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>2016
>October-December
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27920881
The Importance of Clinical Trials in Drug Development
<p>While preclinical research answers basic questions about a drug’s safety, it is not a substitute for studies of ways the drug will interact with the human body. “Clinical research” refers to studies, or trials, that are done in people <sup>1,2</sup>. As the developers design the clinical study, they will consider what they want to accomplish for each of the different Clinical Research Phases and begin the Investigational New Drug Process, a process they must go through before clinical research begins. The ultimate goal of drug development is to bring a new compound with proven therapeutic effect to the market. In this context, the transition from preclinical research to clinical stages marks a critical turning point, as it nears the new medicinal product to the market <sup>3,4</sup>. With the promise of marketing authorization, though far ahead in the road, hanging on the horizon, the approval of a clinical trial usually attracts investors and leads to a respectable rise of the company shares. However, everything comes at a price. Clinical trials are not without risks, and while the perspective of success is encouraging, the crude reality is that most compounds fail before reaching the market. As explained in previous entries, despite higher R&D expenditures, attrition rates are high and, what is worse, on the rise. Data collected between 1990 and 2004 show that the number of unsuccessful clinical trials has been steadily increasing during the last years: from 30% to 50% at Phase 1, from 40% to 70% at Phase 2 and from 20% to nearly 50% at Phase 3 <sup>3,4</sup>. As a result, less than 10% of the drugs that enter clinical trials end up being approved by regulatory agencies. Clinical trials are only a small part of the research that goes into developing a new treatment. Drugs of the future, for example, first have to be discovered or created, purified, described, and tested in labs (in cell and animal studies) before ever reaching human clinical trials. Of all the substances that are tested in these early stages, very few are promising enough to be tested in humans. Drug development is the process of bringing a new pharmaceutical drug to the market once a lead compound has been identified through the process of drug discovery. It includes pre-clinical research on microorganisms and animals, filing for regulatory status, such as via the United States Food and Drug Administration for an investigational new drug to initiate clinical trials on humans, and may include the step of obtaining regulatory approval with a new drug application to market the drug. Indeed, of every 5,000 cancer molecules identified in the laboratory, about 250 will enter pre-clinical testing. Of this 250, fewer than 10 are tested in clinical trials and on average only one will be approved by regulatory authorities. The process of bringing a new treatment from the research stage (laboratory) to clinic is estimated to take between 10-13 years<sup> 5</sup>.</p>
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https://www.ajmb.org/En/Article.aspx?id=249
https://www.ajmb.org/PDF/En/FullText/249.pdf
ShahinAkhondzadehPsychiatric Research Center, Roozbeh Hospital, South Kargar Street, Tehran, Iran739
en
27920882
Paroxetine Can Enhance Neurogenesis During Neurogenic Differentiation of Human Adipose-derived Stem Cells
<p>Background: Some antidepressant drugs can promote neuronal cell proliferation <em>in vitro</em> as well as hippocampal neurogenesis in human and animal models. Furthermore, adipose tissue is an available source of adult stem cells with the ability to differentiate in to multiple lineages. Therefore, human Adipose-Derived Stem Cells (hADSCs) may be a suitable source for regenerative medical applications. Since there is no evidence for the effect of Paroxetine as the most commonly prescribed antidepressant drug for neurogenic potential of hADSCs, an attempt was made to determine the effect of Paroxetine on proliferation and neural differentiation of hADSCs.<br />
Methods: ADSCs were isolated from human abdominal fat. These cells differentiated to neuron-like cells and were treated with Paroxetine. 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide (MTT) assay and immunofluorescence technique were used for assessment of cell proliferation and neurogenic differentiation potential of induced cells, respectively.<br />
Results: MTT assay analysis showed that Paroxetine significantly increased the proliferation rate of induced hADSCs (p<0.05), while immunofluorescent staining indicated that Paroxetine treatment during neurogenic differentiation could enhance the mean percentage of Nestin and MAP2 (Microtubule-associated protein-2) positive cells but the mean percentage of GFAP (Glial acidic fibrillary protein) positive cells significantly decreased relative to control group (p<0.05).<br />
Conclusion: Our results provide evidence that Paroxetine can promote proliferation and differentiation rate during neurogenic differentiation of hADSCs. Moreover, Paroxetine can reduce gliogenesis of induced hADSCs during neurogenic differentiation.</p>
Antidepressant drugs, Neurogenic differentiation, Paroxetine, Proliferation, Stem cells
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158
https://www.ajmb.org/En/Article.aspx?id=251
https://www.ajmb.org/PDF/En/FullText/251.pdf
MalihehJahromiDepartment of Anatomical Sciences, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran1024
ShahnazRazaviDepartment of Anatomical Sciences, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran1025
NushinAmirpourDepartment of Anatomical Sciences, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran1026
ZahraKhosravizadehDepartment of Anatomical Sciences and Molecular Biology, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran1027
en
27920883
Exogenous Secreted Frizzled-Related Protein-4 Modulates Steroidogenesis of Rat Granulosa Cells Through Wnt/Bcatenin and PI3K/AKT Signaling Pathways
<p>Background: It has been reported that secreted frizzled-related protein-4 known as an antagonist of Wnt signaling pathway plays a role in luteinization process of rodent granulosa cells. The purpose of this study was twofold: 1) to determine whether recombinant human secreted frizzled-related protein-4 (rhSFRP-4) could directly induce terminal differentiation of rat Granulosa Cells (GCs) and 2) to understand how the modulation of β-catenin and Protein Kinase B (PKB)/AKT activity by exogenous SFRP-4 could be involved in steroidogenesis.<br />
Methods: GCs were firstly stimulated with Follicle-Stimulating Hormone (FSH) named as FSH-primed cells then were treated with luteinizing hormone (LH). Then estradiol (E<sub>2</sub>) and progesterone (P<sub>4</sub>) production levels were assessed in the absence or presence of rhSFRP-4 treatment. The expression levels of activated β-catenin, pAKTser<sup>473</sup>, pGSK3βser<sup>9</sup> were assessed by western blot or immuno-fluoresence.<br />
Results: In the presence of rhSFRP-4, there was 38% decreased E<sub>2</sub> levels compared to untreated FSH-primed cells (p<0.05), and P<sub>4</sub> production subsequently decreased. However, in GCs pre-treated with rhSFRP-4 prior to addition of FSH, P<sub>4</sub> levels increased 2-fold compared with untreated cells (p<0.05). Unexpectedly, treatment with rhSFRP-4 prior to LH stimulation inhibited LH-induced P<sub>4</sub> secretion. Treatment with low (0.5 <em>ng/ml</em>) but not high (50 <em>ng/ml</em>) concentrations of rhSFRP-4 led to significantly increased levels of pGSK3βser<sup>9</sup> (1.6-fold) and nuclear active β-catenin (2.8-fold) in GCs compared with untreated cells. Interestingly, pre-treating GCs with rhsFPR4 prior to LH stimulation resulted in a 38% decrease in pAKTser<sup>473</sup> levels compared with those in LH-treated cells (p<0.05).<br />
Conclusion: Taken together, our results showed that rhSFRP-4 could directly induce terminal differentiation in GCs via the modulation of β-catenin and PKB/AKT pathways and that it does so in a dose-dependent manner.</p>
Active β-catenin, GSK3β, PKB/AKT, Rat granulosa cell, Secreted frizzled-related protein-4 (SFRP-4)
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https://www.ajmb.org/En/Article.aspx?id=252
https://www.ajmb.org/PDF/En/FullText/252.pdf
GhamartajHosseinDepartment of Animal Physiology, Developmental Biology Laboratory, Faculty of Biology, University College of Science, University of Tehran, Tehran, Iran313
ManijehKhanmohammadiReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran1028
ParissaSahranavard FardDepartment of Stem Cells & Developmental Biology at Cell Science Research Center, Royan Institute for Stem Cell Biology & Technology, ACECR, Tehran, Iran315
YasamanHeidarianDepartment of Animal Physiology, Developmental Biology Laboratory, Faculty of Biology, University College of Science, University of Tehran, Tehran, Iran1030
SomaiehKazemnejadReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran64
Mohammad MehdiAkhondiReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran13
en
27920884
Novel Combinations of Synthesized ZnO NPs and Ceftazidime: Evaluation of their Activity Against Standards and New Clinically Isolated Pseudomonas aeruginosa
<p>Background: Antibiotic resistant bacteria can be considered as a main problem in infection management. Zinc oxide nanoparticles (ZnO NPs), individually or in combination with antibiotics, can be considered as good candidates for struggling against drug resistant bacteria.<br />
Methods: In this study, Zinc oxide nanoparticles were synthesized using sol-gel method in low temperature as a cost effective procedure and characterized by X-ray diffraction and Scanning Electron Microscopy. Antibacterial activity of 9 new combinations of Zinc oxide nanoparticles and ceftazidime was assessed against standards and new clinically isolated multi drug resistant <em>Pseudomonas aeruginosa (P. aeruginosa)</em>, in order to evaluate enhancement effect of synthesized Zinc oxide nanoparticles on antibacterial activity of ceftazidime.<br />
Results: The results indicated that desirable effects can be seen at 6 and 7 <em>mM</em> of Zinc oxide nanoparticles (60 to 100% inhibition). Moreover, after evaluation of 9 new combinations with various concentrations of both components, it was demonstrated that Zinc oxide nanoparticles can enhance the antibacterial activity of ceftazidime, against some bacterial strains of <em>P. aeruginosa</em>. The highest activity was observed with the concentration of 20 <em>μg/ml</em> ceftazidime in the presence of 5, 6 or 7 <em>mM</em> of Zinc oxide nanoparticles.<br />
Conclusion: Zinc oxide nanoparticles in appropriate concentrations can be proposed as new and promising candidates for overcoming bacterial resistance.</p>
Antibiotic resistance, Ceftazidime, Pseudomonas aeruginosa, ZnO NPs
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https://www.ajmb.org/En/Article.aspx?id=253
https://www.ajmb.org/PDF/En/FullText/253.pdf
ElhamIsaeiDepartment of Microbiology, Afzalipour Faculty of Medicine, Kerman University of Medical Sciences, Kerman, Iran1031
ShahlaMansouriDepartment of Microbiology, Afzalipour Faculty of Medicine, Kerman University of Medical Sciences, Kerman, Iran1032
FereshtehMohammadiNanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran1033
SadeghTaheritarighDepartment of Plant Breeding and Biotechnology, Faculty of Plant Production, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran1123
ZohrehMohammadiNanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran1034
en
27920885
Cloning and Expression of Soluble Recombinant HIV-1 CRF35 Protease-HP Thioredoxin Fusion Protein
<p>Background: As a drug target and an antigenic agent, HIV-1 protease (HIV-1 PR) is at the center of attention for designing anti-AIDS inhibitors and diagnostic tests. In previous studies, the production of the recombinant protease has been faced with several difficulties; therefore, the aims of this study were the easy production, purification of the soluble form of protease in <em>E. coli</em> and investigation of its immunoreactivity.<br />
Methods: Protease coding region was isolated from the serum of an infected individual, amplified by RT-PCR and cloned into PTZ57R using TA-cloning. Protease coding frame was isolated by PCR and cloned in pET102/D. TOPO expression vector and cloned protease was expressed in <em>Escherichia coli (E. coli)</em> BL21. Produced recombinant protein was purified by affinity Ni-NTA column and protein concentration was checked by BCA protein assay kit. Subsequently, immunoreactivity of recombinant protease (rPR) was assayed by Western blotting and ELISA.<br />
Results: Cloning of the HIV protease by TOPO cloning system in pET102/D.TOPO was confirmed with PCR and sequencing. The concentration range of purified recombinant protein was 85 to 100<em> μg/ml</em>. Immunogenicity of rPR was confirmed by Western blotting and ELISA.<br />
Conclusion: Soluble production of recombinant HIV-1 protease (HIV-1 rPR) was performed successfully. This recombinant protein disclosed 86% specificity and 90% sensitivity in immunoassay tests.</p>
Human immunodeficiency virus, Molecular cloning, Protease, Recombinant proteins
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https://www.ajmb.org/En/Article.aspx?id=254
https://www.ajmb.org/PDF/En/FullText/254.pdf
AsaadAzarnezhadDepartment of Medical Genetics, Faculty of Medicine, Tehran University of Medical Science, Tehran, Iran1035
ZohrehSharifiBlood Transfusion Research Center, Institute for Research and Education in Transfusion Medicine, Tehran, Iran640
RahmatollahSeyedabadiDepartment of Molecular Medicine and Genetics, Faculty of Medicine, Hamedan University of Medical Sciences, Hamedan, Iran1036
ArshadHosseiniDepartment of Medical Biotechnology, Faculty of Allied Medicine, Iran University of Medical Sciences, Tehran, Iran1037
BehroozJohariDepartment of Medical Biotechnology, Faculty of Allied Medicine, Iran University of Medical Sciences, Tehran, Iran1038
MahsaSobhani FardStudent Research Center, Hamedan University of Medical Sciences, Hamedan, Iran1039
en
27920886
Evaluation of Recombinant Human Growth Hormone Secretion in E. coli using the L-asparaginase II signal Peptide
<p>Background: In the recent years, there has been an increasing interest in secretory production of recombinant proteins, due to its various advantages compared with intracellular expression. Signal peptides play a critical role in prosperous secretion of recombinant proteins. Accordingly, different signal peptides have been assessed for their ability to produce secretory proteins by trial-and-error experiments. The aim of this study was to evaluate the effect of L-asparaginase II signal peptide on the recombinant human Growth Hormone (hGH) protein secretion in the <em>Escherichia coli (E. coli)</em> host.<br />
Methods: Cloning and expression of a synthetic hGH gene, containing L-asparaginase II signal sequence was performed in <em>E. coli</em> BL21 (DE3) using 0.1 <em>mM</em> IPTG as an inducer at 23<sup>o</sup><em>C</em> overnight. Periplasmic protein extraction was performed using three methods, including osmotic shock, osmotic shock in the presence of glycine and combined Lysozyme/EDTA osmotic shock. Afterwards, the hGH expression was determined by SDS-PAGE.<br />
Results: Based on experimentally obtained results, hGH protein is expressed as inclusion body even in the presence of L-asparaginase II signal peptide.<br />
Conclusion: Therefore, this signal peptide is not effective for secretory production of the recombinant hGH.</p>
Escherichia coli (E. coli), Human growth hormone, L-asparaginase II, Recombinant proteins, Signal peptide
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https://www.ajmb.org/En/Article.aspx?id=255
https://www.ajmb.org/PDF/En/FullText/255.pdf
MozhdehZamaniDepartment of Pharmaceutical Biotechnology, Faculty of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran1040
NavidNezafatPharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz, Iran1041
YounesGhasemiDepartment of Pharmaceutical Biotechnology, Faculty of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran1042
en
27920887
Preliminary Study on Cost-effective L-Tryptophan Production from Indole and L-Serine by E. coli Cells
<p>Background: L-tryptophan is used widespread in the pharmaceutical industry. The majority of L-Trp production depends on microbial processes that produce L-tryptophan from indole and L-serine. These processes are very costly due to the costs of precursors, especially L-serine. Use of inexpensive substitutions as the L-serine source of L-tryptophan production enables us to reach a cost-effective process. In this paper, effect of Triton X-100 on L-Trp production and the ability to use Iranian cane molasses as inexpensive L-serine source was investigated.<br />
Methods: <em>Escherichia coli (E. coli)</em> <em>ATCC 11303</em> cells were grown in 10-L fermenter containing minimal medium supplemented with beet molasses as an inexpensive carbon source and indole as tryptophan synthase inducer. Whole cells of stationary phase were used as biocatalyst for L-Trp production. Triton X-100 addition to the production medium as indole reservoir was investigated. Then, cane molasses was used as L-Ser source in L-Trp production medium. Amount of L-Tryptophan and theoretical yield of L-Trp production was determined by HPLC and by a colorimetrically method on the basis of remaining indole assay, respectively.<br />
Results: As a result, triton X-100 increased L-Trp production three times. Also, the result showed that 0.68 <em>mM</em> L-Tryptophan was produced in the presence of cane molasses at 37<sup>o</sup><em>C</em> for 8 <em>hr</em>.<br />
Conclusion: This result showed that cane molasses of Qazvin sugar factory includes significant amounts of L-Ser that makes it a suitable substitution for L-Ser in L-Trp production. Therefore, it has the potential to be used for cost-effective L-Trp production in industrial scale.</p>
Molasses, PLP, Tryptophan synthase, Tryptophanase
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https://www.ajmb.org/En/Article.aspx?id=256
https://www.ajmb.org/PDF/En/FullText/256.pdf
TaherehSadeghiyan-RiziDepartment of Pharmaceutical Biotechnology, Faculty of Pharmacy, Isfahan University of Medical Sciences, Isfahan, Iran1043
JamshidFooladiNational Laboratory of Industrial Microbiology, Department of Biology, Faculty of Science, Alzahra University, Tehran, Iran1044
SimaSadraiDepartment of Pharmaceutics, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran1045
en
27920888
Development of a High-Resolution Melting Analysis Method for CYP2C19*17 Genotyping in Healthy Volunteers
<p>Background: Genetic polymorphisms of drug metabolisms by cytochrome P450 (P450s) could affect drug response, attracting particular interest in the pharm-acogenetics. Due to the importance of CYP2C19* 17 allele and its capability of super- fast metabolism and also lack of information about distribution of the alleles in Iranian population, this research aimed to use High Resolution Melting (HRM) method compared to PCR-RFLP for genotyping healthy Iranian population.<br />
Methods: Blood samples were collected from 100 healthy Iranian volunteers. DNA was extracted by salting out method. Real-time PCR was used for amplification of the CYP2C19 gene and the alleles were identified by HRM. Sequencing was used to confirm the amplified DNA fragments and data were analyzed using SPSS software ver.18.<br />
Results: The frequency of alleles CYP2C19*1/*1, CYP2C19*1/*17 and CYP2C19*17/*17 were estimated as 58.33, 29.1 and 11.1%, respectively. Specificity and sensitivity of HRM method were 90% and 100%, with respect to PCR-RFLP. Also, HRM analysis has been evaluated as a faster and more effective approach.<br />
Conclusion: Comparison of our results based on HRM analysis with PCR-RFLP showed that our developed method is rapid, accurate, fast and economic to study the CYP2C19*17 allele and it is appropriate for other similar population genetic studies.</p>
Cytochrome P-450 CYP2C19, Pharmacogenetics, Real time polymerase chain reaction
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https://www.ajmb.org/En/Article.aspx?id=257
https://www.ajmb.org/PDF/En/FullText/257.pdf
ZahraGhasemiDepartment of Genetics, Tehran Medical Sciences Branch, Islamic Azad University, Tehran, Iran1046
MehrdadHashemiDepartment of Genetics, Tehran Medical Sciences Branch, Islamic Azad University, Tehran, Iran93
MahsaEjabatiDepartment of Biotechnology, Faculty of Pharmacy, Zanjan University of Medical Sciences, Zanjan, Iran1047
Seyyed MeisamEbrahimiDepartment of Medical Surgical Nursing, Abhar Faculty of Nursing, Zanjan University of Medical Sciences, Zanjan, Iran1048
HamidrezaKheiri ManjiliDepartment of Biotechnology, Faculty of Pharmacy, Zanjan University of Medical Sciences, Zanjan, Iran1049
AliSharafiDepartment of Biotechnology, Faculty of Pharmacy, Zanjan University of Medical Sciences, Zanjan, Iran1050
AliRamazaniDepartment of Biotechnology, Faculty of Pharmacy, Zanjan University of Medical Sciences, Zanjan, Iran710
en
27920889
Lack of Association between Interleukin-12 Gene Polymorphisms and Recurrent Aphthous Stomatitis
Interleukin 12, Recurrent aphthous stomatitis, Single nucleotide polymorphisms
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201
https://www.ajmb.org/En/Article.aspx?id=258
https://www.ajmb.org/PDF/En/FullText/258.pdf
IsaacFirouze MoqadamMolecular Medicine Research Center, Hormozgan University of Medical Sciences, Bandar Abbas, Iran1052
ShamsolmouloukNajafiDental Research Center, Faculty of Dentistry, Tehran University of Medical Sciences, Tehran, Iran1053
MahsaMohammadzadehOrthodontic Department, Dental Branch, Islamic Azad University, Tehran, Iran1054
AlirezaZare BidokiMolecular Immunology Research Center, Tehran University of Medical Sciences, Tehran, Iran1055
HilaYousefiResearch Center for Immunodeficiencies, Children's Medical Center, Tehran University of Medical Sciences, Tehran, Iran1056
ElhamFarhadiDepartment of Hematology, Faculty of Allied Medical Sciences, Iran University of Medical Sciences, Tehran, Iran1057
ArghavanTonekaboniDental Research Center, Faculty of Dentistry, Tehran University of Medical Sciences, Tehran, Iran1058
GhasemMeighaniDepartment of Pediatrics, Faculty of Dentistry, Tehran University of Medical Sciences, Tehran, Iran1059
MohsenMohammadzadehDepartment of Ophthalmology, Khatam Hospital, Mashhad University of Medical Sciences, Mashhad, Iran1124
Ali AkbarAmirzargarMolecular Immunology Research Center, Tehran University of Medical Sciences, Tehran, Iran179
NimaRezaeiNetwork of Immunity in Infection, Malignancy and Autoimmunity (NIIMA), Universal Scientific Education and Research Network (USERN), Boston, USA186