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Genotoxicity test study of bioactive protein fraction from Lumbricus rubellus, DLBS1033 2 2 Introduction: Genotoxicity is one of the safety concerns that can lead to damage at the genetic level. The observation of DLBS1033 using AMES and micronucleus tests was conducted to ensure the safety precautions. Methods: Ames test was done using 3 strains of Salmonella typhimurium (TA98, TA100, and TA1535) with or without metabolic activation (S9 enzyme). In vitro micronucleus test was performed with prolonged exposure without S9. The variation of DLBS1033 concentrations were 100, 600, and 1200 µg/ml. Results: The results of the Ames test (with/without S9) and in vitro micronucleus (prolonged exposure without S9) showed that DLBS1033 did not have a mutagenic effect. Conclusion: DLBS1033 was safe and did not show any genotoxic activity. 106 110 2023/12/18 1402/9/27 2024/09/14 1403/6/24 Agung Heru Karsono Agung Heru Karsono Dexa Laboratories of Biomolecular Sciences, PT Dexa Medica, Industri Selatan V Block PP No. 7, Jababeka Industrial Estate II, Cikarang, West Java, 17550, Indonesia Indonesia agung.karsono@dexa-medica.com Lidwina Tri Kristanti Setiawan Lidwina Tri Kristanti Setiawan Dexa Laboratories of Biomolecular Sciences, PT Dexa Medica, Industri Selatan V Block PP No. 7, Jababeka Industrial Estate II, Cikarang, West Java, 17550, Indonesia Indonesia lidwina.setiawan@dexa-medica.com Raymond Rubianto Tjandrawinata Raymond Rubianto Tjandrawinata Center for Pharmaceutical and Nutraceutical Research and Policy, Atma Jaya Catholic University of Indonesia, Jakarta Selatan 12930, DKI Jakarta, Indonesia Indonesia raymond@dexa-medica.com Ames test Genotoxicity Lumbricus rubellus Micronucleus test Agoncillo A. Meta-analysis on the safety and efficacy of Lumbrokinase in peripheral arterial disease. Eur Heart J Acute Cardiovasc Care. 2021;10(Suppl 1).##Biran A, Yagur-Kroll S, Pedahzur R, et al. Bacterial genotoxicity bioreporters. Microb Biotechnol 2010; 3: 412-427.##Christy G, Tjandrawinata R R, Rachmawati H. Tabletting process of pellets containing bioactive fraction DLBS1033F isolated from lumbricus rubellus: Challenge and strategy. Biol Pharm Bull 2015; 38: 1843-1849.##Gayatri A, Nafrialdi N, Setiabudy R D, Tjandrawinata R R, Rachman A, Louisa M. A clinical trial on biological half life of bioactive protein from Lumbricus rubellus, DLBS1033 in healthy volunteers. Acta Med Indones 2018; 50: 208-214.##Hamel A, Roy M, Proudlock R. Chapter 4 - the bacterial reverse mutation test. In: Proudlock R, editor. Genetic toxicology testing. Boston: Academic Press, 2016, p. 79-138.##ICH. S2(R1): Guidance on genotoxicity testing and data interpretation for pharmaceuticals intended for human use. ICH; 2011.##Levy D D, Zeiger E, Escobar P A, Hakura A, B-JM v d L, Kato M, et al. Recommended criteria for the evaluation of bacterial mutagenicity data (Ames test). Mutation research/genetic toxicology and environmental. Mutagenesis 2019; 848: 403074.##OECD. Test No. 471: bacterial reverse mutation test. OECD guidelines for the testing of chemicals, section 4. Paris: Organization for Economic Cooperation & Development; 1997.##Phillips D H, Arlt V M. Genotoxicity: damage to DNA and its consequences. EXS. 2009; 99: 87-110.##Pinzon R T, Tjandrawinata R R, Wijaya V O, Veronica V. Effect of DLBS1033 on functional outcomes for patients with acute ischemic stroke: A randomized controlled trial. Stroke Res Treat 2021; 2021: 5541616.##Rong P, Zhang Z J, He R Q. Earthworm protease. Appl Environ Soil Sci 2010; 2010: 1-13.##Sommer S, Buraczewska I, Kruszewski M. Micronucleus assay: The state of art, and future directions. Int J Mol Sci 2020; 21:1534.##Stephani L, Rahayu P, Retnoningrum D, Suhartono MT, Rachmawati H, Tjandrawinata RR. Purification and proteomic analysis of potent fibrinolytic enzymes extracted from Lumbricus rubellus. Proteome Sci. 2023;21(1):8.##Sukandar E Y, Anggadireja K, Sigit J I, Adnyana I K, Tjandrawinata R R. Toxicity studies of a bioactive protein with antithrombotic-thrombolytic activity, DLBS1033. Drug Chem Toxicol 2014; 37: 8-16.##Tjandrawinata R R, Trisina J, Rahayu P, Prasetya L A, Hanafiah A, Rachmawati H. Bioactive protein fraction DLBS1033 containing lumbrokinase isolated from Lumbricus rubellus: ex vivo, in vivo, and pharmaceutic studies. Drug Des Devel Ther 2014; 8: 1585-1593.##Tjandrawinata R R, stu Yunaidi D A, Susanto L W. The safety and tolerability of lumbrokinase DLBS1033 in healthy adult subjects. Drug Res (Stuttg) 2016; 66: 293-299.##Trisina J, Sunardi F, Suhartono M T, Tjandrawinata R R. DLBS1033, a protein extract from Lumbricus rubellus, possesses antithrombotic and thrombolytic activities. J Biomed Biotechnol 2011; 2011: 519652.## ##

Enhancing Clinical Outcomes and Survival in Hospitalized Multiple Sclerosis Patients with COVID-19: Challenges of Antiviral Therapy 2 2 Introduction: This study aimed to evaluate the efficacy and safety concerns of remdesivir and type 1 interferons on the clinical outcomes of hospitalized multiple sclerosis patients with COVID-19. Methods: Using electronic health records systems; this is a cross-sectional study of two years of hospital admissions in terms of COVID-19 in Iran from March 2019 to August 2021. The severities of COVID-19 outcomes were ICU admission, hospitalization days, and 30-day survival rates. Results: A total of 993 hospitalized multiple sclerosis patients with a confirmed diagnosis of COVID-19 based on PCR testing were recorded in the electronic health systems. Nearly half of these patients (50.3%) had received treatment with an anti-CD20 agent (rituximab or ocrelizumab) at the hospital admission. This group exhibited higher mortality rates, increased need for ICU admission, and longer hospitalization (p<0.05). There was a significant association between taking interferon-β1 alone (adjusted IRR=1.21, 95% CI 1.32 to 1.42) or in combination with remdesivir (adjusted IRR=1.30, 95% CI 1.18 to 1.5042) and longer hospitalization. There were no significant associations between antiviral treatment (remdesivir alone, interferon-β1- β1 alone, and interferon-β1plus remdesivir) and ICU admission (p>0.2), the in-hospital mortality rate (p>0.2), or 30-day survival rate (p>0.2). The results were similar in patients who did or did not receive anti-CD20 agents. These results were consistent among patients regardless of whether they received anti-CD20 agents. Conclusion: Our data suggest that remdesivir, interferon-β1, or a combination of both does not benefit hospitalized MS patients with COVID-19. 111 122 2023/12/182024/09/29 1403/7/8 2024/09/142024/12/8 1403/9/18 Zhila Maghbooli Zhila Maghbooli Neuroscience Institute, Multiple Sclerosis Research Center, Tehran University of Medical Sciences, Tehran, Iran Iran zhilayas@gmail.com Amir Kasaeian Amir Kasaeian Liver and Pancreatobiliary Diseases Research Center, Digestive Diseases Research Institute, Tehran University of Medical Sciences, Tehran, Iran Iran amir_kasaeian@yahoo.com Mohammad Reza Fattahi Mohammad Reza Fattahi Neuroscience Institute, Multiple Sclerosis Research Center, Tehran University of Medical Sciences, Tehran, Iran Iran mrfatahi93@gmail.com Tarlan Varzandi Tarlan Varzandi Neuroscience Institute, Multiple Sclerosis Research Center, Tehran University of Medical Sciences, Tehran, IRAN Iran t.varzandi@gmail.com Sara Hamtaeigashi Sara Hamtaeigashi Neuroscience Institute, Multiple Sclerosis Research Center, Tehran University of Medical Sciences, Tehran, IRAN Iran Sarah.Hamtaee@yahoo.com Sara Mohammadnabi Sara Mohammadnabi Neuroscience Institute, Multiple Sclerosis Research Center, Tehran University of Medical Sciences, Tehran, IRAN Iran semagh62@gmail.com Mohammad Ali Sahraian Mohammad Ali Sahraian Neuroscience Institute, Multiple Sclerosis Research Center, Tehran University of Medical Sciences, Tehran, IRAN Iran ms.genelab@gmail.com Multiple sclerosis COVID-19 Remdesivir Interferon Anti-CD20 agents Baldi F, Dentone C, Mikulska M, Fenoglio D, Mirabella M, Magnè F, et al. Case report: Sotrovimab, remdesivir and nirmatrelvir/ritonavir combination as salvage treatment option in two immunocompromised patients hospitalized for COVID-19. Frontiers in Medicine 2022; 9: 1062450.##Bellucci G, Albanese A, Rizzi C, Rinaldi V, Salvetti M, Ristori G. The value of Interferon β in multiple sclerosis and novel opportunities for its anti-viral activity: a narrative literature review. Frontiers in Immunology 2023; 14: 1161849.##Chisari C G, Sgarlata E, Arena S, Toscano S, Luca M, Patti F. Rituximab for the treatment of multiple sclerosis: a review. Journal of Neurology 2022; 269: 159-183.##Cragg M S, Walshe C A, Ivanov A O, Glennie M J. The biology of CD20 and its potential as a target for mAb therapy. Current directions in autoimmunity 2005; 8: 140-174.##D’Abramo A, Vita S, Maffongelli G, Mariano A, Agrati C, Castilletti C, et al. Prolonged and severe SARS-CoV-2 infection in patients under B-cell-depleting drug successfully treated: A tailored approach. Int J Infect Dis 2021; 107: 247-250.##Davoudi-Monfared E, Rahmani H, Khalili H, Hajiabdolbaghi M, Salehi M, Abbasian L, et al. A Randomized clinical trial of the efficacy and safety of interferon β-1a in treatment of severe Covid-19. Antimicrob Agents Chemother 2020; 64.##De Wit E, Van Doremalen N, Falzarano D, Munster V J. SARS and MERS: recent insights into emerging coronaviruses. Nature reviews microbiology 2016; 14: 523-534.##Furlan A, Forner G, Cipriani L, Vian E, Rigoli R, Gherlinzoni F, et al. Covid-19 in B cell-depleted patients after rituximab: A diagnostic and therapeutic challenge. Frontiers in Immunology 2021; 12: 763412.##Haddad F, Dokmak G, Karaman R. A Comprehensive review on the efficacy of several pharmacologic agents for the treatment of Covid-19. Life 2022; 12: 1758.##Hartinger J M, Kratky V, Hruskova Z, Slanar O, Tesar V. Implications of rituximab pharmacokinetic and pharmacodynamic alterations in various immune-mediated glomerulopathies and potential anti-CD20 therapy alternatives. Frontiers in Immunology 2022; 13: 1024068.##Hashemi-Meshkini A, Koochak R, Nikfar S, Rezaei-Darzi E, Yaghoubifard S. Evaluation of Covid-19 treatments in iran in comparison with local therapeutic recommendations: A population-level study on utilization and costs of prescription drugs. Journal of Research in Pharmacy Practice 2022; 11: 1-7.##Hawker K, O’Connor P, Freedman M S, Calabresi P A, Antel J, Simon J, et al. Rituximab in patients with primary progressive multiple sclerosis: results of a randomized double-blind placebo-controlled multicenter trial. Annals of Neurology 2009; 66: 460-471.##Hollen C, Bernard J. Multiple sclerosis management during the Covid-19 pandemic. Current Neurology and Neuroscience Reports 2022; 22: 537-543.##https://www.covid19treatmentguidelines.nih.gov/. Covid-19 treatment guidelines panel. coronavirus disease 2019 (Covid-19) treatment guidelines. National Institutes of Health.##Hueso T, Pouderoux C, Péré H, Beaumont A L, Raillon L A, Ader F, et al. Convalescent plasma therapy for B-cell-depleted patients with protracted COVID-19. Blood 2020; 136: 2290-2295.##Hughes R, Whitley L, Fitovski K, Schneble H-M, Muros E, Sauter A, et al. COVID-19 in ocrelizumab-treated people with multiple sclerosis. Multiple Sclerosis and Related Disorders 2021; 49: 102725.##Jakimovski D, Kolb C, Ramanathan M, Zivadinov R, Weinstock-Guttman B. Interferon β for multiple sclerosis. Cold Spring Harbor Perspectives in Medicine 2018; 8.##Januel E, Hajage D, Labauge P, Maillart E, De Sèze J, Zephir H, et al. Association between anti-CD20 therapies and Covid-19 severity among patients with relapsing-remitting and progressive multiple sclerosis. JAMA Network Open 2023; 6: e2319766-e2319766.##Jelcic I, Al Nimer F, Wang J, Lentsch V, Planas R, Jelcic I, et al. Memory B cells activate brain-homing, autoreactive CD4(+) T cells in multiple sclerosis. Cell 2018; 175: 85-100.##Kado R, Sanders G, McCune W J. Suppression of normal immune responses after treatment with rituximab. Current Opinion in Rheumatology 2016; 28: 251-258.##Kalil A C, Mehta A K, Patterson T F, Erdmann N, Gomez C A, Jain M K, et al. Efficacy of interferon beta-1a plus remdesivir compared with remdesivir alone in hospitalised adults with COVID-19: a double-bind, randomised, placebo-controlled, phase 3 trial. The Lancet Respiratory Medicine 2021; 9: 1365-1376.##Kintrilis N, Gkinos C P, Galinos I. Prolonged Covid-19 in a multiple sclerosis patient treated with rituximab. Cureus 2022; 14: e32523.##Lassmann H. Pathogenic mechanisms associated with different clinical courses of multiple sclerosis. Frontiers in Immunology 2018; 9: 3116.##Lisak R P, Benjamins J A, Nedelkoska L, Barger J L, Ragheb S, Fan B, et al. Secretory products of multiple sclerosis B cells are cytotoxic to oligodendroglia in vitro. Journal of Neuroimmunology 2012; 246: 85-95.##Maghbooli Z, Hosseinpour H, Fattahi M R, Varzandi T, Hamtaeigashi S, Mohammad-Nabi S, et al. Association between disease-modifying therapies and adverse clinical outcomes in multiple sclerosis patients with Covid-19 infection. Multiple Sclerosis and Related Disorders 2022; 67: 104067.##Mary A, Hénaut L, Macq P Y, Badoux L, Cappe A, Porée T, et al. Rationale for Covid-19 treatment by nebulized interferon-β-1b-literature review and personal preliminary experience. Frontiers in Pharmacology 2020; 11: 592543.##McLaughlin P, Grillo-López A J, Link B K, Levy R, Czuczman M S, Williams M E, et al. Rituximab chimeric anti-CD20 monoclonal antibody therapy for relapsed indolent lymphoma: half of patients respond to a four-dose treatment program. Journal of Clinical Oncology 1998; 16: 2825-2833.##Mehta P, Porter J C, Chambers R C, Isenberg D A, Reddy V. B-cell depletion with rituximab in the COVID-19 pandemic: where do we stand? Lancet Rheumatol 2020; 2: e589-e590.##Okoli G N, Reddy V K, Lam O L, Askin N, Rabbani R. Update on efficacy of the approved remdesivir regimen for treatment of Covid-19: a systematic review with meta-analysis and trial sequential analysis of randomized controlled trials. Current Medical Research and Opinion 2024; 40: 1277-1287.##Pan H, Peto R, Henao-Restrepo A M, Preziosi M P, Sathiyamoorthy V, Abdool Karim Q, et al. Repurposed antiviral drugs for Covid-19 - interim WHO solidarity trial results. The New England Journal of Medicine 2021; 384: 497-511.##Schreiber G. The Role of type I interferons in the pathogenesis and treatment of Covid-19. Frontiers in Immunology 2020; 11: 595739.##Spinner C D, Gottlieb R L, Criner G J, Arribas López J R, Cattelan A M, Soriano Viladomiu A, et al. Effect of remdesivir vs standard care on clinical status at 11 days in patients with moderate Covid-19: A randomized clinical trial. Jama 2020; 324: 1048-1057.##van der Kolk L E, Baars J W, Prins M H, van Oers M H. Rituximab treatment results in impaired secondary humoral immune responsiveness. Blood 2002; 100: 2257-2259.## ##

Evaluation of bronchial response among occupationally exposed vulcanizers in calabar municipality, cross river state -nigeria 2 2 Introduction: Histamine directly interacts with airways smooth muscles to initiate contraction. This occurs even at low concentrations among individuals predisposed to increased airway inflammation and asthma. Methods: A digital vitalograph, nebulizer, histamine, and pulse oximeter were used in this research. A total of one hundred and sixty subjects were selected and categorized into two groups; Group 1 (Test subject) and Group 2 (Control), each group contained eighty (80) subjects. Group 1 comprised occupationally exposed vulcanizers and Group 2 comprised selected students from the University of Calabar as control. Both groups comprised young male subjects of equal age (20-40 years), devoid of pre-existing respiratory disorders or on any medication(s). Demographic parameters were used to obtain sufficient information for overall analysis. Lung parameters (FVC, FEV1, PEFR, and FEV1%) were evaluated using a Vitalograph Spirometer. Results: Basal measurement showed that Group 1 had a moderate decrease in lung volumes (FVC, FEV1, PEFR, and FEV1%) of 65 subjects (representing 81.25%) and a mild decrease in 15 subjects (18.75%); all the subjects in Group 2 showed a mild decrease in lung volumes. Inhalation of histamine mist of different concentrations showed that Group 1 had a severe decrease in lung volumes of 70 subjects (87.5%) and a mild decrease in 10 subjects (12.5%). All Subjects in Group 2 showed a mild decrease in lung volumes. Conclusion: Histamine provocative test helps to evaluate asthma signs and symptoms. Airways Hyper reactiv-ity is associated with inflammation as a consequence of airway hyperreflexia. It is evidenced that associated pollutants and their chronic inhalation by the vulcanizers have a retrogressive impact on their airways which manifests with hyper response an indication for asthma signs. 123 134 2023/12/182024/09/292023/10/28 1402/8/6 2024/09/142024/12/82024/10/5 1403/7/14 Utionkpan Linna Paul Utionkpan Linna Paul Human Physiology, University of Calabar, Calabar, Nigeria Nigeria linns.paul@yahoo.com Helyn A Ikem Helyn A Ikem Human Physiology, University of Calabar, Calabar, Nigeria Nigeria Cecilia Peter Cecilia Peter Human Physiology, University of Calabar, Calabar, Nigeria Nigeria Solomon Beshel Solomon Beshel Zoology and Environmental Biology, University of Calabar, Calabar, Nigeria Nigeria Spirometric evaluation Vulcanizer Histamine Bronchial responses Akiba M, Hashim A S. Vulcanization and crosslinking in elastomers. Progress in Polymer Science 1997; 22: 475-521.##Anderson S D, Brannan J D. Methods for “indirect” challenge tests including exercise, eucapnic voluntary hyperpnea, and hypertonic aerosols. Clinical Reviews in Allergy & Immunology 2003; 24: 27-54.##Araujo Z T d S, Holanda G. Does the BODE index correlate with quality of life in patients with COPD? Jornal Brasileiro de Pneumologia 2010; 36: 447-452.##Beydon N, Davis S D, Lombardi E, Allen J L, Arets H G, Aurora P, et al. An official American Thoracic Society/European Respiratory Society statement: pulmonary function testing in preschool children. American Journal of Respiratory and Critical Care Medicine 2007; 175: 1304-1345.##Bimestral P. Diretrizes da Sociedade Brasileira de Pneumologia e Tisiologia para o manejo da asma-2012. Jornal Brasileiro de Pneumologia 2012; 38.##Borges M, Ferraz E, Terra-Filho J, Vianna E. Protective effect of bronchial challenge with hypertonic saline on nocturnal asthma. Brazilian Journal of Medical and Biological Research 2008; 41: 209-214.##Borges M d C, Ferraz E, Vianna E O. Bronchial provocation tests in clinical practice. Sao Paulo Medical Journal 2011; 129: 243-249.##Boulet L-P, Laviolette M, Turcotte H, Cartier A, Dugas M, Malo J-L, et al. Bronchial subepithelial fibrosis correlates with airway responsiveness to methacholine. Chest 1997; 112: 45-52.##Brannan J, Gulliksson M, Anderson S, Chew N, Kumlin M. Evidence of mast cell activation and leukotriene release after mannitol inhalation. European Respiratory Journal 2003; 22: 491-496.##Brudno D S, Wagner J M, Rupp N T. Length of postexercise assessment in the determination of exercise-induced bronchospasm. Annals of Allergy 1994; 73: 227-231.##Caldeira R D, Bettiol H, Barbieri M A, Terra-Filho J, Garcia C A, Vianna E O. Prevalence and risk factors for work related asthma in young adults. Occupational and Environmental Medicine 2006; 63: 694-699.##Cockcroft D W. Bronchoprovocation methods: direct challenges. Clinical reviews in allergy & immunology 2003; 24: 19-26.##Cockcroft D W, Berscheid B A, Murdock K Y. Unimodal distribution of bronchial responsiveness to inhaled histamine in a random human population. Chest 1983; 83: 751-4.##Cockcroft D W, Davis B E. Mechanisms of airway hyperresponsiveness. Journal of allergy and clinical immunology 2006; 118: 551-559.##Córdova-Guerrero I, San Andrés L, Leal-Orozco A E, Padrón J M, Cornejo-Bravo J, León F. New strategy toward the diverted synthesis of oxidized abietane diterpenes via oxidation of 6, 7-dehydroferruginol methyl ether with dimethyldioxirane. Tetrahedron Letters 2013; 54: 4479-4482.##Curry J J. The effect of antihistamine substances and other drugs on histamine bronchoconstriction in asthmatic subjects. The Journal of Clinical Investigation 1946; 25: 792-799.##De Meer G, Marks G, Postma D. Review direct or indirect stimuli for bronchial challenge testing: what is the relevance for asthma epidemiology? Clinical & Experimental Allergy 2004; 34.##Echazarreta A, Gomez F, Ribas J, Sala E, Barbera J, Roca J, et al. Pulmonary gas exchange responses to histamine and methacholine challenges in mild asthma. European Respiratory Journal 2001; 17: 609-614.##Ferraz E, Borges M C, Terra-Filho J, Martinez J A, Vianna E O. Comparison of 4 AM and 4 PM bronchial responsiveness to hypertonic saline in asthma. Lung 2006; 184: 341-346.##Fitch K D, Sue-Chu M, Anderson S D, Boulet L-P, Hancox R J, McKenzie D C, et al. Asthma and the elite athlete: summary of the International Olympic Committee’s consensus conference, Lausanne, Switzerland, January 22-24, 2008. Journal of Allergy and Clinical Immunology 2008; 122: 254-260. e7.##Gauderman W J, Vora H, McConnell R, Berhane K, Gilliland F, Thomas D, et al. Effect of exposure to traffic on lung development from 10 to 18 years of age: a cohort study. The Lancet 2007; 369: 571-577.##Gotshall R W. Exercise-induced bronchoconstriction. Drugs 2002; 62: 1725-1739.##Holzer K, Anderson S D, Douglass J. Exercise in elite summer athletes: challenges for diagnosis. Journal of allergy and clinical immunology 2002; 110: 374-380.##Jensen L K, Larsen A, Mølhave L, Hansen M K, Knudsen B. Health evaluation of volatile organic compound (VOC) emissions from wood and wood-based materials. Archives of Environmental Health: An International Journal 2001; 56: 419-432.##Joos G F. Bronchial hyperresponsiveness: too complex to be useful? Current Opinion in Pharmacology 2003; 3: 233-238.##Joos G F, O’Connor B, Anderson S D, Chung F, Cockcroft D W, Dahlén B, et al. Indirect airway challenges. European Respiratory Journal 2003; 21: 1050-1068.##Knapp M, Bunn W, Stave G. Adult respiratory distress syndrome from sulfuric acid fume inhalation. Southern Medical Journal 1991; 84: 1031-1033.##Marone G, Gentile M, Petraroli A, De Rosa N, Triggiani M. Histamine-induced activation of human lung macrophages. 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American journal of respiratory and critical care medicine 2001; 163: 1256-1276.##Pogson Z E, Antoniak M D, Pacey S J, Lewis S A, Britton J R, Fogarty A W. Does a low sodium diet improve asthma control? A randomized controlled trial. American Journal of Respiratory and Critical Care Medicine 2008; 178: 132-138.##Rafferty P, Beasley R, Holgate S T. The contribution of histamine to immediate bronchoconstriction provoked by inhaled allergen and adenosine 5’monophosphate in atopic asthma. American Review of Respiratory Disease 1987; 136: 369-373.##Randolph C. An update on exercise-induced bronchoconstriction with and without asthma. Current Allergy and Asthma Reports 2009; 9: 433-438.##Ro C. Guidelines for methacholine and exercise challenge testing-1999. This official statement of the American Thoracic Society was adopted by the ATS Board of Directors, July 1999. 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Standardized challenge testing with pharmacological, physical and sensitizing stimuli in adults. Report Working Party Standardization of Lung Function Tests, European Community for Steel and Coal. Official Statement of the European Respiratory Society. European Respiratory Journal 1993; 16: 53-83.##Svedahl S, Svendsen K, Qvenild T, Sjaastad A K, Hilt B. Short term exposure to cooking fumes and pulmonary function. Journal of Occupational Medicine and Toxicology 2009; 4: 1-8.##TsangKW T. Bronchiectasis; notanorphandisease intheEast. The International Journal of Tuberculosis and Lung Disease 2004; 8: 691-702.##Van der lende R. Long-term variability of bronchial responsiveness to histamine in a random population sample of adults. American Review of Respiratory Disease 1993; 148: 944-949.##Van Schoor J, Joos G, Pauwels R. Indirect bronchial hyperresponsiveness in asthma: mechanisms, pharmacology and implications for clinical research. 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Expression of Apolipoprotein C-I (Apo C-I) in oral squamous cell carcinoma 2 2 Introduction: One of the most common cancers of the oral cavity is oral squamous cell carcinoma (OSCC). Previous studies have provided evidence that apolipoprotein C-I (Apo C-I) has an oncogenic role beyond its classical function. Although it has been investigated in prostate, breast, and lung cancer, there is no information available regarding Apo C-I expression in tumor and pri-tumor tissues of OSCC patients. Thus, the goal of the present study was to unravel the expression of Apo C-I in OSCC and investigate its correlation with the survival and grade of OSCC patients. Methods: The Apo C-I mRNA level was measured in tumor and pri-tumor tissues of 16 OSCC patients. In addition, the 34 paraffin-embedded tissues of OSCC patients and IHC technique were used to analyze the association of Apo C-I protein with the survival of OSCC patients.  Results: The mRNA (P=0.0386) gene expression of Apo C-I showed a statistically significant difference between tumor and pri-tumor tissues of OSCC patients. It seems that a high protein level of Apo C-I is related with poor survival of OSCC patients (P=0.04). The Apo C-I protein was positively correlated with the tumor site (Pearson r= 0.485, P=0.0036) but not the grade (Pearson r= 0.2295, P=0.1917). Our data showed that high Apo C-I protein levels might be correlated with poor survival of OSCC patients. Conclusion: Our findings suggest that patients with high protein levels of Apo C-I may have lower survival, making it a potential prognostic factor for OSCC. However, further investigation is necessary to establish this concept. 135 145 2023/12/182024/09/292023/10/282024/07/1 1403/4/11 2024/09/142024/12/82024/10/52024/09/25 1403/7/4 Kurosh Hamzanloui Moghadam Kurosh Hamzanloui Moghadam Department of Clinical Biochemistry, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran Iran kuroshmqdm75@gmail.com Walla Alazzawi Walla Alazzawi Department of Clinical Biochemistry, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran Iran hussain89abd@gmail.com Samira Derakhshan Samira Derakhshan Oral and Maxillofacial Pathology Department, School of Dentistry, Tehran University of Medical Sciences, Tehran, Iran Iran samderakhshan.den@gmail.com Abbas Karimi Abbas Karimi Oral and Maxillofacial Surgery Department, School of Dentistry, Tehran University of Medical Sciences, Tehran, Iran Iran abkarimi@tums.ac.ir Zahra Shahsavari Zahra Shahsavari Department of Clinical Biochemistry, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran Iran shahsavari.z@gmail.com Afsaneh Goudarzi Afsaneh Goudarzi Department of Clinical Biochemistry, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran Iran afsaneh.goudarzi@hotmail.com Apolipoprotein C-I Oral Squamous Cell Carcinoma Survival Argiris A, Karamouzis M V, Raben D, Ferris R L. 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Kisspeptin-13 attenuates the rewarding but not the reinstatement effects of methamphetamine in the conditioned place preference test in rats 2 2 Introduction: Methamphetamine (MA) addiction is a major global public health concern, yet there is currently no approved medication that effectively treats this addiction. Kisspeptin is a neuropeptide that has a role in the reproductive system, metabolism and energy balance, and metastasis suppression in different types of cancers. The kisspeptin receptors, GPR54 are widely distributed in the brain’s memory-related structures. Previous studies have revealed that the opioid system contributes to the addictive effects of MA. Additionally, preclinical studies have shown that a derivative of kisspeptin possesses anti-opioid properties. This study aimed to clarify the role of kisspeptin-13 (KP-13) on reward and reinstatement-related memory associated with MA in the conditioned place preference test. Methods: We evaluated pre-treatment with intracerebroventricular KP-13 for 3 consecutive days (2.5 μl/nostril, once a day) in a conditioned place preference test induced by MA. MA was administered intraperitoneally at a dose ranging from 1 to 7 mg/kg, once daily, beginning at 1 mg/kg on day one and increasing by 1 mg/kg per day up to the day 7. Results: We found that KP-13 suppresses the reward behavior in MA-treated rats, while it has no significant effect on reinstatement behavior after one week of MA cessation, which could be attributed to the ineffective dose of KP-13. Conclusion: The findings indicate that KP-13 alters the rewarding and motivating effects of MA. Further research involving both multiple and single administrations of KP-13 before the reinstatement test is necessary to throw light on its impact on withdrawal-related reinstatement. 146 154 2023/12/182024/09/292023/10/282024/07/12024/01/16 1402/10/26 2024/09/142024/12/82024/10/52024/09/252024/09/7 1403/6/17 Somayeh Nazari Somayeh Nazari Student Research Committee, School of Advanced Technologies in Medicine, Mazandaran University of Medical Sciences, Sari, Iran Iran snazari70@yahoo.com Mobina Gheibi Mobina Gheibi Student Research Committee, Mazandaran University of Medical Sciences, Sari, Iran Iran mobinagheibi78@gmail.com Hamed Ghazvini Hamed Ghazvini Department of Neuroscience, School of Advanced Technologies in Medicine, Mazandaran University of Medical Sciences, Sari, Iran Iran hghazvini1@gmail.com Raheleh Rafaiee Raheleh Rafaiee Department of Neuroscience, School of Advanced Technologies in Medicine, Mazandaran University of Medical Sciences, Sari, Iran Iran rachel.rafaie@yahoo.com seyedeh masoumeh Seyedhosseini Tamijani seyedeh masoumeh Seyedhosseini Tamijani Department of Neuroscience, School of Advanced Technologies in Medicine, Mazandaran University of Medical Sciences, Sari, Iran Iran m.seyedhoseini@mazums.ac.ir Kisspeptin-13 Methamphetamine Reward Reinstatement Condition place preference Anooshe M, Nouri K, Karimi-Haghighi S, Mousavi Z, Haghparast A. Cannabidiol efficiently suppressed the acquisition and expression of methamphetamine-induced conditioned place preference in the rat. Behavioural Brain Research 2021; 404: 113158.##Chiu C-T, Ma T, Ho I K. Methamphetamine-induced behavioral sensitization in mice: alterations in μ-opioid receptor. Journal of biomedical science 2006; 13: 797-811.##Delmas S, Porteous R, Bergin D H, Herbison A E. Altered aspects of anxiety-related behavior in kisspeptin receptor-deleted male mice. Scientific Reports 2018; 8: 2794.##Ebrahimi Z, Kahvandi N, Komaki A, Karimi S A, Naderishahab M, Sarihi A. The role of mGlu4 receptors within the nucleus accumbens in acquisition and expression of morphine-induced conditioned place preference in male rats. BMC neuroscience 2021; 22: 1-10.##Ekhtiari H, Nasseri P, Yavari F, Mokri A, Monterosso J. Neuroscience of drug craving for addiction medicine: From circuits to therapies. Progress in brain research 2016; 223: 115-141.##Everitt B J. Neural and psychological mechanisms underlying compulsive drug seeking habits and drug memories-indications for novel treatments of addiction. Eur J Neurosci 2014; 40: 2163-2182.##Gibula-Bruzda E, Marszalek-Grabska M, Gawel K, Trzcinska R, Silberring J, Kotlinska J H. The new kisspeptin derivative-kissorphin (KSO)-attenuates acute hyperlocomotion and sensitization induced by ethanol and morphine in mice. Alcohol 2017; 64: 45-53.##Gibula-Tarlowska E, Grochecki P, Silberring J, Kotlinska J H. The kisspeptin derivative kissorphin reduces the acquisition, expression, and reinstatement of ethanol-induced conditioned place preference in rats. Alcohol 2019a; 81: 11-19.##Gibula-Tarlowska E, Kedzierska E, Piechura K, Silberring J, Kotlinska J. The influence of a new derivate of kisspeptin-10-Kissorphin (KSO) on the rewarding effects of morphine in the conditioned place preference (CPP) test in male rats. Behavioural Brain Research 2019b; 372: 112043.##Ibos K E, Bodnár É, Bagosi Z, Bozsó Z, Tóth G, Szabó G, et al. Kisspeptin-8 induces anxiety-like behavior and hypolocomotion by activating the HPA axis and increasing GABA release in the nucleus accumbens in rats. Biomedicines 2021; 9: 112.##Jiang J, He Z, Peng Y, Jin W, Wang Z, Han R, et al. Kisspeptin-13 enhances memory and mitigates memory impairment induced by Aβ1-42 in mice novel object and object location recognition tasks. Neurobiology of learning and memory 2015; 123: 187-195.##Khodamoradi M, Tirgar F, Ghazvini H, Rafaiee R, Tamijani S M S, Karimi N, et al. Role of the cannabinoid CB1 receptor in methamphetamine-induced social and recognition memory impairment. Neuroscience letters 2022; 779: 136634.##Khonacha S E, Janahmadi M, Motamedi F. Kisspeptin-13 improves spatial memory consolidation and retrieval against amyloid-β pathology. Iran J Pharm Res 2019; 18: 169.##Kotlinska J, Pachuta A, Dylag T, Silberring J. 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Can the kisspeptin help us in the understanding of pathology of some neurodegenerative brain diseases? Folia Morphologica 2021; 80: 756-765.##Mills E G, Dhillo W S, Comninos A N. Kisspeptin and the control of emotions, mood and reproductive behaviour. Journal of Endocrinology 2018; 239: R1-R12.##Mills E G, O’Byrne K T, Comninos A N. Kisspeptin as a behavioral hormone. Journal 2019; 37: 056-063.##Milton N G, Chilumuri A, Rocha-Ferreira E, Nercessian A N, Ashioti M. Kisspeptin prevention of amyloid-β peptide neurotoxicity in vitro. ACS chemical neuroscience 2012; 3: 706-719.##Moenk M D, Matuszewich L. Juvenile but not adult methamphetamine exposure improves performance in the Morris Water Maze in male rats. Int J Dev Neurosci 2012; 30: 325-331.##Oishi S, Misu R, Tomita K, Setsuda S, Masuda R, Ohno H, et al. Activation of neuropeptide FF receptors by kisspeptin receptor ligands. ACS Medicinal Chemistry Letters 2011; 2: 53-57.##Park S W, He Z, Shen X, Roman R J, Ma T. 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Identification of endometriosis molecular regulatory axes through bioinformatics analysis: insights into lncRNAs, miRNAs, and mRNAs 2 2 Introduction: Endometriosis (EMS) is a highly prevalent gynecological disorder with substantial health consequences, affecting as many as 10% of women in their reproductive years. Although EMS is widespread, its intricate origin and pathophysiology are not well understood. This study investigated the molecular characteristics of EMS by examining the expression patterns of long noncoding RNAs (lncRNAs), microRNAs (miRNAs), and genes linked to the condition. Methods: The study used publicly accessible datasets, namely RNA-seq data for long non-coding RNAs (GSE105764), miRNA expression data (GSE105765), and gene expression data (GSE12768), to compare samples of ectopic and eutopic endometrial samples. The study conducted differential expression analysis, pathway enrichment analyses, protein-protein interaction network creation, and hub gene identification to uncover the molecular markers linked to EMS.  Results: The investigation identified substantial dysregulation of lncRNAs, miRNAs, and genes in endometriotic tissues compared with the control eutopic endometrium. Hub genes, including CDH1, ESR1, GATA4, PGR, FOXA2, EPCAM, APOA1, BDNF, FGFR2, and PAX2, dominate the molecular landscape. miR-6500-3p has been identified as a pivotal regulator that influences the expression of seven essential genes. Pathway enrichment analysis identified biological processes, including embryonic organ morphogenesis, immunological response, and medication metabolism. Exploring lncRNA-mediated regulatory axes, particularly H19 and MIR600HG, suggests complex networks that may clarify the molecular underpinnings of EMS. Conclusion: This thorough investigation offers significant insight into the molecular pathways underlying EMS. Hub genes and regulatory miRNAs, including miR-6500-3p, have been identified as prospective targets for therapeutic interventions. Pathway enrichment studies provide a more comprehensive understanding of the biological processes implicated in EMS, indicating new possibilities for therapeutic interventions. 155 170 2023/12/182024/09/292023/10/282024/07/12024/01/162024/04/27 1403/2/8 2024/09/142024/12/82024/10/52024/09/252024/09/72024/08/31 1403/6/10 Shahrzad Zhaeentan Shahrzad Zhaeentan Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran Iran zhaeentan.shahrzad@yahoo.com Fardin Amidi Fardin Amidi Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran Iran amidifardin@yahoo.com Ashraf Moini Ashraf Moini Department of Gynecology and Obstetrics, Arash Women’s Hospital, Tehran University of Medical Sciences, Tehran, Iran Iran moeinias@tums.ac.ir Seyed Danial Mohammadi Seyed Danial Mohammadi Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran Iran drsdmohammadi@gmail.com Aligholi Sobhani Aligholi Sobhani Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran Iran sobhania@tums.ac.ir Masoomeh Nataj Majd Masoomeh Nataj Majd Department of Infertility, Arash Women’s Hospital, Tehran University of Medical Sciences, Tehran, Iran Iran mnataj54@yahoo.com Mahshad khodarahmian Mahshad khodarahmian Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran Iran ma.khodarahmian@gmail.com Endometriosis lncRNAs miRNAs Hub Genes Bioinformatics The Gene Ontology Resource: 20 years and still GOing strong. 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Caffeine and exercise training alter expression of genes involved in inflammation and the browning of adipose tissue in High-Fat Diet-Fed Rats 2 2 Introduction: Caffeine, as a popular drink, along with exercise training, may help restore altered gene expres-sion in high-fat diet (HFD)-induced obesity. This study examined the effects of exercise training, caffeine consumption, and their interaction on inflammation and genes involved in metabolism in rats fed an HFD. Methods: Eighty male Wistar rats were separated into two groups: HFD and normal diet (ND). Each group was subsequently divided into four groups: sedentary, caffeine-only, exercise, and caffeine-plus-exercise. For eight weeks, the animals in the training groups engaged in aerobic exercise on a mo-torized treadmill for 60 minutes, five times per week. Animals in the caffeine group ingested a solution containing caffeine daily (6 mg/kg/bw). The expression of Peroxisome proliferator-acti­vated receptor-gamma coactivator 1 alpha (PGC-1α) and fibronectin type III domain-containing (FNDC5) genes in the calf muscle, uncoupling protein-1 (UCP1) in subcutaneous adipose tissue, NF-KB and TLR4 in vis-ceral adipose tissue, and fetuin-A (Fet-A) in the liver were investigated. Results: The findings demonstrated that HFD significantly elevated the NF-Kβ gene and downregulated the skeletal muscle PGC-1α and FNDC5 genes, as well as serum fetuin-A. UCP-1 (366% vs. 56%), FNDC5 (26% vs. 54%), and PGC-1α (40% vs. 1700%) genes were all considerably elevated by exercise training and caffeine supplementation, respectively. Additionally, exercise training reduced TLR4 and NF-Kβ expression in visceral adipose tissue and liver Fet-A gene expression. Furthermore, following HFD, when compared to the sedentary group, exercise training with and without caffeine consumption decreased the NF-Kβ gene and liver Fet-A and increased PGC1-α, FNDC5, UCP1, and serum Fet-A. Conclusion: These findings support the idea that exercise and caffeine may reduce inflammation by downregulating genes involved in inflammation and adipose tissue browning. 171 183 2023/12/182024/09/292023/10/282024/07/12024/01/162024/04/272024/04/26 1403/2/7 2024/09/142024/12/82024/10/52024/09/252024/09/72024/08/312024/09/25 1403/7/4 Mohammad Rahman Rahimi Mohammad Rahman Rahimi Department of Exercise Physiology, University of Kurdistan, Sanandaj, Iran Iran r.rahimi@uok.ac.ir Sanaz Mahmazi Sanaz Mahmazi Department of Genetics, Faculty of Basic Sciences, Islamic Azad University, Zanjan Branch, Zanjan, Iran Iran sanazmahmazi@yahoo.com Mehdi Abbaspoor Mehdi Abbaspoor Department of Physical Education and Sport Sciences, Shahid Bahonar University of Kerman, Kerman, Iran Iran m.abbaspoor@uk.ac.ir Mehdi Rahnema Mehdi Rahnema Department of Genetics, Faculty of Basic Sciences, Islamic Azad University, Zanjan Branch, Zanjan, Iran Iran meh_rahnema@yahoo.com Dara Latif Saifalddin Dara Latif Saifalddin Department of Physical Education and Sport Sciences, University of Halabja, Kurdistan region, Iraq Iraq dara.saifaddin@uoh.edu.iq Nasser Ghanbari Nasser Ghanbari Department of Exercise Physiology, University of Kurdistan, Sanandaj, Iran Iran naser2617@gmail.com Sarkawt Kolahdouzi Sarkawt Kolahdouzi Department of Exercise Physiology, Faculty of Sport Sciences, University of Mazandaran, Babolsar, Iran Iran skolahdouzi@yahoo.com Hadi Golpasandi Hadi Golpasandi Department of Exercise Physiology, University of Kurdistan, Sanandaj, Iran Iran hadi.golpasandi@uok.ac.ir Exercise training Caffeine Supplementation Brown Adipose Tissue Uncoupling Protein Inflammation Blumenthal J B, Gitterman A, Ryan A S, Prior S J. 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The effect of Safranal on histological damages and oxidative stress induced by ischemia-reperfusion in adult rat ovaries 2 2 Introduction: Ovarian torsion is an emergency condition that occurs when the adnexa undergoes complete or partial rotation. Following ovarian torsion, damage caused by ischemia/reperfusion can impact fertility and sex hormone secretion. Our study aims to investigate the role of safranal in preventing ovarian ischemia-reperfusion injury in rats. Methods: Animal subgroups included: sham, ovarian torsion/detorsion (OT), ovarian torsion/detorsion with the treatment of safranal (OTS 0.1, 0.5), and safranal group (OS 0.1, 0.5). Ovarian torsion was induced in the left ovary of rats for 2 hours. Intraperitoneal treatment with safranal at 0.1 and 0.5 mg/kg doses was performed 30 minutes before the detorsion operation. 48 h after detorsion, ovarian tissue was collected to evaluate the histopathological scores and apoptosis gene expression of Bcl-2, caspase 3, and Bax. Blood samples were collected to measure plasma estradiol levels and oxidative stress parameters. Results: Analyses demonstrated that ovarian follicular damage was accompanied by increased serum malondialdehyde (MDA) levels and increased expression of Bcl2 and caspase-3 genes. Additionally, serum levels of glutathione peroxidase (GPX), estrogen, and superoxide dismutase (SOD) decreased in the OT group. However, treatment with safranal 0.5 mg/kg was accompanied by an improvement in the histopathological score of the ovarian tissue and a reduction in apoptosis and oxidative stress. Conclusion: Safranal exhibits antioxidant properties in a rat model of ovarian torsion and could be considered a cost-effective therapeutic option for ovarian detorsion in gynecological clinics.  184 193 2023/12/182024/09/292023/10/282024/07/12024/01/162024/04/272024/04/262024/02/19 1402/11/30 2024/09/142024/12/82024/10/52024/09/252024/09/72024/08/312024/09/252024/10/5 1403/7/14 Marzieh Ebrahimi Marzieh Ebrahimi Student Research Committee, Gonabad University of Medical Sciences, Gonabad, Iran Iran Marzieh.ebr74@gmail.com Masoumeh Fani Masoumeh Fani Department of Anatomical Sciences, School of Medicine, Gonabad University of Medical Sciences, Gonabad, Iran Iran m.fani2017@yahoo.com Seyed-Hosein Abtahi-Evari Seyed-Hosein Abtahi-Evari Department of Biochemistry, School of Medicine, Gonabad University of Medical Sciences, Gonabad, Iran Iran Abtahi.h@gmu.ac.ir Balal Brazvan Balal Brazvan Department of Anatomical Sciences, School of Medicine, Gonabad University of Medical Sciences, Gonabad, Iran Iran bbarazvan@gmail.com Sohrab Azin Sohrab Azin Student Research Committee, Gonabad University of Medical Sciences, Gonabad, Iran Iran Sohrab.azin.75119@gmail.com Majid Shokoohi Majid Shokoohi Department of Anatomical Sciences, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran Iran a.shokoohy@yahoo.com Sajjad Abbasi Sajjad Abbasi Student Research Committee, Gonabad University of Medical Sciences, Gonabad, Iran Iran Sajjadabbasi75@gmail.com vida Alikhani vida Alikhani Department of Physiology, School of Medicine, Gonabad University of Medical Sciences, Gonabad, Iran Iran vd.alikhani@gmail.com Amir- hosein Ebadi Amir- hosein Ebadi Student Research Committee, Gonabad University of Medical Sciences, Gonabad, Iran Iran ebadi.amirhossein@hotmail.com Malihe soltani Malihe soltani Department of Anatomical Sciences, School of Medicine, Gonabad University of Medical Sciences, Gonabad, Iran Iran soltani.m@gmu.ac.ir Maryam Moghimian Maryam Moghimian Department of Physiology, School of Medicine, Gonabad University of Medical Sciences, Gonabad, Iran Iran moghimian.m@gmu.ac.ir Safranal Oxidative Stress Ischemia-reperfusion Ovaries Abali R, Tasdemir N, Yuksel M A, Guzel S, Oznur M, Nalbantoglu B, et al. 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Regular exercise and enriched environment ameliorate neuroinflammation in pilocarpineinduced epileptic rats by antioxidant activity and NLRP3/caspase-1 pathway inhibition 2 2 Introduction: As a chronic neurological disorder, epilepsy is affected by social stress, which is one of the numerous complications in societies. In addition to medication, enriched environment (EE) and exercise are among the complementary strategies in the treatment of epilepsy. Oxidative stress, which potentially can activate the inflammatory pathways, is one of the causes of this disorder. So, we tried to examine thoroughly the beneficial impacts of EE and exercise on neuroinflammation in epileptic rats. Methods: Male Wistar rats were divided into five groups of twelve rats each, including: a control group, a group induced with pilocarpine to simulate epilepsy, an epileptic group subjected to social stress, an epileptic group placed in an enriched environment, and an epileptic group subjected to an exercise regimen. The impact of social stress, enriched environment, and exercise on oxidative stress biomarkers was investigated through TBARS  spectrophotometric test   and the gene expression of NLRP3, Caspase-1, IL18, and IL1β were evaluated through real-time PCR method. Results: Epilepsy and social stress caused a reduction in superoxide dismutase (SOD) and glutathione peroxidase (GPx) (p<0.05). Moreover, they resulted in an enhancement of plasma malondialdehyde (MDA), NLRP3, Caspase-1, interleukin-18 (IL18), and IL1β gene expression (p<0.05). Exercise increased the GPx and diminished the expression of Caspase-1 and IL-18 inflammatory genes (p<0.05). Accordingly, EE enhanced the SOD and GPx antioxidant indicators and reduced proinflammatory gene expression. Conclusion: In this research, social stress resulted in elevated levels of oxidative markers and upregulation of inflammatory gene expression. EE and regular exercise improved the situation. 194 204 2023/12/182024/09/292023/10/282024/07/12024/01/162024/04/272024/04/262024/02/192024/04/23 1403/2/4 2024/09/142024/12/82024/10/52024/09/252024/09/72024/08/312024/09/252024/10/52024/08/19 1403/5/29 Fatemeh Moradi Fatemeh Moradi Department of Physiology, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran Iran f.moradi@zums.ac.ir Mahsa Jazini Mahsa Jazini Department of Physiology, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran Iran mahsa.jazini92@gmail.com Hadi Feizi Hadi Feizi Department of Physiology, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran Iran hfeizik@zums.ac.ir Mahin Ganjkhani Mahin Ganjkhani Department of Physiology, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran Iran mganjkhani@zums.ac.ir Temporal lobe epilepsy social stress enriched environment exercise NLRP3 pathway Abdanipour A, Tiraihi T, Mirnajafi-Zadeh J. 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Conditioned medium of Wharton’s jelly-derived mesenchymal stem cells restores spermatogenesis in scrotal hyperthermia-induced azoospermia 2 2 Introduction: Azoospermia is a common cause of male infertility. The latest hope for more affordable and safer therapies comes from stem cell research. In this study, the effect of mesenchymal stem cells derived from Wharton’s jelly on spermatogenesis in azoospermic mice with scrotal hyperthermia was investigated. Methods: For the experimental study, 24 adult male mice were divided into four groups: 1. control (Cont), 2. scrotal hyperthermia (Hyp), 3. scrotal hyperthermia + DMEM (10μl) (Hyp/DMEM), 4. scrotal hyperthermia + conditioned medium (10 μl) (Hyp/CM). A temperature of 43 °C was applied for 20 minutes every other day for two weeks to induce hyperthermia. The mice in the experimental group received 10 µl of human Jelly Wharton’s Stem Cell-Conditioned Medium (hJWSC-CM) intraperitoneally every other day for 35 days after hyperthermia. Animals were sacrificed after the experiments to perform additional molecular, biochemical, and stereological analyses. Results: The results showed that CM significantly increased both the total amount of sperm and the number of testicular cells, including spermatids, primary spermatocytes, spermatogonia, Leydig cells, and Sertoli cells, compared with Hyp and Hyp/DMEM; in addition, the biochemical characteristics of testicular tissue were significantly higher in the Hyp/CM group compared with the hyperthermia-induced groups. Further analysis revealed that when the Hyp/CM group was compared with the Hyp and Hyp/DMEM groups, Sox9 protein expression and proliferative gene expression increased significantly and the percentage of apoptotic testicular cells and pro-inflammatory gene expression decreased significantly. Conclusion: These results suggest that realistic therapeutic approaches for reproductive and regenerative medicine could benefit from hWJSC-CM. 205 218 2023/12/182024/09/292023/10/282024/07/12024/01/162024/04/272024/04/262024/02/192024/04/232024/01/27 1402/11/7 2024/09/142024/12/82024/10/52024/09/252024/09/72024/08/312024/09/252024/10/52024/08/192024/09/7 1403/6/17 Atefeh Shirazi tehrani Atefeh Shirazi tehrani Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Tehran, Iran Iran atefeh.shirazi@yahoo.com Hojjat-Allah Abbaszadeh Hojjat-Allah Abbaszadeh Laser Applications in Medical Sciences Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Tehran, Iran Iran hoomanabs@gmail.com Abbas Aliaghaei Abbas Aliaghaei Hearing Disorders Research Center, Loghman Hakim Hospital, Shahid Beheshti University of Medical Sciences, Tehran, Tehran, Iran Iran aghaei60@gmail.com Hamid Nazarian Hamid Nazarian Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Tehran, Iran Iran hamid.nazarian@gmail.com Azar Afshar Azar Afshar Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Tehran, Iran Iran azar.afshar8894@gmail.com Fakhroddin Aghajanpour Fakhroddin Aghajanpour Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Tehran, Iran Iran fakhroddin72@gmail.com Reza Soltani Reza Soltani Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Tehran, Iran Iran rezasoltanikazemii@gmail.com Pourya Raee Pourya Raee Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Tehran, Iran Iran pourayarayi3@gmail.com Fatemeh Fadaei Fathabadi Fatemeh Fadaei Fathabadi Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Tehran, Iran Iran mrs.tehrani92@yahoo.com Mohammad-Amin Abdollahifar Mohammad-Amin Abdollahifar Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Tehran, Iran Iran m_amin58@yahoo.com Scrotal hyperthermia Conditioned medium Wharton's jelly Spermatogenesis Azoospermia Afshar A, Aliaghaei A, Nazarian H, Abbaszadeh H-A, Naserzadeh P, Fathabadi F F, et al. Curcumin-loaded iron particle improvement of spermatogenesis in azoospermic mouse induced by long-term scrotal hyperthermia. Reproductive Sciences 2021; 28: 371-380.##Almeria C, Weiss R, Roy M, Tripisciano C, Kasper C, Weber V, et al. Hypoxia conditioned mesenchymal stem cell-derived extracellular vesicles induce increased vascular tube formation in vitro. Frontiers in Bioengineering and Biotechnology 2019; 7: 292.##Bazoobandi S, Tanideh N, Rahmanifar F, Zare S, Koohi-Hosseinabadi O, Razeghian-Jahromi I, et al. Preventive effects of intrauterine injection of bone marrow-derived mesenchymal stromal cell-conditioned media on uterine fibrosis immediately after endometrial curettage in rabbit. Stem Cells International 2020; 2020: 8849537.##Boni R. Heat stress, a serious threat to reproductive function in animals and humans. Molecular Reproduction and Development 2019; 86: 1307-1323.##Boroujeni M E, Simani L, Bluyssen H A R, Samadikhah H R, Zamanlui Benisi S, Hassani S, et al. Inflammatory response leads to neuronal death in human post-mortem cerebral cortex in patients with COVID-19. ACS Chemical Neuroscience 2021; 12: 2143-2150.##Chudickova M, Vackova I, Machova Urdzikova L, Jancova P, Kekulova K, Rehorova M, et al. The effect of Wharton jelly-derived mesenchymal stromal cells and their conditioned media in the treatment of a rat spinal cord injury. International Journal of Molecular Sciences 2019; 20: 4516.##Durairajanayagam D, Agarwal A, Ong C. Causes, effects and molecular mechanisms of testicular heat stress. Reproductive Biomedicine Online 2015; 30: 14-27.##Galipeau J, Sensébé L. Mesenchymal stromal cells: Clinical challenges and therapeutic opportunities. Cell Stem Cell 2018; 22: 824-833.##Ghaem Maghami R, Mirzapour T, Bayrami A. Differentiation of mesenchymal stem cells to germ-like cells under induction of Sertoli cell-conditioned medium and retinoic acid. Andrologia 2018; 50: e12887.##Ghanbari A, Moradi M, Raoofi A, Falahi M, Seydi S. Tribulus terrestris hydroalcoholic extract administration effects on reproductive parameters and serum level of glucose in diabetic male rats. International Journal of Morphology 2016; 34: 796-803.##Halder S, Sarkar M, Dey S, Bhunia S K, Koley A R, Giri B. Protective effects of red grape (Vitis vinifera) juice through restoration of antioxidant defense, endocrine swing and Hsf1, Hsp72 levels in heat stress induced testicular dysregulation of Wister rat. Journal of Thermal Biology 2018; 71: 32-40.##Ilacqua A, Izzo G, Emerenziani G P, Baldari C, Aversa A. Lifestyle and fertility: the influence of stress and quality of life on male fertility. Reproductive Biology and Endocrinology 2018; 16: 1-11.##Ilkhani S, Moradi A, Aliaghaei A, Norouzian M, Abdi S, Rojhani E, et al. Spatial arrangement of testicular cells disrupted by transient scrotal hyperthermia and subsequent impairment of spermatogenesis. Andrologia 2020; 52: e13664.##Jahromi I R, Mehrabani D, Mohammadi A, Seno M M G, Dianatpour M, Zare S, et al. Emergence of signs of neural cells after exposure of bone marrow-derived mesenchymal stem cells to fetal brain extract. Iranian Journal of Basic Medical Sciences 2017; 20: 301.##Khosravi A, Hasani A, Behnam P, Piryaei A, Pirani M, Aliaghaei A, et al. An effective method for establishing animal models of azoospermia and oligospermia. Andrologia 2021; 53: e14095.##Kumar Roy V, Marak T R, Gurusubramanian G. Alleviating effect of Mallotus roxburghianus in heat-induced testicular dysfunction in Wistar rats. Pharmaceutical Biology 2016; 54: 9-18.##Li X, Tian E, Wang Y, Wen Z, Lei Z, Zhong Y, et al. Stem Leydig cells: Current research and future prospects of regenerative medicine of male reproductive health. Seminars in Cell and Developmental Biology 2022; 121: 63-70.##Mahiddine F Y, Kim J W, Qamar A Y, Ra J C, Kim S H, Jung E J, et al. Conditioned medium from canine amniotic membrane-derived mesenchymal stem cells improved dog sperm post-thaw quality-related parameters. Animals 2020; 10: 1899.##Moghimi N, Eslami Farsani B, Ghadipasha M, Mahmoudiasl G-R, Piryaei A, Aliaghaei A, et al. COVID-19 disrupts spermatogenesis through the oxidative stress pathway following induction of apoptosis. Apoptosis 2021; 26: 415-430.##Moscatelli N, Lunetti P, Braccia C, Armirotti A, Pisanello F, De Vittorio M, et al. Comparative proteomic analysis of proteins involved in bioenergetics pathways associated with human sperm motility. International Journal of Molecular Sciences 2019; 20: 3000.##Mozafar A, Mehrabani D, Vahdati A, Hosseini E, Forouzanfar M. Histomorphometric evaluation of allogeneic transplantation of bone marrow mesenchymal stem cells in azoospermic mice model. The Indonesian Biomedical Journal 2018; 10: 171-178.##Panahi S, Karamian A, Sajadi E, Aliaghaei A, Nazarian H, Abdi S, et al. Sertoli cell-conditioned medium restores spermatogenesis in azoospermic mouse testis. Cell and Tissue Research 2020; 379: 577-587.##Park K-S, Bandeira E, Shelke G V, Lässer C, Lötvall J. Enhancement of therapeutic potential of mesenchymal stem cell-derived extracellular vesicles. Stem Cell Research & Therapy 2019; 10: 1-15.##Park S G, Yon J M, Lin C, Gwon L W, Lee J G, Baek I J, et al. Capsaicin attenuates spermatogenic cell death induced by scrotal hyperthermia through its antioxidative and anti-apoptotic activities. Andrologia 2017; 49: e12656.##Payehdar A, Hosseini E, Mehrabani D, Forouzanfar M. Healing effect of conditioned medium of adipose tissue-derived mesenchymal stem cells on histomorphometric changes of mice testis in busulfan induced-azoospermia model. Internal Medicine Today 2017; 23: 23-24.##Pirani M, Novin M G, Abdollahifar M-A, Piryaei A, Kuroshli Z, Mofarahe Z S. Protective effects of fisetin in the mice induced by long-term scrotal hyperthermia. Reproductive Sciences 2021; 28: 3123-3136.##Ranjbaran H, Abediankenari S, Mohammadi M, Jafari N, Khalilian A, Rahmani Z, et al. Wharton’s Jelly derived-mesenchymal stem cells: isolation and characterization. Acta Medica Iranica 2018: 28-33.##Rao M, Xia W, Yang J, Hu L X, Hu S F, Lei H, et al. Transient scrotal hyperthermia affects human sperm DNA integrity, sperm apoptosis, and sperm protein expression. Andrology 2016; 4: 1054-1063.##Rao M, Zhao X-L, Yang J, Hu S-F, Lei H, Xia W, et al. Effect of transient scrotal hyperthermia on sperm parameters, seminal plasma biochemical markers, and oxidative stress in men. Asian Journal of Andrology 2015; 17: 668-675.##Shah S M, Saini N, Singh M K, Manik R, Singla S K, Palta P, et al. Testicular cell-conditioned medium supports embryonic stem cell differentiation toward germ lineage and to spermatocyte- and oocyte-like cells. Theriogenology 2016; 86: 715-729.##Shen C, Lie P, Miao T, Yu M, Lu Q, Feng T, et al. Conditioned medium from umbilical cord mesenchymal stem cells induces migration and angiogenesis. Molecular Medicine Reports 2015; 12: 20-30.##Shirazi R, Zarnani A H, Soleimani M, Nayernia K, Ragerdi Kashani I. Differentiation of bone marrow-derived stage-specific embryonic antigen 1 positive pluripotent stem cells into male germ cells. Microscopy Research and Technique 2017; 80: 430-440.##Takashima S, Kanatsu-Shinohara M, Tanaka T, Morimoto H, Inoue K, Ogonuki N, et al. Functional differences between GDNF-dependent and FGF2-dependent mouse spermatogonial stem cell self-renewal. Stem Cell Reports 2015; 4: 489-502.##Vij S C, Sabanegh E, Jr., Agarwal A. Biological therapy for non-obstructive azoospermia. Expert Opinion on Biological Therapy 2018; 18: 19-23.##Vizoso F J, Eiro N, Cid S, Schneider J, Perez-Fernandez R. Mesenchymal stem cell secretome: Toward cell-free therapeutic strategies in regenerative medicine. 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Stevia and especially Nano-Stevia induced anti diabetic, antidepressant, and analgesic effects in streptozotocin-induced diabetic rats 2 2 Introduction: Diabetes mellitus is a metabolic disease. This disease is almost associated with depression and impaired pain sensation. Stevia rebaudiana Bertoni (stevia) has a sweet taste and a useful effect in blood glucose control. This study was designed to evaluate the effect of Stevia and Nano-stevia on psychological problems (e.g. depression and pain) induced by diabetes. Methods: By a single intraperitoneal administration of streptozotocin, diabetes mellitus was induced in male Wistar rats. Stevia (20 mg/dl per day for 30 days) and Nano-stevia (20 mg/dl per day for 30 days) were administrated intra-gastrically in diabetic rats. Using the forced swim test (FST) and formalin test, depression and pain behaviors were measured, respectively.  Results: The results exhibited that male diabetic rats showed hyperglycemia, depressant-related behavior, and hyperalgesic response. On the other hand, intragastric usage of Stevia (20 mg/dl per day for 30 days) and Nano-stevia (20 mg/dl per day for 30 days) modified the hyperglycemia, depressant-related response, and hyperalgesic effect in diabetic rats. Nano-stevia indicated the maximum significant response rather than Stevia. Conclusion: The results of this study indicate that Stevia and particularly Nano-stevia could be beneficial in managing diabetes and its associated behavioral complications. 219 231 2023/12/182024/09/292023/10/282024/07/12024/01/162024/04/272024/04/262024/02/192024/04/232024/01/272023/12/30 1402/10/9 2024/09/142024/12/82024/10/52024/09/252024/09/72024/08/312024/09/252024/10/52024/08/192024/09/72024/09/25 1403/7/4 Neda Mousavi-Niri Neda Mousavi-Niri Department of Biotechnology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran Iran Neda.mousaviniri@gmail.com Maryam Naseroleslami Maryam Naseroleslami Department of Cellular and Molecular Biology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran Iran m.naseroleslami@iautmu.ac.ir Elham Ghanimati Elham Ghanimati Department of Cellular and Molecular Biology, Faculty of Advanced Science and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran Iran lham74gh@gmail.com Marzieh Moheb-Alian Marzieh Moheb-Alian Department of Cellular and Molecular Biology, Faculty of Advanced Science and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran. 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