Inflammation-Schizophrenia: A Bidirectional Causal Association Mediated by Cytokines 2 2 No Abstract 1 2 Ahmad Shamabadi School of Medicine, Tehran University of Medical Sciences School of Medicine, Tehran University of Medical Sciences Iran Shahin Akhondzadeh Immune responseMental disorderNeuroimmunomodulationPathophysiologyPsychiatryPsychotic disorderRefractory schizophreniaSchizophrenic disorder 60522.pdf Julayanont P, Suryadevara U. Psychosis. Continuum (Minneap Minn) 2021;27(6):1682-711.##Samaei A, Moradi K, Bagheri S, Ashraf-Ganjouei A, Alikhani R, Mousavi SB, et al. Resveratrol Adjunct Therapy for Negative Symptoms in Patients With Stable Schizophrenia: A Double-Blind, Randomized Placebo-Controlled Trial. Int J Neuropsychopharmacol 2020;23(12):775-82.##Iranpour N, Zandifar A, Farokhnia M, Goguol A, Yekehtaz H, Khodaie-Ardakani MR, et al. The effects of pioglitazone adjuvant therapy on negative symptoms of patients with chronic schizophrenia: a double-blind and placebo-controlled trial. Hum Psychopharmacol 2016;31(2):103-12.##Upthegrove R, Manzanares-Teson N, Barnes NM. Cytokine function in medication-naive first episode psychosis: a systematic review and meta-analysis. Schizophr Res 2014;155(1-3):101-8.##Upthegrove R, Khandaker GM. Cytokines, oxidative stress and cellular markers of inflammation in schizophrenia. Curr Top Behav Neurosci 2020;44:49-66.##Dunleavy C, Elsworthy RJ, Upthegrove R, Wood SJ, Aldred S. Inflammation in first-episode psychosis: The contribution of inflammatory biomarkers to the emergence of negative symptoms, a systematic review and meta-analysis. Acta Psychiatr Scand 2022;146(1):6-20.##Miller BJ, Buckley P, Seabolt W, Mellor A, Kirkpatrick B. Meta-analysis of cytokine alterations in schizophrenia: clinical status and antipsychotic effects. Biol Psychiatry 2011;70(7):663-71.##Goldsmith DR, Rapaport MH, Miller BJ. A meta-analysis of blood cytokine network alterations in psychiatric patients: comparisons between schizophrenia, bipolar disorder and depression. Mol Psychiatry 2016;21(12):1696-709.##Miller BJ, Culpepper N, Rapaport MH. C-reactive protein levels in schizophrenia: a review and meta-analysis. Clin Schizophr Relat Psychoses 2014;7(4):223-30.##Park S, Miller BJ. Meta-analysis of cytokine and C-reactive protein levels in high-risk psychosis. Schizophr Res 2020;226:5-12.##Wang AK, Miller BJ. Meta-analysis of cerebrospinal fluid cytokine and tryptophan catabolite alterations in psychiatric patients: Comparisons between schizophrenia, bipolar disorder, and depression. Schizophr Bull 2018;44(1):75-83.##Arabzadeh S, Ameli N, Zeinoddini A, Rezaei F, Farokhnia M, Mohammadinejad P, et alS. Celecoxib adjunctive therapy for acute bipolar mania: a randomized, double-blind, placebo-controlled trial. Bipolar Disord 2015;17(6):606-14.##Rabbani B, Nakaoka H, Akhondzadeh S, Tekin M, Mahdieh N. Next generation sequencing: implications in personalized medicine and pharmacogenomics. Mol Biosyst 2016;12(6):1818-30.##

Opportunistic Challenges of Computer-aided Drug Discovery of Lipopeptides: New Insights for Large Molecule Therapeutics 2 2 Computer-aided drug designing is a promising approach to defeating the dry pipeline of drug discovery. It aims at reduced experimental efforts with cost-effectiveness. Naturally occurring large molecules with molecular weight higher than 500 Dalton such as cationic peptides, cyclic peptides, glycopeptides and lipopeptides are a few examples of large molecules which have successful applications as the broad spectrum antibacterial, anticancer, antiviral, antifungal and antithrombotic drugs. Utilization of microbial metabolites as potential drug candidates incur cost effectiveness through large scale production of such molecules rather than a synthetic approach. Computational studies on such compounds generate tremendous possibilities to develop novel leads with challenges to handle these complex molecules with available computational tools. The opportunities begin with the desired structural modifications in the parent drug molecule. Virtual modifications followed by molecular interaction studies at the target site through molecular modeling simulations and identification of structure-activity relationship models to develop more prominent and potential drug molecules. Lead optimization studies to develop novel compounds with increased specificity and reduced off targeting is a big challenge computationally for large molecules. Prediction of optimized pharmacokinetic properties facilitates development of a compound with lower toxicity as compared to the natural compounds. Generating the library of compounds and studies for target specificity and ADMET (Absorption, Distribution, Metabolism, Excretion and Toxicity) for large molecules are laborious and incur huge cost and chemical wastage through in-vitro methods. Hence, computational methods need to be explored to develop novel compounds from natural large molecules with higher specificity. This review article is focusing on possible challenges and opportunities in the pathway of computer-aided drug discovery of large molecule therapeutics. 3 13 Manisha Yadav Department of Biotechnology, National Institute of Technology Raipur Department of Biotechnology, National Institute of Technology Raipur India J. Satya Eswari Antifungal agentsCyclic peptidesDrug discoveryGlycopeptidesLipopeptides 60523.pdf Cragg GM, Newman DJ. Natural products: a continuing source of novel drug leads. Biochim Biophys Acta 2013 Jun 1;1830(6):3670-95.##Pham JV, Yilma MA, Feliz A, Majid MT, Maffetone N, Walker JR, et al. A review of the microbial production of bioactive natural products and biologics. Front Microbiol 2019 Jun 20;10:1404.##Bondaryk M, Staniszewska M, Zielińska P, Urbańczyk-Lipkowska Z. Natural antimicrobial peptides as inspiration for design of a new generation antifungal compounds. J Fungi (Basel) 2017 Aug 26;3(3):46##Balleza D, Alessandrini A, Beltrán García MJ. Role of lipid composition, physicochemical interactions, and membrane mechanics in the molecular actions of microbial cyclic lipopeptides. J Membr Biol 2019 Jun;252(2):131-57.##Ullman MA, Rotschafer JC. Glycopeptides, Lipopeptides, and Lipoglycopeptides. In: Drug Interactions in Infectious Diseases. Humana Press; 2011. P. 333–53.##Cochrane SA, Vederas JC. Lipopeptides from Bacillus and Paenibacillus spp.: a gold mine of antibiotic candidates. Med Res Rev 2016 Jan;36(1):4-31.##Inès M, Dhouha G. Lipopeptide surfactants: production, recovery and pore forming capacity. Peptides 2015 Sep 1;71:100-12.##Meena KR, Kanwar SS. Lipopeptides as the antifungal and antibacterial agents: applications in food safety and therapeutics. BioMed Res Int 2015 Oct;2015.##Caulier S, Nannan C, Gillis A, Licciardi F, Bragard C, Mahillon J. Overview of the antimicrobial compounds produced by members of the Bacillus subtilis group. Front Microbiol 2019 Feb 26;10:302.##Deleu M, Paquot M, Nylander T. Effect of fengycin, a lipopeptide produced by Bacillus subtilis, on model biomembranes. Biophys J 2008 Apr 1;94(7):2667-79.##Hue N, Serani L, Laprévote O. Structural investigation of cyclic peptidolipids from Bacillus subtilis by high‐energy tandem mass spectrometry. Rapid Commun Mass Spectrom 2001 Feb 15;15(3):203-9.##Kowall M, Vater J, Kluge B, Stein T, Franke P, Ziessow D. Separation and characterization of Surfactin isoforms produced byBacillus subtilisOKB 105. J Colloid Interface Sci 998 Aug 1;204(1):1-8.##Sliwoski G, Kothiwale S, Meiler J, Lowe EW. Computational methods in drug discovery. Pharmacol Rev 2014 Jan 1;66(1):334-95.##Lionta E, Spyrou G, K Vassilatis D, Cournia Z. Structure-based virtual screening for drug discovery: principles, applications and recent advances. Curr Top Med Chem 2014 Aug 1;14(16):1923-38.##Lavecchia A, Di Giovanni C. Virtual screening strategies in drug discovery: a critical review. Curr Med Chem 2013 Aug 1;20(23):2839-60.##Lau JL, Dunn MK. Therapeutic peptides: Historical perspectives, current development trends, and future directions. Bioorg Med Chem 2018 Jun 1;26(10):2700-7.##Farhadi T, Hashemian SM. Computer-aided design of amino acid-based therapeutics: A review. Drug Des Devel Ther 2018;12:1239.##Lee AC, Harris JL, Khanna KK, Hong JH. A comprehensive review on current advances in peptide drug development and design. Int J Mol Sci 2019 May 14;20(10):2383.##An S, Fu L. Small-molecule PROTACs: An emerging and promising approach for the development of targeted therapy drugs. EBio Medicine 2018 Oct;36:553-562.##Mojsoska B, Jenssen H. Peptides and peptidomimetics for antimicrobial drug design. Pharmaceuticals 2015 Jul 13;8(3):366-415.##Trosset JY, Carbonell P. Synthetic biology for pharmaceutical drug discovery. Dove Press 2015;6285-302.##Iwatsuki M, Kinoshita Y, Niitsuma M, Hashida J, Mori M, Ishiyama A, et al. Antitrypanosomal peptaibiotics, trichosporins B-VIIa and B-VIIb, produced by Trichoderma polysporum FKI-4452. J Antibiot (Tokyo) 2010 Jun;63(6):331-3.##Eswari SJ, Dhagat S, Yadav M. Computer-Aided Design of Antimicrobial Lipopeptides as Prospective Drug Candidates. CRC Press LLC; 2019. 146 p.##Peypoux F, Bonmatin JM, Wallach J. Recent trends in the biochemistry of surfactin. Appl Microbiol Biotechnol 1999 May;51(5):553-63.##Eswari JS, Dhagat S, Kaser S, Tiwari A. Homology modeling and molecular docking studies of Bacillomycin and Iturin synthetases with novel ligands for the production of therapeutic lipopeptides. Curr Drug Discov Technol 2018 Jun 1;15(2):132-41.##Gu Q, Yang Y, Yuan Q, Shi G, Wu L, Lou Z, et al. Bacillomycin D produced by Bacillus amyloliquefaciens is involved in the antagonistic interaction with the plant-pathogenic fungus Fusarium graminearum. Appl Environ Microbiol 2017 Oct 1;83(19):e01075-17.##Pirri G, Giuliani A, Nicoletto S, Pizzuto L, Rinaldi A. Lipopeptides as anti-infective: a practical perspective. Open Life Sciences 2009;4(3):258-73.##Shebley M, Polepally A, Nader A , Ng JW, Winzenborg I, Klein CE, et al. Clinical pharmacology of elagolix: an oral gonadotropin- releasing hormone receptor antagonist for endometriosis. Clin Pharmacokinet 2020 Mar;59(3):297-309.##Craik DJ, Fairlie DP, Liras S, Price D. The future of peptide-based drugs. Chem Biol Drug Des 2013 Jan;81(1):136-47.##Vlieghe P, Lisowski V, Martinez J, Khrestchatisky M. Synthetic therapeutic peptides: science and market. Drug Discov Today 2010 Jan;15(1-2):40-56.##Zhao L, Ren TH, Wang DD. Clinical pharmacology considerations in biologics development. Acta Pharmacol Sin 2012 Nov;33(11):1339-47.##O'Brien-Simpson NM, Hoffmann R, Chia CB, Wade JD. Antimicrobial and anticancer peptides. Front Chem 2018 Feb 6;6:13.##Jin G, Weinberg A. Human antimicrobial peptides and cancer. Semin Cell Dev Biol 2019 Apr;88:156-162.##Karpiński TM, Adamczak A. Anticancer activity of bacterial proteins and peptides. Pharmaceutics 2018 Apr 30;10(2):54.##Usmani S, Bedi G, Samuel JS, Singh S, Kalra S, Kumar P, et al. THPdb: Database of FDA-approved peptide and protein therapeutics. PLoS One 2017 Jul 31;12(7):e0181748.##Salmaso V, Sturlese M, Cuzzolin A, Moro S. Exploring protein-peptide recognition pathways using a supervised molecular dynamics approach. Structure 2017 Apr 4;25(4):655-662.e2.##Diller DJ, Swanson J, Bayden AS, Jarosinski M, Audie J. Rational, computer-enabled peptide drug design: Principles, methods, applications and future directions. Future Med Chem 2015;7(16):2173-93.##Verschueren E, Vanhee P, Rousseau F, Schymkowitz J, Serrano L. Protein-peptide complex prediction through fragment interaction patterns. Structure 2013 May 7;21(5):789-97.##Fair RJ, Tor Y. Antibiotics and bacterial resistance in the 21st century. Perspect Medicin Chem 2014 Aug 28;6:25-64.##Hancock RE. Cationic peptides: Effectors in innate immunity and novel Antimicrobials. Lancet Infectious Diseases 2001;1(3):156-64.##Kuppusamy R, Willcox M, Black DSC, Kumar N. Short cationic peptidomimetic antimicrobials. Antibiotics (Basel) 2019 Apr 18;8(2):44.##Jensen KJ. Peptide and protein design for biopharmaceutical applications. Wiley; 2009. 306 p.##Xu WY, Tang J, Ji CJ, He WJ, Tan NH. Application of a TLC chemical method to detection of cyclotides in plants. Chinese Science Bulletin 2008;53(11): 1671-4.##Namjoshi S, Benson HAE. Cyclic peptides as potential therapeutic agents for skin disorders. Biopolymers 2010;94(5):673-80. ##Veber DF, Johnson SR, Cheng HY, Smith BR, Ward KW, Kopple KD. Molecular properties that influence the oral bioavailability of drug candidates. J Med Chem 2002 Jun 6;45(12):2615-23. ##Renukuntla J, Vadlapudi AD, Patel A, Boddu SHS, Mitra AK. Approaches for enhancing oral bioavailability of peptides and proteins. Int J Pharm 2013 Apr 15;447(1-2):75-93.##Lawson ADG, Maccoss M, Heer JP. Importance of rigidity in designing small molecule drugs to tackle protein-protein interactions (PPIs) through stabilization of desired conformers. J Med Chem 2018 May 24;61(10):4283-9. ##Lagassé HD, Alexaki A, Simhadri VL, Katagiri NH, Jankowski W, Sauna ZE, Kimchi-Sarfaty C. Recent advances in (therapeutic protein) drug development. F1000Res 2017;6. ##Malanovic N, Lohner K. Antimicrobial peptides targeting gram-positive bacteria. Pharmaceuticals (Basel) 2016 Sep 20;9(3):59. ##Martínez de Tejada G, Sánchez-Gómez S, Rázquin-Olazaran I, Kowalski I, Kaconis Y, Heinbockel L, et al. Bacterial cell wall compounds as promising targets of antimicrobial agents I. Antimicrobial peptides and lipopolyamines. Curr Drug Targets 2012 Aug;13(9):1121-30. ##Greber KE, Ciura K, Belka M, Kawczak P, Nowakowska J, Bączeket T, al. Characterization of antimicrobial and hemolytic properties of short synthetic cationic lipopeptides based on QSAR/QSTR approach. Amino Acids 2018 Apr;50(3-4):479-485. ##Maget-Dana R, Thimon L, Peypoux F, Ptak M. Surfactin/iturin A interactions may explain the synergistic effect of surfactin on the biological properties of iturin A. Biochimie 1992 Dec;74(12):1047-51. ##Maget-Dana R, Peypoux F Iturins, a special class of pore-forming lipopeptides: biological and physicochemical properties. Toxicology 1994 Feb 28;87(1-3):151-74. ##Huttner B, Jones M, Rubin MA, Neuhauser MM, Gundlapalli A, Samore M. Drugs of last resort? The use of polymyxins and tigecycline at US veterans affairs medical centers, 2005–2010. PLoS One 2012;7(5):e36649. ##Zhao H, Yan L, Xu X, Jiang C, Shi J, Zhang Y, et al. Potential of Bacillus subtilis lipopeptides in anti-cancer I: induction of apoptosis and paraptosis and inhibition of autophagy in K562 cells. AMB Express2018 May 9;8(1):78.##Eswari JS, Dhagat S, Yadav M. Computer-Aided Design of Antimicrobial Lipopeptides as Prospective Drug Candidates. CRC Press. 2019.##Krogsgaard-Larsen P, Strømgaard K, Madsen U. Textbook of drug design and discovery. CRC Press; 2022. 452 p.##Ghasemi F, Fassihi A, Pérez-Sánchez H, Mehri Dehnavi A. The role of different sampling methods in improving biological activity prediction using deep belief network. J Comput Chem 2017 Feb 5;38(4):195-203. ##Merz KM, Ringe D, Reynolds CH. Drug design: structure- and ligand-based approaches. 1st ed. Cambridge, UK: Cambridge University Press, 2010. ##Fosgerau K, Hoffmann T. Peptide therapeutics: current status and future directions. Drug Disc Today 2015 Jan 1;20(1):122-8.##Lewis RA. The Development of Molecular Modelling Programs: The Use and Limitations of Physical Models. In: Livingstone DJ, Davis AM. Drug Design Strategies: Quantitative Approaches. 2011.p. 88-107.##Wu YS, Ngai SC, Goh BH, Chan KG, Lee LH, Chuah LH. Anticancer activities of surfactin potential application of nanotechnology assisted surfactin delivery. Front Pharmacol 2017 Oct 26;8:761.##Seydlová G, Svobodová J. Review of surfactin chemical properties and the potential biomedical applications. Cent Eur J Med 2008;3(2):123-33.##Neves BJ, Braga RC, Melo-Filho CC, Moreira-Filho JT, Muratov EN, Andrade CH. QSAR-based virtual screening: advances and applications in drug discovery. Front Pharmacol 2018 Nov 13;9:1275.##Greber KE, Zielińska J, Nierzwicki L, Ciura K, Kawczak P, Nowakowska J, et al. Are the short cationic lipopeptides bacterial membrane disruptors? Structure-Activity Relationship and molecular dynamic evaluation. Biochim Biophys Acta Biomembr 2019 Jan;1861(1):93-9.##Anderson AC. The process of structure-based drug design. Chem Biol 2003 Sep 1;10(9):787-97.##Dixon SJ, Stockwell BR. Identifying druggable disease-modifying gene products. Curr Opin Chem Biol 2009 Dec;13(5-6):549-55.##Imming P, Sinning C, MeyerA. Drugs, their targets and the nature and number of drug targets. Nat Rev Drug Discov 2006 Oct;5(10):821-34.##Chartier M, Najmanovich R. Detection of binding site molecular interaction field similarities. J Chem Inf Model 2015 Aug 24;55(8):1600-15.##Rishton GM. Nonleadlikeness and leadlikeness in biochemical screening. Drug Discov Today 2003 Jan 15;8(2):86-96.##Kirchmair J. Drug metabolism prediction. Wiley-VCH; 2014.##Walters WP. Going further than Lipinski’s rule in drug design. Expert Opin Drug Discov 2012 Feb;7(2):99-107.##Wermuth CG, Aldous D, Raboisson P, Rognan D. The Practice of Medicinal Chemistry. 4th ed. Elsevier; 2015.##Nicolaou CA, Brown N. Multi-objective optimization methods in drug design. Drug Discov Today Technol 2013 Sep;10(3):e427-35. ##Ban TA. The role of serendipity in drug discovery. Dialogues in clinical Neuroscience. Dialogues Clin Neurosci 2006;8(3):335–44. ##Vamathevan J, Clark D, Czodrowski P, Dunham I, Ferran E, Lee G, et al. Applications of machine learning in drug discovery and development. Nat Rev Drug Discov 2019 Jun;18(6):463-77. ##Aungst BJ. Optimizing oral bioavailability in drug discovery: an overview of design and testing strategies and formulation options. Journal of pharmaceutical sciences 2017;106(4):921-9. ##Wermuth CG. Analogues as a Means of Discovering New Drugs. In: Fischer J, Ganellin CR. Analogue-Based Drug Discovery. Wiley; 2006.##IUPAC, Fischer J, Ganellin CR. Analogue-Based Drug Discovery. Wiley; 2006. 575 p.##Ciemny M, Kurcinski M, Kamel K, Kolinski A, Alam N, Furman OS, et al. Protein–peptide docking: opportunities and challenges. Drug Discov Today 2018 Aug;23(8):1530-7.##Liu T, Pan X, Chao L, Tan W, Qu S, Yang L, et al. Subangstrom accuracy in pHLA-I modeling by rosetta FlexPepDock refinement protocol. J Chem Inf Model 2014 Aug 25;54(8):2233-42. ##Lima AH, dos Santos AM, Alves CN, Lameira J. Computed insight into a peptide inhibitor preventing the induced fit mechanism of MurA enzyme from Pseudomonas aeruginosa. Chem Biol Drug Des 2017 Apr;89(4):599-607. ##Marcu O, Dodson EJ, Alam N, Sperber M, Kozakov D, Lensink MF, et al. FlexPepDock lessons from CAPRI peptide–protein rounds and suggested new criteria for assessment of model quality and utility. Proteins 2017 Mar;85(3):445-62. ##Shan Y, Kim ET, Eastwood MP, Dror RO, Seeliger MA, Shaw DE. How does a drug molecule find its target binding site? J Am Chem Soc 2011 Jun 22;133(24):9181-3. ##Durrant JD, Keränen H, Wilson BA, McCammon JA. Computational identification of uncharacterized cruzain binding sites. PLoS Negl Trop Dis 2010 May 11;4(5):e676. ##Hazuda DJ, Anthony NJ, Gomez RP, Jolly SM, Wai JS, Zhuang L, et al. A naphthyridine carboxamide provides evidence for discordant resistance between mechanistically identical inhibitors of HIV-1 integrase. Proc Natl Acad Sci USA 2004 Aug 3;101(31):11233-8. ##Durrant JD, McCammon JA. Molecular dynamics simulations and drug Discovery. BMC Biol 2011 Oct 28;9:71. ##Li T, Froeyen M, Herdewijn P. Computational alanine scanning and free energy decomposition for E. coli type I signal peptidase with lipopeptide inhibitor Complex. J Mol Graph Model 2008 Jan;26(5):813-23.##Sur S, Romo TD, Grossfield A. Selectivity and mechanism of fengycin, an antimicrobial lipopeptide, from molecular dynamics. J Phys Chem B 2018 Mar 1;122(8):2219-26.##Singh SS. Preclinical pharmacokinetics: an approach towards safer and efficacious drugs. Curr Drug Metab 2006 Feb;7(2):165-82.##

An Anti-TAZ Monoclonal Antibody Recognizing Cell Surface Expressed TAZ Protein in Human Tumor Cells 2 2 Background: WWTR1 or TAZ is a transcriptional co-activator protein expressed in cytoplasm which functions as the main downstream effector of the Hippo signaling pathway. This pathway is an evolutionally conserved signal cascade, which plays a pivotal role in organ size control and tumorigenesis. Ectopic expression of TAZ has already been observed in many malignancies, while the ectopic localization of TAZ is reported for the first time. The aim of this study was to produce a specific monoclonal antibody (mAb) against a synthetic peptide derived from WWTR1 protein to be used as a research tool in human carcinomas. Methods: A 21-mer synthetic peptide (derived from human TAZ protein) was used for immunization of BALB/c mice after conjugation with Keyhole Limpet Haemocyanin (KLH) using hybridoma technology. The generated mAb reacted with the immunizing peptide employing ELISA assay. The reactivity of the antibody with native TAZ protein was assessed through Western blot, immunocytochemistry, and flow cytometry using different cancer cell lines. Results: The produced mAb could recognize the immunizing peptide in ELISA and Kaff was 0.6×10-9 M. The produced anti-TAZ mAb unlike available commercial anti-TAZ antibody, was capable of specifically recognizing cell surface TAZ in human carcinoma cell lines including MCF-7, Raji, and A431 in Western blot, immunocytochemistry, and flow cytometry assays. As expected, no reactivity was observed using normal Peripheral Blood Mononuclear Cell (PBMC) from healthy donors. Conclusion: Based on the results, TAZ is ectopically expressed on the surface of tumor cell lines which is not the case in normal cells. The generated mAb has a potential to be used as a research tool in studying the expression of TAZ in human carcinomas in different applications. 14 20 Mozhan Haji Ghaffari Department of Medical Biotechnology, School of Advanced Science in Medicine, Tehran University of Medical Science Department of Medical Biotechnology, School of Advanced Science in Medicine, Tehran University of Medical Science Iran Mahsa Mohammadzadeh Department of Genetics, Islamic Azad University, Tehran Medical Sciences Branch Department of Genetics, Islamic Azad University, Tehran Medical Sciences Branch Iran Miganoosh Simonian Department of Medical Biotechnology, School of Advanced Science in Medicine, Tehran University of Medical Science Department of Medical Biotechnology, School of Advanced Science in Medicine, Tehran University of Medical Science Iran Mehrdad Hashemi Department of Genetics, Islamic Azad University, Tehran Medical Sciences Branch Department of Genetics, Islamic Azad University, Tehran Medical Sciences Branch Iran Niloufar Sadeghi Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Babak Negahdari Department of Medical Biotechnology, School of Advanced Science in Medicine, Tehran University of Medical Science Department of Medical Biotechnology, School of Advanced Science in Medicine, Tehran University of Medical Science Iran Mohammadali Mazloomi Hodjattallah Rabbani CarcinomaHumansMonoclonal antibodyTAZ (WWTR1) 60524.pdf Available from: https://www.who.int/news-room/fact-sheets/detail/cancer.##Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 2021;71(3):209-49.##Henry NL, Hayes DF. Cancer biomarkers. Mol Oncol 2012;6(2):140-6.##Wang K, Degerny C, Xu M, Yang XJ. YAP, TAZ, and Yorkie: a conserved family of signal-responsive transcriptional coregulators in animal development and human disease. Biochem Cell Biol 2009;87(1):77-91.##Kanai F, Marignani PA, Sarbassova D, Yagi R, Hall RA, Donowitz M, et al. TAZ: a novel transcriptional co-activator regulated by interactions with 14-3-3 and PDZ domain proteins. EMBO J 2000;19(24):6778-91.##Liu H, Du S, Lei T, Wang H, He X, Tong R, et al. Multifaceted regulation and functions of YAP/TAZ in tumors. Oncol Rep 2018;40(1):16-28.##Noguchi S, Saito A, Horie M, Mikami Y, Suzuki HI, Morishita Y, et al. An integrative analysis of the tumorigenic role of TAZ in human non–small cell lung cancer. Clin Cancer Res 2014;20(17):4660-72.##Maugeri-Saccà M, Barba M, Pizzuti L, Vici P, Di Lauro L, Dattilo R, et al. The Hippo transducers TAZ and YAP in breast cancer: oncogenic activities and clinical implications. Expert Rev Mol Med 2015;17:e14.##Wang L, Shi S, Guo Z, Zhang X, Han S, Yang A, et al. Overexpression of YAP and TAZ is an independent predictor of prognosis in colorectal cancer and related to the proliferation and metastasis of colon cancer cells. PloS One 2013;8(6):e65539.##Zhang S, Zhou D. Role of the transcriptional coactivators YAP/TAZ in liver cancer. Curr Opin Cell Biol 2019;61:64-71.##Köhler G, Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 1975;256(5517):495-7.##Bayat AA, Sadeghi N, Salimi A, Fazli G, Nowroozi MR, Moghadam SO, et al. The association of cell surface fibromodulin expression and bladder carcinoma. Urol J 2021;19(3):189-95.##Farahi L, Ghaemimanesh F, Milani S, Razavi SM, Bayat AA, Rabbani H, et al. Monoclonal and polyclonal antibodies specific to human fibromodulin. Iran J Biotechnol 2019 11;17(1):e2277.##Farahi L, Ghaemimanesh F, Milani S, Razavi SM, Hadavi R, Bayat AA, et al. GPI-anchored fibromodulin as a novel target in chronic lymphocytic leukemia: diagnostic and therapeutic implications. Iran J Immunol 2019;16(2):127-41.##Lu YP, Ishiwata T, Kawahara K, Watanabe M, Naito Z, Moriyama Y, et al. Expression of lumican in human colorectal cancer cells. Pathol Int 2002;52(8):519-26.##Pobbati AV, Hong W. A combat with the YAP/TAZ-TEAD oncoproteins for cancer therapy. Theranostics 2020;10(8):3622-35.##Zhang H, Liu CY, Zha Z-Y, Zhao B, Yao J, Zhao S, et al. TEAD transcription factors mediate the function of TAZ in cell growth and epithelial-mesenchymal transition. J Biol Chem 2009;284(20):13355-62.##

Proteome Analysis of Adult Acute Lymphoblastic Leukemia by Two-dimensional Blue Native/Sodium Dodecyl Sulfate Gel Electrophoresis 2 2 Background: Despite the significant progress in the treatment of Acute Lymphoblastic Leukemia (ALL) in children, it still remains as one of the most challenging malignancies in adults. Identification of new biomarkers may improve the management of adult ALL. Proteins expressed on the cell surface can be considered as disease-associated biomarkers with potential for diagnosis and targeted therapies. Thus, membrane proteome studies give essential information about the disease-related biomarkers. Methods: We applied 2-dimensional blue-native SDS-PAGE technique followed by MALDI-TOF/TOF-mass spectrometry to study the cell membrane proteome of peripheral blood mononuclear cells of adult B-ALL patients in comparison to that of the healthy controls. Results: Sixty seven differentially expressed protein spots were detected, among them 52 proteins were found to be up-regulated but the other 15 proteins were down-regulated in B-ALL. Five differentially expressed proteins, involved in energy metabolism pathways, were detected in B-ALL patients compared to the healthy control group. Conclusion: Differentially expressed proteins provide an insight into the molecular biology of B-ALL. Further studies must be done to confirm our data to be considered as potential targets for detection and treatment of B-ALL. 21 27 Servin Bagheralmoosavi Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences Iran Parastou Gholami Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences Iran Mahdi Amini Integrative Oncology Department, Breast Cancer Research Center, Motamed Cancer Institute, ACECR Integrative Oncology Department, Breast Cancer Research Center, Motamed Cancer Institute, ACECR Iran Mahdi Alizadeh Integrative Oncology Department, Breast Cancer Research Center, Motamed Cancer Institute, ACECR Integrative Oncology Department, Breast Cancer Research Center, Motamed Cancer Institute, ACECR Iran Marjan Yaghmaei Hematology, Oncology and Stem Cell Transplantation Research Center, Tehran University of Medical Sciences Hematology, Oncology and Stem Cell Transplantation Research Center, Tehran University of Medical Sciences Iran Sahar Tavakkoli Hematology, Oncology and Stem Cell Transplantation Research Center, Tehran University of Medical Sciences Hematology, Oncology and Stem Cell Transplantation Research Center, Tehran University of Medical Sciences Iran Sina Salari HSCT Research Center, Shahid Beheshti University of Medical Sciences HSCT Research Center, Shahid Beheshti University of Medical Sciences Iran Mahmood Jeddi-Tehrani Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Alireza Ghasempour Integrative Oncology Department, Breast Cancer Research Center, Motamed Cancer Institute, ACECR Integrative Oncology Department, Breast Cancer Research Center, Motamed Cancer Institute, ACECR Iran Kambiz Gilany Mahdi Shabani Acute lymphoblastic leukemiaBiomarkersChildMassMatrix-assisted laser desorption-ionizationMolecular biologyProteomeSpectrometry 60526.pdf Terwilliger T, Abdul-Hay M. Acute lymphoblastic leukemia: a comprehensive review and 2017 update. Blood Cancer J 2017;7(6):e577.##Siegel RL, Miller KD, Jemal A. Cancer statistics, 2019. CA Cancer J Clin 2019 Jan;69(1):7-34.##Harrison CJ. Acute lymphoblastic leukemia. Clin Lab Med 2011;31(4):631-47 (2011).##Campo E, Swerdlow SH, Harris NL, Pileri S, Stein H, Jaffe ES. The 2008 WHO classification of lymphoid neoplasms and beyond: evolving concepts and practical applications. Blood 2011 May 12;117(19):5019-32.##Samra B, Jabbour E, Ravandi F, Kantarjian H, Short NJ. Evolving therapy of adult acute lymphoblastic leuke-mia: state-of-the-art treatment and future directions. J He-matol Oncol 2020;13:70.##Short NJ, Kantarjian H, Jabbour E. Optimizing the treatment of acute lymphoblastic leukemia in younger and older adults: new drugs and evolving paradigms. Leukemia 2021 Nov;35(11):3044-58.##Elschenbroich S, Kim Y, Medin JA, Kislinger T. Isolation of cell surface proteins for mass spectrometry-based proteomics. Expert Rev Proteomics 2010 Feb 1;7(1):141-54.##Wu CC, Yates JR. The application of mass spectrometry to membrane proteomics. Nat Biotechnol 2003 Mar;21(3):262-7.##Saha S, Banerjee S, Banerjee D, Chandra S, Chakrabarti A. 2DGE and DIGE based proteomic study of malignant B-cells in B-cell acute lymphoblastic leukemia. EuPA Open Proteomics 2014 Jun 1;3:13-26.##Wang D, Lv YQ, Liu YF, Du XJ, Li B. Differential protein analysis of lymphocytes between children with acute lymphoblastic leukemia and healthy children. Leuk Lymphoma 2013 Feb 1;54(2):381-6.##Hu J, Lin M, Liu T, Li J, Chen B, Chen Y. DIGE-based proteomic analysis identifies nucleophosmin/B23 and nucleolin C23 as over-expressed proteins in relapsed/refractory acute leukemia. Leuk Res 2011 Aug 1;35(8):1087-92.##Cui JW, Wang J, He K, Jin BF, Wang HX, Li W, et al. Proteomic analysis of human acute leukemia cells: insight into their classification. Clin Cancer Res 2004 Oct 15;10(20):6887-96.##Braoudaki M, Lambrou GI, Vougas K, Karamolegou K, Tsangaris GT, Tzortzatou-Stathopoulou F. Protein biomarkers distinguish between high-and low-risk pediatric acute lymphoblastic leukemia in a tissue specific manner. J Hematol Oncol 2013 Dec;6(1):1-20.##Alberts B. Molecular biology of the cell. WW Norton & Company; 2017 Aug 7.##Schägger H, von Jagow G. Blue native electrophoresis for isolation of membrane protein complexes in enzymatically active form. Anal Biochem 1991 Dec 1;199(2):223-31.##Wittig I, Schägger H. Features and applications of blue‐native and clear‐native electrophoresis. Proteomics 2008 Oct;8(19):3974-90.##Burns CP, Armitage JO, Frey AL, Dick FR, Jordan JE, Woolson RF. Analysis of the presenting features of adult acute leukemia: The French‐American‐British classification. Cancer 1981 May 15;47(10):2460-9.##Wittig I, Braun HP, Schägger H. Blue native PAGE. Nature Protoc 2006 Jun;1(1):418-28.##Schägger H. Tricine–sds-page. Nature Protoc 2006 Jun;1(1):16-22.##Shevchenko A, Tomas H, Havli J, Olsen JV, Mann M. In-gel digestion for mass spectrometric characterization of proteins and proteomes. Nat Protoc 2006 Dec;1(6):2856-60.##Koppenol WH, Bounds PL, Dang CV. Otto Warburg's contributions to current concepts of cancer metabolism. Nat Rev Cancer 2011 May;11(5):325-37.##Denko NC. Hypoxia, HIF1 and glucose metabolism in the solid tumour. Nat Rev Cancer 2008 Sep;8(9):705-13.##DeBerardinis RJ, Chandel NS. We need to talk about the Warburg effect. Nat Metab 2020 Feb;2(2):127-9.##Boag JM, Beesley AH, Firth MJ, Freitas JR, Ford J, Hoffmann K, et al. Altered glucose metabolism in childhood pre-B acute lymphoblastic leukaemia. Leukemia 2006 Oct;20(10):1731-7.##Miccheli A, Tomassini A, Puccetti C, Valerio M, Peluso G, Tuccillo F, et al. Metabolic profiling by 13C-NMR spectroscopy: glucose reveals a heterogeneous metabolism in human leukemia T cells. Biochimie 2006 May;88(5):437-48.##Tiefenthaler M, Amberger A, Bacher N, Hartmann BL, Margreiter R, Kofler R, Konwalinka G. Increased lactate production follows loss of mitochondrial membrane potential during apoptosis of human leukaemia cells. Br J Haematol 2001 Sep;114(3):574-80.##Handschuh L, Kaźmierczak M, Milewski MC, Góralski M, Łuczak M, Wojtaszewska M, et al. Gene expression profiling of acute myeloid leukemia samples from adult patients with AML-M1 and -M2 through boutique microarrays, real-time PCR and droplet digital PCR. Int J Oncol 2018 Mar;52(3):656-78.##Altman BJ, Stine ZE, Dang CV. From Krebs to clinic: glutamine metabolism to cancer therapy. Nat Rev Cancer 2016 Nov;16(11):749.##Hosios A, Hecht VC, Danai LV, Johnson MO, Rathmell JC, Steinhauser ML, et al. Amino acids rather than glucose account for the majority of cell mass in proliferating mammalian cells. Dev Cell 2016 Mar 7;36(5):540-9.##Craze ML, El-Ansari R, Aleskandarany MA, Cheng KW, Alfarsi L, Masisi B, et al. Glutamate dehydrogenase (GLUD1) expression in breast cancer. Breast Cancer Res Treat 2019 Feb;174(1):79-91.##Pajic T, Cernelc P, Briski AS, Lejko-Zupanc T, Malesic I. Glutamate dehydrogenase activity in lymphocytes of B-cell chronic lymphocytic leukaemia patients. Clin Biochem 2009 Nov;42(16-17):1677-84.##Wang Y, Yu L, Ding J, Chen Y. Iron metabolism in cancer. Int J Mol Sci 2018 Dec 27;20(1):95.##Carpentieri U, Myers J, Thorpe L, Daeschner III CW, Haggard ME. Copper, zinc, and iron in normal and leukemic lymphocytes from children. Cancer Res 1986 Feb;46(2):981-4.##Smilevska T, Stamatopoulos K, Samara M, Belessi C, Tsompanakou A, Paterakis G, et al. Transferrin receptor-1 and 2 expression in chronic lymphocytic leukemia. Leuk Res 2006 Feb;30(2):183-9.##Tei I, Makino Y, Sakagami H, Kanamaru I, Konno K. Decrease of transferrin receptor during mouse myeloid leukemia (Ml) cell differentiation. Biochem Biophys Res Commun 1982 Aug 31;107(4):1419-24.##Hagag AA Badraia IM, Abdelmageed MM, Hablas NM, Hazzaa SME, Nosair NA. Prognostic value of transferrin receptor-1 (CD71) expression in acute lymphoblastic leukemia study of beta globin gene mutations in Egyptian children with β-thalassemia. Endocr Metab Immune Disord Drug Targets 2018;18(6):610-617.##Zhang X, Long Q. Elevated serum plasma fibrinogen is associated with advanced tumor stage and poor survival in hepatocellular carcinoma patients. Medicine (Baltimore) 2017 Apr;96(17):e6694.##Luo Y, Kim HS, Kim M, Lee M, Song YS. Elevated plasma fibrinogen levels and prognosis of epithelial ovarian cancer: A cohort study and meta-analysis. J Gynecol Oncol 2017 May;28(3):e36.##Yu X, Hu F, Yao Q, Li C, Zhang H, Xue Y. Serum fibrinogen levels are positively correlated with advanced tumor stage and poor survival in patients with gastric cancer undergoing gastrectomy: A large cohort retrospective study. BMC Cancer 2016 Jul 14;16:480.##Wang P, Meng Z , Song HW, Yao K , Duan ZJ, Li SW, et al. Higher plasma fibrinogen levels are associated with malignant phenotype and worse survival in patients with glioblastomas. J Cancer 2018 Apr 30;9(11):2024-2029.##Hunault-Berger M, Chevallier P, Delain M, Bulabois CE, Bologna S, Bernard M, et al. Changes in antithrombin and fibrinogen levels during induction chemotherapy with L-asparaginase in adult patients with acute lymphoblastic leukemia or lymphoblastic lymphoma. Use of supportive coagulation therapy and clinical outcome: the CAPELAL study. Haematologica 2008 Oct;93(10):1488-94.##

Optimization of Degenerate PCR Conditions for Reducing Error Rates in Detection of PKS and NRPS Gene groups in Actinomycetes 2 2 Background: The screen of Polyketide Synthase (PKS) and Nonribosomal Peptide Synthetase (NRPS) gene groups is a quick way to discover new therapeutic agents. However, errors in laboratory techniques cause a loss of touch with reality. This study aimed to evaluate the presence of PKS and NRPS gene groups in previously isolated strains by optimizing their specialized amplification by degenerate primers and indicating the evolutionary relationships with reference strains. Methods: PKS-I, II, and NRPS genes PCR amplification was performed using three degenerate primer sets for 22 actinomycete strains with antibacterial activity. Annealing temperature and the amount of template DNA and primers were optimized. PCR products of PKS-I, II, and NRPS from three strains were sequenced after TA cloning. Besides, strains with high antibacterial activity were identified by biochemical features and partial 16S rDNA sequencing and hypothetically classified by a phylogenetic tree. Results: High frequencies of PKS-I (86.4%), PKS-II (81.8%), and NRPS (95.4%) genes were found among the strains after optimization. Fourteen strains (64%) contained all of the genes, and 100% of strains had at least two genes. These numbers are pretty distinct in comparison with the previous researches. All of the sequenced strains were members of Streptomyces genus. Conclusion: Our research showed that degenerate PCR strongly depends on the variation of annealing temperature and primer concentration, resulting in an unexpected shift in PCR outputs. The sequencing results confirmed the optimized conditions for specialized PCR of PKS-I, PKS-II, and NRPS gene groups. 28 37 Sara Ghashghaei Department of Cell and Molecular Biology & Microbiology, Faculty of Biological Science and Technology, University of Isfahan Department of Cell and Molecular Biology & Microbiology, Faculty of Biological Science and Technology, University of Isfahan Iran Zahra Etemadifar Manoochehr Tavassoli Department of Cell and Molecular Biology & Microbiology, Faculty of Biological Science and Technology, University of Isfahan, Isfahan Department of Cell and Molecular Biology & Microbiology, Faculty of Biological Science and Technology, University of Isfahan, Isfahan Iran Mohammad Reza Mofid Department of Biochemistry, Isfahan Pharmaceutical Sciences Research Center and Bioinformatics Research Center, School of Pharmacy, Isfahan University of Medical Sciences Department of Biochemistry, Isfahan Pharmaceutical Sciences Research Center and Bioinformatics Research Center, School of Pharmacy, Isfahan University of Medical Sciences Iran Antimicrobial peptidesBiological productsPCRPolyketide synthaseStreptomyces 60528.pdf Lee LH, Zainal N, Azman AS, Eng SK, Goh BH, Yin WF, et al. Diversity and antimicrobial activities of Actinobacteria isolated from tropical mangrove sediments in Malaysia. ScientificWorldJournal 2014;2014:698178.##Li J, Zhao GZ, Chen HH, Wang HB, Qin S, Zhu WY, et al. Antitumour and antimicrobial activities of endophytic streptomycetes from pharmaceutical plants in rainforest. Lett Appl Microbiol 2008;47(6):574-80.##Gao P, Huang Y. Detection, distribution, and organohalogen compound discovery implications of the reduced flavin adenine dinucleotide-dependent halogenase gene in major filamentous actinomycete taxonomic groups. Appl Environ Microbiol 2009;75(14):4813-20.##Ehrenreich IM, Waterbury JB, Webb EA. Distribution and diversity of natural product genes in marine and freshwater cyanobacterial cultures and genomes. Appl Environ Microbiol 2005;71(11):7401-13.##Wang H, Fewer DP, Holm L, Rouhiainen L, Sivonen K. Atlas of nonribosomal peptide and polyketide biosynthetic pathways reveals common occurrence of nonmodular enzymes. Proc Natl Acad Sci USA (PNAS) 2014;111(25):9259-9264.##Amoutzias GD, Chaliotis A, Mossialos D. Discovery strategies of bioactive compounds synthesized by nonribosomal peptide synthetases and type-I polyketide synthases derived from marine microbiomes. Mar drugs 2016;14(4):80.##Giessen TW, Marahiel MA. Ribosome-independent biosynthesis of biologically active peptides: Application of synthetic biology to generate structural diversity. FEBS Lett 2012;586(15):2065-75.##Felnagle EA, Jackson EE, Chan YA, Podevels AM, Berti AD, McMahon MD, et al. Nonribosomal peptide synthetases involved in the production of medically relevant natural products. Mol Pharm 2008;5(2):191-211.##Shen B. Polyketide biosynthesis beyond the type I, II and III polyketide synthase paradigms. Curr Opin Chem Biol 2003;7(2):285-95.##Seow KT, Meurer G, Gerlitz M, Wendt-Pienkowski E, Hutchinson CR, Davies J. A study of iterative type II polyketide synthases, using bacterial genes cloned from soil DNA: A means to access and use genes from uncultured microorganisms. J Bacteriol 1997;179(23):7360-8.##Yu D, Xu F, Zeng J, Zhan J. Type III polyketide synthases in natural product biosynthesis. IUBMB life 2012;64(4):285-95.##Hillenmeyer ME, Vandova GA, Berlew EE, Charkoudian LK. Evolution of chemical diversity by coordinated gene swaps in type II polyketide gene clusters. Proc Natl Acad Sci USA (PNAS) 2015;112(45):13952-7.##Ghashghaei S, Etemadifar Z, Mofid MR. Studies on the kinetics of antibacterial agent production in two actinomycete strains, F9 and Is5, isolated from soil samples, Iran. Iran J Sci Technol Trans Sci 2018;42(3):1139-47.##Mesapogu S, Jillepalli CM, Arora DK. Microbial DNA extraction, purification, and quantitation. In: Arora DK, Das S, Sukumar M (eds). Analyzing microbes: manual of molecular biology techniques. Berlin Heidelberg: Springer-Verlag; 2013, 1-16.##Barsotti O, Renaud F, Freney J, Benay G, Decoret D, Dumont J. Rapid isolation of DNA from Actinomyces. Ann Inst Pasteur Microbiol 1987;138(5):529-36.##Ayuso-Sacido A, Genilloud O. New PCR primers for the screening of NRPS and PKS-I systems in actinomycetes: detection and distribution of these biosynthetic gene sequences in major taxonomic groups. Microb Ecol 2005;49(1):10-24.##Meklat A, Sabaou N, Zitouni A, Mathieu F, Lebrihi A. Isolation, taxonomy, and antagonistic properties of halophilic actinomycetes in Saharan soils of Algeria. Appl Environ Microbiol 2011;77(18):6710-4.##Li J, Dong JD, Yang J, Luo XM, Zhang S. Detection of polyketide synthase and nonribosomal peptide synthetase biosynthetic genes from antimicrobial coral-associated actinomycetes. Antonie van Leeuwenhoek 2014;106(4):623-35.##Gupta P, Samant K, Sahu A. Isolation of cellulose-degrading bacteria and determination of their cellulolytic potential. Int J Microbiol 2012;2012:578925.##Poosarla A, Ramana LV, Krishna RM. Isolation of potent antibiotic producing actinomycetes from marine sediments of Andaman and Nicobar Marine Islands. J Microbiol Antimicrob 2013;5(1):6-12.##Sujatha P, Bapi Raju KVVSN, Ramana T. Studies on a new marine streptomycete BT-408 producing polyketide antibiotic SBR-22 effective against methicillin resistant Staphylococcus aureus. Microbiol Res 2005;160(2):119-26.##Fulzele R, DeSa E, Yadav A, Shouche Y, Bhadekar R. Characterization of novel extracellular protease produced by marine bacterial isolate from the Indian Ocean. Braz J Microbiol 2011;42(4):1364-73.##Harrigan WF. Laboratory methods in food microbiology. New York: Academic Press; 1998.##Atlas RM. Handbook of media for environmental microbiology. Boca Raton (FL): CRC Press; 2005.##Oceguera-Cervantes A, Carrillo-García A, López N, Bolaños-Nuñez S, Cruz-Gómez MJ, Wacher C, et al. Characterization of the polyurethanolytic activity of two Alicycliphilus sp. strains able to degrade polyurethane, and N-methylpyrrolidone. Appl Environ Microbiol 2007;73(19):6214-23.##Frank JA, Reich CI, Sharma S, Weisbaum JS, Wilson BA, Olsen GJ. Critical evaluation of two primers commonly used for amplification of bacterial 16S rRNA genes. Appl Environ Microbiol 2008;74(8):2461-70.##Méndez-Vilas A. Industrial, medical and environmental applications of microorganisms. Current status and trends. The Netherlands: Wageningen Academic Publishers; 2014.##Gökçekus H, Türker U, LaMoreaux JW. Survival and sustainability: Environmental concerns in the 21st century. Berlin Heidelberg: Springer-Verlag; 2011.##Leroy PD, Sabri A, Heuskin S, Thonart P, Lognay G, Verheggen FJ, et al. Microorganisms from aphid honeydew attract and enhance the efficacy of natural enemies. Nat Commun 2011;2(1):348-54.##Ishikawa J, Tsuchizaki N, Yoshida M, Ishiyama D, Hotta K. Colony PCR for detection of specific DNA sequences in actinomycetes. Actinomycetologica 2000;14(1):1-5.##Ronquist F, Huelsenbeck JP. MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics 2003;19(12):1572-4.##Nylander JAA. MrModeltest, version 2. Program distributed by the author. Evolutionary Biology Centre, Uppsala University; 2004. http://www.abc.se/~nylander/mrmodeltest2/mrmodeltest2.html##Stöver BC, Müller KF. TreeGraph 2: Combining and visualizing evidence from different phylogenetic analyses. BMC Bioinform 2010;11(1):1-9.##Qin S, Li J, Chen HH, Zhao GZ, Zhu WY, Jiang CL, et al. Isolation, diversity, and antimicrobial activity of rare Actinobacteria from medicinal plants of tropical rain forests in Xishuangbanna, China. Appl Environ Microbiol 2009;75(19):6176-86.##Amos GCA, Borsetto C, Laskaris P, Krsek M, Berry AE, Newsham KK, et al. Designing and implementing an assay for the detection of rare and divergent NRPS and PKS clones in European, Antarctic and Cuban soils. PLoS One 2015;10(9):e0138327.##Zhang H, Liu Y, Wang X, Hu R, Xu G, Mao C, et al. Gene sequence diversity of the nonribosomal peptide and polyketide natural products in Changbaishan soil correlates with changes in landscape belts. Ecol Indic 2021;133:108160##

Formulation, Characterization, and Evaluation of Wound Healing Potency of a Novel Mattan tailam Nanogel Based on a Famous Traditional Siddha Formula 2 2 Background: The Mattan tailam mixture has been extensively used to heal ulcerous wounds in traditional Siddha practice. The present study aimed to synthesize a Mattan tailam nanogel and evaluate the enhancement of wound healing potential in an experimental wound model. Methods: Mattan tailam nanogel was synthesized using the high-energy milling approach, and characterization of nanogel and potency of wound healing was investigated. The novelty of this study was the nanogel preparation of Mattan tailam. Results: As expected, a synthesized novel nanogel of Mattan tailam has a distinct, prominent peak with a spherical form, is negatively charged and has an average particle size of 20–30 nm. Mattan tailam nanogel treated rats showed a remarkable reduction (p<0.001) in the wound area. On the 16th day, 10% Mattan tailam nanogel treatment resulted in a higher percentage of wound contraction. The 10% Mattan tailam nanogel group exhibited a faster epithelialization time (14.33 days) and a greater hydroxyproline concentration than the others. The topical application of 10% Mattan tailam nanogel increased tensile strength, signifying a better therapeutic indication. Conclusion: The present findings prove that polyherbal Mattan tailam nanogel formulation significantly improves collagen production, wound contraction, and tensile strength. 38 47 Meenachisundaram Sakthiganapathi Department of Pharmacognosy, College of Pharmacy, Mother Theresa Post Graduate and Research Institute of Health Sciences, A Government of Puducherry Institution, GorimeduSchool of Pharmacy, Sri Balaji Vidyapeeth (Deemed to be University) Department of Pharmacognosy, College of Pharmacy, Mother Theresa Post Graduate and Research Institute of Health Sciences, A Government of Puducherry Institution, GorimeduSchool of Pharmacy, Sri Balaji Vidyapeeth (Deemed to be University) IndiaIndia Gnanakumar Prakash Yoganandam Department of Pharmacognosy, College of Pharmacy, Mother Theresa Post Graduate and Research Institute of Health Sciences, A Government of Puducherry Institution, Gorimedu, Puducherry 605 006 Department of Pharmacognosy, College of Pharmacy, Mother Theresa Post Graduate and Research Institute of Health Sciences, A Government of Puducherry Institution, Gorimedu, Puducherry 605 006 India Venkatachalam Gopal Department of Pharmacognosy, College of Pharmacy, Mother Theresa Post Graduate and Research Institute of Health Sciences, A Government of Puducherry Institution, Gorimedu, Puducherry 605 006 Department of Pharmacognosy, College of Pharmacy, Mother Theresa Post Graduate and Research Institute of Health Sciences, A Government of Puducherry Institution, Gorimedu, Puducherry 605 006 India CollagenHydroxyprolineNanogelsRatsTensile strengthWound healing 60529.pdf Enyedi B, Niethammer P. Mechanisms of epithelial wound detection. Trends Cell Biol 2015 Jul;25(7):398–407.##Leoni G, Neumann PA, Sumagin R, Denning TL, Nusrat A. Wound repair: role of immune–epithelial interactions. Mucosal Immunol 2015 Sep 15;8(5):959–68.##Takeo M, Lee W, Ito M. Wound healing and skin regeneration. Cold Spring Harb Perspect Med 2015 Jan 1;5(1):a023267.##Ellis S, Lin EJ, Tartar D. Immunology of wound healing. Curr Dermatol Rep 2018 Dec 28;7(4):350–8.##Thiruvoth F, Mohapatra D, Sivakumar D, Chittoria R, Nandhagopal V. Current concepts in the physiology of adult wound healing. Plast Aesthet Res 2015;2(5):250-6.##Sorg H, Tilkorn DJ, Hager S, Hauser J, Mirastschijski U. Skin wound healing: An update on the current knowledge and concepts. Eur Surg Res 2017;58(1–2):81–94.##Carlson MA, Longaker MT. The fibroblast-populated collagen matrix as a model of wound healing: a review of the evidence. Wound Repair Regen 2004 Mar;12(2):134–47.##Tracy LE, Minasian RA, Caterson EJ. Extracellular matrix and dermal fibroblast function in the healing wound. Adv Wound Care (New Rochelle) 2016 Mar;5(3):119–36.##Chitturi RT, Balasubramaniam AM, Parameswar RA, Kesavan G, Haris KT, Mohideen K. The role of myofibroblasts in wound healing, contraction and its clinical implications in cleft palate repair. J Int Oral Health 2015;7(3):75-80.##Desmouliere A, Darby IA, Laverdet B, Bonté F. Fibroblasts and myofibroblasts in wound healing. Clin Cosmet Investig Dermatol 2014 Nov;301:301-11.##Georgescu M, Marinas O, Popa M, Stan T, Lazar V, Bertesteanu SV, et al. Natural compounds for wound healing. In: Worldwide Wound Healing - Innovation in Natural and Conventional Methods. InTech; 2016.##Thakur R, Jain N, Pathak R, Sandhu SS. Practices in wound healing studies of plants. Evid Based Complement Alternat Med 2011;2011:438056.##Sindhu RK, Gupta R, Wadhera G, Kumar P. Modern herbal nanogels: formulation, delivery methods, and applications. Gels 2022 Feb 7;8(2):97.##Ansari SH, Islam F, Sameem Mohd. Influence of nanotechnology on herbal drugs: A Review. J Adv Pharm Technol Res 2012;3(3):142-6.##Soni G, Yadav KS. Nanogels as potential nanomedicine carrier for treatment of cancer: A mini review of the state of the art. Saudi Pharm J 2016 Mar;24(2):133–9.##Kabanov AV, Vinogradov SV. Nanogels as pharmaceutical carriers: finite networks of infinite capabilities. Angew Chemie Int Ed Engl 2009 Jul 13;48(30):5418–29.##Soni KS, Desale SS, Bronich TK. Nanogels: An overview of properties, biomedical applications and obstacles to clinical translation. Control Release 2016 Oct;240:109–26.##Sinha A, Simnani FZ, Singh D, Nandi A, Choudhury A, Patel P, et al. The translational paradigm of nanobiomaterials: Biological chemistry to modern applications. Mater Today Bio 2022;17:100463.##Patra JK, Das G, Fraceto LF, Campos EVR, Rodriguez-Torres M del P, Acosta-Torres LS, et al. Nano based drug delivery systems: recent developments and future prospects. J Nanobiotechnology 2018 Dec 19;16(1):71.##Baskar V, Meeran SI, Subramani A, Sruthi, Ali J, Shabeer TK. Historic review on modern herbal nanogel formulation and delivery methods. Int J Pharm Pharm Sci 2018 Oct 1;10(10):1.##Yin Y, Hu B, Yuan X, Cai L, Gao H, Yang Q. Nanogel: A versatile nano-delivery system for biomedical applications. Pharmaceutics 2020;12(3):290.##Arunadevi R, Susila R, Murugammal S, Divya S. Preparation and standardization of Mathan Tailam: A classical Siddha formulation for diabetic ulcerative wound healing. J Ayurveda Integr Med 2020 Jan;11(1):10–5.##Lahorkar P, Ramitha K, Bansal V, Anantha Narayana D. A comparative evaluation of medicated oils prepared using ayurvedic and modified processes. Indian J Pharm Sci 2009;71(6):656-62.##Prasad Yadav T, Manohar Yadav R, Pratap Singh D. Mechanical milling: a top down approach for the synthesis of nanomaterials and nanocomposites. Nanoscience and Nanotechnology 2012 Aug 31;2(3):22–48.##Barkat MA, Harshita, Ahmad I, Ali R, Singh SP, Pottoo FH, et al. Nanosuspension-based aloe vera gel of silver sulfadiazine with improved wound healing activity. AAPS PharmSciTech 2017 Nov 5;18(8):3274–85.##Prince C. In-vitro Evaluation of mutagenic and anti-mutagenic effects of Siddha formulation Mathan thailam. International Journal of Emerging Technologies and Innovative Research 2020 May;7(5):663–6.##Morton JJ, Malone MH. Evaluation of vulneray activity by an open wound procedure in rats. Arch Int Pharmacodyn Ther 1972 Mar;196(1):117–26.##Ehrlich HP, Hunt TK. The effects of cortisone and anabolic steroids on the tensile strength of healing wounds. Ann Surg 1969 Aug;170(2):203–6.##Lee KH. Studies on the Mechanism of Action of Salicylate II. Retardation of wound healing by aspirin. J Pharm Sci 1968 Jun;57(6):1042–3.##Khan I, Saeed K, Khan I. Nanoparticles: Properties, applications and toxicities. Arabian Journal of Chemistry 2019 Nov;12(7):908–31.##Kabanov AV, Gendelman HE. Nanomedicine in the diagnosis and therapy of neurodegenerative disorders. Prog Polym Sci 2007 Aug;32(8–9):1054–82.##Rezaei R, Safaei M, Mozaffari HR, Moradpoor H, Karami S, Golshah A, et al. The role of nanomaterials in the treatment of diseases and their effects on the immune system. Open Access Maced J Med Sci 2019 Jun 16;7(11):1884–90.##Patra JK, Das G, Fraceto LF, Campos EVR, Rodriguez-Torres M del P, Acosta-Torres LS, et al. Nano based drug delivery systems: recent developments and future prospects. J Nanobiotechnology 2018 Dec 19;16(1):71.##Su S, M. Kang P. Recent advances in nanocarrier-assisted therapeutics delivery systems. Pharmaceutics 2020 Sep 1;12(9):837.##Mujahid M, Ahmad L, Ahmad M. Synthesis of ZnO nanogel for the treatment of superficial skin microbial infections. Journal of Drug Delivery and Therapeutics 2017 Mar 15;7(2).##Huang X, Zheng X, Xu Z, Yi C. ZnO-based nanocarriers for drug delivery application: From passive to smart strategies. Int J Pharm 2017 Dec;534(1–2):190–4.##Gushiken LFS, Beserra FP, Bastos JK, Jackson CJ, Pellizzon CH. Cutaneous wound healing: An update from physiopathology to current therapies. Life (Basel) 2021 Jul 7;11(7):665.##Rodrigues M, Kosaric N, Bonham CA, Gurtner GC. Wound healing: A cellular perspective. Physiol Rev 2019 Jan 1;99(1):665–706.##Loretelli C, ben Nasr M, Giatsidis G, Bassi R, Lancerotto L, D’Addio F, et al. Embryonic stem cell extracts improve wound healing in diabetic mice. Acta Diabetol 2020 Jul 2;57(7):883–90.##Elshamy AI, Ammar NM, Hassan HA, El-Kashak WA, Al-Rejaie SS, Abd-ElGawad AM, et al. Topical wound healing activity of myricetin isolated from Tecomaria capensis v. aurea. Molecules 2020 Oct 22;25(21):4870.##Rahman MS, Islam R, Rana MM, Spitzhorn LS, Rahman MS, Adjaye J, et al. Characterization of burn wound healing gel prepared from human amniotic membrane and Aloe vera extract. BMC Complement Altern Med 2019 Dec 3;19(1):115.##Lambebo MK, Kifle ZD, Gurji TB, Yesuf JS. Evaluation of wound healing activity of methanolic crude extract and solvent fractions of the leaves of Vernonia auriculifera Hiern (Asteraceae) in mice. J Exp Pharmacol 2021 Jul;13:677–92.##Andritoiu CV, Andriescu CE, Ibanescu C, Lungu C, Ivanescu B, Vlase L, et al. Effects and characterization of some topical ointments based on vegetal extracts on incision, excision, and thermal wound models. Molecules 2020 Nov 16;25(22):5356.##Preethi KC, Kuttan R. Wound healing activity of flower extract of Calendula officinalis. J Basic Clin Physiol Pharmacol 2009 Jan;20(1):73-9.##Bozkırlı BO, Gündoğdu RH, Ersoy E, Lortlar N, Yıldırım Z, Temel H, et al. Pilot experimental study on the effect of arginine, glutamine, and β-hydroxy β-methylbutyrate on secondary wound healing. JPEN J Parenter Enteral Nutr 2015 Jul 24;39(5):591–7.##Kalavathy N, Rajalakshmi S, Sundharam M, Veluchami G. A siddha remedy for chronic ulcer. Indian Journal of Hospital Pharmacy 1984;21(1):30–5.##Bapat AC, Ubale MR, Gaurea SH, Bapat UC, Kartikeyan S, Malgaonkar AA. In vitro study of anti-dermatophytic activity of select plant extracts. IOSR Journal of Dental and Medical Sciences 2017 Jun;16(06):30–3.##Reid KA, Maes J, Maes A, van Staden J, de Kimpe N, Mulholland DA, et al. Evaluation of the mutagenic and antimutagenic effects of South African plants. J Ethnopharmacol 2006 Jun;106(1):44–50.##Danielsen CC, Gottrup F. Healing of incisional wounds in stomach and duodenum. Eur Surg Res 1981;13(3):194–201.##Oxlund H, Christensen H, Seyer-Hansen M, Andreassen TT. Collagen deposition and mechanical strength of colon anastomoses and skin incisional wounds of rats. J Surg Res 1996 Nov;66(1):25–30.##

MGMT Gene rs1625649 Polymorphism in Iranian Patients with Brain Glioblastoma: A Case Control Study 2 2 Glioblastoma (GBM) is the most common and aggressive primary malignant brain tumor with poor prognosis and high potential of dispersion to other brain tissues in adult. Effective and modern choices of treatment including chemotherapy with alkylating agents marginally extend survival of GBM. However, alkylating agents can lead to highly harmful mismatch during DNA replication causing apoptosis and cell death. Accordingly, O6-Methylguanine-DNA methyltransferase (MGMT) removes alkyl adducts, thereby causing resistance to alkylating drugs. Single-Nucleotide Polymorphisms (SNPs) in MGMT promoter region may play a role in the regulation of MGMT expression and prediction of glioma development risk. In order to evaluate the clinical significance of rs1625649 SNP in the MGMT promoter region of glioblastoma, genomic DNA from a series of 54 patients with GBM and 50 healthy individuals in Iranian population were collected for tetra ARMS PCR amplification. None of the "A" or "C" alleles were associated with tumor occurrence, the "AA" genotype was more frequent in healthy subjects, and the "AC" genotype was 4.6 times more common in patients with GBM. The longest survival time was observed in the "CC" genotype; however, this difference was not statistically significant. On the other hand, homozygous rs1625649 (AA genotype) was significantly associated with a better survival than the cases with heterozygous rs1625649 (CA genotype) or wild type rs1625649 (CC genotype), predicting better response to temozolomide-based chemotherapy. 48 52 Reyhaneh Safaei Department of Pathology, Faculty of Veterinary Medicine, University of Tehran Department of Pathology, Faculty of Veterinary Medicine, University of Tehran Iran Hanieh Mojtahedi Molecular Immunology Research Center, Tehran University of Medical SciencesNetwork of Immunity in Infection, Malignancy and Autoimmunity (NIIMA), Universal Scientific Education and Research Network (USERN) Molecular Immunology Research Center, Tehran University of Medical SciencesNetwork of Immunity in Infection, Malignancy and Autoimmunity (NIIMA), Universal Scientific Education and Research Network (USERN) IranIran Sara Hanaei Department of Neurosurgery, Imam Khomeini Hospital Complex, Tehran University of Medical Sciences Department of Neurosurgery, Imam Khomeini Hospital Complex, Tehran University of Medical Sciences Iran Azadehsadat Razavi Research Center for Immunodeficiencies, Children's Medical Center, Tehran University of Medical Sciences Research Center for Immunodeficiencies, Children's Medical Center, Tehran University of Medical Sciences Iran Marzie Esmaeili Research Center for Immunodeficiencies, Children's Medical Center, Tehran University of Medical Sciences Research Center for Immunodeficiencies, Children's Medical Center, Tehran University of Medical Sciences Iran Maryam Sadr Molecular Immunology Research Center, Tehran University of Medical Sciences Molecular Immunology Research Center, Tehran University of Medical Sciences Iran Arezou Rezaei Research Center for Immunodeficiencies, Children's Medical Center, Tehran University of Medical Sciences Research Center for Immunodeficiencies, Children's Medical Center, Tehran University of Medical Sciences Iran Maryam Edalatfar Department of Neurosurgery, lmam Hossein Hospital, Shahid Beheshti University of Medical Sciences Department of Neurosurgery, lmam Hossein Hospital, Shahid Beheshti University of Medical Sciences Iran Hamidreza Khayat Kashani Department of Neurosurgery, lmam Hossein Hospital, Shahid Beheshti University of Medical Sciences Department of Neurosurgery, lmam Hossein Hospital, Shahid Beheshti University of Medical Sciences Iran Mohsen Sadeghi-Naini Department of Neurosurgery, Lorestan University of Medical Sciences Department of Neurosurgery, Lorestan University of Medical Sciences Iran Farzaneh Darbeheshti Department of Genetics, School of Medicine, Tehran University of Medical Sciences Department of Genetics, School of Medicine, Tehran University of Medical Sciences Iran Jaber Gharehdaghi Legal Medicine Research Center, Legal Medicine Organization Legal Medicine Research Center, Legal Medicine Organization Iran Mehdi Forouzesh Legal Medicine Research Center, Legal Medicine Organization Legal Medicine Research Center, Legal Medicine Organization Iran Abdolali Ebrahimi Department of Pathology, lmam Hossein Hospital, School of Medicine, Shahid Beheshti University of Medical Sciences Department of Pathology, lmam Hossein Hospital, School of Medicine, Shahid Beheshti University of Medical Sciences Iran Nima Rezaei Glioblastoma multiformeMGMTO6-Methylguanine DNA methyltransferaseSNP 60525.pdf Longo Machado de Almeida M, Pires de Aguiar PH, De Gois K, De Sousa Gehrke F, Fonseca F. The importance of MGMT promoter methylation status for glioblastomas prognosis: meta-analysis. JBNC - J Bras Neurocir 2018;29(4):595-611.##Reifenberger G, Hentschel B, Felsberg J, Schackert G, Simon M, Schnell O, et al. Predictive impact of MGMT promoter methylation in glioblastoma of the elderly. Int J Cancer 2012 Sep 15;131(6):1342-50.##Hanif F, Muzaffar K, Perveen K, Malhi SM, Simjee SU. Glioblastoma multiforme: A review of its epidemiology and pathogenesis through clinical presentation and treatment. Asian Pac J Cancer Prev 2017 Jan 1;18(1):3-9.##Tamimi AF, Juweid M. Epidemiology and outcome of glioblastoma. In: Glioblastoma. 2017. [Internet]. Brisbane (AU): Codon Publications; 2017 Sep 27. Chapter 8.##Wilson TA, Karajannis MA, Harter DH. Glioblastoma multiforme: State of the art and future therapeutics. Surg Neurol Int 2014 May 8;5:64.##Ohgaki H, Kleihues P. The definition of primary and secondary glioblastoma. Clin Cancer Res 2013 Feb 15;19(4):764-72.##Louis DN, Perry A, Reifenberger G, von Deimling A, Figarella-Branger D, Cavenee WK, et al. The 2016 World Health Organization Classification of Tumors of the Central Nervous System: a summary. Acta Neuropathologica 2016;131(6):803-20.##Li J, Di C, Mattox AK, Wu L, Adamson DC. The future role of personalized medicine in the treatment of glioblastoma multiforme. Pharmgenomics Pers Med 2010;3:111-27.##Anthony C, Mladkova-Suchy N, Adamson DC. The evolving role of antiangiogenic therapies in glioblastoma multiforme: current clinical significance and future potential. Expert Opin Investig Drugs 2019 Sep;28(9):787-97.##Lv Y, Jia C, Jiang A, Zhang H, Wang Y, Liu F, et al. Analysis of association between MGMT and p53 gene single nucleotide polymorphisms and laryngeal cancer. Anticancer Res 2017 Aug;37(8):4399-4403.##Kristensen LS, Michaelsen SR, Dyrbye H, Aslan D, Grunnet K, Christensen JI, et al. Assessment of quantitative and allelic MGMT methylation patterns as a prognostic marker in glioblastoma. J Neuropathol Exp Neurol 2016 Mar;75(3):246-55.##Leng S, Bernauer AM, Hong C, Do KC, Yingling CM, Flores KG, et al. The A/G allele of Rs16906252 predicts for MGMT methylation and is selectively silenced in premalignant lesions from smokers and in lung adenocarcinomas. Clin Cancer Res 2011 Apr 1;17(7):2014-23.##Sharma S, Salehi F, Scheithauer BW, Rotondo F, Syro L V., Kovacs K. Role of MGMT in tumor development, progression, diagnosis, treatment and prognosis. Anticancer Res 2009 Oct;29(10):3759-68.##Bethke L, Webb E, Murray A, Schoemaker M, Johansen C, Christensen HC, et al. Comprehensive analysis of the role of DNA repair gene polymorphisms on risk of glioma. Hum Mol Genet 2008 Mar 15;17(6):800-5.##Zawlik I, Vaccarella S, Kita D, Mittelbronn M, Franceschi S, Ohgaki H. Promoter methylation and polymorphisms of the MGMT gene in glioblastomas: A population-based study. Neuroepidemiology 2008;32(1):21-9.##Hegi ME, Diserens A-C, Gorlia T, Hamou M-F, de Tribolet N, Weller M, et al. MGMT gene silencing and benefit from temozolomide in glioblastoma. N Engl J Med 2005;352(10):997-1003.##Park JH, Kim NS, Park JY, Chae YS, Kim JG, Sohn SK, et al. MGMT 2535G>T polymorphism is associated with prognosis for patients with metastatic colorectal cancer treated with oxaliplatin-based chemotherapy. J Cancer Res Clin Oncol 2010 Aug;136(8):1135-42.##Hsu CY, Ho HL, Lin SC, Ho TDH, Ho DMT. The MGMT promoter single-nucleotide polymorphism rs1625649 had prognostic impact on patients with MGMT methylated glioblastoma. PLoS One 2017;12(10):e0186430. ##Xu M, Nekhayeva I, Cross CE, Rondelli CM, Wickliffe JK, Abdel-Rahman SZ. Influence of promoter/enhancer region haplotypes on MGMT transcriptional regulation: A potential biomarker for human sensitivity to alkylating agents. Carcinogenesis 2014 Mar;35(3):564-71.##Margison GP, Heighway J, Pearson S, McGown G, Thorncroft MR, Watson AJ, et al. Quantitative trait locus analysis reveals two intragenic sites that influence O6-alkylguanine-DNA alkyltransferase activity in peripheral blood mononuclear cells. Carcinogenesis 2005 Aug;26(8):1473-80.##

Identification of Critical Molecular Factors and Side Effects Underlying the Response to Thalicthuberine in Prostate Cancer: A Systems Biology Approach 2 2 Background: Uncontrolled mitosis of cancer cells and resistance cells to chemotherapy drugs are the challenges of prostate cancer. Thalicthuberine causes a mitotic arrest and a reduction of the effects of drug resistance, resulting in cell death. In this study, we applied bioinformatics and computational biology methods to identify functional pathways and side effects in response to Thalicthuberine in prostate cancer patients. Methods: Microarray data were retrieved from Gene Expression Omnibus (GEO), and protein-protein interactions and gene regulatory networks were constructed, using the Cytoscape software. The critical genes and molecular mechanisms in response to Thalicthuberine and its side effects were identified, using the Cytoscape software and WebGestalt server, respectively. Finally, GEPIA2 was used to predict the relationship between critical genes and prostate cancer. Results: The POLQ, EGR1, CDKN1A, FOS, MDM2, CDC20, CCNB1, and CCNB2 were identified as critical genes in response to this drug. The functional mechanisms of Thalicthuberine include a response to oxygen levels, toxic substances and immobilization stress, cell cycle regulation, regeneration, the p53 signaling pathway, the action of the parathyroid hormone, and the FoxO signaling pathway. Besides, the drug has side effects including muscle cramping, abdominal pains, paresthesia, and metabolic diseases. Conclusion: Our model suggested newly predicted crucial genes, molecular mechanisms, and possible side effects of this drug. However, further studies are required. 53 64 Fatemeh Saberi Student Research Committee, Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical SciencesCellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences Student Research Committee, Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical SciencesCellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences IranIran Zeinab Dehghan Department of Comparative Biomedical Sciences, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences Department of Comparative Biomedical Sciences, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences Iran Effat Noori Student Research Committee, Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical SciencesCellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences Student Research Committee, Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical SciencesCellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences IranIran Zahra Taheri Department of Biology and Biotechnology, Pavia University Department of Biology and Biotechnology, Pavia University Italy Marzieh Sameni Student Research Committee, Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical SciencesCellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences Student Research Committee, Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical SciencesCellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences IranIran Hakimeh Zali BioinformaticsComputational biologyGene expressionGene regulatory networksProstate cancer 60527.pdf Smith-Palmer J, Takizawa C, Valentine W. Literature review of the burden of prostate cancer in Germany, France, the United Kingdom and Canada. BMC Urol 2019 Dec;19(1):1-6.##Hwang C. Overcoming docetaxel resistance in prostate cancer: a perspective review. Ther Adv Med Oncol 2012 Nov;4(6):329-40.##Ilic D, Neuberger MM, Djulbegovic M, Dahm P. Screening for prostate cancer. Cochrane Database Syst Rev 2013(1).##Levrier C, Rockstroh A, Gabrielli B, Kavallaris M, Lehman M, Davis RA, et al. Discovery of thalicthuberine as a novel antimitotic agent from nature that disrupts microtubule dynamics and induces apoptosis in prostate cancer cells. Cell Cycle 2018 Mar 4;17(5):652-68.##Chuang SC, Chen HC, Sun CW, Chen YA, Wang YH, Chiang CJ, et al. Phthalate exposure and prostate cancer in a population-based nested case-control study. Environ Res 2020 Feb 1;181:108902.##Kaler J, Hussain A, Haque A, Naveed H, Patel S. A comprehensive review of pharmaceutical and surgical interventions of prostate cancer. Cureus 2020 Nov 22;12(11).##Pienta KJ, Esper PS. Risk factors for prostate cancer. Ann Intern Med 1993 May 15;118(10):793-803.##Litwin MS, Tan HJ. The diagnosis and treatment of prostate cancer: a review. JAMA 2017 Jun 27;317(24):2532-42.##Nurgali K, Jagoe RT, Abalo R. Editorial: Adverse effects of cancer chemotherapy: Anything new to improve tolerance and reduce sequelae?. Front Pharmacol 2018 Mar 22;9:245.##Zachariou M, Minadakis G, Oulas A, Afxenti S, Spyrou GM. Integrating multi-source information on a single network to detect disease-related clusters of molecular mechanisms. J Proteomics 2018 Sep 30;188:15-29.##Karns R, Tabar S, Bardes EE, Jegga AG, Aronow BJ. How Do Bioinformatics Approaches Apply to the Analysis and Understanding of Disease Pathology?.##Golubnitschaja O, Baban B, Boniolo G, Wang W, Bubnov R, Kapalla M, et al. Medicine in the early twenty-first century: paradigm and anticipation-EPMA position paper 2016. EPMA J 2016 Dec;7(1):1-3.##Sundaresan L, Kumar P, Chatterjee S. Mechanistic insights into the differential effects of thalidomide and lenalidomide in metastatic prostate cancer. Future Oncol 2018 Oct;14(23):2383-401.##Eastham JA, Hall SJ, Sehgal I, Wang J, Timme TL, Yang G, et al. In vivo gene therapy with p53 or p21 adenovirus for prostate cancer. Cancer Res 1995 Nov 15;55(22):5151-5.##Latchman DS. Inhibitory transcription factors. Int J Biochem Cell Biol 1996 Sep 1;28(9):965-74.##Shen-Orr SS, Milo R, Mangan S, Alon U. Network motifs in the transcriptional regulation network of Escherichia coli. Nat Genet 2002 May;31(1):64-8.##Nevedomskaya E, Baumgart SJ, Haendler B. Recent advances in prostate cancer treatment and drug discovery. Int J Mol Sci 2018 May 4;19(5):1359.##Barrett T, Wilhite SE, Ledoux P, Evangelista C, Kim IF, Tomashevsky M, et al. NCBI GEO: archive for functional genomics data sets—update. Nucleic Acids Res 2012 Nov 26;41(D1):D991-5.##Shannon P, Markiel A, Ozier O, Baliga NS, Wang JT, Ramage D, et al. Cytoscape: a software environment for integrated models of biomolecular interaction networks. Genome Res 2003 Nov 1;13(11):2498-504.##Kargar M, An A. The effect of sequence complexity on the construction of protein-protein interaction networks. In: International Conference on Brain Informatics 2010 Aug 28 (pp. 308-319). Springer, Berlin, Heidelberg.##Ma H, Zeng AP. Reconstruction of metabolic networks from genome data and analysis of their global structure for various organisms. Bioinformatics 2003 Jan 22;19(2):270-7.##Bader GD, Hogue CW. An automated method for finding molecular complexes in large protein interaction networks. BMC Bioinformatics 2003 Dec;4(1):1-27.##Dehghan Z, Mohammadi-Yeganeh S, Sameni M, Mirmotalebisohi SA, Zali H, Salehi M. Repurposing new drug candidates and identifying crucial molecules underlying PCOS pathogenesis based on bioinformatics analysis. Daru 2021 Dec;29(2):353-66.##Mirmotalebisohi SA, Sameni M, Dehghan Z, Khazaei-Poul Y, Sepehrizadeh Z, Faramarzi MA, et al. Deciphering the molecular pathogenesis behind neurological manifestations of SARS-CoV-2 and drug repurposing, A systems biology approach. 2021.##Huang HY, Lin YC, Li J, Huang KY, Shrestha S, Hong HC, et al. miRTarBase 2020: updates to the experimentally validated microRNA–target interaction database. Nucleic Acids Res 2020 Jan 8;48(D1):D148-54.##Xiao F, Zuo Z, Cai G, Kang S, Gao X, Li T. miRecords: an integrated resource for microRNA–target interactions. Nucleic Acids Res 2009 Jan 1;37(suppl_1):D105-10.##Wingender E, Dietze P, Karas H, Knüppel R. TRANSFAC: a database on transcription factors and their DNA binding sites. Nucleic Acids Res 1996 Jan 1;24(1):238-41.##Han H, Cho JW, Lee S, Yun A, Kim H, Bae D, et al. TRRUST v2: an expanded reference database of human and mouse transcriptional regulatory interactions. Nucleic Aacids Res 2018 Jan 4;46(D1):D380-6.##Tong Z, Cui Q, Wang J, Zhou Y. TransmiR v2. 0: an updated transcription factor-microRNA regulation database. Nucleic Acids Res 2019 Jan 8;47(D1):D253-8.##Ahnert SE, Fink TM. Form and function in gene regulatory networks: the structure of network motifs determines fundamental properties of their dynamical state space. J R Soc Interface 2016 Jul 31;13(120):20160179.##Defoort J, Van de Peer Y, Vermeirssen V. Function, dynamics and evolution of network motif modules in integrated gene regulatory networks of worm and plant. Nucleic Acids Res 2018 Jul 27;46(13):6480-503.##Wernicke S, Rasche F. FANMOD: a tool for fast network motif detection. Bioinformatics 2006 May 1;22(9):1152-3.##Farahani M, Rezaei–Tavirani M, Zali H, Arefi Oskouie A, Omidi M, Lashay A. Deciphering the transcription factor-microRNA-target gene regulatory network associated with graphene oxide cytotoxicity. Nanotoxicology 2018 Oct 21;12(9):1014-26.##Liao Y, Wang J, Jaehnig EJ, Shi Z, Zhang B. WebGestalt 2019: gene set analysis toolkit with revamped UIs and APIs. Nucleic Acids Res 2019 Jul 2;47(W1):W199-205.##Tang Z, Li C, Kang B, Gao G, Li C, Zhang Z. GEPIA: a web server for cancer and normal gene expression profiling and interactive analyses. Nucleic Acids Research. 2017 Jul 3;45(W1):W98-102.##Roy S, Raj M, Ghosh P, Das SK. Role of motifs in topological robustness of gene regulatory networks. In2017 IEEE International Conference on Communications (ICC) 2017 May 21 (pp. 1-6). IEEE.##Schrempf A, Slyskova J, Loizou JI. Targeting the DNA repair enzyme polymerase θ in cancer therapy. Trends Cancer 2021 Feb 1;7(2):98-111.##Ravindran F, Jain A, Desai S, Menon N, Srivastava K, Bawa PS, et al. Whole-exome sequencing of Indian prostate cancer reveals a novel therapeutic target: POLQ. J Cancer Res Clin Oncol 2022 Jun 23:1-2.##Baron V, Adamson ED, Calogero A, Ragona G, Mercola D. The transcription factor Egr1 is a direct regulator of multiple tumor suppressors including TGFβ1, PTEN, p53, and fibronectin. Cancer Gene Ther 2006 Feb;13(2):115-24.##Li L, Ameri AH, Wang S, Jansson KH, Casey OM, Yang Q, et al. EGR1 regulates angiogenic and osteoclastogenic factors in prostate cancer and promotes metastasis. Oncogene 2019 Aug;38(35):6241-55.##Jain AK, Raina K, Agarwal R. Deletion of p21/Cdkn1a confers protective effect against prostate tumorigenesis in transgenic adenocarcinoma of the mouse prostate model. Cell Cycle 2013 May 15;12(10):1598-604.##Sivoňová MK, Vilčková M, Kliment J, Mahmood S, Jurečeková J, Dušenková S, et al. Association of p53 and p21 polymorphisms with prostate cancer. Biomed Rep 2015 Sep 1;3(5):707-14.##Riedel M, Berthelsen MF, Cai H, Haldrup J, Borre M, Paludan SR, et al. In vivo CRISPR inactivation of Fos promotes prostate cancer progression by altering the associated AP-1 subunit Jun. Oncogene 2021;40(13):2437-47.##Shankar E, Song K, Corum SL, Bane KL, Wang H, Kao HY, et al. A signaling network controlling androgenic repression of c-Fos protein in prostate adenocarcinoma cells. J Biol Chem 2016 Mar 11;291(11):5512-5526.##Zhang Q, Huang H, Liu A, Li J, Liu C, Sun B, et al. Cell division cycle 20 (CDC20) drives prostate cancer progression via stabilization of β-catenin in cancer stem-like cells. EBioMedicine 2019;42:397-407.##Li K, Mao Y, Lu L, Hu C, Wang D, Si-Tu J, et al. Silencing of CDC20 suppresses metastatic castration-resistant prostate cancer growth and enhances chemosensitivity to docetaxel. International Int J Oncol 2016 Oct;49(4):1679-85.##Dai L, Song ZX, Wei DP, Zhang JD, Liang JQ, Wang BB, et al. CDC20 and PTTG1 are important biomarkers and potential therapeutic targets for metastatic prostate cancer. Adv Ther 2021 Jun;38(6):2973-89.##Hou H, Sun D, Zhang X. The role of MDM2 amplification and overexpression in therapeutic resistance of malignant tumors. Cancer Cell Int 2019 Aug 22;19:216.##Feng FY, Zhang Y, Kothari V, Evans JR, Jackson WC, Chen W, et al. MDM2 inhibition sensitizes prostate cancer cells to androgen ablation and radiotherapy in a p53-dependent manner. Neoplasia 2016 Apr;18(4):213-22.##Leite KR, Franco MF, Srougi M, Nesrallah LJ, Nesrallah A, Bevilacqua RG, et al. Abnormal expression of MDM2 in prostate carcinoma. Mod Pathol 2001 May;14(5):428-36.##Huang CG, Li FX, Pan S, Xu CB, Dai JQ, Zhao XH. Identification of genes associated with castration‑resistant prostate cancer by gene expression profile analysis. Mol Med Rep 2017 Nov;16(5):6803-6813.##Gomez LA, de Las Pozas A, Reiner T, Burnstein K, Perez-Stable C. Increased expression of cyclin B1 sensitizes prostate cancer cells to apoptosis induced by chemotherapy. Mol Cancer Ther 2007 May;6(5):1534-43.##Shen H, Guo YL, Li GH, Zhao W, Zhang L. Gene expression analysis reveals key genes and signalings associated with the prognosis of prostate cancer. Comput Math Methods Med 2021 Aug 28;2021:9946015.##de Almeida A, Parthimos D, Dew H, Smart O, Wiltshire M, Errington RJ. Aquaglyceroporin-3’s expression and cellular localization is differentially modulated by hypoxia in prostate cancer cell lines. Cells 2021;10(4):838.##Mamede A, Abrantes AM, Pedrosa L, Casalta-Lopes J, Pires A, Teixo R, et al. Beyond the limits of oxygen: effects of hypoxia in a hormone-independent prostate cancer cell line. ISRN Oncol 2013 Sep 12;2013:918207.##Jing X, Yang F, Shao C, Wei K, Xie M, Shen H, et al. Role of hypoxia in cancer therapy by regulating the tumor microenvironment. Mol Cancer 2019;18(1):157.##Teicher BA, Herman TS, Hopkins RE, Menon K. Effect of oxygen level on the enhancement of tumor response to radiation by perfluorochemical emulsions or a bovine hemoglobin preparation. Int J Radiat Oncol Biol Phys 1991 Sep;21(4):969-74.##Hassan S, Pullikuth A, Nelson KC, Flores A, Karpova Y, Baiz D, et al. β2-Adrenoreceptor signaling increases therapy resistance in prostate cancer by upregulating MCL1. Mol Cancer Res 2020 Dec;18(12):1839-1848.##Watabe T, Lin M, Ide H, Donjacour AA, Cunha GR, Witte ON, et al. Growth, regeneration, and tumorigenesis of the prostate activates the PSCA promoter. Proc Natl Acad Sci USA 2002 Jan 8;99(1):401-6.##Oviedo NJ, Beane WS. Regeneration: The origin of cancer or a possible cure Semin Cell Dev Biol 2009 Jul;20(5):557-64.##Rahman M, Park MN, Hasanur Rahman Md, Mamunur Rashid Md, Islam R, Jamal Uddin Md, et al. p53 Modulation of autophagy signaling in cancer therapies: perspectives mechanism and therapeutic targets. Front Cell Dev Biol 2022 Jan 26;10:761080.##Mathew R, Karantza-Wadsworth V, White E. Role of autophagy in cancer. Nat Rev Cancer 2007 Dec;7(12):961-7.##Li H, Ruan G, Li Z, Liu Z, Zheng X, Cheng G, et al. The calcimimetic R-568 induces apoptotic cell death in prostate cancer cells. J Exp Clin Cancer Res 2009 Jul 14;28(1):100.##Wang L, Xu M, Li Z, Shi M, Zhou X, Jiang X, et al. Calcium and CaSR/IP3R in prostate cancer development. Cell Biosci 2018;8:1-7.##Schwartz GG. Prostate cancer, serum parathyroid hormone, and the progression of skeletal metastases. Cancer Epidemiol Biomarkers Prev 2008;17(3):478-83.##Goltzman D. Physiology of parathyroid hormone. Endocrinol Metab Clin North Am 2018 Dec;47(4):743-758.##Jiramongkol Y, Lam EW-F. FOXO transcription factor family in cancer and metastasis. Cancer Metastasis Rev 2020 Sep;39(3):681-709.##Accili D, Arden KC. FoxOs at the crossroads of cellular metabolism, differentiation, and transformation. Cell 2004;117:421-6.##Farhan M, Wang H, Gaur U, Little PJ, Xu J, Zheng W. FOXO signaling pathways as therapeutic targets in cancer. Int J Biol Sci 2017 Jul 6;13(7):815-27.##

Monkeypox Outbreaks in Non-Endemic Countries: Correspondence 2 2 No Abstract 65 65 Rujittika Mungmunpuntipantip Viroj Wiwanitkit Joseph Ayobabalola University Joseph Ayobabalola University Nigeria Editorial 60530.pdf Shamabadi A, Akhondzadeh S. Monkeypox outbreaks in non-endemic countries: What do we know and what do we need?. Avicenna J Med Biotechnol 2022 Jul-Sep;14(3):186-7.##Joob B, Wiwanitkit V. Monkeypox: Revisit of the old threat and emerging imported cases. Med J DY Patil Vidyapeeth 2022;15:457-9.##Niedrig M, Meyer H, Panning M, Drosten C. Follow-up on diagnostic proficiency of laboratories equipped to perform orthopoxvirus detection and quantification by PCR: the second international external quality assurance study. J Clin Microbiol 2006 Apr;44(4):1283-7.##