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32431789
Repositioning Drugs for Psychiatry
<p>Drug development can be time-consuming and expensive. Recent estimates suggest that, on average, it takes 10 years and at least $1 billion to bring a drug to market. Since last decade, 30-40% of drugs or biologics that were approved or launched for the first time in the US were either drugs repositioned for new indications, reformulations or new combinations of existing drugs. This is the lifecycle business with repositioning as a major contributor, and it is rarely given much attention outside of its practitioners <sup>1,2</sup>.</p>
<p>In general, drug repurposing or drug repositioning alludes to the development of existing drugs or pro-drugs for new indications, not necessarily related to the original disease focus. These drugs have probably failed in late-stage clinical trials by lacking in efficacy or safety, or have problems associated with commercial strategies, patent expiration or geographic expansion. Repositioning existing drug substances for the treatment of different indications can significantly reduce the cost and time required for the development of new medicines. Therefore, drug repurposing brings forth the benefit of quickening patient access to innovative and effective treatment at lower risk and development cost for the industry <sup>1,2</sup>.</p>
<p>There are a number of different definitions of drug repurposing. All of them contain two key elements:</p>
<p>Taking existing scientific or medical knowledge and technology that is "approved" for human use in one disease or condition; and</p>
<p>Applying this knowledge and technology to another disease or condition.</p>
<p>Aspirin, a drug that’s been in use in some form or other for many hundreds of years was originally employed, and indeed still is, as a mild pain-relieving analgesic. But it’s probably more commonly used today as an antiplatelet agent helping to prevent blood clotting that can occur in thromboembolic disease.</p>
<p>Nervous system diseases represent a major health concern worldwide. Although important financial and professional investment, their etiology and pathophysiology still remain mostly elusive. Moreover, the clinical need of disease-modifying therapies is still unmet. In medicine in general and in psychiatry in particular, repositioning was the result of serendipitous but astute clinical observation of an unexpected benefit or expected or unexpected adverse effects <sup>3</sup>. A number of old drugs have been reintroduced for psychiatric indications such as celecoxib for schizophrenia, tamoxifen for mania and scopolamine for depression <sup>4-7</sup>. Drug repurposing has become a new business segment for the life science services industry. In conclusion, drug repurposing emerges as a new value proposition for the industry, patients and payers.</p>
67
67
https://www.ajmb.org/En/Article.aspx?id=20422
https://www.ajmb.org/PDF/En/FullText/20422.pdf
ShahinAkhondzadeh739
en
32431790
Bone Regeneration Using Bio-Nanocomposite Tissue Reinforced with Bioactive Nanoparticles for Femoral Defect Applications in Medicine
<p>Background: In recent years, the method of constructing and evaluating the properties of polymer nanocomposite and bioactive ceramics in tissue engineering such as biocompatible scaffolds was studied by some researchers. </p>
<p>Methods: In this study, the bio-nanocomposite scaffolds of Chitosan (CS)–Hydroxya-patite (HA)–Wllastonite (WS), incorporated with 0, 10, 20 and 30 wt% of zirconium were produced using a freeze-drying method. Also, the phase structure and morphology of scaffolds were investigated using X-ray Diffraction (XRD), Scanning Electron Microscopy (SEM) and Energy Dispersive Spectroscopy (EDS). By analyzing the SEM images, the porosity of the scaffolds was observed in the normal bone area of the body. In the next step, bioactivity and biodegradability tests of the scaffolds were carried out. Due to the presence of hydrophilic components and the high-water absorption capacity of these materials, the bio-nanocomposite scaffolds were able to absorb water properly. After that, the mechanical properties of the scaffolds were studied.</p>
<p>Results: The mechanical test results showed that the preparation of reinforced bio-nanocomposites containing 10 wt% of zirconium presented better properties compared to incorporated bio-nanocomposites with different loadings of zirconium.</p>
<p>Conclusion: According to MTT assay results, the prepared scaffolds did not have cytotoxicity at different concentrations of scaffold extracts. Consequently, the investigated scaffold can be beneficial in bone tissue engineering applications because of its similarity to natural bone structure and its proper porosity.</p>
Bone regeneration, Chitosan, Tissue engineering , Zirconium
68
76
https://www.ajmb.org/En/Article.aspx?id=20406
https://www.ajmb.org/PDF/En/FullText/20406.pdf
Mohammad AliMaghsoudlouDepartment of Pharmacology and Toxicology, AJA University of Medical Sciences, Tehran, Iran31519
EhsanNassireslami31520
SaeedSaber-SamandariNew Technologies Research Center, Amirkabir University of Technology, Tehran, Iran31580
AmirsalarKhandanNew Technologies Research Center, Amirkabir University of Technology, Tehran, Iran31581
en
32431791
Polyaniline Based Electrochemical Sensor for the Detection of Dengue Virus Infection
<p>Background: Dengue burden is increasing day-by-day globally. A rapid, sensitive, cost-effective early diagnosis kit is the need of the hour. In this study, a label-free electrochemical immunosensor was proposed for dengue virus detection. A modified Polyaniline (PANI) coated Glassy Carbon (GC) electrode, immobilized with DENV NS1 antibody was used to detect the circulating DENV NS1 antigen in both spiked and infected sample.</p>
<p>Methods: Cloning, purification and expression of DENV NS1 protein in <em>Escherichia coli</em> (<em>E. coli)</em> was performed and sensor design, PANI modification on GC electrode surface by electrochemical polymerization and immobilization of NS1 antibody on the modified electrode surface was done and finally the analytical performance of the electrochemical immunosensor was done using Cyclic Voltammetry (CV) and Electrochemical Impedance Spectroscopy (EIS).</p>
<p>Results: CV and EIS were used to study and quantitate the circulating DENV antigen. The calibration curve showed wide linearity, good sensitivity (Slope=13.8% IpR/<em>ml.ng</em><sup>-1</sup>) and distribution of data with a correlation coefficient (R) of 0.997. A lower Limit of Detection (LOD) was found to be 0.33 <em>ng.ml</em><sup>-1</sup> which encourages the applicability of the sensor.</p>
<p>Conclusion: Thus, a PANI based new electrochemical immunosensor has been developed which has the potential to be further modified for the development of cost effective, point of care dengue diagnostic kit.</p>
Dengue virus, Dielectric spectroscopy, Electrodes, Polyaniline, Voltammetry
77
84
https://www.ajmb.org/En/Article.aspx?id=10373
https://www.ajmb.org/PDF/En/FullText/10373.pdf
ReshmiDuttaDepartment of Biotechnology, Faculty of Bioengineering, SRM Institute of Science and Technology, Kattankulathur, Tamil Nadu, India31509
KThangapandiDepartment of Biotechnology, National Institute of Technology, Yupia, Papum Pare, Arunachal Pradesh, India31510
SumantraMondalDepartment of Biotechnology, Faculty of Bioengineering, SRM Institute of Science and Technology, Kattankulathur, Tamil Nadu, India31511
AmaleshNandaDepartment of Biotechnology, National Institute of Technology, Yupia, Papum Pare, Arunachal Pradesh, India31512
ShreyosiBoseDepartment of Biotechnology, Faculty of Bioengineering, SRM Institute of Science and Technology, Kattankulathur, Tamil Nadu, India31513
ShaireeSanyalDepartment of Biotechnology, Faculty of Bioengineering, SRM Institute of Science and Technology, Kattankulathur, Tamil Nadu, India31514
SaikatKumar Jana31515
SuvankarGhorai31590
en
32431792
Cysteine/Histidine-Dependent Amidohydrolase/Peptidase (CHAP)-Displayed Nano Phages: Antimicrobial Function against Methicillin-Resistant Staphylococcus aureus (MRSA)
<p>Background: Emergence and prevalence of multi drug resistance strains such as Methicillin-Resistant <em>Staphylococcus aureus</em> (MRSA) call for new antibacterial option. Endolysins as a new option is suggested. The phage display technique is suggested for production of recombinant endolysins. The recombinant endolysins displayed nano phages specifically lysis bacteria, which penetrate to the depth of tissue and the effective dose is reduced.</p>
<p>Methods: <em>CHAPK</em> gene was ligated in T7Select vector arms in T7Select10-3b cloning kit. To produce recombinant nano phages, ligation reaction was added directly to the packaging extract. Recombinant nano phages were amplified by Double Layer Agar assay (DLA). The recombinant nano phages were characterized using TEM. Size of recombinant nano phages was determined using DLS. The spot test was performed to confirm CHAPk -displayed on the surface of nano phages. The turbidimetry was used to investigate lytic activity of recombinant nano phages against MRSA ATCC No. 33591.</p>
<p>Results: The results showed recombinant nano phages belonged to order Caudovirales and family Podoviridae with titer 2×10<sup>7</sup> <em>PFU/ml</em>. According to the results of DLS, size of recombinant nano phages was 71 <em>nm</em>. Formation inhibition zone confirmed the presence of CHAPk on the surface of nano phage phenotypically. The turbidimetry showed lytic activity recombinant nano phages against MRSA after 5 <em>min</em>.</p>
<p>Conclusion: This study suggests that CHAPk -displayed nano phages can be effective in MRSA infections.</p>
Bacteriophages, Endolysin, Methicillin-resistant Staphylococcus aureus
85
90
https://www.ajmb.org/En/Article.aspx?id=20404
https://www.ajmb.org/PDF/En/FullText/20404.pdf
GolnarRahimzadehPediatric Infectious Diseases Research Center, Mazandaran University of Medical Sciences, Sari, Iran31503
PooriaGillNanomedicine Group, Immunogenetics Research Center, Mazandaran University of Medical Sciences, Sari, Iran21547
Mohammad SadeghRezai 31593
en
32431793
Design of Anti-Angiogenic Peptidomimetics and Evaluation their Biological Activity by In Vitro Assays
<p>Background: One of the important therapeutic approaches in cancer field is development of compounds which can block the initial tumor growth and the progression of tumor metastasis with no side effects. Thus, the recent study was carried out to design anti-VEGFR2-peptidomimetics as the most significant factor of angiogenesis process- and evaluate their biological activity by <em>in vitro</em> assays.</p>
<p>Methods: We designed anti-VEGFR2 peptidomimetics with anti-angiogenic activity, including compound P (lactam derivative) and compound T (indole derivative) by using in silico methods. Then, the inhibitory activity on angiogenesis was evaluated by using angiogenesis specific assays such as Human Umbilical Vein Endothelial Cell (HUVEC) proliferation, tube formation in Matrigel, MTT and Real-Time PCR. IC50 values of the compounds were also determined by cytotoxicity plot in MTT assay.</p>
<p>Results: Compounds P and T inhibited HUVEC cell proliferation and viability in a dose-dependent manner. The IC50 for compound T and compound P in HUVEC cell line were 113 and 115 <em>μg/ml</em>, respectively. Tube formation assay revealed that both compounds can inhibit angiogenesis effectively. The results of Real-Time PCR also showed these compounds are able to inhibit the expression of <em>CD31</em> gene in HUVEC cell line.</p>
<p>Conclusion: Our study suggested that compounds P and T may act as therapeutic molecules, or lead compounds for development of angiogenesis inhibitors in VEGF-related diseases.</p>
Angiogenesis inhibitors, Drug design, Peptidomimetics, Vascular endothelial growth factor receptor
91
98
https://www.ajmb.org/En/Article.aspx?id=20409
https://www.ajmb.org/PDF/En/FullText/20409.pdf
MonaGhadamDepartment of Medical Biotechnology, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran31543
SoroushSardari54
Mohammad AliShokrgozar202
MahdiyehSadat MahdaviDepartment of Medical Biotechnology, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran31595
en
32431794
Methylation Analysis of P16, RASSF1A, RPRM, and RUNX3 in Circulating Cell-Free DNA for Detection of Gastric Cancer: A Validation Study
<p>Background: Most of Gastric Cancer (GC) patients are diagnosed at an advanced stage with poor prognosis. Hypermethylations of several tumor suppressor genes in cell-free DNA of GC patients have been previously reported. In this study, an attempt was made to investigate the methylation status of <em>P16</em>, <em>RASSF1A</em>, <em>RPRM</em>, and <em>RUNX3 </em>and their potentials for early diagnosis of GC.</p>
<p>Methods: Methylation status of the four tumor suppressor genes in 96 plasma samples from histopathologically confirmed gastric adenocarcinoma patients (Stage I-IV) and 88 healthy controls was determined using methylation-specific PCR method. Receiver operating characteristic curve analysis was performed and Area Under the Curve (AUC) was calculated. Two tailed p<0.05 were considered statistically significant.</p>
<p>Results: Methylated <em>P16</em>, <em>RASSF1A</em>, <em>RPRM</em>, and <em>RUNX3</em> were significantly higher in the GC patients (41.7, 33.3, 66.7, and 58.3%) compared to the controls (15.9, 0.0, 6.8, and 4.5%), respectively (p<0.001). Stratification of patients showed that <em>RPRM</em> (AUC: 0.70, Sensitivity: 0.47, Specificity: 0.93, and p<0.001) and <em>RUNX3 </em>(AUC: 0.77, Sensitivity: 0.59, Specificity: 0.95, and p<0.001) had the highest performances in detection of early-stage (I+II) GC. The combined methylation of <em>RPRM </em>and <em>RUNX3 </em>in detection of early-stage GC had a higher AUC of 0.88 (SE=0.042; 95% CI:0.793–0.957; p<0.001), higher sensitivity of 0.82 and reduced specificity of 0.89.</p>
<p>Conclusion: Methylation analysis of <em>RPRM </em>and <em>RUNX3 </em>in circulating cell free-DNA of plasma could be suggested as a potential biomarker for detection of GC in early-stages.</p>
Biomarkers, Cell-free DNA, Gastric cancer, DNA methylation
99
106
https://www.ajmb.org/En/Article.aspx?id=10370
https://www.ajmb.org/PDF/En/FullText/10370.pdf
KioomarsSaliminejadReproductive Biotechnology Research Center, Avicenna Research Institute (ACECR), Tehran, Iran393
ShahrzadSoleymani FardHematology, Oncology and Stem Cell Transplantation Research Center, Shariati Hospital, Tehran University of Medical Sciences, Tehran, Iran92
Hamid RezaKhorram KhorshidGenetics Research Center, University of Social Welfare and Rehabilitation Sciences, Tehran, Iran, Tehran, Iran42
MarjanYaghmaeiHematology, Oncology and Stem Cell Transplantation Research Center, Shariati Hospital, Tehran University of Medical Sciences, Tehran, Iran373
HabibollahMahmoodzadehDepartment of Surgery, Cancer Institute, Imam Khomeini Hospital, Tehran, University of Medical Sciences, Tehran, Iran31506
Seyed AsadollahMousaviHematology, Oncology and Stem Cell Transplantation Research Center, Shariati Hospital, Tehran University of Medical Sciences, Tehran, Iran31507
Seyed HamidollahGhaffari31598
en
32431795
Analysis of Glioblastoma Multiforme Tumor Metabolites Using Multivoxel Magnetic Resonance Spectroscopy
<p>Background: Glioblastoma Multiforme (GBM) is the most common and deadly type of primary brain tumor in adults. Magnetic Resonance Spectroscopy (MRS) is a non-invasive imaging technique used to study metabolic changes in the brain tumors. Some metabolites such as Phosphocholine, Creatine, NAA/Cr, and Pcho/Cr have been proven to show a diagnostic role in GBM. The present study was conducted to analyze important metabolites using MRS multivoxel in GBM tumor.</p>
<p>Methods: In this study, information was collected from 8 individuals diagnosed with GBM using Siemens multivoxel MRS with a magnetic field strength of 3 T. Data were obtained by Point-Resolved Spectroscopy (PRESS) protocol with TE=135 <em>ms</em> and TR=1570 <em>ms</em>. NAA, Pcho, Cr, Ala, Gln, Gly, Glu, Lac, NAAG, and Tau metabolites were extracted and evaluated statistically.</p>
<p>Results: Given total number of normal voxels and total number of all voxels, levels of Cr, Glu, NAA, NAAG, and Gly/Tau ratio in healthy voxels were significantly higher than tumoral voxels (p=0.005, p=0.03, p<0.001, p<0.001 and p=0.041, respectively). In contrast, levels of Gly, Gln, Tau, Lac/Cr, Pcho/Cr, Pcho/NAA, Lac/NAA, and Gln/Glu ratios in tumoral voxels were significantly more than healthy voxels (p=0.001, p=0.037, p<0.001, p=0.010, p<0.001, p<0.001, and p=0.024, respectively). However, levels of Lac and Pcho had no significant difference in the two types of voxels.</p>
<p>Conclusion: In summary, compared to patients with glioblastoma with <sup>1</sup>H-MRS, the Pcho/Cr and Pcho/NAA ratios, and NAAG are the most important parameters to differentiate between tumoral and normal voxels.</p>
Glioblastoma multiform, Magnetic resonance spectroscopy, Neurochemical profiles, Voxel
107
115
https://www.ajmb.org/En/Article.aspx?id=20407
https://www.ajmb.org/PDF/En/FullText/20407.pdf
MeysamSiyah Mansoory31599
AyobFaramarziDepartment of Biomedical Engineering, Faculty of Medicine, Kermanshah University of Medical Sciences, Kermanshah, Iran31524
KarimKhoshgardDepartment of Medical Physics, Faculty of Medicine, Kermanshah University of Medical Sciences, Kermanshah, Iran31601
HadiMozafariMedical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran31602
en
32431796
Optimization of Fermentation Conditions to Enhance Cytotoxic Metabolites Production by Bacillus velezensis Strain RP137 from the Persian Gulf
<p>Background: Isolation, introduction and producing bioactive compounds from bacteria, especially marine bacteria, is an attractive research area. One of the main challenges of using these metabolites as drug and their industrialization is the optimization of production conditions.</p>
<p>Methods: In the present study, the response surface methodology was applied to optimize the production of a cytotoxic extract (C-137-R) by <em>Bacillus velezensis</em> (<em>B.</em> <em>velezensis) </em>strain RP137. Initially, among the three carbon and three nitrogen sources, rice starch and potassium nitrate were selected as the best, with cell toxicity equal to IC50=54.4 and 45.1 <em>μg/ml</em> in human lung and liver cancer cell lines, respectively (A549 and HepG2). In the next step, fractional factorial design was performed to survey effect of seven physical and chemical factors on the amount of production, and the most important factors including carbon and nitrogen sources with the positive effect and the sea salt with negative effect were determined. Finally, using the central composite design with 20 experiments, the best concentrations of rice starch and potassium nitrate (1.5%) and sea salt (1%) were obtained.</p>
<p>Results: The average amount of dried extract produced in the optimum conditions was 131.1 <em>mg/L</em> and the best response was 71.45%, which is more than 28-fold better than the pre-optimized conditions.</p>
<p>Conclusion: In general, it can be suggested that the use of modern statistical methods to optimize environmental conditions affecting the growth and metabolism of bacteria can be a highly valuable tool in industrializing the production of bioactive compounds.</p>
A549 cells, Bacillus, Industrial development, Liver neoplasms
116
123
https://www.ajmb.org/En/Article.aspx?id=20405
https://www.ajmb.org/PDF/En/FullText/20405.pdf
RoyaPournejatiMolecular Biotechnology Laboratory, Department of Biology, Faculty of Science, Shiraz University, Shiraz, Iran31517
Hamid RezaKarbalaei-Heidari31518
en
32431797
Investigation of Integron-Associated Resistance Gene Cassettes in Urinary Isolates of Klebsiella pneumoniae in Yasuj, Southwestern Iran during 2015-2016
<p>Background: Growing antibiotic resistance among urinary opportunistic pathogens such as <em>Klebsiella pneumoniae (K. pneumonia)</em> has created a worrisome condition in the treatment of the Urinary Tract Infections (UTIs) in recent years. Integrons play a significant role in the dissemination of antibiotic resistance genes. The present study was conducted to investigate class 1-3 integrons and the corresponding resistance gene cassettes in urinary <em>K. pneumoniae</em> isolates.</p>
<p>Methods: In this study, from December 2015 to September 2016, a total of 196 <em>K. pneumoniae</em> isolates were collected from the patients with UTI referred to medical diagnostic laboratories in Yasouj, Southwestern Iran. Antibiotic susceptibility patterns of isolates were determined using 12 antibiotics by the disc diffusion method. Polymerase Chain Reaction (PCR) was used for detection of integron genes (<em>intI1</em>, <em>intI2</em>, and <em>intI3</em>). The variable regions of integrons were amplified by PCR and sequenced to identify the corresponding gene cassettes.</p>
<p>Results: Thirty-nine different antibiotic resistance profiles were observed among <em>K. pneumoniae</em> isolates. Only 12.2% of <em>K. pneumoniae</em> isolates were found to harbor the <em>intI1</em> gene. While 17 (60.7%) out of 28 Multidrug Resistance (MDR) <em>K. pneumoniae </em>isolates carried the <em>intI1</em> gene, only 4.2% of non-MDR isolates harbored <em>intI1</em> gene. Totally 7 different gene cassette arrays were found in the <em>intI1</em> gene of <em>K. pneumoniae</em> isolates. The <em>aadA1 </em>was the most prominent gene cassette. Also, high frequency of <em>dfrA</em> containing gene cassettes was observed.</p>
<p>Conclusion: Continuous monitoring and characterization of integrons and their associated gene cassettes could be helpful in controlling the rising rate of antibiotic resistance.</p>
Antibiotic resistance, Integrons, Iran, Klebsiella pneumoniae
124
131
https://www.ajmb.org/En/Article.aspx?id=20408
https://www.ajmb.org/PDF/En/FullText/20408.pdf
FaribaJahanbinDepartment of Basic Sciences, Islamic Azad University, Yasuj Branch, Yasuj, Iran31536
MasoudMarashifardTreatment Management of Social Security Organization of Kohgiluyeh and Boyer-Ahmad Province, Yasuj, Iran31537
SanazJamshidiDepartment of Basic Sciences, Islamic Azad University, Yasuj Branch, Yasuj, Iran31538
MaryamZamanzadehDepartment of Basic Sciences, Islamic Azad University, Yasuj Branch, Yasuj, Iran31539
MasumehDehshiriCellular and Molecular Research Center, Yasuj University of Medical Sciences, Yasuj, Iran31540
Seyed Ali AsgharMalek HosseiniStudent Research Committee, Yasuj University of Medical Sciences, Yasuj, Iran31541
Seyed SajjadKhoramroozDepartment of Microbiology, Faculty of Medicine, Yasuj University of Medical Sciences, Yasuj, Iran1064
en
32431798
Gene Expression and Levels of TGF-B in PBMC is Associated with Severity of Symptoms in Chronic Heart Failure
<p>Background: TGF-β1 is known to promote cardiac remodeling and fibrosis during Congestive Heart Failure (CHF). In this study, an attempt was made to investigate expression of Transforming Growth Factor beta1 (TGF-β1) and relative expansion or contraction of regulatory T-cell (Tregs) population in peripheral blood of patients with Chronic Heart Failure (CHF).</p>
<p>Methods: Real-time PCR assay was used to investigate expression and post-stimulation levels of TGF-β1 in cell culture supernatant of Peripheral Blood Mononuclear Cells (PBMC) of 42 patients with CHF and 42 controls. Flow cytometry was used to identify relative counts of CD4<sup>+</sup>CD25<sup>+</sup>FoxP3<sup>+</sup> Tregs.</p>
<p>Results: PBMCs in patients with CHF expressed higher levels of TGF-β1 compared to controls. Post-stimulation levels of TGF-β1 expression were significantly higher in New York Heart Association (NYHA) functional class IV patients compared to stage I patients. Tregs were significantly expanded in PBMC in CHF, while the CD4<sup>+­</sup> helper T-cells were unchanged. Treg expansion was more significant in NYHA functional class I patients compared to class IV patients.</p>
<p>Conclusion: Expansion of Treg population in CHF provides an extrinsic source for TGF-β1 production to induce reactive fibrosis and cardiac remodeling. Relative decrease in Treg population at advanced stages of CHF is indicative of a loss of regulatory characteristics in these cells and unopposed proinflammatory milieu. </p>
Cell culture techniques, Chronic heart failure, T-lymphocytes, Transforming growth factor beta1
132
134
https://www.ajmb.org/En/Article.aspx?id=20410
https://www.ajmb.org/PDF/En/FullText/20410.pdf
SamanehSaadatiDepartment of Immunology, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran31545
VajihehEskandariCellular and Molecular Research Center, Faculty of Medicine, Guilan University of Medical Science, Rasht, Iran31546
FarzanehRahmaniNeuroImaging Network (NIN), Universal Scientific Education and Research Network (USERN), Tehran, Iran21511
Mohammad JafarMahmoudiDepartment of Cardiology, Amir Alam Hospital, Tehran University of Medical Sciences, Tehran, Iran1230
ZahraRahnemoonCardiac Heart Center, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran31547
ZahraRahmatiDepartment of Immunology, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran31548
FatemehGorzinDepartment of Immunology, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran31549
MonaHedayatDivision of Immunology, Boston Children's Hospital, Harvard Medical School, Boston, Boston, USA1229
Ali AkbarAmirzargar179
NimaRezaeiNetwork of Immunity in Infection, Malignancy and Autoimmunity (NIIMA), Universal Scientific Education and Research Network (USERN), , Boston, USA186
en
32431799
Construction, Cloning, and Expression of CagA Recombinant Protein of Helicobacter pylori
<p>Background: This study aimed to assess construction and expression of CagA recombinant protein of <em>Helicobacter pylori (H. pylori)</em> in <em>Escherichia coli</em> <em>(E. coli)</em> BL21.</p>
<p>Methods: Bioinformatics was used in designing the desired gene by Gene Runner. Next, the construct was subcloned to pET21b vector and this process was confirmed by Polymerase Chain Reaction (PCR), enzyme digestion and sequencing techniques. Then, it was cloned in the Escherichia coli BL21 as an expression host. Expression of protein was verified using sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting technique. For purification of the protein, the Ni-NTA column was used. Protein concentration was determined by the Bicinchoninic Acid Protein Assay Kit (Parstoos). Finally, Western blotting was performed using CagA antibodies and normal human serum for determining immunogenicity feature with human antiserum.</p>
<p>Results: According to the results of the present study, CagA construct was cloned into the pET21b vector and after confirmation and cloning in host expression, recombinant protein with the size of 38 <em>kDa</em> was successfully expressed and purified. The recombinant CagA protein showed immunogenicity characteristics with human antiserum.</p>
<p>Conclusion: In conclusion, only 5′-end of recombinant protein CagA with high immunogenicity effects was successfully constructed, cloned and expressed. Also, CagA recombinant protein showed good immunogenicity activity with human antiserum.</p>
CagA, Helicobacter pylori, Recombinant proteins, Vaccine candidate
135
138
https://www.ajmb.org/En/Article.aspx?id=10375
https://www.ajmb.org/PDF/En/FullText/10375.pdf
AbbasShapouri MoghaddamDepartment of Immunology, BuAli Research Institute, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran31527
ShamseddinMansouriAntimicrobial Resistance Research Center, Department of Microbiology, Ghaem Hospital, Mashhad University of Medical Sciences, Mashhad, Iran31528
AlirezaNeshaniAntimicrobial Resistance Research Center, Department of Microbiology, Ghaem Hospital, Mashhad University of Medical Sciences, Mashhad, Iran31529
FarzanehFiroozehDepartment of Microbiology, Faculty of Medicine, Alborz University of Medical Sciences, Karaj, Iran31530
AzadeMatinpurDepartment of Microbiology and Immunology, Faculty of Medicine, Kashan University of Medical Sciences, Kashan, Iran31533
AzadKhaledi31534
MehranGhazalibinaDepartment of Microbiology, Faculty of Public Health, Tehran University of Medical Sciences, Tehran, Iran31535