The Need for Serious Support for Basic Medical Science in Iran 2 2 No Abstract 136 136 Shahin Akhondzadeh Editorial 60578.pdf Akhondzadeh S. Hippocampal synaptic plasticity and cognition. J Clin Pharm Ther 1999 Aug;24(4):241-8.##Noorbala AA, Akhondzadeh S, Davari-Ashtiani R, Amini-Nooshabadi H. Piracetam in the treatment of schizophrenia: implications for the glutamate hypothesis of schizophrenia. J Clin Pharm Ther 1999; 24(5):369-74.##Akhondzadeh S, Ahmadi-Abhari SA, Assadi SM, Shabestari OL, Kashani AR, Farzanehgan ZM. Double-blind randomized controlled trial of baclofen vs. clonidine in the treatment of opiates withdrawal. J Clin Pharm Ther 2000; 25(5):347-53.##Akhondzadeh S. The 5-HT hypothesis of schizophrenia. IDrugs 2001;4(3):295-300.##Kahbazi M, Ghoreishi A, Rahiminejad F, Mohammadi MR, Kamalipour A, Akhondzadeh S. A randomized, double-blind and placebo-controlled trial of modafinil in children and adolescents with attention deficit and hyperactivity disorder. Psychiatry Res 2009;168(3):234-7.##Abbasi SH, Behpournia H, Ghoreshi A, Salehi B, Raznahan M, Rezazadeh SA, Rezaei F, Akhondzadeh S. The effect of mirtazapine add on therapy to risperidone in the treatment of schizophrenia: a double-blind randomized placebo-controlled trial. Schizophr Res 2010;116(2-3):101-6.##Milajerdi A, Jazayeri S, Hashemzadeh N, Shirzadi E, Derakhshan Z, Djazayeri A, Akhondzadeh S. The effect of saffron (Crocus sativus L.) hydroalcoholic extract on metabolic control in type 2 diabetes mellitus: A triple-blinded randomized clinical trial. J Res Med Sci 2018;23:16.##

Role of Recombinant Proteins for Treating Rheumatoid Arthritis 2 2 Rheumatoid Arthritis (RA) is an autoimmune disease and chronic inflammatory disorder that affects joints and causes inflammation, pain, stiffness, and eventually progressive joint destruction. Approximately 1% of the world's population is estimated to suffer from RA, and if this disease is left untreated, it can lead to severe disability. Despite all the efforts and advances made by professionals in the field, there is currently no definitive treatment for RA, and most treatment strategies are aimed at relieving symptoms and improving patients' quality of life. One of the most promising current approaches is the use of recombinant proteins that target specific signaling pathways involved in the development of RA to alleviate symptoms and slow the progression of the disease. This article discusses the genetic and immunological factors that influence the development of RA, recombinant proteins, methods of using these proteins, approved drugs, and side effects associated with treating RA. 137 145 Mahboubeh Soleimani Sasani Yeganeh Moradi Department of Biotechnology Engineering, Faculty of Medical Engineering, Shahab Danesh University Department of Biotechnology Engineering, Faculty of Medical Engineering, Shahab Danesh University Iran Autoimmune diseasesDrug therapyRecombinant proteinsRheumatoid arthritisTherapeutic use 60579.pdf Veale DJ, Orr C, Fearon U. Cellular and molecular perspectives in rheumatoid arthritis. Semin Immunopathol 2017 Jun;39(4):343-354.##Zhao J, Guo S, Schrodi SJ, He D. Molecular and cellular heterogeneity in rheumatoid arthritis: mechanisms and clinical implications. Front Immunol 2021 Nov 25;12:790122.##Ishikawa Y, Terao C. The impact of cigarette smoking on risk of rheumatoid arthritis: a narrative review. Cells 2020 Feb 19;9(2):475.##Smolen JS, Landewé R, Breedveld FC, Dougados M, Emery P, Gaujoux-Viala C, et al. EULAR recommendations for the management of rheumatoid arthritis with synthetic and biological disease-modifying antirheumatic drugs. Ann Rheum Dis 2010 Jun;69(6):964-75.##West SC. Molecular views of recombination proteins and their control. Nat Rev Mol Cell Biol 2003 Jun 1;4(6):435-45.##Blair HA, Deeks ED. Abatacept: a review in rheumatoid arthritis. Drugs 2017;77(11):1221-33.##Klareskog L, Stolt P, Lundberg K, Källberg H, Bengtsson C, Grunewald J, et al. A new model for an etiology of rheumatoid arthritis: smoking may trigger HLA–DR (shared epitope)–restricted immune reactions to autoantigens modified by citrullination. Arthritis Rheum 2006 Jan;54(1):38-46.##Okada Y, Wu D, Trynka G, Raj T, Terao C, Ikari K, et al. Genetics of rheumatoid arthritis contributes to biology and drug discovery. Nature 2014 Feb;506(7488):376-81.##Eyre S, Bowes J, Diogo D, Lee A, Barton A, Martin P, et al. High-density genetic mapping identifies new susceptibility loci for rheumatoid arthritis. Nat Genet 2012 Dec;44(12):1336-40.##Glyn-Jones S, Palmer AJR, Agricola R, Price AJ, Vincent TL, Weinans H, et al. Osteoarthritis. Lancet 2015;386(9991):376-87. ##Sakaguchi S, Yamaguchi T, Nomura T, Ono M. Regulatory T cells and immune tolerance. Cell 2008 May 30;133(5):775-87. ##Cope AP, Schulze-Koops H, Aringer M. The central role of T cells in rheumatoid arthritis. Clin Exp Rheumatol 2007 Sep 1;25(5):S4.##Weyand CM, Bryl E, Goronzy JJ. The role of T cells in rheumatoid arthritis. Autoimmunity 2001:183-95.##Edwards JC, Cambridge G. B-cell targeting in rheumatoid arthritis and other autoimmune diseases. Nat Rev Immunol 2006 May 1;6(5):394-403.##Cassese G, Arce S, Hauser AE, Lehnert K, Moewes B, Mostarac M, et al. Plasma cell survival is mediated by synergistic effects of cytokines and adhesion-dependent signals. J Immunol 2003 Aug 15;171(4):1684-90.##Berek C. [Do B cells play an important role in the pathogenesis of rheumatoid arthritis?] Z Rheumatol 2005 Sep;64(6):383-8. German.##Schröder AE, Greiner A, Seyfert C, Berek C. Differentiation of B cells in the nonlymphoid tissue of the synovial membrane of patients with rheumatoid arthritis. Proc Natl Acad Sci USA 1996 Jan 9;93(1):221-5.##McInnes IB, Schett G. The pathogenesis of rheumatoid arthritis. N Engl J Med 2011 Dec 8;365(23):2205-19.##Gavrilă BI, Ciofu C, Stoica V. Biomarkers in rheumatoid arthritis, what is new?. J Med Life 2016 Apr;9(2):144.##Aletaha D, Neogi T, Silman AJ, Funovits J, Felson DT, Bingham III CO, et al. 2010 rheumatoid arthritis classification criteria: an American College of Rheumatology/European League Against Rheumatism collaborative initiative. Arthritis Rheum 2010 Sep;62(9):2569-81.##Hirano T. Interleukin 6 in autoimmune and inflammatory diseases: a personal memoir. Proc Jpn Acad Ser B Phys Biol Sci 2010;86(7):717-30. ##Mohan C, Assassi S. Biomarkers in rheumatic diseases: how can they facilitate diagnosis and assessment of disease activity?. BMJ 2015 Nov 26;351.##van der Helm-van Mil AH, Verpoort KN, Breedveld FC, Toes RE, Huizinga TW. Antibodies to citrullinated proteins and differences in clinical progression of rheumatoid arthritis. Arthritis Res Ther 2005 Sep;7:1-0.##Welsing PM, Van Gestel AM, Swinkels HL, Kiemeney LA, Van Riel PL. The relationship between disease activity, joint destruction, and functional capacity over the course of rheumatoid arthritis. Arthritis Rheum 2001 Sep;44(9):2009-17. ##Van Der Pouw Kraan TC, Wijbrandts CA, Van Baarsen LG, Voskuyl AE, Rustenburg F, Baggen JM, et al. Rheumatoid arthritis subtypes identified by genomic profiling of peripheral blood cells: assignment of a type I interferon signature in a subpopulation of patients. Ann Rheum Dis 2007 Aug 1;66(8):1008-14.##Klareskog L, van der Heijde D, de Jager JP, Gough A, Kalden J, Malaise M, et al. Therapeutic effect of the combination of etanercept and methotrexate compared with each treatment alone in patients with rheumatoid arthritis: double-blind randomised controlled trial. Lancet 2004 Feb 28;363(9410):675-81.##Miriovsky BJ, Michaud K, Thiele GM, O'Dell JR, Cannon GW, Kerr G, et al. Anti-CCP antibody and rheumatoid factor concentrations predict greater disease activity in men with rheumatoid arthritis. Ann Rheum Dis 2010 Jul 1;69(7):1292-7.##Di Matteo A, Mankia K, Azukizawa M, Wakefield RJ. The role of musculoskeletal ultrasound in the rheumatoid arthritis continuum. Curr Rheumatol Rep 2020 Aug;22:1-2.##Stergaard M, Emery P, Conaghan PG, Fleischmann R, Hsia EC, Xu W, et al. Significant improvement in synovitis, osteitis, and bone erosion following golimumab and methotrexate combination therapy as compared with methotrexate alone: a magnetic resonance imaging study of 318 methotrexate‐naive rheumatoid arthritis patients. Arthritis Rheum 2011 Dec;63(12):3712-22. ##Quartuccio L, Fabris M, Pontarini E, Salvin S, Zabotti A, Benucci M, et al. The 158VV Fcgamma receptor 3A genotype is associated with response to rituximab in rheumatoid arthritis: results of an Italian multicentre study. Ann Rheum Dis 2014 Apr 1;73(4):716-21. ##Narváez J, Magallares B, Torné CD, Hernández MV, Reina D, Corominas H, et al. Predictive factors for induction of remission in patients with active rheumatoid arthritis treated with tocilizumab in clinical practice. Semin Arthritis Rheum 2016 Feb;45(4):386-90. ##Aletaha D, Smolen JS. The simplified disease activity index (SDAI) and clinical disease activity index (CDAI) to monitor patients in standard clinical care. Best Pract Res Clin Rheumatol 2007 Aug;21(4):663-75.##Saleem B, Keen H, Goeb V, Parmar R, Nizam S, Hensor EM, Churchman SM, Quinn M, Wakefield R, Conaghan PG, Ponchel F. Patients with RA in remission on TNF blockers: when and in whom can TNF blocker therapy be stopped?. Ann Rheum Dis 2010 Sep 1;69(9):1636-42. ##Sivasankaran R, Mallesh P, Chikkaiah PB, Zuhadulla M, Bhagvath B. Association of serum interleukin 6 levels with clinical outcome of COVID-19 associated mucormycosis. Int J Adv Med 2021;8:1319-22.##Emery P, Breedveld FC, Hall S, Durez P, Chang DJ, Robertson D, et al. Comparison of methotrexate monotherapy with a combination of methotrexate and etanercept in active, early, moderate to severe rheumatoid arthritis (COMET): a randomised, double-blind, parallel treatment trial. Lancet 2008 Aug 2;372(9636):375-82. ##Fraenkel L, Bathon JM, England BR, St. Clair EW, Arayssi T, Carandang K, et al. 2021 American College of Rheumatology guideline for the treatment of rheumatoid arthritis. Arthritis Rheum 2021 Jul;73(7):1108-23.##Rigopoulos D, Korfitis C, Gregoriou S, Katsambas AD. Infliximab in dermatological treatment: beyond psoriasis. Expert Opin Biol Ther 2008 Jan 1;8(1):123-33. ##Klotz U, Teml A, Schwab M. Clinical pharmacokinetics and use of infliximab. Clin Pharmacokinet 2007 Aug;46:645-60. ##Lim H, Lee SH, Lee HT, Lee JU, Son JY, Shin W, et al. Structural biology of the TNFα antagonists used in the treatment of rheumatoid arthritis. Int J Mol Sci 2018 Mar 7;19(3):768. ##Oriama R, Mudida R, Burger-Helmchen T. A Multi-level Perspective to Biosimilars Development: Pathways Towards Incremental Innovation in the Health Bioeconomy. In: Transdisciplinarity 2022 Nov 14 (pp. 249-266). Cham: Springer International Publishing.##Fleischmann RM, Alten R, Pileckyte M, Lobello K, Hua SY, Cronenberger C, et al. A comparative clinical study of PF-06410293, a candidate adalimumab biosimilar, and adalimumab reference product (Humira®) in the treatment of active rheumatoid arthritis. Arthritis Res Ther 2018 Dec;20:1-2.##Houel S, Hilliard M, Yu YQ, McLoughlin N, Martin SM, Rudd PM, et al. N-and O-glycosylation analysis of etanercept using liquid chromatography and quadrupole time-of-flight mass spectrometry equipped with electron-transfer dissociation functionality. Anal Chem 2014 Jan 7;86(1):576-84.##Smola MG, Soyer HP, Scharnagl E. Surgical treatment of dermatofibrosarcoma protuberans. A retrospective study of 20 cases with review of literature. Eur J Surg Oncol 1991 Oct 1;17(5):447-53.##Belei O, Dobrescu A, Iacob ER, Iacob D, Amaricai E, Marginean O. The role of molecular typing for DQ2 and DQ8 alleles using polymerase chain reaction amplification in children with autoimmune conditions. Rev Chim 2020;71(6):212-21.##Moreland LW. Clinical and pharmacological experience with etanercept. Expert Opin Investig Drugs 1999 Sep 1;8(9):1443-51. ##Zhou SY, Shu C, Korth‐Bradley J, Raible D, Palmisano M, Wadjula J, et al. Integrated population pharmacokinetics of etanercept in healthy subjects and in patients with rheumatoid arthritis ankylosing spondylitis. J Clin Pharmacol 2011 Jun;51(6):864-75.##Smolen JS, Beaulieu A, Rubbert-Roth A, Ramos-Remus C, Rovensky J, Alecock E, et al. Effect of interleukin-6 receptor inhibition with tocilizumab in patients with rheumatoid arthritis (OPTION study): a double-blind, placebo-controlled, randomised trial. Lancet 2008 Mar 22;371(9617):987-97.##Zulkiply NH, Ramachandran V, Muthukumarasamy R. A narrative review on the effectiveness of tocilizumab in reducing the mortality risk in covid-19 patients. Malaysian Journal of Medicine and Health Sciences 2021:268-79.##Abdallah H, Hsu JC, Lu P, Fettner S, Zhang X, Douglass W, et al. Pharmacokinetic and pharmacodynamic analysis of subcutaneous tocilizumab in patients with rheumatoid arthritis from 2 randomized, controlled trials: SUMMACTA and BREVACTA. J Clin Pharmacol 2017 Apr;57(4):459-68.##Boyce EG, Rogan EL, Vyas D, Prasad N, Mai Y. Sarilumab: review of a second IL-6 receptor antagonist indicated for the treatment of rheumatoid arthritis. Ann Pharmacother 2018 Aug;52(8):780-791.##Wells AF, Parrino J, Mangan EK, Paccaly A, Lin Y, Xu C, et al. Immunogenicity of sarilumab monotherapy in patients with rheumatoid arthritis who were inadequate responders or intolerant to disease-modifying antirheumatic drugs. Rheumatol Ther 2019 Sep 1;6:339-52.##Moretz D, Page J. Drug Class Update: Biologics for Autoimmune Conditions.##Fleischmann RM, Schechtman J, Bennett R, Handel ML, Burmester GR, Tesser J, et al. Anakinra, a recombinant human interleukin‐1 receptor antagonist (r‐metHuIL‐1ra), in patients with rheumatoid arthritis: a large, international, multicenter, placebo‐controlled trial. Arthritis Rheum 2003 Apr;48(4):927-34.##Cavalli G, Dinarello CA. Anakinra therapy for non-cancer inflammatory diseases. Front Pharmacol 2018 Nov 6:9:1157.##Negi K, Agarwal M, Pahuja I, Bhardwaj B, Rawat M, Bhaskar A, et al. Combating the challenges of COVID-19 pandemic: Insights into molecular mechanisms, immune responses and therapeutics against SARS-CoV-2. Oxf Open Immunol 2023 Jan 1;4(1):iqad001.##Emery P, Deodhar A, Rigby WF, Isaacs JD, Combe B, Racewicz AJ, et al. Efficacy and safety of different doses and retreatment of rituximab: a randomised, placebo-controlled trial in patients who are biological naive with active rheumatoid arthritis and an inadequate response to methotrexate (Study Evaluating Rituximab's Efficacy in MTX iNadequate rEsponders (SERENE)). Ann Rheum Dis 2010 Sep 1;69(9):1629-35.##Smolen JS, Landewé R, Bijlsma J, Burmester G, Chatzidionysiou K, Dougados M, et al. EULAR recommendations for the management of rheumatoid arthritis with synthetic and biological disease-modifying antirheumatic drugs: 2016 update. Ann Rheum Dis 2017 Jun 1;76(6):960-77.##Beck A, Diemer H, Ayoub D, Debaene F, Wagner-Rousset E, Carapito C, et al. Analytical characterization of biosimilar antibodies and Fc-fusion proteins. TrAC Trends in Analytical Chemistry 2013 Jul 1;48:81-95. ##Amin-Anaraki H, Kabiri-Samani S. Treatment of rheumatoid arthritis based on personalized medicine: a new approach in rheumatology. Personalized Medicine Journal 2023 Mar 1;8(28):35-45. ##Scheinfeld N. A comprehensive review and evaluation of the side effects of the tumor necrosis factor alpha blockers etanercept, infliximab and adalimumab. J Dermatolog Treat 2004 Sep 1;15(5):280-94.##Scheinfeld N. Adalimumab: a review of side effects. Expert Opin Drug Saf 2005 Jul 1;4(4):637-41. ##Preuss CV, Anjum F. Tocilizumab. I In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2024 Jan. 2022 Sep 21.##Bhandari S, Baral MR, Barbery M, Rudinskaya A, Sostin O. Hematologic side effects of biologics and kinase inhibitors used in rheumatologic diseases: a review of the current evidence. Ann Hematol 2022 Sep;101(9):1897-904.##Fleischmann RM, Tesser J, Schiff MH, Schechtman J, Burmester GR, Bennett R, et al. Safety of extended treatment with anakinra in patients with rheumatoid arthritis. Ann Rheum Dis 2006 Aug 1;65(8):1006-12.##Onrust SV, Lamb HM, Barman Balfour JA. Rituximab. Drugs 1999 Jul;58:79-88. ##

Exploring the Molecular Underpinnings of Skin Regeneration and Wound Healing: The Role of Renin Angiotensin 2 2 The aim of this study is to review the role of renin-angiotensin in skin regeneration and wound healing with a focus on molecular mechanisms. Angiotensin receptor type 1 (AT1R) are abundant in the wounded area, and thus, lead to the activation of ERK, STAT1, and STAT3 which can lead to epidermal self-renewal. The expression of Renin Angiotensin System (RAS) components was significantly lower in wounds caused by burning, rather than intact skin, noting that RAS is involved in the re-epithelialization of skin. ERK, STAT and STAT3 are the targets of Ang II, indicating that RAS active components are involved in fibroblast, stem cells and keratinocyte migration. The effect of inhibiting the RAS on wound healing is context-dependent. On one hand, it is suggested that inhibiting RAS during this phase may slow down wound healing speed. On the other hand, studies have shown that RAS inhibition in this phase can lead to α-SMA activation, ultimately accelerating the wound healing process. Most of the investigations indicate that the inhibition of RAS with Angiotensin Receptor Blockers (ARBs) and Angiotensin Converting Enzyme (ACE) plays a significant role in tissue remodeling in the last phase of wound healing. It has been shown that the inhibition of RAS can inhibit scar formation and fibrosis through the downregulation of inflammatory and fibrogenic agents, such as TGF-β, SMAD2/3, and TAK1, PDGF-BB, and HSP47. To sum up, that local administration of RAS regulators might lead to less scar formation and inflammation in the last phase of wound closure. 146 155 Seyedeh Hoda Qoreishi Faculty of Medicine, Mazandaran University of Medical SciencesUSERN Office, Mazandaran University of Medical Sciences Faculty of Medicine, Mazandaran University of Medical SciencesUSERN Office, Mazandaran University of Medical Sciences IranIran Mohammad Amin Khazeei Tabari USERN Office, Mazandaran University of Medical SciencesStudent Research Committee, Mazandaran University of Medical Sciences USERN Office, Mazandaran University of Medical SciencesStudent Research Committee, Mazandaran University of Medical Sciences IranIran Mihnea-Alexandru Găman Faculty of Medicine, “Carol Davila” University of Medicine and Pharmacy, 050474Department of Hematology, Center of Hematology and Bone Marrow Transplantation, Fundeni Clinical Institute, 022328 Faculty of Medicine, “Carol Davila” University of Medicine and Pharmacy, 050474Department of Hematology, Center of Hematology and Bone Marrow Transplantation, Fundeni Clinical Institute, 022328 RomaniaRomania Armaghan Kazeminejad Department of Dermatology, Antimicrobial Resistance Research Center, Communicable Diseases institute, Mazandaran University of Medical Sciences Department of Dermatology, Antimicrobial Resistance Research Center, Communicable Diseases institute, Mazandaran University of Medical Sciences Iran AngiotensinAngiotensin-converting enzyme inhibitorsCicatrixDown-regulationFibroblastsInflammationKeratinocytesReceptorsRe-EpithelializationStem cells 60580.pdf Krafts KP. Tissue repair: The hidden drama. Organogenesis 2010;6(4):225-33. ##Eming SA, Martin P, Tomic-Canic M. Wound repair and regeneration: mechanisms, signaling, and translation. Sci Transl Med 2014;6(265):265sr6.##Gonzalez AC, Costa TF, Andrade ZA, Medrado AR. Wound healing - A literature review. An Bras Dermatol 2016;91(5):614-20. ##Cowled P, Fitridge R. Ch18 Mechanisms of Vascular Disease: University of Adelaide Press; 2011.##Diller RB, Tabor AJ. The role of the extracellular matrix (ECM) in wound healing: A review. Biomimetics 2022;7(3):87. ##Akita S. Wound repair and regeneration: mechanisms, signaling. MDPI; 2019. p. 6328.##Reinke J, Sorg H. Wound repair and regeneration. Eur Surg Res 2012;49(1):35-43. ##Martyniak A, Tomasik PJ. A new perspective on the renin-angiotensin system. Diagnostics (Basel) 2023;13(1):16.##Fountain JH, Kaur J, Lappin SL. Physiology, Renin Angiotensin System. StatPearls. Treasure Island (FL): StatPearls Publishing Copyright © 2023, StatPearls Publishing LLC.; 2023.##Touyz R, Berry C. Recent advances in angiotensin II signaling. Braz J Med Biol Res 2002;35:1001-15.##Dzau VJ, Gibbons G, Pratt R. Molecular mechanisms of vascular renin-angiotensin system in myointimal hyperplasia. Hypertension 1991;18(4_supplement):II100-5. ##Ardaillou R. Angiotensin II receptors. J Am Soc Nephrol 1999;10:S30-9.##Weber KT. Extracellular matrix remodeling in heart failure: a role for de novo angiotensin II generation. Circulation 1997;96(11):4065-82.##Rodgers K, Xiong S, Felix J, Roda N, Espinoza T, Maldonado S, et al. Development of angiotensin (1‐7) as an agent to accelerate dermal repair. Wound Repair Regen 2001;9(3):238-47.##Silva IMS, Assersen KB, Willadsen NN, Jepsen J, Artuc M, Steckelings UM. The role of the renin‐angiotensin system in skin physiology and pathophysiology. Exp Dermatol 2020;29(9):891-901. ##Kawaguchi Y, Takagi K, Hara M, Fukasawa C, Sugiura T, Nishimagi E, et al. Angiotensin II in the lesional skin of systemic sclerosis patients contributes to tissue fibrosis via angiotensin II type 1 receptors. Arthritis Rheuma 2004;50(1):216-26.##Wagenaar LJ, Voors AA, Buikema H, van Gilst WH. Angiotensin receptors in the cardiovascular system. Can J Cardiol 2002 Dec;18(12):1331-9.##Ren M, Yang C, Zhu P, Cheng H, Yan L. Activation of skin renin–angiotensin system in diabetic rats. Endocrine 2011;39(3):242-50. ##Bernasconi R, Nyström A. Balance and circumstance: the renin angiotensin system in wound healing and fibrosis. Cell Signal 2018;51:34-46.##Aleksiejczuk M, Gromotowicz-Poplawska A, Marcinczyk N, Przylipiak A, Chabielska E. The expression of the renin-angiotensin-aldosterone system in the skin and its effects on skin physiology and pathophysiology. J Physiol Pharmacol 2019;70(3). ##Silva IMS, Assersen KB, Willadsen NN, Jepsen J, Artuc M, Steckelings UM. The role of the renin‐angiotensin system in skin physiology and pathophysiology. Exp Dermatol 2020;29(9):891- 901.##Hedayatyanfard K, Haddadi NS, Ziai SA, Karim H, Niazi F, Steckelings UM, et al. The renin‐angiotensin system in cutaneous hypertrophic scar and keloid formation. Exp Dermatol 2020;29(9):902-9.##Sano S, Itami S, Takeda K, Tarutani M, Yamaguchi Y, Miura H, et al. Keratinocyte-specific ablation of Stat3 exhibits impaired skin remodeling, but does not affect skin morphogenesis. EMBO J 1999;18(17):4657-68. ##Medley SC, Rathnakar BH, Georgescu C, Wren JD, Olson LE. Fibroblast-specific Stat1 deletion enhances the myofibroblast phenotype during tissue repair. Wound Repair Regen 2020;28(4):448-59. ##Reudelhuber TL. The renin-angiotensin system: peptides and enzymes beyond angiotensin II. Curr Opin Nephrol Hypertens 2005;14(2):155-9.##Balakumar P, Jagadeesh G. A century old renin-angiotensin system still grows with endless possibilities: AT1 receptor signaling cascades in cardiovascular physiopathology. Cell Signal 2014;26(10):2147-60. ##Karnik SS, Unal H, Kemp JR, Tirupula KC, Eguchi S, Vanderheyden PM, et al. International Union of Basic and Clinical Pharmacology. XCIX. Angiotensin receptors: interpreters of pathophysiological angiotensinergic stimuli. Pharmacol Rev 2015;67(4):754-819. ##De Gasparo M, Catt K, Inagami T, Wright J, Unger T. International union of pharmacology. XXIII. The angiotensin II receptors. Pharmacol Rev 2000;52(3):415-72.##Sumners C, Peluso AA, Haugaard AH, Bertelsen JB, Steckelings UM. Anti‐fibrotic mechanisms of angiotensin AT2‐receptor stimulation. Acta Physiologica 2019;227(1):e13280.##Rompe F, Unger T, Steckelings UM. The angiotensin AT2 receptor in inflammation. Drug News Perspect 2010;23(2):104-11.##Saavedra JM. Brain angiotensin II receptor subtypes. Angiotensin receptors: Springer; 1994. p. 151-75.##Steckelings UM, Wollschläger T, Peters J, Henz BM, Hermes B, Artuc M. Human skin: source of and target organ for angiotensin II. Exp Dermatol 2004;13(3):148-54. ##Jadhav SS, Sharma N, Meeks CJ, Mordwinkin NM, Espinoza TB, Roda NR, et al. Effects of combined radiation and burn injury on the renin–angiotensin system. Wound Repair Regen 2013;21(1):131-40.##Wu H-J, Liu H-W, Cheng B, Gu Y-F, Xie B, Xiao L-L, et al. The change of angiotensin II production and its receptor expression during wound healing: possible role of angiotensin II in wound healing. Zhonghua Zheng Xing Wai Ke Za Zhi 2011;27(2):124-8. [Chinese]##Herrera M, Sparks MA, Alfonso-Pecchio AR, Harrison-Bernard LM, Coffman TM. Lack of specificity of commercial antibodies leads to misidentification of angiotensin type 1 receptor protein. Hypertension 2013;61(1):253-8.##Hafko R, Villapol S, Nostramo R, Symes A, Sabban EL, Inagami T, et al. Commercially available angiotensin II At2 receptor antibodies are nonspecific. PloS One 2013;8(7):e69234. ##Jiang X, Wu F, Xu Y, Yan JX, Wu YD, Li SH, et al. A novel role of angiotensin II in epidermal cell lineage determination: Angiotensin II promotes the differentiation of mesenchymal stem cells into keratinocytes through the p38 MAPK, JNK and JAK 2 signalling pathways. Exp Dermatol 2019;28(1):59-65. ##Selvarajah V, Connolly K, McEniery C, Wilkinson I. Skin sodium and hypertension: a paradigm shift? Curr Hypertens Rep 2018;20(11):94.##Opländer C, Volkmar CM, Paunel-Görgülü A, van Faassen EE, Heiss C, Kelm M, et al. Whole body UVA irradiation lowers systemic blood pressure by release of nitric oxide from intracutaneous photolabile nitric oxide derivates. Circ Res 2009;105(10):1031-40. ##Cowburn AS, Takeda N, Boutin AT, Kim J-W, Sterling JC, Nakasaki M, et al. HIF isoforms in the skin differentially regulate systemic arterial pressure. Proc Natl Acad Sci USA 2013;110(43):17570-5. ##Huang H, Fan Y, Gao Z, Wang W, Shao N, Zhang L, et al. HIF-1α contributes to Ang II-induced inflammatory cytokine production in podocytes. BMC Pharmacol Toxicol 2019;20(1):59.##Liu C, Zhang JW, Hu L, Song YC, Zhou L, Fan Y, et al. Activation of the AT1R/HIF-1 α /ACE axis mediates angiotensin II-induced VEGF synthesis in mesenchymal stem cells. Biomed Res Int 2014;2014:627380.##Nangaku M, Fujita T. Activation of the renin-angiotensin system and chronic hypoxia of the kidney. Hypertens Res 2008;31(2):175-84.##Díaz-García D, Filipová A, Garza-Veloz I, Martinez-Fierro ML. A beginner's introduction to skin stem cells and wound healing. Int J Mol Sci 2021;22(20):11030. ##Mentz RJ, Bakris GL, Waeber B, McMurray JJ, Gheorghiade M, Ruilope LM, et al. The past, present and future of renin–angiotensin aldosterone system inhibition. Int J Cardiol 2013;167(5):1677-87.##Scholzen TE, Luger TA. Neutral endopeptidase and angiotensin‐converting enzyme–key enzymes terminating the action of neuroendocrine mediators. Exp Dermatol 2004;13:22-6. ##Liu HW, Cheng B, Li JF, Wu HJ, Li KY, Sun TZ, et al. Characterization of angiotensin‐converting enzyme expression during epidermis morphogenesis in humans: a potential marker for epidermal stem cells. Br J Dermatol 2009;160(2):250-8.##Liao X, Xiao J, Li S-H, Xiao L-L, Cheng B, Fu X-B, et al. Critical role of the endogenous renin-angiotensin system in maintaining self-renewal and regeneration potential of epidermal stem cells. Biochim Biophys Acta Mol Basis Dis 2019;1865(10):2647-56. ##Singh S, Young A, McNaught C-E. The physiology of wound healing. Surgery (Oxford) 2017;35(9):473-7.##Kurosaka M, Suzuki T, Hosono K, Kamata Y, Fukamizu A, Kitasato H, et al. Reduced angiogenesis and delay in wound healing in angiotensin II type 1a receptor-deficient mice. Biomed Pharmacother 2009;63(9):627-34.##Khazeei Tabari MA, Mirjalili R, Khoshhal H, Shokouh E, Khandan M, Hasheminasabgorji E, et al. Nature against diabetic retinopathy: A review on antiangiogenic, antioxidant, and anti-Inflammatory phytochemicals. Evid Based Complement Alternat Med 2022;2022:4708527.##Vaduganathan M, Butler J, Pitt B, Gheorghiade M. Contemporary drug development in heart failure: call for hemodynamically neutral therapies. Circ Heart Fail 2015;8(4):826-31. ##Kamber M, Papalazarou V, Rouni G, Papageorgopoulou E, Papalois A, Kostourou V. Angiotensin II inhibitor facilitates epidermal wound regeneration in diabetic mice. Front Physiol 2015;6:170. ##Lapray M, Petit J-M, Fourmont C, Rouland A, Vergès B, Bouillet B. Healing of diabetic foot ulcers is independently associated with the use of angiotensin receptor blockers but not with those of diuretics and angiotensin conversion enzyme inhibitors. Diabetes Metab 2022;48(4):101336. ##Jadhav SS, Sharma N, Meeks CJ, Mordwinkin NM, Espinoza TB, Roda NR, et al. Effects of combined radiation and burn injury on the renin-angiotensin system. Wound Repair Regen 2013;21(1):131-40.##Darby IA, Desmoulière A. Scar formation: cellular mechanisms. In: Textbook on Scar Management: State of the Art Management and Emerging Technologies [Internet]. Cham (CH): Springer; 2020. Chapter 3.##Cabral-Pacheco GA, Garza-Veloz I, Castruita-De la Rosa C, Ramirez-Acuna JM, Perez-Romero BA, Guerrero-Rodriguez JF, et al. The roles of matrix metalloproteinases and their inhibitors in human diseases. Int J Mol Sci 2020;21(24):9739.##Spielman AF, Griffin MF, Parker J, Cotterell AC, Wan DC, Longaker MT. Beyond the scar: A basic science review of wound remodeling. Adv Wound Care (New Rochelle) 2023;12(2):57-67.##Zhang T, Wang X-F, Wang Z-C, Lou D, Fang Q-Q, Hu Y-Y, et al. Current potential therapeutic strategies targeting the TGF-β/Smad signaling pathway to attenuate keloid and hypertrophic scar formation. Biomed Pharmacother 2020;129:110287. ##Ogawa R, Akaishi S. Endothelial dysfunction may play a key role in keloid and hypertrophic scar pathogenesis–keloids and hypertrophic scars may be vascular disorders. Med Hypotheses 2016;96:51-60.##Amadeu T, Braune A, Mandarim-de-Lacerda C, Porto LC, Desmoulière A, Costa A. Vascularization pattern in hypertrophic scars and keloids: a stereological analysis. Pathol Res Pract 2003;199(7):469-73.##Takeda H, Katagata Y, Hozumi Y, Kondo S. Effects of angiotensin II receptor signaling during skin wound healing. Am J Pathol 2004;165(5):1653-62.##Steckelings UM, Henz B, Wiehstutz S, Unger T, Artuc M. Differential expression of angiotensin receptors in human cutaneous wound healing. Br J Dermatol 2005;153(5):887-93.##Morihara K, Takai S, Takenaka H, Sakaguchi M, Okamoto Y, Morihara T, et al. Cutaneous tissue angiotensin–converting enzyme may participate in pathologic scar formation in human skin. J Am Acad Dermatol 2006;54(2):251-7.##Akershoek JJ, Vlig M, Brouwer K, Talhout W, Beelen RH, Middelkoop E, et al. The presence of tissue renin-angiotensin system components in human burn wounds and scars. Burns Open 2018;2(3):114-21.##Niazi F, Hooshyar SH, Hedayatyanfard K, Ziai SA, Doroodgar F, Niazi S, et al. Detection of angiotensin II and AT1 receptor concentrations in keloid and hypertrophic scar. J Clin Aesthet Dermatol 2018;11(10):36-9.##Hedayatyanfard K, Ziai SA, Niazi F, Habibi I, Habibi B, Moravvej H. Losartan ointment relieves hypertrophic scars and keloid: A pilot study. Wound Rep Regen 2018;26(4):340-3. ##Hu YY, Fang QQ, Wang XF, Zhao WY, Zheng B, Zhang DD, et al. Angiotensin-converting enzyme inhibitor and angiotensin II type 1 receptor blocker: Potential agents to reduce post-surgical scar formation in humans. Basic Clin Pharmacol Toxicol 2020;127(6):488-94. ##Faghih M, Hosseini SM, Smith B, Ansari AM, Lay F, Ahmed AK, et al. Knockout of Angiotensin AT2 receptors accelerates healing but impairs quality. Aging (Albany NY) 2015;7(12):1185-97. ##Liao X, Xiao J, Li SH, Xiao LL, Cheng B, Fu XB, et al. Critical role of the endogenous renin-angiotensin system in maintaining self-renewal and regeneration potential of epidermal stem cells. Biochim Biophys Acta Mol Basis Dis 2019;1865(10):2647-56.##Rha EY, Kim JW, Kim JH, Yoo G. Angiotensin-converting enzyme inhibitor, captopril, improves scar healing in hypertensive rats. Int J Med Sci 2021;18(4):975-83. ##Zheng B, Fang QQ, Wang XF, Shi BH, Zhao WY, Chen CY, et al. The effect of topical Ramipril and Losartan cream in inhibiting scar formation. Biomed Pharmacother 2019;118:109394. ##Ohishi T, Saito H, Tsusaka K, Toda K, Inagaki H, Hamada Y, et al. Anti-fibrogenic effect of an angiotensin converting enzyme inhibitor on chronic carbon tetrachloride-induced hepatic fibrosis in rats. Hepatol Res 2001;21(2):147-58.##Mezzano SA, Ruiz-Ortega M, Egido J. Angiotensin II and renal fibrosis. Hypertension 2001;38(3 Pt 2):635-8.##Tan WQ, Fang QQ, Shen XZ, Giani JF, Zhao TV, Shi P, et al. Angiotensin‐converting enzyme inhibitor works as a scar formation inhibitor by down‐regulating Smad and TGF‐β‐activated kinase 1 (TAK1) pathways in mice. Br J Pharmacol 2018;175(22):4239-52. ##Fang Q-Q, Wang X-F, Zhao W-Y, Ding S-L, Shi B-H, Xia Y, et al. Angiotensin-converting enzyme inhibitor reduces scar formation by inhibiting both canonical and noncanonical TGF-β1 pathways. Sci Rep 2018;8(1):3332.##Chen J, Zhao S, Liu Y, Cen Y, Nicolas C. Effect of captopril on collagen metabolisms in keloid fibroblast cells. ANZ J Surg 2016;86(12):1046-51.##Suzuki Y, Ruiz-Ortega M, Egido J. Angiotensin II: a double-edged sword in inflammation. J Nephrol 2000;13 Suppl 3:S101-10.##Yang L, Qiu CX, Ludlow A, Ferguson MW, Brunner G. Active transforming growth factor-β in wound repair: determination using a new assay. Am J Pathol 1999;154(1):105-11. ##Murphy A, LeVatte T, Boudreau C, Midgen C, Gratzer P, Marshall J, et al. Angiotensin II type I receptor blockade is associated with decreased cutaneous scar formation in a rat model. Plast Reconstr Surg 2019;144(5):803e-13e.##

Exploring the Potential of In vitro Maturation (IVM) of Oocytes: Indications, Applications, and Treatment Protocols 2 2 This review addresses the current understanding of In Vitro Maturation (IVM) treatment, including indications and effective treatment protocols influencing oocyte developmental competence. A comprehensive literature search was performed to gather relevant studies, clinical trials, and reviews related to IVM. Databases such as PubMed, MEDLINE, and pertinent medical journals were searched. The selected literature was analyzed and synthesized to offer a comprehensive overview. IVM has emerged as a promising technique for inducing maturation in immature oocytes across various developmental stages. Its applications extend to areas utilizing In Vitro Fertilization (IVF), gaining traction as a treatment option for Polycystic Ovary Syndrome (PCOS) and fertility preservation in cancer patients. Recent advancements have led to improved global pregnancy rates, resulting in successful births. IVM also holds potential in reducing risks associated with conventional IVF, including ovarian hyperstimulation syndrome and multiple pregnancies. Despite these advantages, IVM adoption in clinical practice remains limited. Ongoing research aims to refine therapeutic protocols and expand clinical indications. IVM holds promise in assisted reproductive technology, spanning applications from cancer patient fertility preservation to addressing PCOS. Enhanced pregnancy rates highlight efficacy, while risk reduction compared to IVF underscores its importance. Further research is needed for optimal use across patient groups, emphasizing protocol refinement and expanded applications. 156 164 Hossein Torkashvand Department of Anatomical Science, School of Medicine, Iran University of Medical SciencesEndometrium and Endometriosis Research Centre, Hamadan University of Medical Sciences Department of Anatomical Science, School of Medicine, Iran University of Medical SciencesEndometrium and Endometriosis Research Centre, Hamadan University of Medical Sciences IranIran Ronak Shabani Reproductive Sciences and Technology Research Center, Department of Anatomy, Iran University of Medical Sciences Reproductive Sciences and Technology Research Center, Department of Anatomy, Iran University of Medical Sciences Iran Iraj Amiri Endometrium and Endometriosis Research Centre, Hamadan University of Medical Sciences Endometrium and Endometriosis Research Centre, Hamadan University of Medical Sciences Iran Roya Darakhshan Endometriosis Research Center, Iran University of Medical Sciences Endometriosis Research Center, Iran University of Medical Sciences Iran Behnam Maleki Infertility Center, Department of Obstetrics and Gynecology, Mazandaran University of Medical Sciences Infertility Center, Department of Obstetrics and Gynecology, Mazandaran University of Medical Sciences Iran Mohammad Reza Doostabadi Department of Anatomy, School of Medicine, Birjand University of Medical Sciences Department of Anatomy, School of Medicine, Birjand University of Medical Sciences Iran Mehdi Mehdizadeh Clinical protocolFertility preservationIn vitro maturation oocyte (IVM) 60581.pdf Li H, Chian R-C. Follicular development and oocyte growth. In: Chian, RC., Nargund, G., Huang, J. (eds) Development of In Vitro Maturation for Human Oocytes. Springer, Cham. 2017;37-57.##Steptoe PC, Edwards RG. Birth after the reimplantation of a human embryo. Lancet 1978;312(8085):366.##Lunenfeld B. Gonadotropin stimulation: past, present and future. Reprod Med Biol 2012;11(1):11-25.##Lunenfeld B, Bilger W, Longobardi S, Alam V, D’Hooghe T, Sunkara SK. The development of gonadotropins for clinical use in the treatment of infertility. Front Endocrinol (Lausanne) 2019;10:429.##Timmons D, Montrief T, Koyfman A, Long B. Ovarian hyperstimulation syndrome: A review for emergency clinicians. Am J Emerg Med 2019;37(8):1577-84. ##Tarlatzi TB, Venetis CA, Devreker F, Englert Y, Delbaere A. What is the best predictor of severe ovarian hyperstimulation syndrome in IVF? A cohort study. J Assist Reprod Genet 2017;34(10):1341-51. ##Choi S-H, Shapiro H, Robinson GE, Irvine J, Neuman J, Rosen B, et al. Psychological side-effects of clomiphene citrate and human menopausal gonadotrophin. J Psychosom Obstet Gynecol 2005;26(2):93-100.##Yang S-H, Patrizio P, Yoon S-H, Lim J-H, Chian R-C. Comparison of pregnancy outcomes in natural cycle IVF/M treatment with or without mature oocytes retrieved at time of egg collection. Syst Biol Reprod Med 2012;58(3):154-9.##Edwards RG. Meiosis in ovarian oocytes of adult mammals. Nature 1962;196(4853):446-50.##Braam SC, Consten D, Smeenk JMJ, Cohlen BJ, Curfs MHJM, Hamilton CJCM, et al. In vitro maturation of oocytes in women at risk of ovarian hyperstimulation syndrome-a prospective multicenter cohort study. Int J Fertil Steril 2019;13(1):38-44.##De Vos M, Smitz J, Thompson JG, Gilchrist RB. The definition of IVM is clear—variations need defining. Hum Reprod 2016;31(11):2411-5. ##Tabatabaie M, Amiri S, Golestan Jahromi M, Sene AA, Zandieh Z, Mehdizadeh M, et al. The effect of Myo-Inositol supplement on molecular regulation of folliculogenesis, steroidogenesis, and assisted reproductive technique outcomes in patients with polycystic ovarian syndrome. Mol Biol Rep 2022;49(2):875-84. ##Dahan MH, Tan SL, Chung J, Son W-Y. Clinical definition paper on in vitro maturation of human oocytes. Hum Reprod 2016;31(7):1383-6.##La X, Zhao J, Wang Z. Clinical Application of In Vitro Maturation of Oocytes. In: Bin Wu and Huai L. Feng (eds) Embryology-Theory and Practice. IntechOpen; 2019.##Coticchio G. IVM in need of clear definitions. Hum Reprod 2016;31(7):1387-9. ##Govahi A, Nasr-Esfahani M-H, Amjadi F, Mahdevar M, Mehdizadeh R, Mehdizadeh M. Cutting-edge techniques provide insights regarding repeated implantation failure patients. Reprod Biomed Online 2023;46(4):687-96.##Rodrigues P, Marques M, Manero JA, Marujo MD, Carvalho MJ, Plancha CE. In vitro maturation of oocytes as a laboratory approach to polycystic ovarian syndrome (PCOS): From oocyte to embryo. WIREs Mech Dis 2023;15(3):e1600. ##Hourvitz A, Maman E, Brengauz M, Machtinger R, Dor J. In vitro maturation for patients with repeated in vitro fertilization failure due to “oocyte maturation abnormalities.” Fertil Steril 2010;94(2):496-501.##Chang EM, Song HS, Lee DR, Lee WS, Yoon TK. In vitro maturation of human oocytes: Its role in infertility treatment and new possibilities. Clin Exp Reprod Med 2014;41(2):41-6. ##Yalçınkaya E, Çalışkan E, Budak Ö. In vitro maturation may prevent the cancellation of in vitro fertilization cycles in poor responder patients: A case report. J Turkish Ger Gynecol Assoc 2013;14(4):235-7.##Jie H, Zhao M, Alqawasmeh OAM, Chan CPS, Lee TL, Li T, et al. In vitro rescue immature oocytes–a literature review. Hum Fertil (Camb) 2022;25(4):640-50. ##Gong X, Li H, Zhao Y. The improvement and clinical application of human oocyte in vitro maturation (IVM). Reprod Sci 2022;29(8):2127-35. ##Kornilov NV, Pavlova MN, Yakovlev PP. The live birth in a woman with resistant ovary syndrome after in vitro oocyte maturation and preimplantation genetic testing for aneuploidy. J Assist Reprod Genet 2021;38(6):1303-9. ##Edwards RG, Bavister BD, Steptoe PC. Early stages of fertilization in vitro of human oocytes matured in vitro. Nature 1969;221(5181):632-5. ##Amjadi F, Zandieh Z, Mehdizadeh M, Ajdary M, Aghamajidi A, Raoufi E, et al. Molecular signature of immunological mechanism behind impaired endometrial receptivity in polycystic ovarian syndrome. Arch Endocrinol Metab 2022;66(3):303-11. ##Trounson A, Wood C, Kausche A. In vitro maturation and the fertilization and developmental competence of oocytes recovered from untreated polycystic ovarian patients. Fertil Steril 1994;62(2):353-62. ##Child TJ, Abdul-Jalil AK, Gulekli B, Tan SL. In vitro maturation and fertilization of oocytes from unstimulated normal ovaries, polycystic ovaries, and women with polycystic ovary syndrome. Fertil Steril 2001;76(5):936-42. ##Seok HH, Song H, Lyu SW, Kim YS, Lee DR, Lee WS, et al. Application of serum anti-Müllerian hormone levels in selecting patients with polycystic ovary syndrome for in vitro maturation treatment. Clin Exp Reprod Med 2016 Jun 23;43(2):126-32. ##Gremeau A-S, Andreadis N, Fatum M, Craig J, Turner K, Mcveigh E, et al. In vitro maturation or in vitro fertilization for women with polycystic ovaries? A case–control study of 194 treatment cycles. Fertil Steril 2012;98(2):355-60. ##Siristatidis C, Sergentanis TN, Vogiatzi P, Kanavidis P, Chrelias C, Papantoniou N, et al. In vitro maturation in women with vs. without polycystic ovarian syndrome: a systematic review and meta-analysis. PLoS One 2015;10(8):e0134696. ##Yoon H-G, Yoon S-H, Son W-Y, Lee S-W, Park S-P, Im K-S. Clinical assisted reproduction: pregnancies resulting from in vitro matured oocytes collected from women with regular menstrual cycle. J Assist Reprod Genet 2001;18(6):325-9. ##Demirtas E, Elizur SE, Holzer H, Gidoni Y, Son W-Y, Chian R-C, et al. Immature oocyte retrieval in the luteal phase to preserve fertility in cancer patients. Reprod Biomed Online 2008;17(4):520-3.##Fadini R, Renzini MM, Dal Canto M, Epis A, Crippa M, Caliari I, et al. Oocyte in vitro maturation in normo-ovulatory women. Fertil Steril 2013;99(5):1162-9. ##Fadini R, Dal Canto MB, Renzini MM, Brambillasca F, Comi R, Fumagalli D, et al. Effect of different gonadotrophin priming on IVM of oocytes from women with normal ovaries: a prospective randomized study. Reprod Biomed Online 2009;19(3):343-51. ##Braga DP de AF, Figueira R de CS, Ferreira RC, Pasqualotto FF, Iaconelli Jr A, Borges Jr E. Contribution of in-vitro maturation in ovarian stimulation cycles of poor-responder patients. Reprod Biomed Online 2010;20(3):335-40. ##Li Y, Pan P, Yuan P, Qiu Q, Yang D. Successful live birth in a woman with resistant ovary syndrome following in vitro maturation of oocytes. J Ovarian Res 2016;9(1):54. ##Lim KS, Chae SJ, Choo CW, Ku YH, Lee HJ, Hur CY, et al. In vitro maturation: Clinical applications. Clin Exp Reprod Med 2013;40(4):143-7. ##Lopata A, Johnston IWH, Hoult IJ, Speirs AI. Pregnancy following intrauterine implantation of an embryo obtained by in vitro fertilization of a preovulatory egg. Fertil Steril 1980;33(2):117-20.##Grynberg M, Peltoketo H, Christin-Maître S, Poulain M, Bouchard P, Fanchin R. First birth achieved after in vitro maturation of oocytes from a woman endowed with multiple antral follicles unresponsive to follicle-stimulating hormone. J Clin Endocrinol Metab 2013;98(11):4493-8.##Flageole C, Toufaily C, Bernard DJ, Ates S, Blais V, Chénier S, et al. Successful in vitro maturation of oocytes in a woman with gonadotropin-resistant ovary syndrome associated with a novel combination of FSH receptor gene variants: a case report. J Assist Reprod Genet 2019;36(3):425-32.##Galvao A, Segers I, Smitz J, Tournaye H, De Vos M. In vitro maturation (IVM) of oocytes in patients with resistant ovary syndrome and in patients with repeated deficient oocyte maturation. J Assist Reprod Genet 2018;35(12):2161-71. ##de Ziegler D, Streuli I, Gayet V, Frydman N, Bajouh O, Chapron C. Retrieving oocytes from small non-stimulated follicles in polycystic ovary syndrome (PCOS): in vitro maturation (IVM) is not indicated in the new GnRH antagonist era. Fertil Steril 2012;98(2):290-3.##Lindenberg S. New approach in patients with polycystic ovaries, lessons for everyone. Fertil Steril 2013;99(5):1170-2.##Pak KA, Lee WD, Lim JH. Optimal size of the dominant follicle on HCG injection day of hyperresponder who received controlled ovarian hyperstimulation followed by in vitro maturation. Fertil Steril 2009;92(3):S253. ##Prasath EB, Chan MLH, Wong WHW, Lim CJW, Tharmalingam MD, Hendricks M, et al. First pregnancy and live birth resulting from cryopreserved embryos obtained from in vitro matured oocytes after oophorectomy in an ovarian cancer patient. Hum Reprod 2014;29(2):276-8.##Jeruss JS, Woodruff TK. Preservation of fertility in patients with cancer. N Engl J Med 2009;360(9):902-11.##Walls ML, Hart RJ. In vitro maturation. Best Pract Res Clin Obstet Gynaecol 2018;53:60-72. ##Creux H, Monnier P, Son W-Y, Buckett W. Thirteen years’ experience in fertility preservation for cancer patients after in vitro fertilization and in vitro maturation treatments. J Assist Reprod Genet 2018;35:583-92.##Oktay K, Buyuk E, Libertella N, Akar M, Rosenwaks Z. Fertility preservation in breast cancer patients: a prospective controlled comparison of ovarian stimulation with tamoxifen and letrozole for embryo cryopreservation. J Clin Oncol 2005;23(19):4347-53. ##Shalom-Paz E, Almog B, Shehata F, Huang J, Holzer H, Chian R-C, et al. Fertility preservation for breast-cancer patients using IVM followed by oocyte or embryo vitrification. Reprod Biomed Online 2010;21(4):566-71. ##Gulekli B, Kovali M, Aydiner F, Dogan S, Dogan SS. IVM is an alternative for patients with PCO after failed conventional IVF attempt. J Assist Reprod Genet 2011;28(6):495-9. ##Hatirnaz S, Hatirnaz E, Çelik S, Çalışkan CS, Tinelli A, Malvasi A, et al. Unraveling the puzzle: Oocyte maturation abnormalities (OMAS). Diagnostics 2022;12(10): 2501.##Edward RG. Are minimal stimulation IVF and IVM set to replace routine IVF? Reprod Biomed Online 2007;14(2):267-70.##Christopikou D, Karamalegos C, Doriza S, Argyrou M, Sisi P, Davies S, et al. Spindle and chromosome configurations of human oocytes matured in vitro in two different culture media. Reprod Biomed Online 2010;20(5):639-48. ##Hatırnaz Ş, Ata B, Hatırnaz ES, Dahan MH, Tannus S, Tan J, et al. Oocyte in vitro maturation: A sytematic review. Turkish J Obstet Gynecol 2018;15(2):112-25. ##Zhang XY, Ata B, Son W-Y, Buckett WM, Tan S-L, Ao A. Chromosome abnormality rates in human embryos obtained from in-vitro maturation and IVF treatment cycles. Reprod Biomed Online 2010;21(4):552-9.##Ao A, Zhang XY, Tan SL. First successful pregnancy following PGD for chromosome translocation on embryos generated from in-vitro matured oocytes: a case report. Reprod Biomed Online 2011;22(4):371-5.##Das M, Son W-Y. In vitro maturation (IVM) of human immature oocytes: is it still relevant? Reprod Biol Endocrinol 2023;21(1):110.##Liu J, Lu G, Qian Y, Mao Y, Ding W. Pregnancies and births achieved from in vitro matured oocytes retrieved from poor responders undergoing stimulation in in vitro fertilization cycles. Fertil Steril 2003;80(2):447-9.##Li J, Xu Y, Zhou G, Guo J, Xin N. Natural cycle IVF/IVM may be more desirable for poor responder patients after failure of stimulated cycles. J Assist Reprod Genet 2011;28:791-5. ##Hatırnaz Ş, Akarsu S, Hatırnaz ES, Işık AZ, Dahan MH. The use of in vitro maturation in stimulated antagonist in vitro fertilization cycles of normo-hyperresponder women due to arrested follicular development: A rescue procedure. Turkish J Obstet Gynecol 2018;15(3):141-6.##Jie H, Zhao M, Alqawasmeh OAM, Chan CPS, Lee TL, Li T, et al. In vitro rescue immature oocytes – a literature review. Hum Fertil Hum Fertil (Camb) 2022 Oct;25(4):640-50.##De Vos M, Grynberg M, Ho TM, Yuan Y, Albertini DF, Gilchrist RB. Perspectives on the development and future of oocyte IVM in clinical practice. J Assist Reprod Genet 2021;38(6):1265-80.##Gong X, Li H, Zhao Y. The improvement and clinical application of human oocyte in vitro maturation (IVM). Reprod Sci 2021;29(8):2127-2135. ##Ho VNA, Pham TD, Le AH, Ho TM, Vuong LN. Live birth rate after human chorionic gonadotropin priming in vitro maturation in women with polycystic ovary syndrome. J Ovarian Res 2018;11(1):70. ##Licht P, Fluhr H, Neuwinger J, Wallwiener D, Wildt L. Is human chorionic gonadotropin directly involved in the regulation of human implantation? Mol Cell Endocrinol 2007;269(1-2):85-92.##De Vos M, Ortega-Hrepich C, Albuz FK, Guzman L, Polyzos NP, Smitz J, et al. Clinical outcome of non–hCG-primed oocyte in vitro maturation treatment in patients with polycystic ovaries and polycystic ovary syndrome. Fertil Steril 2011;96(4):860-4. ##Makrigiannakis A, Vrekoussis T, Zoumakis E, Kalantaridou SN, Jeschke U. The role of HCG in implantation: a mini-review of molecular and clinical evidence. Int J Mol Sci 2017;18(6):1305. ##Chian R-C, Gülekli B, Buckett WM, Tan S-L. Priming with human chorionic gonadotropin before retrieval of immature oocytes in women with infertility due to the polycystic ovary syndrome. N Engl J Med 1999;341(21):1624-6. ##Grynberg M, Poulain M, Le Parco S, Sifer C, Fanchin R, Frydman N. Similar in vitro maturation rates of oocytes retrieved during the follicular or luteal phase offer flexible options for urgent fertility preservation in breast cancer patients. Hum Reprod 2016;31(3):623-9. ##Reavey J, Vincent K, Child T, Granne IE. Human chorionic gonadotrophin priming for fertility treatment with in vitro maturation. Cochrane Database Syst Rev 2016;11(11): CD008720.##Ochin H, Ma X, Wang L, Li X, Song J, Meng Y, et al. Low dose clomiphene citrate as a mild stimulation protocol in women with unsuspected poor in vitro fertilization result can generate more oocytes with optimal cumulative pregnancy rate. J Ovarian Res 2018;11(1):37.##Pacchiarotti A, Aragona C, Gaglione R, Selman H. Efficacy of a combined protocol of urinary and recombinant follicle-stimulating hormone used for ovarian stimulation of patients undergoing ICSI cycle. J Assist Reprod Genet 2007;24(9):400-5. ##Kaçar MT, Biler A, Köse C, Güleç EŞ, Demir A. Letrozole as an adjunct treatment in antagonist cycles that previously failed in poor responders. Ginekol Pol 2022;93(4):267-72.##Sakaguchi K, Nagano M. Follicle priming by FSH and pre-maturation culture to improve oocyte quality in vivo and in vitro. Theriogenology 2020;150:122-9. ##Wynn P, Picton HM, Krapez JA, Rutherford AJ, Balen AH, Gosden RG. Pretreatment with follicle stimulating hormone promotes the numbers of human oocytes reaching metaphase II by in-vitro maturation. Hum Reprod 1998;13(11):3132-8. ##Vuong LN, Ho VNA, Ho TM, Dang VQ, Phung TH, Giang NH, et al. In-vitro maturation of oocytes versus conventional IVF in women with infertility and a high antral follicle count: a randomized non-inferiority controlled trial. Hum Reprod 2020;35(11):2537-47. ##Vitek WS, Witmyer J, Carson SA, Robins JC. Estrogen-suppressed in vitro maturation: a novel approach to in vitro maturation. Fertil Steril 2013;99(7):1886-90. ##Rose BI, Laky DC, Rose SD. A comparison of the use of clomiphene citrate and letrozole in patients undergoing IVF with the objective of producing only one or two embryos. Facts Views Vis ObGyn 2015;7(2):119-26. ##Thomas RE, Thompson JG, Armstrong DT, Gilchrist RB. Effect of specific phosphodiesterase isoenzyme inhibitors during in vitro maturation of bovine oocytes on meiotic and developmental capacity. Biol Reprod 2004;71(4):1142-9. ##Nogueira D, Cortvrindt R, De Matos DG, Vanhoutte L, Smitz J. Effect of phosphodiesterase type 3 inhibitor on developmental competence of immature mouse oocytes in vitro. Biol Reprod 2003;69(6):2045-52. ##Sanchez F, Le AH, Ho VNA, Romero S, Van Ranst H, De Vos M, et al. Biphasic in vitro maturation (CAPA-IVM) specifically improves the developmental capacity of oocytes from small antral follicles. J Assist Reprod Genet 2019;36(10):2135-44. ##Sánchez F, Lolicato F, Romero S, De Vos M, Van Ranst H, Verheyen G, et al. An improved IVM method for cumulus-oocyte complexes from small follicles in polycystic ovary syndrome patients enhances oocyte competence and embryo yield. Hum Reprod 2017;32(10):2056-68. ##Kirillova A, Bunyaeva E, Van Ranst H, Khabas G, Farmakovskaya M, Kamaletdinov N, et al. Improved maturation competence of ovarian tissue oocytes using a biphasic in vitro maturation system for patients with gynecological malignancy: a study on sibling oocytes. J Assist Reprod Genet 2021;38(6):1331-40. ##

Evaluation of the Spatial Arrangement of Rabbit Hepatocytes Based on Voronoi Tessellation Following Exposure to Zinc Oxide Nanoparticles 2 2 Background: Zinc oxide nanoparticles have been utilized in different fields over the last decades. These nanoparticles can pose significant risks to various organs such as the liver. This study aimed to evaluate the effects of zinc oxide nanoparticles on liver histology, serum biochemistry, and spatial arrangement of the hepatocytes in the female New Zealand white rabbit. Methods: The rabbits received 1, 5, and 10 mg/kg of the zinc oxide nanoparticles (ZnO NPs) intraperitoneally once every 3 days for 28 days. The serum levels of the aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase and zinc were assessed 48 hr following the first administration. The histopathological changes and Voronoi tessellation were evaluated after the last administration. Results: Our findings showed that the ZnO NPs significantly increased the serum levels of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, and zinc. The histopathological findings showed ballooning degeneration, and sinusoidal congestion in ZnO NPs administrated groups. The Voronoi tessellation diagrams also confirmed that ZnO NPs changed the regular spatial arrangement of hepatocytes to random and cluster patterns. Conclusion: In conclusion, the ZnO NPs alter the liver spatial arrangement and induce hepatic pathological changes that may affect liver function in rabbits. 165 173 Zia Moasses Department of Anatomical Sciences, School of Medicine, Shiraz University of Medical Sciences Department of Anatomical Sciences, School of Medicine, Shiraz University of Medical Sciences Iran Arefeh Aryan Department of Anatomical Sciences, School of Medicine, Lorestan University of Medical Sciences Department of Anatomical Sciences, School of Medicine, Lorestan University of Medical Sciences Iran Fakhroddin Mesbah Esmaeil Mirzaei Department of Medical Nanotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical SciencesNanomedicine and Nanobiology Research Center, Shiraz University of Medical Sciences Department of Medical Nanotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical SciencesNanomedicine and Nanobiology Research Center, Shiraz University of Medical Sciences IranIran LiverNanoparticlesRabbitsZinc oxide 60582.pdf Boey A, Ho HK. All roads lead to the liver: metal nanoparticles and their implications for liver health. Small 2020;16(21):2000153.##Yao Y, Zang Y, Qu J, Tang M, Zhang T. The toxicity of metallic nanoparticles on liver: the subcellular damages, mechanisms, and outcomes. Int J Nanomedicine 2019;14:8787-804. ##Messelmani T, Morisseau L, Sakai Y, Legallais C, Le Goff A, Leclerc E, Jellali R. Liver organ-on-chip models for toxicity studies and risk assessment. Lab Chip 2022 Jun 28;22(13):2423-2450. ##de Andrade KQ, Moura FA, dos Santos JM, de Araújo OR, de Farias Santos JC, Goulart MO. Oxidative stress and inflammation in hepatic diseases: therapeutic possibilities of N-acetylcysteine. Int J Mol Sci 2015 Dec 18;16(12):30269-308. ##Sruthi S, Ashtami J, Mohanan P. Biomedical application and hidden toxicity of Zinc oxide nanoparticles. Materials Today Chemistry 2018;10:175-86.##Rana A, Yadav K, Jagadevan S. A comprehensive review on green synthesis of nature-inspired metal nanoparticles: Mechanism, application and toxicity. Journal of Cleaner Production 2020;272:122880.##Bhatia S. Nanoparticles types, classification, characterization, fabrication methods and drug delivery applications. Natural Polymer Drug Delivery Systems: Nanoparticles, Plants, and Algae 2016:33-93.##Kuang H, Yang P, Yang L, Aguilar ZP, Xu H. Size dependent effect of ZnO nanoparticles on endoplasmic reticulum stress signaling pathway in murine liver. J Hazard Mater. 2016 Nov 5;317:119-126. ##Pei X, Jiang H, Xu G, Li C, Li D, Tang S. Lethality of zinc oxide nanoparticles surpasses conventional zinc oxide via oxidative stress, mitochondrial damage and calcium overload: a comparative hepatotoxicity study. Int J Mol Sci 2022;23(12):6724. ##Vandebriel RJ, De Jong WH. A review of mammalian toxicity of ZnO nanoparticles. Nanotechnol Sci Appl 2012;5:61. ##Sirelkhatim A, Mahmud S, Seeni A, Kaus NHM, Ann LC, Bakhori SKM, Hasan H, et al. Review on zinc oxide nanoparticles: antibacterial activity and toxicity mechanism. Nanomicro Lett 2015;7(3):219-42.##Sharma V, Singh P, Pandey AK, Dhawan A. Induction of oxidative stress, DNA damage and apoptosis in mouse liver after sub-acute oral exposure to zinc oxide nanoparticles. Mutat Res 2012;745(1-2):84-91.##Almansour MI, Alferah MA, Shraideh ZA, Jarrar BM. Zinc oxide nanoparticles hepatotoxicity: Histological and histochemical study. Environ Toxicol Pharmacol 2017 Apr;51:124-130. ##Hansen C, Zidowitz S, Hindennach M, Schenk A, Hahn H, Peitgen HO. Interactive determination of robust safety margins for oncologic liver surgery. Int J Comput Assist Radiol Surg 2009 Sep;4(5):469-74. ##Wojciechowska M, Malacrino S, Garcia Martin N, et al., editors. Early Detection of Liver Fibrosis Using Graph Convolutional Networks. International Conference on Medical Image Computing and Computer-Assisted Intervention; 2021: Springer.##Lin L, Wang X, Zeng X. Geometrical modeling of cell division and cell remodeling based on Voronoi tessellation method. CMES: Computer Modeling in Engineering & Sciences. 2014;98(2):203-20.##Lau C, Kalantari B, Batts KP, Ferrell LD, Nyberg SL, Graham RP, Moreira RK. The Voronoi theory of the normal liver lobular architecture and its applicability in hepatic zonation. Sci Rep 2021 Apr 29;11(1):9343.##Pranjali P, Meher MK, Raj R, Prasad N, Poluri KM, Kumar D, et al. Physicochemical and antibacterial properties of PEGylated zinc oxide nanoparticles dispersed in peritoneal dialysis fluid. ACS Omega 2019;4(21):19255-64.##Singh SP, Kumari M, Kumari SI, Rahman MF, Mahboob M, Grover P. Toxicity assessment of manganese oxide micro and nanoparticles in Wistar rats after 28 days of repeated oral exposure. J Appl Toxicol 2013;33(10):1165-79.##Greenfield EA. Sampling and preparation of rabbit serum. Cold Spring Harbor Protocols 2018;2018(12). ##Gürsel FE, Ateş A, Bilal T, Altiner A. Effect of dietary Garcinia cambogia extract on serum essential minerals (calcium, phosphorus, magnesium) and trace elements (iron, copper, zinc) in rats fed with high-lipid diet. Biol Trace Elem Res 2012;148(3):378-82. ##Foley JF, Collins JB, Umbach DM, Grissom S, Boorman GA, Heinloth AN. Optimal sampling of rat liver tissue for toxicogenomic studies. Toxicol Pathol 2006;34(6):795-801. ##Sotoudeh N, Namavar MR, Zarifkar A, Heidarzadegan AR. Age-dependent changes in the medial prefrontal cortex and medial amygdala structure, and elevated plus-maze performance in the healthy male Wistar rats. IBRO Rep 2020 Aug 13;9:183-94. ##Moroni RF, Inverardi F, Regondi MC, Panzica F, Spreafico R, Frassoni C. Altered spatial distribution of PV‐cortical cells and dysmorphic neurons in the somatosensory cortex of BCNU‐treated rat model of cortical dysplasia. Epilepsia 2008;49(5):872-87. ##Lau C, Kalantari B, Batts KP, Ferrell LD, Nyberg SL, Graham RP, Moreira RK. The Voronoi theory of the normal liver lobular architecture and its applicability in hepatic zonation. Sci Rep 2021 Apr 29;11(1):9343.##Sarkala HB, Jahanshahi M, Dolatabadi LK, Namavar MR. Effect of G-CSF on the spatial arrangement of CA1 hippocampal pyramidal neurons after brain ischemia in the male rats. J Chem Neuroanat 2019 Jul;98:80-86.##Owjfard M, Taghadosi Z, Bigdeli MR, Safari A, Zarifkar A, Borhani-Haghighi A, et al. Effect of nicorandil on the spatial arrangement of primary motor cortical neurons in the sub-acute phase of stroke in a rat model. J Chem Neuroanat 2021 Nov;117:102000. ##Peirouvi T, Aliaghaei A, Eslami Farsani B, Ziaeipour S, Ebrahimi V, Forozesh M, et al. COVID-19 disrupts the blood-testis barrier through the induction of inflammatory cytokines and disruption of junctional proteins. Inflamm Res 2021 Dec;70(10-12):1165-75. ##Pardeshi P, Nawale AB, Mathe VL, Lahir YK, Dongre PM. Effects of zinc oxide nanoparticles on the hepatic tissue of chicken embryo: a histopathological approach. Bionano Frontier 2015;7:176-80.##Hsiao IL, Huang YJ. Effects of various physicochemical characteristics on the toxicities of ZnO and TiO2 nanoparticles toward human lung epithelial cells. Sci Total Environ 2011;409(7):1219-28. ##Yao Y, Zang Y, Qu J, Tang M, Zhang T. The toxicity of metallic nanoparticles on liver: the subcellular damages, mechanisms, and outcomes. Int J Nanomedicine 2019 Nov 7;14:8787-8804. ##Schrand AM, Rahman MF, Hussain SM, Schlager JJ, Smith DA, Syed AF. Metal-based nanoparticles and their toxicity assessment. Wiley Interdiscip Rev Nanomed Nanobiotechnol 2010 Sep-Oct;2(5):544-68.##Ribut SH, Abdullah CAC, Mustafa M, Mohd Yusoff MZB. Influence of pH variations on zinc oxide nanoparticles and their antibacterial activity. Materials Research Express 2018;6(2):025016.##Lu PJ, Fu WE, Huang SC, Lin CY, Ho ML, Chen YP, Cheng HF. Methodology for sample preparation and size measurement of commercial ZnO nanoparticles. J Food Drug Anal 2018 Apr;26(2):628-636.##Krobthong S, Rungsawang T, Wongrerkdee S. Comparison of ZnO nanoparticles prepared by precipitation and combustion for UV and sunlight-driven photocatalytic degradation of methylene blue. Toxics 2023;11(3):266.##Watson CY, Molina RM, Louzada A, Murdaugh KM, Donaghey TC, Brain JD. Effects of zinc oxide nanoparticles on Kupffer cell phagosomal motility, bacterial clearance, and liver function. Int J Nanomedicine 2015:4173-84. ##Almansour M, Sajti L, Melhim W, Jararr B. Ultrastructural hepatic alterations induced by 35 nm zinc oxide nanoparticles. Nanoscience and Nanotechnology Letters 2015;7(9):763-9.##Cornu R, Béduneau A, Martin H. Influence of nanoparticles on liver tissue and hepatic functions: A review. Toxicology 2020;430:152344.##Li CH, Shen CC, Cheng YW, Huang SH, Wu CC, Kao CC, et al. Organ biodistribution, clearance, and genotoxicity of orally administered zinc oxide nanoparticles in mice. Nanotoxicology 2012;6(7):746-56.##Al Shoyaib A, Archie SR, Karamyan VT. Intraperitoneal route of drug administration: should it be used in experimental animal studies? Pharm Res 2020;37:12. ##Srivastav AK, Kumar M, Ansari NG, Jain AK, Shankar J, Arjaria N, Jagdale P, Singh D. A comprehensive toxicity study of zinc oxide nanoparticles versus their bulk in Wistar rats: Toxicity study of zinc oxide nanoparticles. Hum Exp Toxicol 2016 Dec;35(12):1286-304. ##Kausar S, Jabeen F, Latif MA, Asad M. Characterization, dose dependent assessment of hepatorenal oxidative stress, hematological parameters and histopathological divulging of the hepatic damages induced by Zinc oxide nanoparticles (ZnO-NPs) in adult male Sprague Dawley rats. Saudi J Biol Sci. 2023 Sep;30(9):103745. ##Hong TK , Tripathy N , Son HJ , Ha KT , Jeong HS , Hahn YB . A comprehensive in vitro and in vivo study of ZnO nanoparticles toxicity. J Mater Chem B 2013 Jun 21;1(23):2985-92.##Hegazy AA, Ahmed MM, Shehata MA, Abdelfattah MM. Changes in rats’ liver structure induced by zinc oxide nanoparticles and the possible protective role of vitamin E. International Journal of Human Anatomy 2018;1(3):1-16.##Sudbø J, Marcelpoil R, Reith A. New algorithms based on the Voronoi Diagram applied in a pilot study on normal mucosa and carcinomas. Anal Cell Pathol 2000;21(2):71-86. ##Duyckaerts C, Godefroy G, Hauw J-J. Evaluation of neuronal numerical density by Dirichlet tessellation. J Neurosci Methods 1994;51(1):47-69. ##

Studying the Effect of Magnetron Copper Deposition on the Surface Topography of Biodegradable Antibacterial Coating 2 2 Background: The surface properties of the materials used significantly influence the success and longevity of medical implants. Increasing surface roughness promotes osteoblast activity and osseointegration, while biodegradable materials such as copper have shown potential for antimicrobial applications. However, the effect of coating parameters on surface topography is not well investigated. Methods: Sputtering of copper was performed using EPOS-PVD-440 system (Zelenograd, Russia). The samples were examined by Scanning Electron Microscopy (SEM) with subsequent image processing in Mountains software (Digital Surf). Antibacterial efficacy was evaluated against Staphylococcus aureus by measuring the zone of inhibition. Additionally, copper ion release was monitored over time to assess its correlation with changes in surface topography. Results: Higher sputtering currents increased surface roughness and particle size, with a significant release of copper ions within the first 24 hr of immersion. Samples sputtered at higher currents exhibited coarser grain structures. The release of copper ions in the simulated biological environment led to further changes in surface topography, highlighting the critical influence of sputtering parameters on coating properties. Conclusion: Optimizing magnetron copper deposition parameters enhances the surface topography and antibacterial effectiveness of biodegradable coatings on implants. 174 179 Maratuly Bauyrzhan Bagdat Nurlanovich Azamatov School of Digital Technologies and Artificial Intelligence, D. Serikbayev East Kazakhstan Technical University School of Digital Technologies and Artificial Intelligence, D. Serikbayev East Kazakhstan Technical University Kazakhstan Alexey Vladimirovich Jes D. Serikbayev East Kazakhstan Technical University D. Serikbayev East Kazakhstan Technical University Kazakhstan Antibacterial agentBiofilmCopperStaphylococcus aureusSurface properties 60583.pdf Barfeie A, Wilson J, Rees J. Implant surface characteristics and their effect on osseointegration. Br Dent J 2015 Mar 13;218(5):E9. ##Boyan BD, Lohmann CH, Dean DD, Sylvia VL, Cochran DL, Schwartz Z. Mechanisms involved in osteoblast response to implant surface morphology. Annual Review of Materials Research 31(1):357-71.##Blatt S, Pabst AM, Schiegnitz E, Hosang M, Ziebart T, Walter C, et al. Early cell response of osteogenic cells on differently modified implant surfaces: Sequences of cell proliferation, adherence and differentiation. J Craniomaxillofac Surg 2018 Mar 1;46(3):453-60. ##Wen X, Wang X, Zhang N. Microrough surface of metallic biomaterials: a literature review. Vol. 6, Bio-Medical Materials and Engineering. lOS Press; 1996. ##Bharadishettar N, Bhat K U, Panemangalore DB. Coating technologies for copper based antimicrobial active surfaces: A perspective review. Metals 2021;11(5):711.##Ebothé J, Nzoghe-Mendome L, Aloufy AK. Back interface effect on the topography and magnetism relationship studied from Ni nano-coatings: Role of ITO and Cu substrates. Surface Topography Metrology and Properties 2015 Dec 1;3(4). ##Caio F, Moreau C. Influence of substrate shape and roughness on coating microstructure in Suspension Plasma Spray. Coatings 2019;9(11):746. ##Kumar S, Roy DN, Dey V. A comprehensive review on techniques to create the anti-microbial surface of biomaterials to intervene in biofouling. Vol. 43, Colloids and Interface Science Communications. Elsevier B.V.; 2021. ##Kelly PJ, Arnell RD. Magnetron sputtering: a review of recent developments and applications. Vacuum March 2000;56(3):159-72.##Chinsakolthanakorn S, Buranawong A, Witit-Anun N, Chaiyakun S, Limsuwan P. Characterization of nanostructured TiZrN thin films deposited by reactive DC magnetron co-sputtering. In: Procedia Engineering. Elsevier Ltd; 2012. p. 571–6. ##Mech K, Kowalik R, Zabiński P. Cu thin films deposited by DC magnetron sputtering for contact surfaces on electronic components. In: Archives of Metallurgy and Materials. 2011. p. 903–8. ##Balázsi K. Magnetron sputtered TiC/a:C nanocomposite thin films: Deposition parameters vs. properties. Vacuum 2019 Jun 1;164:121-5. ##Ferreira F, Cavaleiro A, Oliveira J. Effect of Peak Power in Deep Oscillation Magnetron Sputtering on Film Properties. J Mater Eng Perfor 2021 Jun 1;30(6):3912-24. ##Huang J, Zou S, Xiao W, Liu X, Tang D. Sputtering parameters effect on microstructural parameters of TiN coating via the Williamson-Hall analysis. Mater Res Express 2020 Oct 1;7(10). ##Yahya KA, Rasheed BF. Effects of discharge current and target thickness in DC-magnetron sputtering on grain size of copper deposited samples. Baghdad Science Journal 2019;16(1):84-7. ##Čekada M, Radić N, Jerčinović M, Panjan M, Panjan P, Drnovšek A, et al. Growth defects in magnetron sputtered PVD films deposited in UHV environment. Vacuum 2017 Apr 1;138:213-7. ##Barb AM, Garcia N, Miranda R, Vhzquez L, Aparicio C, Oliv6 " J, et al. Characterization of surface roughness in titanium dental implants measured with scanning tunnelling microscopy at atmospheric pressure. Biomaterials 1986 Nov;7(6):463-6. ##Turkin AA, Pei YT, Shaha KP, Chen CQ, Vainshtein DI, De Hosson JTM. On the evolution of film roughness during magnetron sputtering deposition. J Appl Phys 2010 Nov 1;108(9). ##

The Phenotypic and Molecular Identification of Phyllospheric Bacteria Possessing Antimicrobial Activity from Funtumia elastica (Preuss) Stapf. 2 2 Background: Unlike plant phytochemicals, little has been done to explore the metabolites from phyllosphere bacterial flora, some of which enabled them to survive interspecific competition through amensalism. This study evaluated the antimicrobial activity of metabolites from Phyllospheric Bacteria (PB) isolated from Funtumia elastica (FE), against selected bacterial and fungal pathogens. Phenotypic and molecular methods were used to identify the isolated phyllo-microbiota. Methods: The PB were aseptically isolated by sonication. Their metabolites were obtained from the fresh overnight culture of the organisms. The cell-free supernatants containing the metabolites were used for antimicrobial assays against the pathogens. The DNA of the bacterial isolates were isolated using a NIMR-BIOTECH DNA extraction kit, while their 16S rRNA was amplified with the primer: 799F 5'-AACACGGATTA GATACC-3', 1193R 5'- ACGTCATCCCCACCTTCC-3', using SolisFast* Master Mix, (Solis Biodyne-Estonia). The BLAST of the sequence was done from the NCBI Genbank. The PB strains identified were submitted to NCBI and accession numbers were assigned to them. Results: The phyllosphere of FE yielded 21 bacterial isolates: 7 Gram-positives and 14 Gram-negatives. The metabolites from these isolates showed varying degrees of bioactivity against Staphylococcus aureus (ATCC29213), Escherichia coli (ATCC 25922) Klebsiella pneumoniae (ATCC 35659); Trychophyton rubrum, Candida albicans and Microsporum canis. Fifteen bioactive isolates sequenced yielded four genera, Enterobacter (E. hormaechei 98.44%), Bacillus (B. cereus 100%), Pontoea (P. dispersa 99.72%), Staphylococcus (S. arlettae 99.72%). Conclusion: Bacteria from FE phyllosphere, produced metabolites antagonistic (cidal) to some human pathogens.  This has great potential for possible drug discovery. 180 186 Bolanle A. Adeniyi Mercy Ogunlana Department of Pharmaceutical Microbiology, University of Ibadan Department of Pharmaceutical Microbiology, University of Ibadan Nigeria Christopher O. Igbokwe Department of Pharmaceutical Microbiology, University of Ibadan Department of Pharmaceutical Microbiology, University of Ibadan Nigeria Bamidele Tajudeen Nigerian Institute of Medical Research Nigerian Institute of Medical Research Nigeria Gail B. Mahady Department Pharmacy Practice, College of Pharmacy, University of Illinois Department Pharmacy Practice, College of Pharmacy, University of Illinois United States of America 16s rRNABiotechnologyDNADrug discoveryEnterobacterMicrobiotaPhytochemicals 60584.pdf Mintah SO, Asafo-Agyei T, Archer MA, Junior PA, Boamah D, Kumadoh D, et al. Medicinal plants for treatment of prevalent diseases. Pharmacognosy-Medicinal Plants 2019 Jan 17:1-9.##Saxena M, Saxena J, Nema R, Singh D. Gupta A. Phytochemistry of medicinal plants. Journal of Pharmacognosy and Phytochemistry 2013;1(6):168-82.##Ugboko HU, Nwinyi OC, Oranusi SU, Fatoki TH, Omonhinmin CA. Antimicrobial importance of medicinal plants in Nigeria. ScientificWorldJournal 2020 Sep 22:2020:7059323. ##Manandhar S, Luitel S, Dahal RK. In vitro antimicrobial activity of some medicinal plants against human pathogenic bacteria. J Trop Med 2019 Apr 2:2019:1895340. ##Khameneh B, Iranshahy M, Soheili V, Fazly Bazzaz BS. Review on plant antimicrobials: a mechanistic viewpoint. Antimicrob Resist Infect Control 2019 Jul 16:8:118. ##Bringel F, Couée I. Pivotal roles of phyllosphere microorganisms at the interface between plant functioning and atmospheric trace gas dynamics. Front Microbiol 2015 May 22:6:486. ##Vorholt JA. Microbial life in the phyllosphere. Nat Rev Microbiol 2012;10(12):828-40. ##Sivakumar N, Sathishkumar R, Selvakumar G, Shyamkumar R, Arjunekumar K. Phyllospheric microbiomes: diversity, ecological significance, and biotechnological applications. Plant Microbiomes for Sustainable Agriculture 2020;25:113-72. ##Kumar J, Singh D, Ghosh P, Kumar A. Endophytic and epiphytic modes of microbial interactions and benefits. Plant-Microbe Interactions in Agro-Ecological Perspectives 2017.p.227-253.##Lindow SE, Brandl MT. Microbiology of the phyllosphere. Apple Environ Microbiol 2003;69(4):1875-83. ##Thompson IP, Bailey MJ, Fenlon JS, Fermor TR, Lilley AK, Lynch JM, McCormack PJ, McQuilken MP, Purdy KJ, Rainey PB, Whipps JM. Quantitative and qualitative seasonal changes in the microbial community from the phyllosphere of sugar beet (Beta vulgaris). Plant and Soil 1993 Mar;150:177-91.##Clarridge JE 3rd. Impact of 16S rRNA gene sequence analysis for identification of bacteria on clinical microbiology and infectious diseases. Clin Microbiol Rev 2004;17(4):840-62. ##Jia T, Yao Y, Wang R, Wu T, Chai B. Dynamics relationship of phyllosphere and rhizosphere bacterial communities during the development of Bothriochloa ischaemum in copper tailings. Front Microbiol 2020;11:869. ##Zirihi GN, Grellier P, Guédé-Guina F, Bodo B, Mambu L. Isolation, characterization and antiplasmodial activity of steroidal alkaloids from Funtumia elastica (Preuss) Stapf. Bioorg Med Chem Lett 2005 May 16;15(10):2637-40. ##Agyare C, Koffuor GA, Boakye YD, Mensah KB. Antimicrobial and anti-inflammatory properties of Funtumia elastica. Pharm Biol 2013;51(4):418-25. ##Adekunle AA, Ikumapayi AM. Antifungal property and phytochemical screening of the crude extracts of Funtumia elastica and Mallotus oppositifolius. West Indian Med J 2006;55(4):219-23. ##Adeniyi BA, Odelola HA, Oso BA. Antimicrobial potentials of Diospyros mespiliformis (Ebenaceae). Afr J Med Med Sci 1996;25(3):221-4.##Adeniyi BA, Onwubuc he BC, Anyiam FM, Ekundayo O, Mahady GB. Anti- Helicobacter pylori activities of Eucalyptus grandis: Effects on susceptibility, urease activity and cell surface hydrophobicity. Pharm Biol 2009;47(1):13-7.##Yashiro E, Spear RN, McManus PS. Culture‐dependent and culture‐independent assessment of bacteria in the apple phyllosphere. J Appl Microbiol 2011 May 1;110(5):1284-96. ##Goryluk A, Rekosz-Burlaga H, Blaszczyk M. Isolation and characterization of bacterial endophytes of Chelidonium majus L. Pol J Microbiol 2009;58(4):355-61. ##Mazinani Z, Zamani M, Sardari, S. Isolation and identification of phyllospheric bacteria possessing antimicrobial activity from Astragalus obtusifolius, Prosopis juliflora, Xanthium strumatium and Hippocrepis unisiliqousa. Avicenna J Med Biotechnol 2017;9(1):31-7. ##Ali M, Walait S, Farhan Ul Haque M, Mukhtar S. Antimicrobial activity of bacteria associated with the rhizosphere and phyllosphere of Avena fatua and Brachiaria reptans. Environ Sci Pollut Res Int 2021;28:68846-61. ##Stoltzfus J, So R, Malarvithi P, Ladha J, Bruijn F. Isolation of endophytic bacteria from rice and assessment of their potential for supplying rice with biologically fixed nitrogen. Plant Soil 1997;194:25-36.##Thapa S, Prasanna R. Prospecting the characteristics and significance of the phyllosphere microbiome. Annals of Microbiology 2018;68:229-45. ##Yurimoto H, Shiraishi K, Sakai Y. Physiology of methylotrophs living in the phyllosphere. Microorganisms 2021;9(4):809. ##Teng T, Li X, Zhang L, Li Y. Identification and Characterization of pantocin wh-1 a novel cyclic polypeptide produced by Pantoea dispersa W18. Molecules 2020;25(3):485. ##

Bactericidal Activity of Serum by Brucella Abortus RB51 Outer Membrane Protein’s Combined by Brucella Abortus S99 Lipopolysaccharide Induction 2 2 Background: Brucellosis vaccines are designed to induce cellular immunity. An effective brucellosis vaccine could induce both cellular and humoral immunity. Serum Bactericidal Assay (SBA) is an important method for determining vaccine humoral immunity. This study is the first to observe humoral immunity in brucellosis by SBA. Methods: Extracted Brucella abortus (B. abortus) Lipopolysaccharide (LPS) and Outer Membrane Proteins (OMPs) were injected into rabbits. Group 1 was injected with 25 µg of LPS, Group 2 was injected with 50 µg of OMPs, and Group 3 was injected with 1 ml of combined vaccine, 3 times every 2 weeks. The groups were challenged with B. abortus 544 in the second injection. Sera were separated 2 weeks after the last injection. SBA was performed, and each well was streak-cultured into a plate of Brucella agar. A colony count was done for each plate. Results: Results have shown, the third injection of the combined vaccine had the highest titer of 1/64, and the efficacy of the vaccine was 87.71%. Conclusion: As a conclusion, the results of this study showed that LPS and OMP's from B. abortus can provide acceptable immunity. 187 192 Behnam Hajizadeh Sisakht Department of Bacteriology, Faculty of Veterinary, Islamic Azad University, Science & Research Campus Department of Bacteriology, Faculty of Veterinary, Islamic Azad University, Science & Research Campus Iran Mansoor Khaledi Department of Microbiology and Immunology, School of Medicine, Shahrekord University of Medical Sciences Department of Microbiology and Immunology, School of Medicine, Shahrekord University of Medical Sciences Iran Hamed Afkhami Department of Microbiology, Faculty of Medicine, Shahed University Department of Microbiology, Faculty of Medicine, Shahed University Iran Saber Rouhi Resident of Large Animal Internal Medicine, Department of Clinical Sciences, School of Veterinary Medicine, Shiraz University Resident of Large Animal Internal Medicine, Department of Clinical Sciences, School of Veterinary Medicine, Shiraz University Iran Saeed Sepehrnia Department of Immunology, Faculty of Medicine, Shahed University Department of Immunology, Faculty of Medicine, Shahed University Iran Vahideh Fanaee Department of Microbiology and Immunology, School of Medicine, Shahrekord University of Medical Sciences Department of Microbiology and Immunology, School of Medicine, Shahrekord University of Medical Sciences Iran Hannaneh Karimi Department of Microbiology, Rasht Islamic Azad University Department of Microbiology, Rasht Islamic Azad University Iran Yalda Malekzadegan Javad Fathi Mahdi S. Sadati AgarBrucellosisCellularHumoralImmunityLipopolysaccharidesVaccines 60585.pdf Schurig GG, Sriranganathan N, Corbel MJ. Brucellosis vaccines: past, present and future. Vet Microbiol 2002 Dec 20;90(1-4):479-96.##Ashford DA, di Pietra J, Lingappa J, Woods C, Noll H, Neville B, Weyant R, Bragg SL, Spiegel RA, Tappero J, Perkins BA. Adverse events in humans associated with accidental exposure to the livestock brucellosis vaccine RB51. Vaccine 2004 Sep 3;22(25-26):3435-9. ##Banai M. Control of small ruminant brucellosis by use of Brucella melitensis Rev.1 vaccine: laboratory aspects and field observations. Vet Microbiol 2002 Dec 20;90(1-4):497-519. ##Olsen S, Tatum F. Bovine brucellosis. Vet Clin North Am Food Anim Pract. 2010 Mar;26(1):15-27, table of contents.##Cloeckaert A, Jacques I, Grilló MJ, Marín CM, Grayon M, Blasco JM, Verger JM. Development and evaluation as vaccines in mice of Brucella melitensis Rev.1 single and double deletion mutants of the bp26 and omp31 genes coding for antigens of diagnostic significance in ovine brucellosis. Vaccine 2004 Jul 29;22(21-22):2827-35. ##Moriyón I, Grilló MJ, Monreal D, González D, Marín C, López-Goñi I, et al. Rough vaccines in animal brucellosis: structural and genetic basis and present status. Vet Res 2004 Jan-Feb;35(1):1-38. ##Cloeckaert A, Vizcaíno N, Paquet JY, Bowden RA, Elzer PH. Major outer membrane proteins of Brucella spp.: past, present and future. Vet Microbiol 2002 Dec 20;90(1-4):229-47. ##Caroff M, Karibian D, Cavaillon JM, Haeffner-Cavaillon N. Structural and functional analyses of bacterial lipopolysaccharides. Microbes Infect 2002 Jul;4(9):915-26. ##Pasquevich KA, García Samartino C, Coria LM, Estein SM, Zwerdling A, Ibañez AE, et al. The protein moiety of Brucella abortus outer membrane protein 16 is a new bacterial pathogen-associated molecular pattern that activates dendritic cells in vivo, induces a Th1 immune response, and is a promising self-adjuvanting vaccine against systemic and oral acquired brucellosis. J Immunol 2010 May 1;184(9):5200-12. ##Cardoso PG, Macedo GC, Azevedo V, Oliveira SC. Brucella spp noncanonical LPS: structure, biosynthesis, and interaction with host immune system. Microb Cell Fact 2006 Mar 23;5:13. ##Balouiri M, Sadiki M, Ibnsouda SK. Methods for in vitro evaluating antimicrobial activity: A review. J Pharm Anal 2016 Apr;6(2):71-79. ##Burgess DS, Hall RG 2nd. In vitro killing of parenteral beta-lactams against standard and high inocula of extended-spectrum beta-lactamase and non-ESBL producing Klebsiella pneumoniae. Diagn Microbiol Infect Dis 2004 May;49(1):41-6.##Malerczyk C, Kolbert M, Kinzig-Schippers M, Sörgel F, Machka K, Shah PM. Serum bactericidal activity of gemifloxacin versus clarithromycin against Streptococcus pneumoniae with different susceptibility to quinolones. J Chemother 2004 Feb;16(1):56-61. ##Paulsen VS, Blencke HM, Benincasa M, Haug T, Eksteen JJ, Styrvold OB, Scocchi M, Stensvåg K. Structure-activity relationships of the antimicrobial peptide arasin 1 - and mode of action studies of the N-terminal, proline-rich region. PLoS One 2013;8(1):e53326. ##Garin-Bastuji B, Bowden RA, Dubray G, Limet JN. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting analysis of smooth-lipopolysaccharide heterogeneity among Brucella biovars related to A and M specificities. J Clin Microbiol 1990 Oct;28(10):2169-74. ##Kielkopf, C.L., W. Bauer, and I.L. Urbatsch, Bradford assay for determining protein concentration. Cold Spring Harbor Protocols, 2020. 2020(4): p. pdb. prot102269.##Pakzad, I., et al., Novel approach of vaccination against Brucella abortus 544 based on a combination of fusion proteins, human serum albumin and Brucella abortus lipopolysaccharides. Journal of Biological Sciences, 2010. 10(8): p. 767-772.##Handayani, T., D. Priyoatmojo, and A.C. Trinugraha. Outer Membrane Protein (OMP) Profiles of Brucella abortus Local Isolate by SDS-PAGE Procedure. in International Conference on Improving Tropical Animal Production for Food Security (ITAPS 2021). 2022. Atlantis Press.##Jacques I, Cloeckaert A, Limet JN, Dubray G. Protection conferred on mice by combinations of monoclonal antibodies directed against outer-membrane proteins or smooth lipopolysaccharide of Brucella. J Med Microbiol. 1992 Aug;37(2):100-3. ##Doyle AG, Halliday WJ, Barnett CJ, Dunn TL, Hume DA. Effect of recombinant human macrophage colony-stimulating factor 1 on immunopathology of experimental brucellosis in mice. Infect Immun 1992 Apr;60(4):1465-72.##Ghasemi A, Salari MH, Zarnani AH, Pourmand MR, Ahmadi H, Mirshafiey A, Jeddi-Tehrani M. Immune reactivity of Brucella melitensis-vaccinated rabbit serum with recombinant Omp31 and DnaK proteins. Iran J Microbiol 2013 Mar;5(1):19-23.##Estein SM, Cheves PC, Fiorentino MA, Cassataro J, Paolicchi FA, Bowden RA. Immunogenicity of recombinant Omp31 from Brucella melitensis in rams and serum bactericidal activity against B. ovis. Vet Microbiol 2004 Sep 8;102(3-4):203-13. ##Campbell GA, Adams LG, Sowa BA. Mechanisms of binding of Brucella abortus to mononuclear phagocytes from cows naturally resistant or susceptible to brucellosis. Vet Immunol Immunopathol. 1994 Jun;41(3-4):295-306. ##Diaz R, Jones LM, Leong D, Wilson JB. Surface antigens of smooth brucellae. J Bacteriol 1968 Oct;96(4):893-901.##Biron CA. Initial and innate responses to viral infections--pattern setting in immunity or disease. Curr Opin Microbiol. 1999 Aug;2(4):374-81. ##Briones G, Iñón de Iannino N, Roset M, Vigliocco A, Paulo PS, Ugalde RA. Brucella abortus cyclic beta-1,2-glucan mutants have reduced virulence in mice and are defective in intracellular replication in HeLa cells. Infect Immun 2001 Jul;69(7):4528-35. ##Izadjoo MJ, Bhattacharjee AK, Paranavitana CM, Hadfield TL, Hoover DL. Oral vaccination with Brucella melitensis WR201 protects mice against intranasal challenge with virulent Brucella melitensis 16M. Infect Immun 2004 Jul;72(7):4031-9. ##Rittig MG, Alvarez-Martinez MT, Porte F, Liautard JP, Rouot B. Intracellular survival of Brucella spp. in human monocytes involves conventional uptake but special phagosomes. Infect Immun 2001 Jun;69(6):3995-4006.##

Lippia multiflora Leaves Extracts Enhance Cefotaxime Bactericidal Effects and Quench the Biofilm Formation in Methicillin-Resistant Staphylococcus aureus ATCC 43300 2 2 Background: The emergence of the multidrug-resistant bacteria strain has become a global world crisis. This study was designed to evaluate the antibiofilm and synergistic effects of Lippia multiflora (L. multiflora) leaf extracts on the activity of cefotaxime against the methicillin-resistant Staphylococcus aureus (S. aureus). Methods: The synergistic effect of methanol and dichloromethane extracts on the bactericidal activity of cefotaxime was determined by using the antibiotic susceptibility test on agar medium. The antibiofilm activity of the extracts was measured by using the crystal violet method. The antioxidant potential of the extracts was assessed by using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and Ferric Reduction Activity Potential (FRAP) methods. The main secondary metabolites groups were analyzed by using different standard analytical tests. The total phenolics and total flavonoids were quantified spectrophotometrically. Results: The methanol extract (final concentration of 100 µg/ml) inhibited the formation of bacterial biofilm more than salicylic acid (p<0.05). All extracts combined with cefotaxime (20 µg and 200 µg) showed good synergistic bactericidal effect on S. aureus with inhibitory diameters of up to 40 mm. The methanol extract showed higher total phenolics (462.20±10.90 mg EAG/g) and total flavonoids (26.20±0.20 mg EQ/g) contents than the dichloromethane extract (96.70±1.70 mg EAG/g and 8.00±1.20 mg EQ/g). Moreover, the methanol extract showed a higher FRAP reducing power (353.6± 4.17 mmol EQ/g) than the dichloromethane extract (385.3±7.01 mmol EQ/g). Qualitative phytochemical analysis showed the presence of tannins, flavonoids, terpenes and sterols in both extracts. Conclusion: These data showed that L. multiflora leaves contain effective antibacterial phytomolecules for combating bacterial resistance. 193 199 Ablassé Rouamba Ecole Normale Supérieure, 01 BP 1757 Ecole Normale Supérieure, 01 BP 1757 Burkina Faso Djaouratou Badini Laboratory of Applied Biochemistry and Chemistry, Department of Biochemistry-Microbiology, UFR SVT, Université Joseph Ki-Zerbo, 03 BP 7021 Laboratory of Applied Biochemistry and Chemistry, Department of Biochemistry-Microbiology, UFR SVT, Université Joseph Ki-Zerbo, 03 BP 7021 Burkina Faso Eli Compaoré Laboratory of Applied Biochemistry and Chemistry, Department of Biochemistry-Microbiology, UFR SVT, Université Joseph Ki-Zerbo, 03 BP 7021 Laboratory of Applied Biochemistry and Chemistry, Department of Biochemistry-Microbiology, UFR SVT, Université Joseph Ki-Zerbo, 03 BP 7021 Burkina Faso Vincent Ouédraogo Laboratory of Applied Biochemistry and Chemistry, Department of Biochemistry-Microbiology, UFR SVT, Université Joseph Ki-Zerbo, 03 BP 7021 Laboratory of Applied Biochemistry and Chemistry, Department of Biochemistry-Microbiology, UFR SVT, Université Joseph Ki-Zerbo, 03 BP 7021 Burkina Faso Martin Kiendrebeogo Laboratory of Applied Biochemistry and Chemistry, Department of Biochemistry-Microbiology, UFR SVT, Université Joseph Ki-Zerbo, 03 BP 7021 Laboratory of Applied Biochemistry and Chemistry, Department of Biochemistry-Microbiology, UFR SVT, Université Joseph Ki-Zerbo, 03 BP 7021 Burkina Faso Antibacterial agentsAntioxidantsBiofilmsLippiaMethicillin-resistant Staphylococcus aureusMethylene chloridePlant extracts 60586.pdf Meinen A, Tomczyk S, Noelle F, Sin MA, Eckmanns T. Antimicrobial resistance in Germany and Europe – A systematic review on the increasing threat accelerated by climate change. J Health Monit 2023;8(Suppl 3):93-108.##Sagar KP, Sharma P, Singh R. Anti-quorum sensing and anti-biofilm activity of Ginger (Zingiber officinale) rhizomes against multidrug-resistant clinical isolates of Pseudomonas aeruginosa. Avicenna J Med Biotechnol 2024;16(1):49-56. ##Gherardi G. Staphylococcus aureus infection : pathogenesis and antimicrobial resistance. Int J Mol Sci. 2023;24(9):8182.##Arunachalam K, Pandurangan P, Shi C, Lagos R. Regulation of Staphylococcus aureus virulence and application of nanotherapeutics to eradicate S . aureus infection. Pharmaceutics 2023;15(2):310.##Talapan D, Sandu AM, Rafila A. Antimicrobial resistance of Staphylococcus aureus isolated between 2017 and 2022 from infections at a tertiary care hospital in Romania. Antibiotics (Basel) 2023;12(6):974.##Nikolic P, Mudgil P. The cell wall , cell membrane and virulence factors of Staphylococcus aureus and their role in antibiotic resistance. Microorganisms 2023;11(2):259. ##Khodabux RMJ, Mariappan S, Sekar U. Spectrum of virulence factors in clinical isolates of Staphylococcus aureus and prevalence of SCCmec types in methicillin-resistant Staphylococcus aureus in a tertiary care center. J Lab Physicians 2023;15(3):450-61. ##Jean-pierre V, Boudet A, Sorlin P, Menetrey Q, Chiron R, Lavigne JP, et al. Biofilm formation by Staphylococcus aureus in the specific context of cystic fibrosis. Int J Mol Sci 2022;24(1):597.##Balducci E, Papi F, Capialbi DE, Bino L Del. Polysaccharides ’ structures and functions in biofilm architecture of antimicrobial-resistant ( AMR ) pathogens. Int J Mol Sci 2023;24(4):4030.##François P, Schrenzel J, Götz F. Biology and regulation of Staphylococcal biofilm. Int J Mol Sci 2023;24(6):5218.##Lade H, Kim JS. Molecular determinants of β -lactam resistance in methicillin-resistant Staphylococcus aureus (MRSA): an updated review. Antibiotics (Basel) 2023;12(9):1362. ##Gandonou DC, Ahissou H, Tokoudagba JM, Dansou C. Ethnobotanical , phytochemical and toxicity analysis of a Beninese antihypertensive plant : Lippia multiflora. Int J Biol Chem Sci 2017;11(4):1816-28. ##Mpiana PT. Traditional uses, physical properties, phytochemistry and bioactivity of Lippia multiflora moldenke (Verbenaceae): a mini-review. Discovery Phytomedicine-Journal of Natural Products Research and Ethnopharmacology 2020 Jan 15;7(1):19-26. ##Muharrami LK, Munawaroh F, Ersam T, Santoso M. Phytochemical screening of ethanolic extract: a preliminary test on five medicinal plants on Bangkalan. J Pena Sains 2020;7(2):96-102. ##Lucas BN, Nora DMF, Boeira CP, Verruck S, Rosa SC. Determination of total phenolic compounds in plant extracts via Folin-Ciocalteu ’ s method adapted to the usage of digital images. Food Sci Technol 2022;42:e35122. ##Hilma R, Hilma A, Almurdani M. Determination of total phenolic, flavonoid content and free radical scavenging activity of ethanol extract Sawo stem bark (Manilkara Zapota ( L .). CelSci Tech-UMRI 2018;3:62–68. ##Borges L da S, Lima PPG, Artés F, Souza ME De, Freitas LDS, Jesus HI De, et al. Efficiency of DPPH and FRAP assays for estimating antioxidant activity and separation of organic acids and phenolic compounds by liquid chromatography in fresh-cut nectarine. Aust J Crop Sci 2019;13(07):1053-60. ##Rodríguez-Melcón C, Alonso-calleja C, García-Fernández C, Carballo J, Capita R. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) for twelve antimicrobials (biocides and antibiotics) in fight strains of Listeria monocytogenes. Biology (Basel) 2022;11(1):46.##Kamimura R, Kanematsu H, Ogawa A, Kogo T, Miura H, Kawai R, et al. Quantitative analyses of biofilm by using crystal violet staining and optical reflection. Materials (Basel) 2022;15(19):6727. ##Okasha H, Asser S, Kholeif D. Rapid disc diffusion antibiotic susceptibility testing for gram negative non- fermenters : Pseudomonas and Acinetobacter. Egypt J Med Microbiol 2023;32(1):77-86. ##Parastan R, Kargar M, Solhjoo K, Kafilzadeh F. Staphylococcus aureus biofilms : structures, antibiotic resistance , inhibition and vaccines. Gene Reports 2020;20:1–8. ##Lopes AAL, Rodrigues BJ dos S, Magnani M, Souza EL De, Siqueira-júnior JP De. Inhibitory effects of flavonoids on biofilm formation by Staphylococcus aureus that overexpresses efflux protein genes. Microb Pathog 2017;107:194-7. ##Matilla-Cuenca L, Gil C, Cuesta S, Rapún-Araiz B, Žiemytė M, Mira A, et al. Antibiofilm activity of flavonoids on staphylococcal biofilms through targeting BAP amyloids. Sci Rep 2020;10(1):18968.##Kauffmann AC, Castro VS. Phenolic compounds in bacterial inactivation : a Perspective from Brazil. Antibiotics (Basel) 2023;12(4):1-24.##Payne DE, Martin NR, Parzych KR, Rickard AH, Underwood A, Boles BR. Tannic acid inhibits Staphylococcus aureus s https://pubmed.ncbi.nlm.nih.gov/23208606/urface colonization in an IsaA-dependent manner. Infect Immun 2013;81(2):496–504. ##Junior DAM, Edzang RWN, Catto AL, Raimundo JM. Quinones as an efficient molecular scaffold in the antibacterial / antifungal or antitumoral arsenal. Int J Mol Sci 2022;23(22):14108.##Salinas C, Florentin G, Rodriguez F, Alvarenga N, Guillen R. Terpenes combinations inhibit biofilm formation in Staphyloccocus aureus by interfering with initial adhesion. Microorganismes 2022;10(8):1527.##Chen H, Zhang J, He Y, Lv Z, Liang Z, Chen J, et al. Exploring the role of Staphylococcus aureus in inflammatory diseases. Toxins (Basel) 2022;14(7):464. ##Munita JM, Arias CA. Mechanisms of antibiotic resistance. Microbiol Spectr 2016 Apr;4(2):10. ##Bush K, Bradford PA. B-Lactams and B-Lactamase inhibitors: an overview. Cold Spring Harb Perspect Med 2016;6(8):1-22. ##Torimiro N, Moshood A, Eyiolawi S. Analysis of Beta-lactamase production and antibiotics resistance in Staphylococcus aureus strains. J Infect Dis Immun 2013;5(3):24-8. ##Dawan J, Ahn J. Assessment of β -lactamase inhibitor potential of medicinal plant extracts against antibiotic-resistant Staphylococcus aureus. Korean J Plant Res 2020;33(6):578-85. ##