammad Foad HeidariDepartment of Medical Laboratory Sciences, School of Allied Health Medicine, AJA University of Medical Sciences, Tehran, IranTaher ElmiDepartment of Laboratory Science, Babol Branch, Islamic Azad University, Babol, Iran0000-0002-4247-4445Fereshteh AlimohamadiSchool of Medicine, AJA University of Medical Sciences, Tehran, Iran0000-0001-6862-6972Reza HeidariCancer Epidemiology Research and Treatment Center, AJA University of Medical Sciences, Tehran, Iran0000-0003-2985-8396Journal Article20211006<span class="fontstyle0"><strong>Introduction:</strong> </span><span class="fontstyle2">The genus </span><span class="fontstyle2"><em>Exiguobacterium</em> </span><span class="fontstyle2">includes psychrotrophic, mesophilic, and moderate thermophilic strains and species that have been isolated from extreme environments, including very cold or hot environments. The genus </span><span class="fontstyle2"><em>Exiguobacterium</em> </span><span class="fontstyle2">has received much attention in biotechnology, and related industries due to secretory enzymes with high enzymatic activity and consequently stable enzymes capable of tolerating extreme conditions. The aim of the present study was to introduce and describe the phenotypic and the genotypic characterization of a novel species of genus <em>Exiguobacterium </em>isolated from the Ilam mountains, Iran.<br /></span><span class="fontstyle0"><strong>Materials and Methods:</strong> </span><span class="fontstyle2">Genotypic features were analyzed using universal genes <em>(</em></span><span class="fontstyle2"><em>gyrB, hsp70, rpoB,</em> and <em>citC</em></span><span class="fontstyle2"><em>)</em> belonging to the </span><span class="fontstyle2"><em>Exiguobacterium</em> </span><span class="fontstyle2">genus. Also, </span><span class="fontstyle2"><em>cspC1, nusA,</em> </span><span class="fontstyle2">and </span><span class="fontstyle2"><em>dnaJ</em> </span><span class="fontstyle2">genes were used to confirm the profile of new psychrotrophic strains. The distinctive phenotypic characteristics of the new strain with the most famous strains of the genus </span><span class="fontstyle2"><em>Exiguobacterium</em> </span><span class="fontstyle2">were investigated. All statistical analyses were conducted using R Packages for data visualization and exploratory data.<br /></span><span class="fontstyle0"><strong>Results:</strong> </span><span class="fontstyle2">A motile, Gram-stain-positive, rod-shaped, non-sporing, tolerant up to 5% NaCl, grew at 0-25</span><span class="fontstyle3"> </span><span class="fontstyle2">°C and pH 6 and 9, designated </span><span class="fontstyle2"><em>Exiguobacterium</em> </span><span class="fontstyle2">sp. HA2 was isolated from the soil. The major isoprenoid quinone is MK-7 and in the smaller amount are MK-6 and MK-8. The major cellular fatty acids (>10 %) were isoC</span><span class="fontstyle2">13:0</span><span class="fontstyle2">, isoC</span><span class="fontstyle2">15:0, </span><span class="fontstyle2">and C</span><span class="fontstyle2">16:0</span><span class="fontstyle2">. To adapt to low temperatures, </span><span class="fontstyle2"><em>Exiguobacterium</em> </span><span class="fontstyle2">sp. HA2 upregulated expresses cold shock response including cold shock protein (</span><span class="fontstyle2">CSP</span><span class="fontstyle2">) and transcription elongation protein </span><span class="fontstyle2">NusA</span><span class="fontstyle2"><em>.</em> Also, downregulated expression of heat shock protein </span><span class="fontstyle2">DnaJ </span><span class="fontstyle2">domain protein.<br /></span><span class="fontstyle0"><strong>Conclusions:</strong> </span><span class="fontstyle2">In the current study we investigated the difference in the genotype, phenotypic, and functional characteristics of </span><span class="fontstyle2"><em>Exiguobacterium</em> </span><span class="fontstyle2">sp. HA2</span><span class="fontstyle2">. </span><span class="fontstyle2">It can be regarded as representing a novel psychrotrophic species within the genus </span><em><span class="fontstyle2">Exiguobacterium.</span></em>