Monoclonal Antibody a Promising Treatment for Depression 2 2 Major depressive disorder (MDD) is one of the prominent causes of disability in the world, affecting 15-20% of people over the period of a lifetime. Patients commonly experience continuous remaining symptoms, functional impairment, and diminished health 1. Treatment of MDD is still far from optimal and drug resistance in MDD is still considered a serious clinical challenge. Only about one-third of patients completely respond to their first antidepressant medication, first with an approximate lag time of 2 months. This notable lag time for onset of therapeutic efficacy is associated with significant morbidity and suicidal risk. As a result, there is a widely accepted need for fast acting antidepressants. Another downside of the currently available treatments is their side effects which are documented in large proportion of patients 1,2. Patients who do not respond to their first-line medication are generally treated by either switching to another treatment or with augmentation therapy to achieve favorable response. Combination therapy from start of treatment has been suggested recently to gain quicker and better response and remission rates, however, not all studies have supported this opinion. Most MDD pathophysiology etiological theories used to focus on brain modulatory monoamine systems (dopamine, serotonin and norepinephrine) 3,4. A more recent line of evidence points to glutamate, the brain’s principal excitatory neurotransmitter, as playing a role in MDD’s pathophysiology. Additionally, glutamate dysregulation is known to cause impairments in structural plasticity and cellular resilience, which seems to be implicated in mood disorders as well. It is therefore reasonable to hypothesize that medications which reduce glutamatergic tone may be able to play a role in treatment of depression 5. Depression is demonstrated to be accompanied with parallel increases in the immuno-inflammatory biomarkers including significantly higher levels of pro-inflammatory cytokines interleukin (IL)-1, IL-6, tumor necrosis factor (TNF)-α, and C-reactive protein (CRP) in depressed patients compared to normal individuals. The dynamic interaction between pro-inflammatory cytokines, prostaglandin (PG)-E2 synthesis and depression has led to suggestions that anti-inflammatory agents could be useful for treatment of depression 6,7. Celecoxib, a nonsteroidal anti-inflammatory drug that acts via the selective inhibition of cyclooxygenase (COX)-2, has shown promising outcomes in several psychiatric diorders, including autistic disorder, schizophrenia, and depression. Antidepressive effects of celecoxib is suggested to be largely attributed to its inhibitory effect on the levels of pro-inflammatory cytokines. In line with this idea, baseline levels of serum IL-6 is demonstrated to be significantly correlated with the Hamilton Depression Rating Scale (HDRS) score. In the same article, adjunctive therapy with celecoxib, as an antidepressant, resulted in significant decreases in both serum IL-6 levels and HDRS scores. Although most of the previous research has focused on the antidepressant effects of celecoxib as an add-on treatment, few reports have investigated the safety and efficacy of celecoxib as a monotherapy 8-10. In clinical trials, two new classes of anti-inflammatory drugs-anti-cytokine monoclonal antibodies and cytokine inhibitors have been shown to reduce inflammation in a range of autoimmune diseases, and these drugs have already started to be administered to patients who do not respond to standard treatments 11,12. 1 2 Shahin Akhondzadeh Psychiatric Research Center, Roozbeh Hospital, Tehran University of Medical Sciences Psychiatric Research Center, Roozbeh Hospital, Tehran University of Medical Sciences Iran Editorial 10336.pdf Ma L, Xu Y, Wang G, Li R. What do we know about sex differences in depression: A review of animal models and potential mechanisms. Prog Neuropsychopharmacol Biol Psychiatry 2019;89:48-56.##Johnson D, Dupuis G, Piche J, Clayborne Z, Colman I. Adult mental health outcomes of adolescent depression: A systematic review. Depress Anxiety 2018;35(8):700-716.##Sepanjnia K, Modabbernia A, Ashrafi M, Modabbernia MJ, Akhondzadeh S. Pioglitazone adjunctive therapy for moderate-to-severe major depressive disorder: randomized double-blind placebo-controlled trial. Neuropsychopharmacology 2012;37(9):2093-2100.##Modabbernia A, Sohrabi H, Nasehi AA, Raisi F, Saroukhani S, Jamshidi A, et al. Effect of saffron on fluoxetine-induced sexual impairment in men: randomized double-blind placebo-controlled trial. Psychopharmacology (Berl) 2012;223(4):381-388.##Khajavi D, Farokhnia M, Modabbernia A, Ashrafi M, Abbasi SH, Tabrizi M, et al. Oral scopolamine augmentation in moderate to severe major depressive disorder: a randomized, double-blind, placebo-controlled study. J Clin Psychiatry 2012;73(11):1428-1433.##Abbasi SH, Hosseini F, Modabbernia A, Ashrafi M, Akhondzadeh S. Effect of celecoxib add-on treatment on symptoms and serum IL-6 concentrations in patients with major depressive disorder: randomized double-blind placebo-controlled study. J Affect Disord 2012;141(2-3):308-314.##Jafari S, Ashrafizadeh SG, Zeinoddini A, Rasoulinejad M, Entezari P, Seddighi S, et al. Celecoxib for the treatment of mild-to-moderate depression due to acute brucellosis: a double-blind, placebo-controlled, randomized trial. J Clin Pharm Ther 2015;40(4):441-446.##Mohammadinejad P, Arya P, Esfandbod M, Kaviani A, Najafi M, Kashani L, et al. Celecoxib versus diclofenac in mild to moderate depression management among breast cancer patients: A double-blind, placebo-controlled, randomized trial. Ann Pharmacother 2015;49(9):953-961.##Arabzadeh S, Ameli N, Zeinoddini A, Rezaei F, Farokhnia M, Mohammadinejad P, et al. Celecoxib adjunctive therapy for acute bipolar mania: a randomized, double-blind, placebo-controlled trial. Bipolar Disord 2015;17(6):606-614.##Alamdarsaravi M, Ghajar A, Noorbala AA, Arbabi M, Emami A, Shahei F, et al. Efficacy and safety of celecoxib monotherapy for mild to moderate depression in patients with colorectal cancer: A randomized double-blind, placebo controlled trial. Psychiatry Res 2017;255:59-65.##Sun Y, Wang D, Salvadore G, Hsu B, Curran M, Casper C, et al. The effects of interleukin-6 neutralizing antibodies on symptoms of depressed mood and anhedonia in patients with rheumatoid arthritis and multicentric Castleman's disease. Brain Behav Immun 2017;66:156-164.##Zhou AJ, Lee Y, Salvadore G, Hsu B, Fonseka TM, Kennedy SH, et al. Sirukumab: A potential treatment for mood disorders? Adv Ther 2017;34(1):78-90.##

Antibody-Drug Conjugates: Possibilities and Challenges 2 2 The design of Antibody Drug Conjugates (ADCs) as efficient targeting agents for tumor cell is still in its infancy for clinical applications. This approach incorporates the antibody specificity and cell killing activity of chemically conjugated cytotoxic agents. Antibody in ADC structure acts as a targeting agent and a nanoscale carrier to deliver a therapeutic dose of cytotoxic cargo into desired tumor cells. Early ADCs encountered major obstacles including, low blood residency time, low penetration capacity to tumor microenvironment, low payload potency, immunogenicity, unusual off-target toxicity, drug resistance, and the lack of stable linkage in blood circulation. Although extensive studies have been conducted to overcome these issues, the ADCs based therapies are still far from having high-efficient clinical outcomes. This review outlines the key characteristics of ADCs including tumor marker, antibody, cytotoxic payload, and linkage strategy with a focus on technical improvement and some future trends in the pipeline. 3 23 Mohammad Reza Nejadmoghaddam Nanotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical SciencesNanobiotechnology Research Center, Avicenna Research Institute, ACECR Nanotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical SciencesNanobiotechnology Research Center, Avicenna Research Institute, ACECR IranIran Arash Minai-Tehrani Nanobiotechnology Research Center, Avicenna Research Institute, ACECR Nanobiotechnology Research Center, Avicenna Research Institute, ACECR Iran Ramin Ghahremanzadeh Nanobiotechnology Research Center, Avicenna Research Institute, ACECR Nanobiotechnology Research Center, Avicenna Research Institute, ACECR Iran Morteza Mahmoudi Nanotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences Nanotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences Iran Rassoul Dinarvand Nanotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical SciencesDepartment of Pharmaceutics, Faculty of Pharmacy, Tehran University of Medical Sciences Nanotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical SciencesDepartment of Pharmaceutics, Faculty of Pharmacy, Tehran University of Medical Sciences IranIran Amir-Hassan Zarnani Department of Immunology, Faculty of Public Health, Tehran University of Medical SciencesReproductive Immunology Research Center, Avicenna Research Institute, ACECRImmunology Research Center, Iran University of Medical Sciences, IUMS Department of Immunology, Faculty of Public Health, Tehran University of Medical SciencesReproductive Immunology Research Center, Avicenna Research Institute, ACECRImmunology Research Center, Iran University of Medical Sciences, IUMS IranIranIran Antibody-DrugCancer therapyCytotoxic drugsMonoclonal antibodiesNanomedicine 10350.pdf Nejadmoghaddam MR, Zarnani AH, Ghahremanzadeh R, Ghods R, Mahmoudian J, Yousefi M, et al. Placenta-specific1 (PLAC1) is a potential target for antibody-drug conjugate-based prostate cancer immunotherapy. Sci Rep 2017;7(1):13373.##Nejadmoghaddam MR, Babamahmoodi A, Minai-Tehrani A, Zarnani AH, Dinarvand R. The use of objective oriented project planning tools for nanosafety and health concerns: a case study in nanomedicine research project. Eur J Nanomed 2016;8(4):225-231.##Venditto VJ, Szoka FC Jr. Cancer nanomedicines: so many papers and so few drugs! Adv Drug Deliv Rev 2013;65(1):80-88.##Sengupta S, Kulkarni A. Design principles for clinical efficacy of cancer nanomedicine: a look into the basics. ACS Nano 2013;7(4):2878-2882.##Decarvalho S, Rand HJ, Lewis A. Coupling of cyclic chemotherapeutic compounds to immune gamma-globulins. Nature 1964;202(4929):255-258.##Katz J, Janik JE, Younes A. Brentuximab vedotin (SGN-35). Clin Cancer Res 2011;17(20):6428-6436.##Younes A, Kim S, Romaguera J, Copeland A, Farial Sde C, Kwak LW, et al. Phase I multidose-escalation study of the anti-CD19 maytansinoid immunoconjugate SAR3419 administered by intravenous infusion every 3 weeks to patients with relapsed/refractory B-cell lymphoma. J Clin Oncol 2012;30(22):2776-2782.##Erickson HK, Park PU, Widdison WC, Kovtun YV, Garrett LM, Hoffman K, et al. Antibody-maytansinoid conjugates are activated in targeted cancer cells by lysosomal degradation and linker-dependent intracellular processing. Cancer Res 2006;66(8):4426-4433.##Krop IE, Beeram M, Modi S, Jones SF, Holden SN, Yu W, et al. Phase I study of trastuzumab-DM1, an HER2 antibody-drug conjugate, given every 3 weeks to patients with HER2-positive metastatic breast cancer. J Clin Oncol 2010;28(16):2698-2704.##LoRusso PM, Weiss D, Guardino E, Girish S, Sliwkowski MX. Trastuzumab emtansine: a unique antibody-drug conjugate in development for human epidermal growth factor receptor 2–positive cancer. Clin Cancer Res 2011;17(20):6437-6447.##Lewis Phillips GD, Li G, Dugger DL, Crocker LM, Parsons KL, Mai E, et al. Targeting HER2-positive breast cancer with trastuzumab-DM1, an antibody–cytotoxic drug conjugate. Cancer Res 2008;68(22):9280-9290.##García-Alonso S, Ocaña A, Pandiella A. Resistance to antibody-drug conjugates. Cancer Res 2018;78(9):2159-2165.##Jen EY, Ko CW, Lee JE, Del Valle PL, Aydanian A, Jewell C, et al. FDA Approval: Gemtuzumab ozogamicin for the treatment of adults with newly-diagnosed CD33-positive acute myeloid leukemia. Clin Cancer Res 2018. [Epub ahead of print].##Beck A, Reichert JM. Antibody-drug conjugates: present and future. MAbs 2014;6(1):15-17.##Webb S. Pharma interest surges in antibody drug conjugates. Nat Biotechnol 2011;29(4):297-298.##Firth D, Bell L, Squires M, Estdale S, McKee C. A rapid approach for characterization of thiol-conjugated antibody-drug conjugates and calculation of drug–antibody ratio by liquid chromatography mass spectrometry. Anal Biochem 2015;485:34-42.##Ab O, Whiteman KR, Bartle LM, Sun X, Singh R,Tavares D, et al. IMGN853, a folate receptor alpha (FRα)-targeting antibody-drug conjugate, exhibits potent targeted anti-tumor activity against FRα-expressing tumors. Mol Cancer Ther 2015;14(7):1605-1613.##Casi G, Neri D. Antibody-drug conjugates: basic concepts, examples and future perspectives. J Control Release 2012;161(2):422-428.##Leipold D, Mallet WG. Case Study: An antibody-drug conjugate targeting MUC16 for ovarian cancer. In: Phillips GL, editor. Antibody-drug conjugates and immunotoxins. New York: Human Press; 2013. p. 221-239.##Ritchie M, Tchistiakova L, Scott N. Implications of receptor-mediated endocytosis and intracellular trafficking dynamics in the development of antibody drug conjugates. MAbs 2013;5(1):13-21.##Onaga M, Ido A, Hasuike S, Uto H, Moriuchi A, Nagata K, et al. Osteoactivin expressed during cirrhosis development in rats fed a choline-deficient, l-amino acid defined diet, accelerates motility of hepatoma cells. J Hepatol 2003;39(5):779-785.##Williams MD, Esmaeli B, Soheili A, Simantov R, Gombos DS, Bedikian AY, et al. GPNMB expression in uveal melanoma: a potential for targeted therapy. Melanoma Res 2010;20(3):184-190.##Kuan CT, Wakiya K, Dowell JM, Herndon JE 2nd, Reardon DA, Graner MW, et al. Glycoprotein nonmetastatic melanoma protein B, a potential molecular therapeutic target in patients with glioblastoma multiforme. Clin Cancer Res 2006;12(7 Pt 1):1970-1982.##Maric G, Rose AA, Annis MG, Siegel PM. Glycoprotein non-metastatic b (GPNMB): A metastatic mediator and emerging therapeutic target in cancer. Onco Targets Ther 2013;6:839-852.##Qian X, Mills E, Torgov M, LaRochelle WJ, Jeffers M. Pharmacologically enhanced expression of GPNMB increases the sensitivity of melanoma cells to the CR011-vcMMAE antibody-drug conjugate. Mol Oncol 2008;2(1):81-93.##Kovtun YV, Audette CA, Ye Y, Xie H, Ruberti MF, Phinney SJ, et al. Antibody-drug conjugates designed to eradicate tumors with homogeneous and heterogeneous expression of the target antigen. Cancer Res 2006;66(6):3214-3221.##Diamantis N, Banerji U. Antibody-drug conjugates--an emerging class of cancer treatment. Br J Cancer 2016;114(4):362-367.##Golfier S, Kopitz C, Kahnert A, Heisler I, Schatz CA, Stelte-Ludwig B, et al. Anetumab ravtansine: a novel mesothelin-targeting antibody-drug conjugate cures tumors with heterogeneous target expression favored by bystander effect. Mol Cancer Ther 2014;13(6):1537-1548.##Shantha Kumara HM, Grieco MJ, Caballero OL, Su T, Ahmed A, Ritter E, et al. MAGE-A3 is highly expressed in a subset of colorectal cancer patients. Cancer Immun 2012;12:16.##Panowski S, Bhakta S, Raab H, Polakis P, Junutula JR. Site-specific antibody drug conjugates for cancer therapy. MAbs 2014;6(1):34-45.##Sievers EL, Senter PD. Antibody-drug conjugates in cancer therapy. Annu Rev Med 2013;64:15-29.##Gerber HP, Kung-Sutherland M, Stone I, Morris-Tilden C, Miyamoto J, McCormick R, et al. Potent antitumor activity of the anti-CD19 auristatin antibody drug conjugate hBU12-vcMMAE against rituximab-sensitive and-resistant lymphomas. Blood 2009;113(18):4352-4361.##Kahl B, Hamadani M, Caimi PF, Reid EG, Havenith K, He S, et al. First clinical results of ADCT‐402, a novel pyrrolobenzodiazepine-based antibody drug conjugate(ADC), in relapsed/refractory B‐cell linage NHL. Hematol Oncol 2017;35(S2):49-51.##Lazar AC, Wang L, Blättler WA, Amphlett G, Lambert JM, Zhang W. Analysis of the composition of immunoconjugates using size‐exclusion chromatography coupled to mass spectrometry. Rapid Commun Mass Spectrom 2005;19(13):1806-1814.##Naddafi F, Davami F. Anti-CD19 monoclonal antibodies: a new approach to lymphoma therapy. Int J Mol Cell Med 2015;4(3):143-151.##Rao C, Pan C, Vangipuram R, Huber M, Vemuri K, Stevens A, et al. Efficacy and toxicity of an anti-CD19 antibody drug conjugate. American Association for Cancer Research Meeting; 2010; Abstr 2452.##Black J, Menderes G, Bellone S, Schwab CL, Bonazzoli E, Ferrari F, et al. SYD985, a novel duocarmycin-based HER2-targeting antibody-drug conjugate, shows antitumor activity in uterine serous carcinoma with HER2/Neu expression. Mol Cancer Ther 2016;15(8):1900-1909.##Elgersma RC, Coumans RGE, Huijbregts T, Menge WMPB, Joosten JAF, Spijker HJ, et al. Design, synthesis, and evaluation of linker-duocarmycin payloads: toward selection of HER2-Targeting antibody-drug conjugate SYD985. Mol Pharm 2015;12(6):1813-1835.##Menderes G, Bonazzoli E, Bellone S, Black J, Altwerger G, Masserdotti A, et al. SYD985, a novel duocarmycin-based HER2-targeting antibody-drug conjugate, shows promising antitumor activity in epithelial ovarian carcinoma with HER2/Neu expression. Gynecol Oncol 2017;46(1):179-186.##Yurkovetskiy A, Gumerov D, Ter-Ovanesyan E, Conlon P, Devit M, Bu C, et al, editors. Non-clinical pharmacokinetics of XMT-1522, a HER2 targeting auristatin-based antibody drug conjugate. American Association for Cancer Research Annual Meeting; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res July 2017.##Humphreys RC, Kirtely J, Hewit A, Biroc S, Knudsen N, Skidmore L, et al, editors. Site specific conjugation of ARX-788, an antibody drug conjugate (ADC) targeting HER2, generates a potent and stable targeted therapeutic for multiple cancers. 106th Annual Meeting of the American Association for Cancer Research; 2015 April 18-22; Philadelphia, PA. Philadelphia (PA):AACR.##Zammarchi F, Chivers S, Williams DG, Adams L, Mellinas-Gomez M, Tyrer P, et al. ADCT-502, a novel pyrrolobenzodiazepine (PBD)-based antibody–drug conjugate (ADC) targeting low HER2-expressing solid cancers. Eur J Cancer 2016;69:S28.##Gan HK, Papadopoulos KP, Fichtel L, Lassman AB, Merrell R, Van Den Bent MJ, et al. Phase I study of ABT-414 mono- or combination therapy with temozolomide (TMZ) in recurrent glioblastoma (GBM). J Clin Oncol 2015,33(suppl;abstr 2016).##Hamblett KJ, Kozlosky CJ, Siu S, Chang WS, Liu H, Foltz IN, et al. AMG 595, an anti-EGFRvIII antibody-drug conjugate, induces potent antitumor activity against EGFRvIII-Expressing glioblastoma. Mol Cancer Ther 2015;14(7):1614-1624.##Chittenden TD, Setiady YY, Park PU, Ponte JF, Dong L, Skaletskaya A, et al. IMGN289, an EGFR-targeting antibody-maytansinoid conjugate with potent activity against non-small cell lung cancer (NSCLC) regardless of dependency on EGFR pathway. Proceedings: AACR 104th Annual Meeting 2013; Apr 6-10, 2013; Washington, DC.##Calvo E, Cleary JM, Moreno V, et al. Preliminary results from a phase 1 study of the antibody-drug conjugate ABBV-221 in patients with solid tumors likely to express EGFR. J Clin Oncol 2017;35:2510.##Tannir NM, Forero-Torres A, Ramchandren R, Pal SK, Ansell SM, Infante JR, et al. Phase I dose-escalation study of SGN-75 in patients with CD70-positive relapsed/refractory non-Hodgkin lymphoma or metastatic renal cell carcinoma. Invest New Drugs 2014;32(6):1246-1257.##Owonikoko TK, Hussain A, Stadler WM, Smith DC, Sznol M, Molina AM, et al. A phase 1 multicenter openlabel dose-escalation study of BMS-936561 (MDX-1203) in clear cell renal cell carcinoma (ccRCC) and Bcell non Hodgkin lymphoma (B-NHL). American Society of Clinical Oncology meeting abstracts 32;2014:2558.##Sandall SL, McCormick R, Miyamoto J, Biechele T, Law C-L, Lewis TS. SGN-CD70A, a pyrrolobenzodiazepine (PBD) dimer linked ADC, mediates DNA damage pathway activation and G2 cell cycle arrest leading to cell death. Proceedings: AACR 106th Annual Meeting 2015; April 18-22, 2015; Philadelphia, PA.##U.S. National Library of Medicine [Internet]. Bethesda (MD): U.S. National Library of Medicine. 2000 Feb -. AMG 172 First in Human Study in Patients with Kidney Cancer; 2016 March 25 [cited 2018 June 08]; [about 4 screens]. Available from: https://clinicaltrials.gov/ct2/ show/NCT01497821.##Sievers E, Appelbaum FR, Spielberger R, Forman S, Flowers D, Smith F, et al. Selective ablation of acute myeloid leukemia using antibody-targeted chemotherapy: a phase I study of an anti-CD33 calicheamicin immunoconjugate. Blood 1999;93(11):3678-3684.##Kung Sutherland MS, Walter RB, Jeffrey SC, Burke PJ, Yu C, Kostner H, et al. SGN-CD33A: a novel CD33-targeting antibody-drug conjugate using a pyrrolobenzodiazepine dimer is active in models of drug-resistant AML. Blood 2013;122(8):1455-1463.##Lapusan S, Vidriales MB, Thomas X, De Botton S, Vekhoff A, Tang R, et al. Phase I studies of AVE9633, an anti-CD33 antibody-maytansinoid conjugate, in adult patients with relapsed/refractory acute myeloid leukemia. Invest New Drugs 2012;30(3):1121-1131.##Chari RV. Targeted cancer therapy: conferring specificity to cytotoxic drugs. Acc Chem Res 2008;41(1):98-107.##Hughes B. Antibody-drug conjugates for cancer: poised to deliver? Nat Rev Drug Discov 2010;9(9):665-667.##Rudnick SI, Lou J, Shaller CC, Tang Y, Klein-Szanto AJ, Weiner LM, et al. Influence of affinity and antigen internalization on the uptake and penetration of Anti-HER2 antibodies in solid tumors. Cancer Res 2011;71(6):2250-2259.##Bendell J, Blumenschein G, Zinner R, Hong D, Jones S, Infante J, et al. First-in-human phase 1 dose escalation study of a novel anti-mesothelin antibody drug conjugate, BAY 94-9343. patients with advanced solid tumors, American Association of Cancer Research, Washington, DC. 2013.##Zhao XY, Subramanyam B, Sarapa N, Golfier S, Dinter H. Novel antibody therapeutics targeting mesothelin in solid tumors. Clin Cancer Drugs 2016;3(2):76-86.##Weekes CD, Lamberts LE, Borad MJ, Voortman J, McWilliams RR, Diamond JR, et al. A phase I study of DMOT4039A, an antibody-drug conjugate (ADC) targeting mesothelin (MSLN), in patients (pts) with unresectable pancreatic (PC) or platinum-resistant ovarian cancer (OC). American Society of Clinical Oncology; 2014.##Weekes CD, Lamberts LE, Borad MJ, Voortman J, McWilliams RR, Diamond JR, et al. Phase I study of DMOT4039A, an antibody-drug conjugate targeting mesothelin, in patients with unresectable pancreatic or platinum-resistant ovarian cancer. Mol Cancer Ther 2016;15(3):439-447.##Advani A, Coiffier B, Czuczman MS, Dreyling M, Foran J, Gine E, et al. Safety, pharmacokinetics, and preliminary clinical activity of inotuzumab ozogamicin, a novel immunoconjugate for the treatment of B-cell non-Hodgkin's lymphoma: results of a phase I study. J Clin Oncol 2010;28(12):2085-2093.##DiJoseph JF, Armellino DC, Boghaert ER, Khandke K, Dougher MM, Sridharan L, et al. Antibody-targeted chemotherapy with CMC-544: a CD22-targeted immunoconjugate of calicheamicin for the treatment of B-lymphoid malignancies. Blood 2004;103(5):1807-1814.##Dotan E, Cohen SJ, Starodub AN, Lieu CH, Messersmith WA, Simpson PS, et al. Phase I/II trial of labetuzumab govitecan (Anti-CEACAM5/SN-38 antibody-drug conjugate) in patients with refractory or relapsing metastatic colorectal cancer. J Clin Oncol 2017;35(29):3338-3346.##Dotan E, Starodub A, Berlin J, Lieu CH, Guarino MJ, Marshall J, et al. A new anti-CEA-SN-38 antibody-drug conjugate (ADC), IMMU-130, is active in controlling metastatic colorectal cancer (mCRC) in patients (pts) refractory or relapsing after irinotecan-containing chemotherapies: Initial results of a phase I/II study. American Society of Clinical Oncology; 2015.##Govindan SV, Cardillo TM, Rossi EA, Trisal P, McBride WJ, Sharkey RM, et al. Improving the therapeutic index in cancer therapy by using antibody-drug conjugates designed with a moderately cytotoxic drug. Mol Pharm 2014;12(6):1836-1847.##Gazzah A, Stjepanovic N, Ryu M, Tabernero J, Soria J, Bedard P, et al. First-in-human phase I trial of the anti-CEACAM5 antibody-drug conjugate SAR408701 in patients with advanced solid tumors (NCT02187848). Europ J Cancer 2016;69:S14-S5.##Bardia A, Mayer IA, Diamond JR, Moroose RL, Isakoff SJ, Starodub AN, et al. Efficacy and safety of anti-Trop-2 antibody drug conjugate sacituzumab govitecan (IMMU-132) in heavily pretreated patients with metastatic triple-negative breast cancer. J Clin Oncol 2017;35(19):2141-2148.##Goldenberg DM, Cardillo TM, Govindan SV, Rossi EA, Sharkey RM. Trop-2 is a novel target for solid cancer therapy with sacituzumab govitecan (IMMU-132), an antibody-drug conjugate (ADC). Oncotarget 2015;6(26):22496-22512.##Heist RS, Guarino MJ, Masters G, Purcell WT, Starodub AN, Horn L, et al. Therapy of advanced nonsmall-cell lung cancer with an SN-38-anti-trop-2 drug conjugate, sacituzumab govitecan. J Clin Oncol 2017;35(24):2790-2797.##Ocean AJ, Starodub AN, Bardia A, Vahdat LT, Isakoff SJ, Guarino M, et al. Sacituzumab govitecan (IMMU‐132), an anti‐Trop‐2‐SN‐38 antibody‐drug conjugate for the treatment of diverse epithelial cancers: Safety and pharmacokinetics. Cancer 2017;123(19):3843-3854.##Starodub A, Camidge DR, Scheff RJ, Thomas SS, Guarino MJ, Masters GA, et al. Trop-2 as a therapeutic target for the antibody-drug conjugate (ADC), sacituzumab govitecan (IMMU-132), in patients (pts) with previously treated metastatic small-cell lung cancer (mSCLC). American Society of Clinical Oncology meeting Abstract. 2016;34(15 suppl):8559.##Starodub AN, Ocean AJ, Shah MA, Guarino MJ, Picozzi VJ, Vahdat LT, et al. First-in-human trial of a novel anti-Trop-2 antibody-SN-38 conjugate, sacituzumab govitecan, for the treatment of diverse metastatic solid tumors. Clin Cancer Res 2015;21(17):3870-3878.##Strop P, Tran TT, Dorywalska M, Delaria K, Dushin R, Wong OK, et al. RN927C, a site-specific trop-2 antibody–drug conjugate (ADC) with enhanced stability, is highly efficacious in preclinical solid tumor models. Mol Cancer Ther 2016;15(11):2698-2708.##Petrylak DP, Kantoff PW, Mega AE, Vogelzang NJ, Stephenson J, Fleming MT, et al. Prostate-specific membrane antigen antibody drug conjugate (PSMA ADC): A phase I trial in metastatic castration-resistant prostate cancer (mCRPC) previously treated with a taxane. American Society of Clinical Oncology meeting Abstract. 2013;31(Suppl; Abstr 5018).##Wang X, Ma D, Olson WC, Heston WD. In vitro and in vivo responses of advanced prostate tumors to PSMA ADC, an auristatin-conjugated antibody to prostate specific membrane antigen. Mol Cancer Ther 2011;10(9):1728-1739.##Henry MD, Wen S, Silva MD, Chandra S, Milton M, Worland PJ. A prostate-specific membrane antigen-targeted monoclonal antibody-chemotherapeutic conjugate designed for the treatment of prostate cancer. Cancer Res 2004;64(21):7995-8001.##Lammerts van Bueren JJ, Bleeker WK, Bøgh HO, Houtkamp M, Schuurman J, van de Winkel JG, et al. Effect of target dynamics on pharmacokinetics of a novel therapeutic antibody against the epidermal growth factor receptor: implications for the mechanisms of action. Cancer Res 2006;66(15):7630-7638.##Hicks SW, Lai KC, Gavrilescu LC, Yi Y, Sikka S, Shah P, et al. The antitumor activity of IMGN529, a CD37-targeting antibody-drug conjugate, is potentiated by rituximab in non-Hodgkin lymphoma models. Neoplasia 2017;19(9):661-671.##Stathis A, Freedman AS, Flinn IW, Maddocks KJ, Weitman S, Berdeja JG, et al. A phase I study of IMGN529, an antibody-drug conjugate (ADC) targeting CD37, in adult patients with relapsed or refractory b-cell non-hodgkin’s lymphoma (NHL). Blood 2014;124(21):1760.##Doñate F, Yang P, Morrison K, Karki S, Aviña H, Lackey J, et al. Analysis of preclinical and clinical samples after treatment with a CD37 targeting antibody drug conjugate (AGS67E) support a high level of CD37 expression in NHL. Hematol Oncol 2017;35(S2):290-291.##Sawas A, Savage KJ, Perez R, Advani RH, Butturini A, Lackey J, et al. A phase 1 study of the anti-CD37 antibody-drug conjugate AGS67E in advanced lymphoid malignancies. Interim results. Blood 2015;126(23):3976.##Kogawa T, Yonemori K, Naito Y, Noguchi E, Shimizu C, Tamura K, et al. Phase 1/2, multicenter, nonrandomized, open-label, multiple-dose first-in-human study of U3-1402 (anti-HER3 antibody drug conjugate) in subjects with HER3-positive metastatic breast cancer. J Clin Oncol 2017;35:TPS1116.##Saunders LR, Bankovich AJ, Anderson WC, Aujay MA, Bheddah S, Black K, et al. A DLL3-targeted antibody drug conjugate eradicates high-grade pulmonary neuroendocrine tumor-initiating cells in vivo. Sci Transl Med 2015;7(302):302ra136.##Ott P, Pavlick A, Johnson D, Hart L, Infante J, Luke J, et al. A phase 2 study of glembatumumab vedotin (GV), an antibody-drug conjugate (ADC) targeting gpNMB, in advanced melanoma. Ann Oncol 2016;27(suppl 1147:6p).##Rose AA, Biondini M, Curiel R, Siegel PM. Targeting GPNMB with glembatumumab vedotin: current developments and future opportunities for the treatment of cancer. Pharmacol Ther 2017;179:127-141.##Saleh M, Bendell J, Rose A, Siegel P, Hart L, Sirpal S, et al. Correlation of GPNMB expression with outcome in breast cancer (BC) patients treated with the antibody–drug conjugate (ADC), CDX-011 (CR011-vcMMAE). J Clin Oncol 2010;28(15_suppl):1095.##Yardley DA, Melisko ME, Forero A, Daniel BR, Montero AJ, Guthrie TH, et al. METRIC: a randomized international study of the antibody-drug conjugate glembatumumab vedotin (GV or CDX-011) in patients (pts) with metastatic gpNMB-overexpressing triple negative breast cancer (TNBC). J Clin Oncol 2015;33(Suppl; abstr TPS1110).##Palanca-Wessels MC, Czuczman M, Salles G, Assouline S, Sehn LH, Flinn I, et al. Safety and activity of the anti-CD79B antibody-drug conjugate polatuzumab vedotin in relapsed or refractory B-cell non-Hodgkin lymphoma and chronic lymphocytic leukaemia: a phase 1 study. Lancet Oncol 2015;16(6):704-715. ##Almhanna K, Wright D, Mercade TM, Van Laethem JL, Gracian AC, Guillen-Ponce C, et al. A phase II study of antibody-drug conjugate, TAK-264 (MLN0264) in previously treated patients with advanced or metastatic pancreatic adenocarcinoma expressing guanylyl cyclase C. Invest New Drugs 2017;35(5):634-641.##Cruz Zambrano C, Almhanna K, Messersmith WA, Rodon Ahnert J, Ryan DP, Faris JE, et al. MLN0264, an investigational antiguanylyl cyclase C (GCC) antibody-drug conjugate (ADC), in patients (pts) with advanced gastrointestinal (GI) malignancies: Phase I study. J Clin Oncol 2014;32(Suppl; abstr 3456).##Burris HA, Gordon MS, Gerber DE, Spigel DR, Mendelson DS, Schiller JH, et al. A phase I study of DNIB0600A, an antibody-drug conjugate (ADC) targeting NaPi2b, in patients (pts) with non-small cell lung cancer (NSCLC) or platinum-resistant ovarian cancer (OC). J Clin Oncol 2014;32(Suppl; abstr 2504).##Lin K, Rubinfeld B, Zhang C, Firestein R, Harstad E, Roth L, et al. Preclinical development of an anti-NaPi2b (SLC34A2) antibody-drug conjugate as a therapeutic for non-small cell lung and ovarian cancers. Clin Cancer Res 2015;21(22):5139-5150.##Gomez-Roca CA, Boni V, Moreno V, Morris JC, Delord JP, Calvo E, et al. A phase I study of SAR566658, an anti CA6-antibody drug conjugate (ADC), in patients (Pts) with CA6-positive advanced solid tumors (STs)(NCT01156870). J Clin Oncol 2016;34:2511.##Ilovich O, Natarajan A, Hori S, Sathirachinda A, Kimura R, Srinivasan A, et al. Development and validation of an immuno-PET tracer as a companion diagnostic agent for antibody-drug conjugate therapy to target the CA6 epitope. Radiology 2015;276(1):191-198.##Kaufman JL, Niesvizky R, Stadtmauer EA, Chanan‐Khan A, Siegel D, Horne H, et al. Phase I, multicentre, dose‐escalation trial of monotherapy with milatuzumab (humanized anti‐CD74 monoclonal antibody) in relapsed or refractory multiple myeloma. Br J Haematol 2013;163(4):478-486.##Kelly KR, Chanan-Khan A, Heffner LT, Somlo G, Siegel DS, Zimmerman T, et al. Indatuximab ravtansine (BT062) in combination with lenalidomide and lowdose dexamethasone in patients with relapsed and/or refractory multiple myeloma: clinical activity in patients already exposed to lenalidomide and bortezomib. Blood 2014;124(21):4736.##Tai YT, Mayes PA, Acharya C, Zhong MY, Cea M, Cagnetta A, et al. Novel anti-B-cell maturation antigen antibody-drug conjugate (GSK2857916) selectively induces killing of multiple myeloma. Blood 2014;123(20):3128-3138.##Khandelwal A, Saber H, Shapiro MA, Zhao H. Antibody-drug conjugate development. In: Phillips GL, editor. Antibody-drug conjugates and immunotoxins. New York: Human Press; 2013. p. 23-38.##Xu S. Internalization, trafficking, intracellular processing and actions of antibody-drug conjugates. Pharm Res 2015;32(11):3577-3583.##U.S. National Library of Medicine [Internet]. Bethesda (MD): U.S. National Library of Medicine. 2000 Feb -. A Study of DFRF4539A in Patients with Relapsed or Refractory Multiple Myeloma; 2016 November 02 [cited 2018 June 08]; [about 4 screens]. Available from: https://clinicaltrials.gov/ct2/show/NCT01432353.##Gish K, Kim H, Power R, Fox M, Hickson J, McGonigal T, et al., editors. Preclinical evaluation of Abbv-838, a first-in-class anti-Cs1 antibody-drug conjugate for the treatment of multiple myeloma. Haematologica 2016;101:253.##Chanan-Khan A, Wolf JL, Garcia J, Gharibo M, Jagannath S, Manfredi D, et al. Efficacy analysis from phase I study of lorvotuzumab mertansine (IMGN901), used as monotherapy, in patients with heavily pretreated CD56-positive multiple myeloma-a preliminary efficacy analysis. Blood 2010;116(21):1962.##Doñate F, Raitano A, Morrison K, An Z, Capo L, Aviña H, et al. AGS16F is a novel antibody drug conjugate directed against ENPP3 for the treatment of renal cell carcinoma. Clin Cancer Res 2016;22(8):1989-1999.##Thompson JA, Motzer R, Molina AM, Choueiri TK, Heath EI, Kollmannsberger CK, et al. Phase I studies of anti-ENPP3 antibody drug conjugates (ADCs) in advanced refractory renal cell carcinomas (RRCC). J Clin Oncol 2015;33(Suppl 2503).##Lassen U, Hong D, Diamantis N, Subbiah V, Kumar R, Sorensen M, et al. A phase I, first-in-human study to evaluate the tolerability, pharmacokinetics and preliminary efficacy of HuMax-tissue factor-ADC (TFADC) in patients with solid tumors. J Clin Oncol 2015;33(Suppl; Abstr 2570).##Thompson JA, Motzer R, Molina AM, Choueiri TK, Heath EI, Kollmannsberger CK, et al. Phase I studies of anti-ENPP3 antibody drug conjugates (ADCs) in advanced refractory renal cell carcinomas (RRCC). J Clin Oncol 2015;33(Suppl 2503).##Martin L, Konner J, Moore K, Seward S, Matulonis U, Perez R, et al. Characterization of folate receptor alpha (FRα) expression in archival tumor and biopsy samples in a phase I study of mirvetuximab soravtansine, a FRα-targeting antibody-drug conjugate (ADC), in relapsed epithelial ovarian cancer patients. Gynecol Oncol 2017;145:34.##Moore KN, Matulonis UA, O'Malley DM, Konner JA, Martin LP, Perez RP, et al. Mirvetuximab soravtansine (IMGN853), a folate receptor alpha (FRα)-targeting antibody-drug conjugate (ADC), in platinum-resistant epithelial ovarian cancer (EOC) patients (pts): Activity and safety analyses in phase I pooled expansion cohorts. American Society of Clinical Oncology; 2017. J Clin Oncol 2017;35(no. 15):5547-5547.##Moore KN, Ponte J, LoRusso P, Birrer MJ, Bauer TM, Borghaei H, et al. Relationship of pharmacokinetics (PK), toxicity, and initial evidence of clinical activity with IMGN853, a folate receptor alpha (FRa) targeting antibody drug conjugate in patients (Pts) with epithelial ovarian cancer (EOC) and other FRa-positive solid tumors. J Clin Oncol 2014;32(Suppl; Abstr 5571).##O'Malley DM, Moore KN, Vergote I, Martin LP, Gilbert L, Gonzalez Martin A, et al. Safety findings from FORWARD II: A Phase 1b study evaluating the folate receptor alpha (FRα)-targeting antibody-drug conjugate (ADC) mirvetuximab soravtansine (IMGN853) in combination with bevacizumab, carboplatin, pegylated liposomal doxorubicin (PLD), or pembrolizumab in patients (pts) with ovarian cancer. J Clin Oncol 2017;35:5553.##Liu J, Moore K, Birrer M, Berlin S, Matulonis U, Infante J, et al. Targeting MUC16 with the antibody-drug conjugate DMUC5754A in patients with platinum resistant ovarian cancer: A phase I study of safety and pharmacokinetics. AACR Annual Meeting; April 6-10; Washington, DC: 2013.##Goff LW, Papadopoulos K, Posey J, Phan A, Patnaik A, Miller J, et al. A phase II study of IMGN242 (huC242-DM4) in patients with CanAg-positive gastric or gastroesophageal (GE) junction cancer. J Clin Oncol 2009;27(15S):e15625-e.##Hong DS, Garrido-Laguna I, Krop IE, Subbiah V, Werner TL, Cotter CM, et al. First-in-human dose escalation, safety, and PK study of a novel EFNA4-ADC in patients with advanced solid tumors. J Clin Oncol 2015;33(15_suppl):2520.##Annunziata CM, Kohn EC, LoRusso P, Houston ND, Coleman RL, Buzoianu M, et al. Phase 1, open-label study of MEDI-547 in patients with relapsed or refractory solid tumors. Invest New Drugs 2013;31(1):77-84.##Jacobson O, Niu G, Kiesewetter DO, Li Q, Yang G, Cook K, et al. PET-guided evaluation and optimization of internalized antibody-drug conjugates targeting erythropoietin producing hepatoma A2 receptor. J Nucl Med 2017;58(11):1838-1844.##Challita-Eid PM, Satpayev D, Yang P, An Z, Morrison K, Shostak Y, et al. Enfortumab vedotin antibody-drug conjugate targeting nectin-4 is a highly potent therapeutic agent in multiple preclinical cancer models. Cancer Res 2016;76(10):3003-3013.##Rosenberg JE, Heath EI, Van Veldhuizen PJ, Merchan JR, Lang JM, Ruether JD, et al. Anti-tumor activity, safety and pharmacokinetics (PK) of ASG-22CE (ASG-22ME; enfortumab vedotin) in a phase I dose escalation trial in patients (Pts) with metastatic urothelial cancer (mUC). J Clin Oncol 2016;34(15_suppl):4533-4533.##Le PU, Nabi IR. Distinct caveolae-mediated endocytic pathways target the Golgi apparatus and the endoplasmic reticulum. J Cell Sc 2003;116(Pt 6):1059-1071.##Petrylak D, Heath E, Sonpavde G, George S, Morgans A, Eigl B, et al. Interim analysis of a phase I dose escalation trial of the antibody drug conjugate (ADC) AGS15E (ASG-15ME) in patients (Pts) with metastatic urothelial cancer (mUC). Ann Oncol 2016;27(suppl 6):780PD.##Angevin E, Strickler JH, Weekes CD, et al. Phase I study of ABBV-399, a c-Met antibody-drug conjugate (ADC), as monotherapy and in combination with erlotinib in patients (pts) with non-small cell lung cancer (NSCLC). J Clin Oncol 2017;35:2509.##Doronina SO, Toki BE, Torgov MY, Mendelsohn BA, Cerveny CG, Chace DF, et al. Development of potent monoclonal antibody auristatin conjugates for cancer therapy. Nat Biotechnol 2003;21(7):778-784.##Wang J, Anderson MG, Oleksijew A, Vaidya KS, Boghaert ER, Tucker L, et al. ABBV-399, a c-Met antibody-drug conjugate that targets both MET-Amplified and c-Met-Overexpressing tumors, irrespective of MET pathway dependence. Clin Cancer Res 2017;23(4):992-1000.##Wang J, Goetsch L, Tucker L, Zhang Q, Gonzalez A, Vaidya KS, et al. Anti-c-Met monoclonal antibody ABT-700 breaks oncogene addiction in tumors with MET amplification. BMC Cancer 2016;16:105.##Sommer A, Kopitz C, Schatz CA, Nising CF, Mahlert C, Lerchen HG, et al. Preclinical efficacy of the auristatin-based antibody-drug conjugate BAY 1187982 for the treatment of FGFR2-positive solid tumors. Cancer Res 2016;76(21):6331-6339.##Willuda J, Linden L, Lerchen H-G, Kopitz C, Stelte-Ludwig B, Pena C, et al. Preclinical antitumor efficacy of BAY 1129980-a novel auristatin-based anti-C4. 4A (LYPD3) antibody-drug conjugate for the treatment of non-small cell lung cancer. Mol Cancer Ther 2017;16(5):893-904.##Menezes D, Abrams TJ, Karim C, Tang Y, Ying C, Miller K, et al. Development and activity of a novel antibody-drug conjugate for the treatment of P-cadherin expressing cancers. Cancer Res 2015;75(15):1682.##Shapiro GI, Vaishampayan UN, LoRusso P, Barton J, Hua S, Reich SD, et al. First-in-human trial of an anti-5T4 antibody-monomethylauristatin conjugate, PF-06263507, in patients with advanced solid tumors. Invest New Drugs 2017;35(3):315-323.##Danila DC, Fleisher M, Carrasquillo JA, Gilbert H, Morris MJ, Bellomo LP, et al. STEAP1 as a predictive biomarker for antibody-drug conjugate (ADC) activity in metastatic castration resistant prostate cancer (mCRPC). J Clin Oncol 2014;32(Suppl; Abstr 5024).##Danila DC, Scher HI, Szafer-Glusman E, Herkal A, Suttmann R, Fleisher M, et al. Predictive biomarkers of tumor sensitivity to STEAP1 antibody-drug conjugate (ADC) in patients (pts) with metastatic castration resistant prostate cancer (mCRPC). AACR 106th Annual Meeting 2015; April 18-22, 2015; Philadelphia, PA.##Danila DC, Szmulewitz RZ, Baron AD, Higano CS, Scher HI, Morris MJ, et al. A phase I study of DSTP3086S, an antibody-drug conjugate (ADC) targeting STEAP-1, in patients (pts) with metastatic castration-resistant prostate cancer (CRPC). J Clin Oncol 2014; 32(Suppl; abstr 5024).##Williams SP, Ogasawara A, Tinianow JN, Flores JE, Kan D, Lau J, et al. ImmunoPET helps predicting the efficacy of antibody-drug conjugates targeting TENB2 and STEAP1. Oncotarget 2016;7(18):25103-25112.##Damelin M, Bankovich A, Bernstein J, Lucas J, Chen L, Williams S, et al. A PTK7-targeted antibody-drug conjugate reduces tumor-initiating cells and induces sustained tumor regressions. Sci Transl Med 2017;9(372). pii: eaag2611.##Sachdev J, Maitland M, Sharma M, Moreno V, Boni V, Kummar S, et al. A phase 1 study of PF-06647020, an antibody-drug conjugate (ADC) targeting protein tyrosine kinase 7 (PTK7), in patients with advanced solid tumors including platinum resistant ovarian cancer (OVCA). Ann Oncol 2016;27(suppl 6):LBA35.##Damelin M, Bankovich A, Park A, Aguilar J, Anderson W, Santaguida M, et al. Anti-EFNA4 calicheamicin conjugates effectively target triple-negative breast and ovarian tumor-initiating cells to result in sustained tumor regressions. Clin Cancer Res 2015;21(18):4165-4173.##Sussman D, Smith LM, Anderson ME, Duniho S, Hunter JH, Kostner H, et al. SGN–LIV1A: a novel antibody-drug conjugate targeting LIV-1 for the treatment of metastatic breast cancer. Mol Cancer Ther 2014;13(12):2991-3000.##Alley SC, Zhang X, Okeley NM, Anderson M, Law CL, Senter PD, et al. The pharmacologic basis for antibody-auristatin conjugate activity. J Pharmacol Exp Ther 2009;330(3):932-938.##Deckert J, Park PU, Chicklas S, Yi Y, Li M, Lai KC, et al. A novel anti-CD37 antibody-drug conjugate with multiple anti-tumor mechanisms for the treatment of B-cell malignancies. Blood 2013;122(20):3500-3510.##English DP, Bellone S, Schwab CL, Bortolomai I, Bonazzoli E, Cocco E, et al. T-DM1, a novel antibody-drug conjugate, is highly effective against primary HER2 overexpressing uterine serous carcinoma in vitro and in vivo. Cancer Med 2014;3(5):1256-1265.##Boswell CA, Mundo EE, Firestein R, Zhang C, Mao W, Gill H, et al. An integrated approach to identify normal tissue expression of targets for antibody‐drug conjugates: case study of TENB2. Br J Pharmacol 2013;168(2):445-457.##The ASCO Post [Internet]. Huntington: HSP News Service, L.L.C., Phase I Trial of New Antibody-Drug Conjugate Shows Promise Against All Forms of Melanoma; 2014 August 04 [cited 2018 June 08]; [about 2 screens]. Available from: http://www.ascopost.com/News/15102.##Bendell J, Moore K, Qin A, Johnson D, Schindler J, Papadopoulos K, et al. 472 A phase I study of IMGN388, an antibody drug conjugate targeting av integrin, in patients with solid tumors. Eur J Cancer Suppl 2010;8(7):152.##Kelly RK, Olson DL, Sun Y, Wen D, Wortham KA, Antognetti G, et al. An antibody-cytotoxic conjugate, BIIB015, is a new targeted therapy for Cripto positive tumours. Eur J Cancer 2011;47(11):1736-1746.##Coveler AL, Von Hoff DD, Ko AH, Whiting NC, Zhao B, Wolpin BM. A phase I study of ASG-5ME, a novel antibody-drug conjugate, in pancreatic ductal adenocarcinoma. J Clin Oncol 2013;31(Suppl 4; abstr 176).##Modi S, Eder JP, Lorusso P, Weekes C, Chandarlapaty S, Tolaney SM, et al. A phase I study evaluating DLYE5953A, an antibody-drug conjugate targeting the tumor-associated antigen lymphocyte antigen 6 complex locus E (Ly6E), in patients (Pts) with solid tumors. Ann Oncol 2016;27(suppl_6):3570.##Lassen UN, Ramalingam SS, Lopez JS, Harvey RD, Ameratunga M, de Hoon J, et al. GCT1021-01, a firstin-human, open-label, dose-escalation trial with expansion cohorts to evaluate safety of Axl-specific antibody-drug conjugate (HuMax-Axl-ADC) in patients with solid tumors (NCT02988817). J Clin Oncol 2017;35:TPS2605.##Canzonieri V, Gattei V, Spina M, Polizzi-Anselmo A, Attanasio N, Kaplan A, et al. CD205, a target antigen for a novel antibody drug conjugate (ADC): Evaluation of antigen expression on non-Hodgkin lymphoma (NHL). J Clin Oncol 2017;35(15_suppl): published online before print.##Horwitz SM, Fanale MA, Spira AI, Havenith K, He S, Feingold JM, et al. Interim data from the first clinical study of ADCT‐301, a novel pyrrplomenzodiaza-pine‐based antibody drug conjugate, in relapsed/refractory hodgkin/non-hodgkin lymphoma. Hematol Oncol 2017;35(S2):270-271.##Baudat Y, Cameron B, Dabdoubi T, Lefebvre AM, Merino-Trigo A, Thomas C, et al, editors. Characterization of a novel maytansinoid-antibody-drug conjugate targeting LAMP1 expressed at the surface of tumor cells. 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia: American Association for Cancer Research; 2016.##Petrul HM, Schatz CA, Kopitz CC, Adnane L, McCabe TJ, Trail P, et al. Therapeutic mechanism and efficacy of the antibody-drug conjugate BAY 79-4620 targeting human carbonic anhydrase 9. Mol Cancer Ther 2012;11(2):340-349.##Herbst R, Wang Y, Gallagher S, Mittereder N, Kuta E, Damschroder M, et al. B-cell depletion in vitro and in vivo with an afucosylated anti-CD19 antibody. J Pharmacol Exp Ther 2010;335(1):213-222.##Cardarelli PM, Rao-Naik C, Chen S, Huang H, Pham A, Moldovan-Loomis M-C, et al. A nonfucosylated human antibody to CD19 with potent B-cell depletive activity for therapy of B-cell malignancies. Cancer Immunol Immunother 2010;59(2):257-265.##Horton HM, Bernett MJ, Pong E, Peipp M, Karki S, Chu SY, et al. Potent in vitro and in vivo activity of an Fc-engineered anti-CD19 monoclonal antibody against lymphoma and leukemia. Cancer Res 2008;68(19):8049-8057.##Awan FT, Lapalombella R, Trotta R, Butchar JP, Yu B, Benson DM, et al. CD19 targeting of chronic lymphocytic leukemia with a novel Fc-domain–engineered monoclonal antibody. Blood 2010;115(6):1204-1213.##Reichert JM. Bispecific antibodies and ADCs: once and future kings? MAbs 2011;3(4):329-330.##Golay J, D’Amico A, Borleri G, Bonzi M, Valgardsdottir R, Alzani R, et al. A novel method using blinatumomab for efficient, clinical-grade expansion of polyclonal T cells for adoptive immunotherapy. J Immunol 2014;193(9):4739-4747.##Ng G, Spreter T, Davies R, Wickman G. ZW38, a novel azymetric bispecific CD19-directed CD3 T cell engager antibody drug conjugate with controlled T cell activation and improved B cell cytotoxicity. Blood 2016;128(22):1841.##Sellmann C, Doerner A, Knuehl C, Rasche N, Sood V, Krah S, et al. Balancing selectivity and efficacy of bispecific epidermal growth factor receptor (EGFR)× c-MET antibodies and antibody-drug conjugates. J Biol Chem 2016;291(48):25106-25119.##Sierra JR, Tsao MS. c-MET as a potential therapeutic target and biomarker in cancer. Ther Adv Med Oncol 2011;3(1 suppl):S21-S35.##Andreev J, Thambi N, Bay AEP, Delfino F, Martin J, Kelly MP, et al. Bispecific antibodies and antibody-drug conjugates (ADCs) bridging HER2 and prolactin receptor improve efficacy of HER2 ADCs. Mol Cancer Ther 2017;16(4):681-693.##Azvolinsky A. Conjugating antibodies to cytotoxic agents: getting the best of both worlds? J Natl Cancer Inst 2013;105(23):1765-1766.##Chari RJ, Goldmacher VS, Lambert JM, Blattler WA, inventors; Google Patents, assignee. Cytotoxic agents comprising maytansinoids and their therapeutic use. United States patent US 5,416,064 A. 1995 May 16.##Sullivan R, Paré GC, Frederiksen LJ, Semenza GL, Graham CH. Hypoxia-induced resistance to anticancer drugs is associated with decreased senescence and requires hypoxia-inducible factor-1 activity. Mol Cancer Ther 2008;7(7):1961-1973.##Widdison WC, Chari RVJ. Factors involved in the design of cytotoxic payloads for antibody-drug conjugates. In: Phillips G, editor. Antibody-Drug Conjugates and Immunotoxins. New York: Human Press; 2013. p. 93-115.##Vankemmelbeke M, Durrant L. Third-generation antibody drug conjugates for cancer therapy--a balancing act. Ther Deliv 2016;7(3):141-144.##Govindan SV, Goldenberg DM. Designing immuno-conjugates for cancer therapy. Expert Opin Biol Ther 2012;12(7):873-890.##Maurya DK, Ayuzawa R, Doi C, Troyer D, Tamura M. Topoisomerase I inhibitor SN-38 effectively attenuates growth of human non-small cell lung cancer cell lines invitro and in vivo. J Environ Pathol Toxicol Oncol 2011;30(1):1-10.##Cardillo TM, Govindan SV, Sharkey RM, Trisal P, Arrojo R, Chang CH, et al. Sacituzumab govitecan (IMMU-132), an anti-Trop-2/SN-38 antibody-drug conjugate: Characterization and efficacy in pancreatic, gastric, and other cancers. Bioconjug Chem 2015;26(5):919-931.##Cardillo TM, Govindan SV, Sharkey RM, Trisal P, Goldenberg DM. Humanized anti-Trop-2 IgG-SN-38 conjugate for effective treatment of diverse epithelial cancers: preclinical studies in human cancer xenograft models and monkeys. Clin Cancer Res 2011;17(10):3157-3169.##Sharkey RM, Govindan SV, Cardillo TM, Goldenberg DM. Epratuzumab-SN-38: a new antibody-drug conjugate for the therapy of hematologic malignancies. Mol Cancer Ther 2012;11(1):224-234.##Acchione M, Kwon H, Jochheim CM, Atkins WM. Impact of linker and conjugation chemistry on antigen binding, Fc receptor binding and thermal stability of model antibody-drug conjugates. MAbs 2012;4(3):362-372.##Sochaj AM, Świderska KW, Otlewski J. Current methods for the synthesis of homogeneous antibody-drug conjugates. Biotechnol Adv 2015;33(6 Pt 1):775-784.##Wang L, Amphlett G, Blättler WA, Lambert JM, Zhang W. Structural characterization of the maytansinoid monoclonal antibody immunoconjugate, huN901-DM1, by mass spectrometry. Protein Sci 2005;14(9):2436-2446.##Junutula JR, Raab H, Clark S, Bhakta S, Leipold DD, Weir S, et al. Site-specific conjugation of a cytotoxic drug to an antibody improves the therapeutic index. Nat Biotechnol 2008b;26(8):925-932.##Schroeder DD, Tankersky DL, Lundblad JL. A new preparation of modified immune serum globulin (human) suitable for intravenous administration. Vox Sang 1981;40(6):373-382.##Willner D, Trail PA, Hofstead SJ, King HD, Lasch SJ, Braslawsky GR, et al. (6-Maleimidocaproyl)hydrazone of doxorubicin--a new derivative for the preparation of immunoconjugates of doxorubicin. Bioconjug Chem 1993;4(6):521-527.##Junutula JR, Bhakta S, Raab H, Ervin KE, Eigenbrot C, Vandlen R, et al. Rapid identification of reactive cysteine residues for site-specific labeling of antibody-Fabs. J Immunol Methods 2008;332(1-2):41-52.##Hofer T, Skeffington LR, Chapman CM, Rader C. Molecularly defined antibody conjugation through a selenocysteine interface. Biochemistry 2009;48(50):12047-12057.##Axup JY, Bajjuri KM, Ritland M, Hutchins BM, Kim CH, Kazane SA, et al. Synthesis of site-specific antibody-drug conjugates using unnatural amino acids. Proc Natl Acad Sci USA 2012;109(40):16101-16106.##Okeley NM, Toki BE, Zhang X, Jeffrey SC, Burke PJ, Alley SC, et al. Metabolic engineering of monoclonal antibody carbohydrates for antibody-drug conjugation. Bioconjug Chem 2013;24(10):1650-1655.##Zhu Z, Ramakrishnan B, Li J, Wang Y, Feng Y, Prabakaran P, et al. Site-specific antibody-drug conjugation through an engineered glycotransferase and a chemically reactive sugar. MAbs 2014;6(5):1190-200.##Zhou Q, Stefano JE, Manning C, Kyazike J, Chen B, Gianolio DA, et al. Site-specific antibody-drug conjugation through glycoengineering. Bioconjug Chem 2014;25(3):510-520.##Drake PM, Albers AE, Baker J, Banas S, Barfield RM, Bhat AS, et al. Aldehyde tag coupled with HIPS chemistry enables the production of ADCs conjugated site-specifically to different antibody regions with distinct in vivo efficacy and PK outcomes. Bioconjug Chem 2014;25(7):1331-1341.##Dennler P, Chiotellis A, Fischer E, Brégeon D, Belmant C, Gauthier L, et al. Transglutaminase-based chemoenzymatic conjugation approach yields homogeneous antibody-drug conjugates. Bioconjug Chem 2014;25(3):569-578.##Spidel J, Vaessen B, Albone E, Cheng X, Verdi A, Kline JB. Site-Specific conjugation to native and engineered lysines in human immunoglobulins by microbial transglutaminase. Bioconjug Chem 2017;28(9):2471-2484.##Strop P, Liu SH, Dorywalska M, Delaria K, Dushin RG,Tran TT, et al. Location matters: site of conjugation modulates stability and pharmacokinetics of antibody drug conjugates. Chem Biol 2013;20(2):161-167.##Beerli RR, Hell T, Merkel AS, Grawunder U. Sortase enzyme-mediated generation of site-specifically conjugated antibody drug conjugates with high in vitro and in vivo potency. PloS One 2015;10(7):e0131177.##Grünewald J, Klock HE, Cellitti SE, Bursulaya B, McMullan D, Jones DH, et al. Efficient preparation of site-specific antibody-drug conjugates using phosphopantetheinyl transferases. Bioconjug Chem 2015;26(12):2554-2562.##Tang F, Wang LX, Huang W. Chemoenzymatic synthesis of glycoengineered IgG antibodies and glycosite-specific antibody-drug conjugates. Nat Protoc 2017;12(8):1702-1721.##189. McCombs JR, Owen SC. Antibody drug conjugates: design and selection of linker, payload and conjugation chemistry. AAPS J 2015;17(2):339-351.##Ritter A. Antibody-drug conjugates: looking ahead to an emerging class of biotherapeutic. Pharm Technol 2012;36(1):42-47.##Dyba M, Tarasova NI, Michejda CJ. Small molecule toxins targeting tumor receptors. Curr Pharm Des 2004;10(19):2311-2334.##Goldmacher VS, Singh R, Chittenden T, Kovtun Y. Linker technology and impact of linker design on ADC properties. In: Phillips GL, editor. Antibody-drug conjugates and immunotoxins. New York: Human Press; 2013. p. 117-135.##Erickson HK, Lambert JM. ADME of antibody-maytansinoid conjugates. AAPS J 2012;14(4):799-805.##Kovtun YV, Audette CA, Mayo MF, Jones GE, Doherty H, Maloney EK, et al. Antibody-maytansinoid conjugates designed to bypass multidrug resistance. Cancer Res 2010;70(6):2528-2537.##Sun X, Ponte JF, Yoder NC, Laleau R, Coccia J, Lanieri L, et al. Effects of drug-antibody ratio on pharmacokinetics, biodistribution, efficacy, and tolerability of antibodymaytansinoid conjugates. Bioconjug Chem 2017;28(5):1371-1381.##van Geel R, Wijdeven MA, Heesbeen R, Verkade JM, Wasiel AA, van Berkel SS, et al. Chemoenzymatic conjugation of toxic payloads to the globally conserved n-glycan of native mAbs provides homogeneous and highly efficacious antibody-drug conjugates. Bioconjug Chem 2015;26(11):2233-2242.##Zimmerman ES, Heibeck TH, Gill A, Li X, Murray CJ, Madlansacay MR, et al. Production of site-specific antibody-drug conjugates using optimized non-natural amino acids in a cell-free expression system. Bioconjug Chem 2014;25(2):351-361.##

Recovery of MicroRNA from Stored Bone Marrow Aspirate Slides 2 2 Background: Archived bone marrow aspirate slides are almost infinite, readily available resource of biospecimens that enable retrospective molecular investigations of diseases. RNAs obtained from slides has limitations in utility because of their low quality and highly fragmented nature. MicroRNAs are small (<22 nt) noncoding RNAs with various cellular regulatory roles. Due to their small size, microRNAs are less prone to degradation and modification, therefore, can be preserved well in archived tissues. Methods: The current study investigated the efficacy of archived bone marrow aspirate slides for miRNA expression analysis in pediatric leukemia. Total RNA was isolated from air-dried unstained archived slides using High pure miRNA isolation Kit with some modifications and from fresh samples using TRizol. After cDNA synthesis, RT-qPCR was then carried out using specific hsa-miR-326 LNA primers. Finally, statistical analyses were conducted using GraphPad Prism 6 software.  Results: The difference observed in miRNA expression due to disease state was far greater than the differences between archived slides and their matching fresh bone marrow specimens. In fact, the expression of archival slide smears for the miR-326 closely mimicked that of fresh-frozen tissues (0.035±0.04 vs. 0.03±0.04) (Mean±SD, p>0.05). Differential expression of hsa-miR-326 was detected between leukemic and non-leukemic samples from archived slides or fresh frozen bone marrows. Conclusion: The demonstration that archived bone marrow aspirate slides can be utilized for miRNA expression studies offers tremendous potential for future investigations into the role that miRNAs play in the development and long term outcome of hematologic, as well as non-hematologic diseases. 24 27 Elaheh Sadat Ghodousi Department of Biology, Faculty of Science, University of Isfahan Department of Biology, Faculty of Science, University of Isfahan Iran Soheila Rahgozar Department of Biology, Faculty of Science, University of Isfahan Department of Biology, Faculty of Science, University of Isfahan Iran Acute lymphoblastic leukemiaBone marrow specimensMicroRNAsmiR-326 10356.pdf Siebolts U, Varnholt H, Drebber U, Dienes HP, Wickenhauser C, Odenthal M. Tissues from routine pathology archives are suitable for microRNA analyses by quantitative PCR. J Clin Pathol 2009;62(1):84-88.##Zanssen S. Single cell PCR from archival stained bone marrow slides: a method for molecular diagnosis and characterization. J Clin Lab Anal 2004;18(3):176-181.##Schoch R, Jenisch S, Haferlach T, Müller-Ruchholtz W, Gassmann W, Löffler H. Glass slide smears are a suitable source for RT-PCR-based analysis of chromosomal aberrations in leukaemias. Br J Haematol 1996;92(1): 140-142.##Goswami RS, Waldron L, Machado J, Cervigne NK, Xu W, Reis PP, et al. Optimization and analysis of a quantitative real-time PCR-based technique to determine microRNA expression in formalin-fixed paraffin-embedded samples. BMC Biotechnol 2010;10:47.##Seelenfreund E, Robinson SE, Amato CM, Bemis LT, Robinson WA. Recovery of microRNA from stored human peripheral blood samples. Biopreserv Biobank 2011;9(1):29-33.##Borze I, Guled M, Musse S, Raunio A, Elonen E, Saarinen-Pihkala U, et al. MicroRNA microarrays on archive bone marrow core biopsies of leukemias--method validation. Leuk Res 2011;35(2):188-195.##Olive V, Minella AC, He L. Outside the coding genome, mammalian microRNAs confer structural and functional complexity. Sci Signal 2015;8(368):re2.##Hoefig KP, Thorns C, Roehle A, Kaehler C, Wesche KO, Repsilber D, et al. Unlocking pathology archives for microRNA-profiling. Anticancer Res 2008;28(1A):119-123.##Morenos L, Saffery R, Mechinaud F, Ashley D, Elwood N, Craig JM, et al. Evaluation of microRNA expression in patient bone marrow aspirate slides. PloS One 2012;7(8):e42951.##Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 2001;25(4):402-408.##Li J, Smyth P, Flavin R, Cahill S, Denning K, Aherne S, et al. Comparison of miRNA expression patterns using total RNA extracted from matched samples of formalin-fixed paraffin-embedded (FFPE) cells and snap frozen cells. BMC Biotechnol 2007;7:36.##Grünewald K, Lyons J, Hansen-Hagge TE, Janssen JW, Feichtinger H, Bartram CR. Molecular genetic analysis of DNA obtained from fixed, air dried or paraffin embedded sources. Ann Hematol 1991;62(4):108-114.##

Evaluation of Differential Gene Expression during Transdifferentiation of Bone Marrow Stromal Cells to Glial phenotype in the Presence of Cerebrospinal Fluid 2 2 Background: The present study assessed the alteration of gene expression during transdifferentiation of Bone Marrow Stromal Cells (BMSCs) into oligodendrocyte in the presence of Cerebrospinal Fluid (CSF). Methods: BMSCs were collected from female Sprague-Dawley rats and were cultured in DMEM/F12 medium supplemented with Retinoic Acid (RA), basic Fibroblast Growth Factor (bFGF), and Epidermal Growth Factor (EGF). CSF was added daily to the culture media. The oligoprogenitor and oligodendrocyte generation was assessed by immunocytochemistry for Oligo 2, A2B5, CNP and MBP markers.  Results: The mean percentages of immunopositive cells for Olig2 and A2B5 were 52.1±1.74 and 56.34±2.55%, respectively. The number of immunopositive cells for glial markers CNP and MBP were 48.8±3.12 and 40.5±8.92%, respectively. Alteration of gene expression of Oct4, Olig 2, PDGFR-α and PLP were examined by real time PCR during transdifferentiation of BMSC to oligodendrocyte. Immunocytochemical results indicate that oligoprogenitor cells were immunopositive for Oligo2 and A2B5 markers. Also, oligodendrocytes expressed the mature glial markers of CNP and MBP indicating successful differentiation.  Conclusion: In conclusion, CSF promotes the transdifferentiation of BMSC into mature oligodendrocyte via providing an appropriate niche for glial maturation.  28 34 Hatef Ghasemi Hamidabadi Immunogenetic Research Center, Department of Anatomy and Cell Biology, Faculty of Medicine, Mazandaran University of Medical Sciences Immunogenetic Research Center, Department of Anatomy and Cell Biology, Faculty of Medicine, Mazandaran University of Medical Sciences Iran Maryam Nazm Bojnordi Immunogenetic Research Center, Department of Anatomy and Cell Biology, Faculty of Medicine, Mazandaran University of Medical Sciences Immunogenetic Research Center, Department of Anatomy and Cell Biology, Faculty of Medicine, Mazandaran University of Medical Sciences Iran Nourollah Rezaei Immunogenetic Research Center, Department of Anatomy and Cell Biology, Faculty of Medicine, Mazandaran University of Medical Sciences Immunogenetic Research Center, Department of Anatomy and Cell Biology, Faculty of Medicine, Mazandaran University of Medical Sciences Iran Sara Soleimani Department of Anatomy & Cell Biology, Faculty of Medicine, Hamadan University of Medical Sciences Department of Anatomy & Cell Biology, Faculty of Medicine, Hamadan University of Medical Sciences Iran Bone marrow stromal cellsCellsCerebrospinal fluidOligodendrocyteOligoprogenitor 10345.pdf Crawford AH, Stockley JH, Tripathi RB, Richardson WD, Franklin RJ. Oligodendrocyte progenitors: adult stem cells of the central nervous system. Exp Neurol 2014;260:50-55. ##Czepiel ME, Boddeke E, Copray S. Human oligodendro-cytes in remyelination research. Glia 2015;63(4):513-530.##Grade S, Bernardino L, Malva JO. Oligodendrogenesis from neural stem cells: perspective for remyelinating strategies. Int J Dev Neurosci 2013;31(7):692-700.##Miller RH, Bai L. Cellular approaches for stimulating CNS remyelination. Regen Med 2007;2(5):817-829.##Haratizadeh S, Nazm Bojnordi M, Niapour A, Bakhtiari M, Ghasemi Hamidabadi H. [Improvement of neuroglial differentiation from human dental pulp stem cells using CSF]. J Mazandaran Univ Med Sci 2016;26(140):1-14. Persian.##Sher F, Balasubramaniyan V, Boddeke E, Copray S. Oligodendrocyte differentiation and implantation: new insights for remyelinating cell therapy. Curr Opin Neuro-biol 2008;21(5):607-614.##Luessi F, KuhlmannT, Zipp F. Remyelinating strategies in multiple sclerosis. Expert Rev Neurother 2014;14(11): 1315-1334.##Bowles AC, Scruggs BA, Bunnell BA. Mesenchymal stem cell-based therapy in a mouse model of experimental autoimmune encephalomyelitis (EAE). Methods Mol Biol 2014;1213:303-319.##Cohen JA. Mesenchymal stem cell transplantation in multiple sclerosis. J Neurol Sci 2013;5;333(1-2):43-49.##Darlington PJ, Boivin MN, Bar-Or A. Harnessing the therapeutic potential of mesenchymal stem cells in multiple sclerosis. Expert Rev Neurother 2011;11(9):1295-1303.##Gao YJ, Qian W, Wang BH, Lin R, Hou XH. Differentiation potential of bone marrow stromal cells to enteric neurons in vitro. Chin J Dig Dis 2006;7(3):156-163.##Shintani A, Nakao N, Kakishita K, ItakuraT. Protection of dopamine neurons by bone marrow stromal cells. Brain Res 2007;1186:48-55.##Sanchez-Ramos J, Song S, Cardozo-Pelaez F, Hazzi C, Stedeford T, Willing A, et al. Adult bone marrow stromal cells differentiate into neural cells in vitro. Exp Neurol 2000;164(2):247-256.##Kaka GR, Tiraihi T, Delshad A, Arabkheradmand J, Kazemi H. In vitro differentiation of bone marrow stromal cells into oligodendrocyte-like cells using triiodothy-ronine as inducer. Int J Neurosci 2012;122(5):237-247.##Sanchez-Ramos JR. Neural cells derived from adult bone marrow and umbilical cord blood. J Neurosci Res 2002; 69(6):880-893.##Miyan JA, Nabiyouni M, Zendah M. Development of the brain: a vital role for cerebrospinal fluid. Can J Physiol Pharmacol 2003;81(4):317-328.##Johanson CE, Duncan JA 3rd, Klinge PM, Brinker T, Stopa EG, Silverberg GD. Multiplicity of cerebrospinal fluid functions: New challenges in health and disease. Cerebrospinal Fluid Res 2008;5:10.##Woodbury D, Schwarz EJ, Prockopand DJ, Black IB. Adult rat and human bone marrow stromal cells differentiate into neurons. J. Neurosci Res 2000;61(4):364-370.##Jiang Y, Jahagirdar BN, Reinhardt RL, Schwartz RE, Keene CD, Ortiz-Gonzalez XR, et al. Pluripotency of mesenchymal stem cells derived from adult marrow. Nature 2002;418(6893):41-49.##Li Y, Zhang B, Wen W, Liu S, Hao D, Liu M, et al. The comparison of three methods of drawing cerebrospinal fluid in rabbit. J Neurosci Methods 2012;209(2):398-402.##Lehtinen, MK, Bjornsson CS, Dymecki SM, Gilbertson RJ, Holtzman DM, Monuki ES. The choroid plexus and cerebrospinal fluid: Emerging roles in development, disease, and therapy. J Neurosci 2013;33(45):17553-17559.##Alexanian AR. Neural stem cells induce bone-marrow-derived mesenchymal stem cells to generate neural stem-like cells via juxtacrine and paracrine interactions. Exp Cell Res 2005;310(2):383-391.##Wislet-Gendebien S, Hans G, Leprince P, Rigo JM, Mo-onen G, Rogister B. Plasticity of cultured mesenchymal stem cells: switch from nestin-positive to excitable neuron-like phenotype. Stem Cells 2005;23(3):392-402.##Lu J, Moochhala S, Moore XL, Ng KC, Tan MH, Lee LK, et al. Adult bone marrow cells differentiate into neural phenotypes and improve functional recovery in rats following traumatic brain injury. Nerosci Lett 2006;398 (1-2):12-17.##Bojnordi MN, Azizi H, Skutella T, Movahedin M, Pour-abdolhossein F, Shojaei A, et al. Differentiation of spermatogonia stem cells into functional mature neurons characterized with differential gene expression. Mol Neurobiol 2017;54(7):5676-5682.##Bojnordi MN, Movahedin M, Tiraihi T, Javan M. A simple co-culture system for generation of embryonic stemlike cells from testis. Iran Red Crescent Med J 2012;14(12):811-815.##Castells, A, Parvas M, Bueno D. Homeostasis of cerebrospinal fluid has a role in early brain development. Neuroreport 2012;23(16):917-921.##Alonso MI, Martín C, Carnicero E, Bueno D, Gato A. Cerebrospinal fluid control of neurogenesis induced by retinoic acid during early brain development. Dev Dyn 2011;240(7):1650-1659.##Parada C, Gato A, Aparicio M, Bueno D. Proteome analysis of chick embryonic cerebrospinal fluid. Proteomics 2006;6(1):312-320.##Buddensiek J, Dressel A, Kowalski M, Runge U, Schro- eder H, Hermann A, et al. Cerebrospinal fluid promotes survival and astroglial differentiation of adult human neural progenitor cells but inhibits proliferation and neuronal differentiation. BMC Neurosci 2010;8;11:48.##Ye Y, Zeng YM, Wan MR, Lu XF. Induction of human bone marrow mesenchymal stem cells, differentiation into neural-like cells using cerebrospinal fluid. Cell Bio-chem Biophys 2011;59(3):179-184.##Yang HY, Gu R, Wang WJ, Gao ZL. The cerebrospinal fluid promotes the differentiation of neural stem cells after transplantation of bone marrow mesenchymal stem cells. Chinese J Lab Diagn 2009;13(3):300-302.##Shen YX, Wang P, Shi ED. Culturing of bone marrow mesenchymal stem cells in vitro with cerebrospinal fluid. Chinese J Clinical Rehabil Tissue Eng Res 2011;15(36):6802-6806.##

Preconditioning with SDF-1 Improves Therapeutic Outcomes of Bone marrow-derived Mesenchymal Stromal Cells in a Mouse Model of STZ-induced Diabetes 2 2 Background: Nowadays, transplantation of bone marrow-derived Mesenchymal Stromal Cells (BMSCs) is currently an important alternative therapy for patient’s type 1 diabetes mellitus. But a number of critical obstacles lie ahead of this new strategy including reducing stem cell homing to the damaged tissue due to oxidative stress. The purpose of the present study was to investigate whether preconditioning of BMSCs with SDF-1 could enhance their homing to the pancreas and promote regeneration of the pancreatic β cells after being intravenously injected. Methods: Mice BMSCs were isolated and expanded. Cell proliferation was assayed by MTT Assay. Preconditioning was performed with 10 ng/ml SDF-1α for 24 hr. Male NMRI mice were injected with high-dose STZ (150 mg/kg). The preconditioned or unpreconditioned BMSCs at a dose of 1×106 cells were infused via the tail vein. Blood and pancreatic tissue samples were taken from all mice for flow cytometry, biochemical and histological studies. Results: Proliferation and homing of BMSCs to the pancreas were significantly increased in the BMSCs with SDF-1α preconditioning. Differentiation of transplanted BMSCs, were significantly increased in preconditioning group. Although BMSCs without SDF-1 preconditioning exhibited remarkable recovery of pancreatic islets structure but this recovery were significantly increased in the BMSCs with SDF-1α preconditioning. Conclusion: Our results showed the effectiveness of SDF-1αpreconditioning in BMSCs transplantation of STZ induced diabetes mice which might be achieved through improvement of BMSCs homing into the injured pancreas. 35 42 Mohammad Sadegh Gholami Farashah Department of Anatomical Sciences, Faculty of Medicine, Tehran University of Medical Sciences Department of Anatomical Sciences, Faculty of Medicine, Tehran University of Medical Sciences Iran Parichehr Pasbakhsh Department of Anatomical Sciences, Faculty of Medicine, Tehran University of Medical Sciences Department of Anatomical Sciences, Faculty of Medicine, Tehran University of Medical Sciences Iran Ameneh Omidi Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University Iran Saied Nekoonam Department of Anatomical Sciences, Faculty of Medicine, Tehran University of Medical Sciences Department of Anatomical Sciences, Faculty of Medicine, Tehran University of Medical Sciences Iran Roya Aryanpour Department of Anatomical Sciences, Faculty of Medicine, Tehran University of Medical Sciences Department of Anatomical Sciences, Faculty of Medicine, Tehran University of Medical Sciences Iran Iraj Regardi Kashani Department of Anatomical Sciences, Faculty of Medicine, Tehran University of Medical Sciences Department of Anatomical Sciences, Faculty of Medicine, Tehran University of Medical Sciences Iran Bone marrow-derived mesenchymal stromal cellsDiabetes mellitusHomingSDF-1 10337.pdf Cafazzo JA, Casselman M, Hamming N, Katzman DK, Palmert MR. Design of an mHealth app for the self-management of adolescent type 1 diabetes: a pilot study. J Med Internet Res 2012;14(3):e70.##Kawashima H, Ihara T, Ioi H, Oana S, Sato S, Kato N, et al. Enterovirus-related type 1 diabetes mellitus and antibodies to glutamic acid decarboxylase in Japan. J Infect 2004;49(2):147-151.##Klinke DJ 2nd. Extent of beta cell destruction is important but insufficient to predict the onset of type 1 diabetes mellitus. PloS One 2008;3(1):e1374.##Hameed A, Yu T, Yuen L, Lam V, Ryan B, Allen R, et al. Use of the harmonic scalpel in cold phase recovery of the pancreas for transplantation: the west mead technique. Transpl Int 2016;29(5):636-638.##Lu J, Xia Q, Zhou Q. How to make insulin-producing pancreatic beta cells for diabetes treatment. Sci China Life Sci 2017;60(3):239-248.##Mortezaee K, Pasbakhsh P, Ragerdi Kashani I, Sabbaghziarani F, Omidi A, Zendedel A, et al. Melatonin pretreatment enhances the homing of bone marrow-derived mesenchymal stem cells following transplantation in a rat model of liver fibrosis. Iran Biomed J 2016;20(4):207-216.##Gong J, Meng HB, Hua J, Song ZS, He ZG, Zhou B, et al. The SDF-1/CXCR4 axis regulates migration of transplanted bone marrow mesenchymal stem cells towards the pancreas in rats with acute pancreatitis. Mol Med Rep 2014;9(5):1575-1582.##Ratajczak MZ, Suszynska M. Emerging strategies to enhance homing and engraftment of hematopoietic stem cells. Stem Cell Rev 2016;12(1):121-128.##Pourjafar M, Saidijam M, Mansouri K, Ghasemibasir H, Karimi Dermani F, Najafi R. All-trans retinoic acid preconditioning enhances proliferation, angiogenesis and migration of mesenchymal stem cell in vitro and enhances wound repair in vivo. Cell Prolif 2017;50(1).##Marquez-Curtis LA, Janowska-Wieczorek A. Enhancing the migration ability of mesenchymal stromal cells by targeting the SDF-1/CXCR4 axis. Biomed Res Int 2013;2013:561098.##Cheng Z, Ou L, Zhou X, Li F, Jia X, Zhang Y, et al. Targeted migration of mesenchymal stem cells modified with CXCR4 gene to infarcted myocardium improves cardiac performance. Mol Ther 2008;16(3):571-579.##Enzmann V, Lecaudé S, Kruschinski A, Vater A. CXCL12/SDF-1-Dependent retinal migration of endogenous bone marrow-derived stem cells improves visual function after pharmacologically induced retinal degeneration. Stem Cell Rev 2017;13(2):278-286.##Reiter J, Drummond S, Sammour I, Huang J, Florea V, Dornas P, et al. Stromal derived factor-1 mediates the lung regenerative effects of mesenchymal stem cells in a rodent model of bronchopulmonary dysplasia. Respiratory Res 2017;18(1):137.##Liu X, Duan B, Cheng Z, Jia X, Mao L, Fu H, et al. SDF-1/CXCR4 axis modulates bone marrow mesenchymal stem cell apoptosis, migration and cytokine secretion. Protein Cell 2011;2(10):845-854.##Abouzaripour M, Ragerdi Kashani I, Pasbakhsh P, Atlasy N. Intravenous transplantation of very small embryonic like stem cells in treatment of diabetes mellitus. Avicenna J Med Biotechnol 2015;7(1):22-31.##Weir C, Morel-Kopp MC, Gill A, Tinworth K, Ladd L, Hunyor SN, et al. Mesenchymal stem cells: isolation, characterisation and in vivo fluorescent dye tracking. Heart Lung Circ 2008;17(5):395-403.##Sordi V, Pellegrini S, Krampera M, Marchetti P, Pessina A, Ciardelli G, et al. Stem cells to restore insulin production and cure diabetes. Nutr Metab Cardiovasc Dis 2017;27(7):583-600.##Tuan RS, Chen AF, Klatt BA. Cartilage regeneration. J Am Acad Orthop Surg 2013;21(5):303-311.##Pasha Z, Wang Y, Sheikh R, Zhang D, Zhao T, Ashraf M. Preconditioning enhances cell survival and differentiation of stem cells during transplantation in infarcted myocardium. Cardiovasc Res 2008;77(1):134-142.##Zhou JY, Zhang Z, Qian GS. Mesenchymal stem cells to treat diabetic neuropathy: a long and strenuous way from bench to the clinic. Cell Death Discov 2016;2:16055.##Cui X, Chopp M, Zacharek A, Roberts C, Lu M, Savant-Bhonsale S, et al. Chemokine, vascular and therapeutic effects of combination Simvastatin and BMSC treatment of stroke. Neurobiol Dis 2009;36(1):35-41.##Stubbs SL, Hsiao ST, Peshavariya HM, Lim SY, Dusting GJ, Dilley RJ. Hypoxic preconditioning enhances survival of human adipose-derived stem cells and conditions endothelial cells in vitro. Stem Cells Dev 2012;21(11):1887-1896.##De Becker A, Riet IV. Homing and migration of mesenchymal stromal cells: How to improve the efficacy of cell therapy? World J Stem Cells 2016;8(3):73-87.##Li L, Chu L, Fang Y, Yang Y, Qu T, Zhang J, et al. Preconditioning of bone marrow-derived mesenchymal stromal cells by tetramethylpyrazine enhances cell migration and improves functional recovery after focal cerebral ischemia in rats. Stem Cell Res Ther 2017;8(1):112.##Tariq M, Masoud MS, Mehmood A, Khan SN, Riazuddin S. Stromal cell derived factor-1alpha protects stem cell derived insulin-producing cells from glucotoxicity under high glucose conditions in-vitro and ameliorates drug induced diabetes in rats. J Translational Med 2013;11:115.##Si Y, Zhao Y, Hao H, Liu J, Guo Y, Mu Y, et al. Infusion of mesenchymal stem cells ameliorates hyperglycemia in type 2 diabetic rats: identification of a novel role in improving insulin sensitivity. Diabetes 2012;61(6):1616-1625.##Aali E, Mirzamohammadi S, Ghaznavi H, Madjd Z, Larijani B, Rayegan S, et al. A comparative study of mesenchymal stem cell transplantation with its paracrine effect on control of hyperglycemia in type 1 diabetic rats. J Diabetes Metab Disord 2014;13(1):76.##Yaochite JN, Caliari-Oliveira C, de Souza LE, Neto LS, Palma PV, Covas DT, et al. Therapeutic efficacy and biodistribution of allogeneic mesenchymal stem cells delivered by intrasplenic and intrapancreatic routes in streptozotocin-induced diabetic mice. Stem Cell Res Ther 2015;6:31.##Xie QP, Huang H, Xu B, Dong X, Gao SL, Zhang B, et al. Human bone marrow mesenchymal stem cells differentiate into insulin-producing cells upon microenviron-men-tal manipulation in vitro. Differentiation 2009;77(5):483-491.##Sun Y, Chen L, Hou XG, Hou WK, Dong JJ, Sun L, et al. Differentiation of bone marrow-derived mesenchymal stem cells from diabetic patients into insulin-producing cells in vitro. Chin Med J (Engl) 2007;120(9):771-776.##

The Anti-cancer Effects of Capparis spinosa Hydroalcoholic Extract 2 2 Background: Recently, due to the steep increase in cancer lethality statistics, pharmaceutical societies seek approaches for designing drugs with higher efficiency and lower expenses. Plant-based drugs have therefore gained much attention, due to their abundance and ease of accessibility, and their higher effectiveness.  Methods: Wild-grown caper [Capparis spinosa (C. spinosa)] was collected from northern Iran and next 100 g of the powder was added to 300 ml of a solvent (Ethanol 80), the solution was mixed for 72 hr and later filtered via Whatman filter papers. The solvent was taken out under vacuum conditions and extracts were then collected and stored in glass vials. The High Pressure Liquid Chromatography (HPLC) method was used to assay quercetin which consisted of the following specifications: C18 column, UV detector wavelength of 260 nm, mobile phase acetonitrile and water and flow rate of 1 ml/min. In this study, the anti-cancer effects of C. spinosa extract on HeLa, MCF7, Saos and Fibroblast cancer cell lines have been investigated. Results: The amount of quercetin was assessed by HPLC. The anti-tumor activity and the antioxidant level of hydroalcoholic extract of C. spinosa have been evaluated with MTT assay and FRAP technique, respectively. HPLC data showed quercetin form the major component of C. spinosa extract. In addition, FRAP data indicated that C. spinosa extract had high antioxidant activity and MTT assay indicated that C. spinosa extract effectively decreased the cancer cell lines. Conclusion: The quercetin in C. spinosa extract had significant anti-tumor effects and may be regarded as an ideal natural drug for cancer therapy. 43 47 Yasaman Moghadamnia Neuroscience Research Center, Health Research Institute, Babol University of Medical Sciences Neuroscience Research Center, Health Research Institute, Babol University of Medical Sciences Iran Seydeh Narges Mousavi Kani Department of Pharmacology, Faculty of Medicine, Babol University of Medical Sciences Department of Pharmacology, Faculty of Medicine, Babol University of Medical Sciences Iran Maryam Ghasemi-Kasman Neuroscience Research Center, Health Research Institute, Babol University of Medical SciencesCellular and Molecular Biology Research Center, Health Research Institute, Babol University of Medical Sciences Neuroscience Research Center, Health Research Institute, Babol University of Medical SciencesCellular and Molecular Biology Research Center, Health Research Institute, Babol University of Medical Sciences IranIran Mohamad Taghi Kazemi Kani Neuroscience Research Center, Health Research Institute, Babol University of Medical Sciences Neuroscience Research Center, Health Research Institute, Babol University of Medical Sciences Iran Sohrab Kazemi Neuroscience Research Center, Health Research Institute, Babol University of Medical SciencesCellular and Molecular Biology Research Center, Health Research Institute, Babol University of Medical Sciences Neuroscience Research Center, Health Research Institute, Babol University of Medical SciencesCellular and Molecular Biology Research Center, Health Research Institute, Babol University of Medical Sciences IranIran CancerCapparisHigh pressure liquid chromatographyQuercetin 10352.pdf Torre LA, Bray F, Siegel RL, Ferlay J, Lortet‐Tieulent J, Jemal A. Global cancer statistics, 2012. CA Cancer J Clin 2015;65(2):87-108.##Craig WJ. Health-promoting properties of common herbs. Am J Clin Nutr 1999;70(3 Suppl):491S-499S.##Katiyar C, Gupta A, Kanjilal S, Katiyar S. Drug discovery from plant sources: An integrated approach. AYU 2012;33(1):10-19.##Mata TM, Martins AA, Caetano NS. Microalgae for biodiesel production and other applications: a review. Renew Sustain Energy Rev 2010;14(1):217-232.##El-Sayed WS, Akhkha A, El-Naggar MY, Elbadry M. In vitro antagonistic activity, plant growth promoting traits and phylogenetic affiliation of rhizobacteria associated with wild plants grown in arid soil. Front Microbiol 2014;5:651.##Aghel S, Pouramir M, Moghadamnia AA, Moslemi D, Molania T, Ghassemi L, et al. Effect of iranian propolis on salivary total antioxidant capacity in gamma-irradiated rats. J Dent Res Dent Clin Dent Prospects 2014;8(4):235-239.##Shamsedin M, Arash V, Jahromi MB, Moghadamnia AA, Kamel MR, Ezoji F, et al. Efficacy of quercetin flavonoid in recovering the postbleaching bond strength of orthodontic brackets: A preliminary study. J Orthod Sci 2017;6(1):16-21.##Dajas F. Life or death: neuroprotective and anticancer effects of quercetin. J Ethnopharmacol 2012;143(2):383-396.##Vargas AJ, Burd R. Hormesis and synergy: pathways and mechanisms of quercetin in cancer prevention and management. Nutr Rev 2010;68(7):418-428.##Salvamani S, Gunasekaran B, Shaharuddin NA, Ahmad SA, Shukor MY. Antiartherosclerotic effects of plant flavonoids. Biomed Res Int 2014;2014:480258.##Kumar S, Pandey AK. Chemistry and biological activities of flavonoids: an overview. ScientificWorldJournal 2013;2013:162750.##Kulisic-Bilusic T, Schmoller I, Schnabele K, Siracusa L, Ruberto G. The anticarcinogenic potential of essential oil and aqueous infusion from caper (Capparis spinosa L.). Food Chem 2012;132(1):261-267.##Fogh J, Fogh JM, Orfeo T. One hundred and twenty-seven cultured human tumor cell lines producing tumors in nude mice. J Natl Cancer Inst 1977;59(1):221-226.##Pautke C, Schieker M, Tischer T, Kolk A, Neth P, Mutschler W, et al. Characterization of osteosarcoma cell lines MG-63, Saos-2 and U-2 OS in comparison to human osteoblasts. Anticancer Res 2004;24(6):3743-3748.##Germano MP, De Pasquale R, D'Angelo V, Catania S, Silvari V, Costa C. Evaluation of extracts and isolated fraction from Capparis spinosa L. buds as an antioxidant source. J Agric Food Chem 2002;50(5):1168-1171.##Al-Soqeer A. Antioxidant activity and biological evaluation of hot-water extract of Artemisia monosperma and Capparis spinosa against lead contamination. Res J Botany 2011;6(1):11-20.##Farzaei MH, Bahramsoltani R, Rahimi R. Phytochemicals as adjunctive with conventional anticancer therapies. Curr Pharm Des 2016;22(27):4201-4218.##Davatgaran-Taghipour Y, Masoomzadeh S, Farzaei MH, Bahramsoltani R, Karimi-Soureh Z, Rahimi R, et al. Polyphenol nanoformulations for cancer therapy: experimental evidence and clinical perspective. Int J Nanomed 2017;12:2689-2702.##Tajik M, Seifi S, Feizi F, Kazemi S, Moghadamnia AA. Histopathologic evaluation of hydroalcoholic extraction Capparis spinosa on the oral wound healing rats. J Babol Univ Med Sci 2016;18(12):33-39.##Özan F, Polat ZA, Er K, Özan Ü, Değer O. Effect of propolis on survival of periodontal ligament cells: new storage media for avulsed teeth. J Endod 2007;33(5):570-573.##Ji YB, Yu L. N-butanol extract of Capparis spinosa L. induces apoptosis primarily through a mitochondrial pathway involving mPTP open, cytochrome C release and caspase activation. Asian Pac J Cancer Prev 2014;15(21):9153-9157.##Mansour RB, Jilani IB, Bouaziz M, Gargouri B, Elloumi N, Attia H, et al. Phenolic contents and antioxidant activity of ethanolic extract of Capparis spinosa. Cytotechnology 2016;68(1):135-142.##Tulumen T, Ayata A, Ozen M, Sutcu R, Canatan D. The protective effect of Capparis ovata on 6-mercaptopurine-induced hepatotoxicity and oxidative stress in rats. J Pediatr Hematol Oncol 2015;37(4):290-294.##Erdogan MS, Babacan H, Kara MI, Gurler B, Akgul H, Soyler DA. Effect of Capparis spinosa extract on sutural ossification: A stereological study. Arch Oral Biol 2015;60(8):1146-1152.##Lam SK, Ng TB. A protein with antiproliferative, antifungal and HIV-1 reverse transcriptase inhibitory activities from caper (Capparis spinosa) seeds. Phytomedicine 2009;16(5):444-450.##

A Multidisciplinary Study to Evaluate the Anti-quorum Sensing Ability of Phyto-compounds in Ruellia patula Jacq 2 2 Background: Staphylococcus aureus (S. aureus) causing numerous diseases in humans, have become resistant to antibiotics, hence, urging the need for alternative medicines.  Methods: In this study, the Indian medicinal weed, Ruellia patula (R. patula) extracted and fractioned through column chromatography was subjected to antibacterial and anti-quorum sensing activity against S. aureus and Methicillin Resistant Staphylococcus aureus MRSA. Results: The obtained results confirmed fraction F44 to have significant effect as antimicrobial and anti-biofilm agent against both the micro-organism. Therefore, few of such highly active fractions were chemical finger printed using GC-MS and the compounds identified were further docked with DNA binding (LytTR) domain of agrA, which revealed that compounds identified from fraction were interactive to the protein. Conclusion: R. patula is promising antimicrobial and anti-biofilm agent against S. aureus and MRSA. 48 58 P Chemmugil Centre for Bioinformatics, Pondicherry University Centre for Bioinformatics, Pondicherry University India PTV Lakshmi Centre for Bioinformatics, Pondicherry University Centre for Bioinformatics, Pondicherry University India A Annamalai PG and Research Department of Botany, Arignar Anna Government College, Thiruvalluvar University, Villupuram PG and Research Department of Botany, Arignar Anna Government College, Thiruvalluvar University, Villupuram India AntibioticsDNA binding domainMethicillin resistant Staphylococcus aureusQuorum sensingStaphylococcus aureus 10338.pdf Zbikowska B, Franiczek R, Sowa A, Polukord G, Krzyzanowska B, Sroka Z. Antimicrobial and antiradical activity of extracts obtained from leaves of five species of the genus bergenia: identification of antimicrobial compounds. Microb Drug Resist 2017;23(6):771-780.##Nostro A, Guerrini A, Marino A, Tacchini M, Di Giulio M, Grandini A, et al. In vitro activity of plant extracts against biofilm-producing food-related bacteria. Int J Food Microbiol 2016;238:33-39.##Yadav S, Arya V, Kumar S, Yadav JP. Anti-inflammatory activity of root, leaves and stem of Dipteracanthus patulus (Jacq.) Nees (Acanthaceae). Asian Pac J Trop Biomed 2012;2(1):S187-S191.##Ramadevi S, Kaleeswaran B, Natarajan P. Phytochemicals analysis and antimicrobial activity of Ruellia patula L. against pathogenic microorganisms. South Indian J Biol Sci 2016;2(2):306-313.##Lakshmi P. Exploring the Phyto-Constituents of Ruellia patula (Acanthaceae) as Antibacterial Agent. EC Microbiol 2017;7:133-148.##Lacey KA, Geoghegan JA, McLoughlin RM. The role of Staphylococcus aureus virulence factors in skin infection and their potential as vaccine antigens. Pathogens 2016;5(1). pii: E22.##Meng Y, Hou X, Lei J, Chen M, Cong S, Zhang Y, et al. Multi-functional liposomes enhancing target and antibacterial immunity for antimicrobial and anti-biofilm against methicillin-resistant staphylococcus aureus. Pharm Res 2016;33(3):763-775.##Thomer L, Schneewind O, Missiakas D. Pathogenesis of Staphylococcus aureus bloodstream infections. Annu Rev Pathol 2016;11:343-364.##Kashef N, Akbarizare M, Razzaghi MR. In vitro activity of Linezolid in combination with photodynamic inactivation against staphylococcus aureus biofilms. Avicenna J Med Biotechnol 2017;9(1):44-48.##Vuong C, Yeh AJ, Cheung GY, Otto M. Investigational drugs to treat methicillin-resistant Staphylococcus aureus. Expert Opin Investig Drugs 2016;25(1):73-93.##Van Hal SJ, Jensen SO, Vaska VL, Espedido BA, Paterson DL, Gosbell IB. Predictors of mortality in Staphylococcus aureus bacteremia. Clin Microbiol Rev 2012;25(2):362-386.##den Reijer PM, Haisma EM, Lemmens-den Toom NA, Willemse J, Koning RI, Demmers JA, et al. Detection of alpha-toxin and other virulence factors in biofilms of Staphylococcus aureus on polystyrene and a human epidermal model. PloS One 2016;11(1):e0145722.##Lina G, Jarraud S, Ji G, Greenland T, Pedraza A, Etienne J, et al. Transmembrane topology and histidine protein kinase activity of AgrC, the agr signal receptor in Staphylococcus aureus. Mol Microbiol 1998;28(3):655-662.##Yarwood JM, Schlievert PM. Quorum sensing in Staphylococcus infections. J Clin Invest 2003;112(11):1620-1625.##Kong KF, Vuong C, Otto M. Staphylococcus quorum sensing in biofilm formation and infection. Int J Med Microbiol 2006;296(2-3):133-139.##Rajasree K, Fasim A, Gopal B. Conformational features of the Staphylococcus aureus AgrA-promoter interactions rationalize quorum-sensing triggered gene expression. Biochem Biophys Rep 2016;6:124-134.##Galperin MY. Telling bacteria: do not LytTR. Structure 2008;16(5):657-659.##Komape NP, Bagla VP, Kabongo-Kayoka P, Masoko P. Anti-mycobacteria potential and synergistic effects of combined crude extracts of selected medicinal plants used by Bapedi traditional healers to treat tuberculosis related symptoms in Limpopo Province, South Africa. BMC Complement Altern Med 2017;17(1):128.##Zhang WM, Wang W, Zhang JJ, Wang ZR, Wang Y, Hao WJ, et al. Antibacterial constituents of hainan morinda citrifolia (Noni) Leaves. J Food Sci 2016;81 (5):M1192-1196.##Rosas-Burgos EC, Burgos-Hernandez A, Noguera-Artiaga L, Kacaniova M, Hernandez-Garcia F, Cardenas-Lopez JL, et al. Antimicrobial activity of pomegranate peel extracts as affected by cultivar. J Sci Food Agric 2017;97(3):802-810.##Su PW, Yang CH, Yang JF, Su PY, Chuang LY. Antibacterial activities and antibacterial mechanism of polygonum cuspidatum extracts against nosocomial drug-resistant pathogens. Molecules 2015;20(6):11119-11130.##Gorle AK, Li X, Primrose S, Li F, Feterl M, Kinobe RT, et al. Oligonuclear polypyridylruthenium (II) complexes: selectivity between bacteria and eukaryotic cells. J Antimicrob Chemother 2016;71(6):1547-1555.##Bazargani MM, Rohloff J. Antibiofilm activity of essential oils and plant extracts against Staphylococcus aureus and Escherichia coli biofilms. Food Control 2016;61:156-164.##Leonard PG, Bezar IF, Sidote DJ, Stock AM. Identification of a hydrophobic cleft in the LytTR domain of AgrA as a locus for small molecule interactions that inhibit DNA binding. Biochemistry 2012;51(50):10035-10043.##Trott O, Olson AJ. AutoDock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading. J Computat Chem 2010;31(2):455-461.##Morris GM, Goodsell DS, Halliday RS, Huey R, Hart WE, Belew RK, et al. Automated docking using a Lamarckian genetic algorithm and an empirical binding free energy function. J Comput Chem 1998;19:1639-1662.##Laskowski RA, Swindells MB. LigPlot+: multiple ligand-protein interaction diagrams for drug discovery. J Chem Inf Model 2011;51(10):2778-2786.##Mhalla D, Bouaziz A, Ennouri K, Chawech R, Smaoui S, Jarraya R, et al. Antimicrobial activity and bioguided fractionation of Rumex tingitanus extracts for meat preservation. Meat Sci 2017;125:22-29.##Akinpelu DA, Odewade JO, Aiyegoro OA, Ashafa AO, Akinpelu OF, Agunbiade MO. Biocidal effects of stem bark extract of Chrysophyllum albidium G. Don on vancomycin-resistant Staphylococcus aureus. BMC Complement Altern Med 2016;16:105.##Desbois AP, Smith VJ. Antibacterial free fatty acids: activities, mechanisms of action and biotechnological potential. Appl Microbiol Biotechnol 2010;85(6):1629-1642.##Negi PS. Plant extracts for the control of bacterial growth: efficacy, stability and safety issues for food application. Int J Food Microbiol 2012;156(1):7-17.##El-kalyoubi S, Fatmah Agili F, Youssif S. Novel 2-thioxanthine and dipyrimidopyridine derivatives: synthesis and antimicrobial activity. Molecules 2015;20 (10):19263-19276.##Sanchez E, Rivas Morales C, Castillo S, Leos-Rivas C, Garcia-Becerra L, Ortiz Martinez DM. Antibacterial and antibiofilm activity of methanolic plant extracts against nosocomial microorganisms. Evid Based Complement Alternat Med. 2016;2016:1572697.##Aqil F, Ahmad I, Owais M. Evaluation of anti-methicillin-resistant Staphylococcus aureus (MRSA) activity and synergy of some bioactive plant extracts. Biotechnol J 2006;1(10):1093-1102.##Betoni JE, Mantovani RP, Barbosa LN, Di Stasi LC, Fernandes Junior A. Synergism between plant extract and antimicrobial drugs used on Staphylococcus aureus diseases. Mem Inst Oswaldo Cruz 2006;101(4):387-390.##Klastersky J, Husson M. Bactericidal activity of the combinations of gentamicin with clindamycin or chloramphenicol against species of Escherichia coli and Bacteroides fragilis. Antimicrob Agents Chemother 1977;12(2):135-138.##Jawetz E, Gunnison J, Speck R, Coleman V. Studies on antibiotic synergism and antagonism: the interference of chloramphenicol with the action of penicillin. AMA Arch Intern Med 1951;87(3):349-359.##Nemeth J, Oesch G, Kuster SP. Bacteriostatic versus bactericidal antibiotics for patients with serious bacterial infections: systematic review and meta-analysis. J Antimicrob Chemother 2015;70(2):382-395.##Soriano A, Marco F, Martinez JA, Pisos E, Almela M, Dimova VP, et al. Influence of vancomycin minimum inhibitory concentration on the treatment of methicillin-resistant Staphylococcus aureus bacteremia. Clin Infect Dis 2008;46(2):193-200.##Tsuchiya H, Sato M, Miyazaki T, Fujiwara S, Tanigaki S, Ohyama M, et al. Comparative study on the antibacterial activity of phytochemical flavanones against methicillin-resistant Staphylococcus aureus. J Ethnopharmacol 1996;50(1):27-34.##Cushnie TT, Lamb AJ. Recent advances in understanding the antibacterial properties of flavonoids. Int J Antimicrob Agents 2011;38(2):99-107.##Ohta S, Shiomi Y, Kawashima A, Aozasa O, Nakao T, Nagate T, et al. Antibiotic effect of linolenic acid from Chlorococcum strain HS-101 and Dunaliella primolecta on methicillin-resistant Staphylococcus aureus. J Appl Phycol 1995;7(2):121-127.##Huang CB, Alimova Y, Myers TM, Ebersole JL. Short-and medium-chain fatty acids exhibit antimicrobial activity for oral microorganisms. Arch Oral Biol 2011;56 (7):650-654.##Borges A, Abreu A, Malheiro J, Saavedra MJ, SimC5es M. Biofilm prevention and control by dietary phytochemicals. Microbial pathogens and strategies for combating them: science, technology and education. Formatex Research Center, Badajoz. 2013:32-41.##

Statistical Optimization of Process Parameters by Central Composite Design (CCD) for an Enhanced Production of L-asparaginase by Myroides gitamensis BSH-3, a Novel Species 2 2 Background: The present study focused on the production of L-asparaginase using Solid State Fermentation (SSF) by Myroides gitamensis.  Methods: Initially, five significant parameters (Carbon source; Nitrogen source, temperature, pH and incubation period) were identified that affect the production process of L-asparaginase using Classical One Factor at a Time (OFAT) optimization. An optimized L-asparaginase specific activity obtained by OFAT was recorded as 85.7 IU. Central Composite Design (CCD) was also employed successively to optimize the multiple parameters at a time and their results were compared.  Results: Maximum L-asparaginase enzyme specific activity obtained by CCD method was 295.6 IU under the hold values of carbon source (wheat bran) 12 g/L, nitrogen source (yeast extract) 7 g/L, temperature 37oC, pH=7.5 and incubation period 47 hr. Upon validation, the obtained results proved that there was a good relation existing between the experimental and the predicted model (p<0.05). L-asparaginase activity was enhanced in statistical method up to 3.4 folds compared to that of classical method.  Conclusion: Utilization of wheat bran as a low cost carbon source in SSF for the production of L-asparaginase enzyme makes the process economical and in turn reduces the environmental pollution by biotransformation to commercially useful bio product. 59 66 VSSL Talluri Department of Biotechnology, College of Natural and Computational Science, University of GondarDepartment of Biotechnology, GITAM Institute of Science, GITAM University, Visakhapatnam- 530 045Department of Biotechnology, University of Chemistry and Technology Prague, Technicka 5 Department of Biotechnology, College of Natural and Computational Science, University of GondarDepartment of Biotechnology, GITAM Institute of Science, GITAM University, Visakhapatnam- 530 045Department of Biotechnology, University of Chemistry and Technology Prague, Technicka 5 EthiopiaIndiaCZ 16628 Prague Sri Lanka Department of Biotechnology, GITAM Institute of Science, GITAM University, Visakhapatnam- 530 045 Department of Biotechnology, GITAM Institute of Science, GITAM University, Visakhapatnam- 530 045 India SV Rajagopal Department of Biotechnology, GITAM Institute of Science, GITAM University, Visakhapatnam- 530 045 Department of Biotechnology, GITAM Institute of Science, GITAM University, Visakhapatnam- 530 045 India BiotransformationBranEnvironmental pollutionFermentationL-asparaginaseMyroides gitamensisWheat 10355.pdf Mishra A. Production of L-asparaginase, an anticancer agent from Aspergillus niger using agricultural waste in solid state fermentation. Appl Biochem Biotechnol 2006;135(1):33-42.##Verma N, Kumar K, Kaur G, Anand S. L-asparaginase: a promising chemotherapeutic agent. Crit Rev Biotechnol 2007;27(1):45-62.##Teodor E, Litescu SC, Lazar V, Somoghi R. Hydrogel-magnetic nanoparticles with immobilized L-asparaginase for biomedical applications. J Mater Sci Mater Med 2009;20(6):1307-1314.##Narta UK, Kanwar SS, Azmi W. Pharmacological and clinical evaluation of l-asparaginase in the treatment of leukemia. Crit Rev Oncol Hematol 2007;61(3):208-221.##Pedreschi F, Kaack K, Granby K. The effect of asparaginase on acrylamide formation in French fries. Food Chem 2008;109(2):386-392.##Offman MN, Krol M, Patel N, Krishnan S, Liu J, Saha V, et al. Rational engineering of L-asparaginase reveals importance of dual activity for cancer cell toxicity. Blood 2011;117(5):1614-1621.##Savitri N, Asthana N, Azmi W. Microbial L-asparaginase: a potent antitumour enzyme. Indian J Biotechnol 2003;2:184-194.##El-Shishtawy RM, Mohamed SA, Asiri AM, Gomaa AB, Ibrahim IH, Al-Talhi HA. Solid fermentation of wheat bran for hydrolytic enzymes production and saccharification content by a local isolate Bacillus megatherium. BMC Biotechnol 2014;14:29.##Stevenson L, Phillips F, O’sullivan K, Walton J. Wheat bran: its composition and benefits to health, a European perspective. Int J Food Sci Nutr 2012;63(8):1001-1013.##Irfan M, Nadeem M, Syed Q. One-factor-at-a-time (OFAT) optimization of xylanase production from Trichoderma viride-IR05 in solid-state fermentation. J Radiation Res Applied Sci 2014;7(3):317-326.##Talluri VP, Bhavana M, Siva Kumar K, Anil Kumar P, Rajagopal SV. Myroides gitamensis sp.nov., L-asparaginase producing bacteria isolated from slaughter house soil sample in Visakhapatnam, India. J Microb Biochem Technol 2014;6(3):144-147.##Gulati R, Saxena RK, Gupta R. A rapid plate assay for screening L-asparaginase producing micro-organisms. Lett Appl Microbiol 1997;24(1):23-26.##Mashburn LT, Wriston JC Jr. Tumor inhibitory effect of Lasparaginase from E.coli. Arch Biochem Biophys 1964;105:450-452.##Lowry OH, Rosebrough NL, Farr AL Randall RJ. Protein measurement with the folin reagent. J Biol Chem 1951;193(1):265-275.##Zhang H, Wang Z, Xu SY. Optimization of processing parameters for cloudy ginkgo (Ginkgo biloba Linn.) juice. J Food Eng 2007;80(4):1226-1232.##Charles P, Devanathan V, Anbu P, Ponnuswamy MN, Kalaichelvan PT, Hur BK. Purification, characterization and crystallization of an extracellular alkaline protease from Aspergillus nidulans HA-10. J Basic Microbiol 2008;48:347-352.##Adinarayana K, Ellaiah P, Prasad DS. Purification and partial characterization of thermostable serine alkaline protease from a newly isolated Bacillus subtilis PE-11. AAPS PharmSciTech 2003;4(4):E56.##Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970;227(5259):680-685.##Shahnoor HM, Abul Kalam A, Abu Sayem SM, Mostafa G, Md. Mozammel Hoq. Production and partial characterization of feather-degrading keratinolytic serine protease from Bacillus licheniformis MZK-3. J Biol Sci 2007;7(4):599-606.##Rajesh MJ, Leelavathy R, Vanapalli VS, Thirumurugan G, Rajaram MS. Effect of inducers and physical parameters on the production of L-asparaginase using Aspergillusterrus. J Bioprocess Biotech 2011;1:1-6.##Manna S, Sinha A, Sadhukhan R, Chakrabarty SL. Purification, characterization and antitumor activity of L-asparaginase isolated from Pseudomonas stutzeri MB-405. Curr Microbiol 1995;30(5):291-298.##Moorthy V, Ramalingam A, Sumantha A, Shankaranaya RT. Production, purification and characterisation of extracellular L-Asparaginase from a soil isolate of Bacillus sp. Afr J Microbiol Res 2010;4(18):1862-1867.##Hosamani R, Kaliwal BB. Isolation, molecular identification and optimization of fermentation parameters for the production of L-asparaginase, an anticancer agent by Fusarium equiseti. Int J Microbiol Res 2011;3(2):108-119.##Hymavathi M, Sathish T, Subba Rao Ch, Prakasham RS. Enhancement of L-asparaginase production by isolated Bacillus circulans (MTCC 8574) using response surface methodology. Appl Biochem Biotechnol 2009;159(1):191-198.##Rani GB, Chiranjeevi T, Chandel AK, Satish T, Radhika K, Narasu ML, et al. Optimization of selective production media for enhanced production of xylanases in submerged fermentation by Thielaviopsis basicola MTCC 1467 using L16 orthogonal array. J Food Sci Technol 2014;51(10):2508-2516.##El-Naggar Nel-A, Moawad H, El-Shweihy NM, El-Ewasy SM. Optimization of culture conditions for production of the anti-Leukemic glutaminase free L-Asparaginase by newly isolated Streptomyces olivaceus NEAE-119 using response surface methodology. Biomed Res Int 2015;2015:627031.##Potumarthi R, Jacques L, Harry W, Michael D. Surface immobilization of Rhizopus oryzae (ATCC 96382) for enhanced production of lipase enzyme by multiple responses optimization. Asia Pac J Chem Eng 2012;7(S3):S285-S295.##Madruga MS, Camara FS. The chemical composition of Multimistura as a food supplement. Food Chem 2000;68(1):41-44.##Beg QK, Bhushan B, Kapoor M, Hoondal GS. Enhanced production of thermostable xylanase from Streptomyces sp. QG11-3 and its application in biobleaching of eucalyptus kraft pulp. Enzyme Microb Technol 2000;27(7):459-466.##Ellaiah P, Srinivasulu B, Adinarayana K. Optimization studies on neomycin production by a mutant strain of Streptomyces marinensis in solid state fermentation. Process Biochem 2004;39(5):529-534.##Krishna C. Solid-state fermentation systems-an overview. Crit Rev Biotechnol 2005;25(1-2):1-30.##

The Effect of TAX-1 Gene of Human T-cell Leukemia Virus Type -1 on the Expression of CCR5 in K562 Cell Line 2 2 Background: Tax-1 protein of Human T-cell Leukemia Virus type 1(HTLV-1) serves as a key transcriptional regulatory gene product and has a crucial role in transactivating genes of infected cells by employing their transcriptional factors. This modulation includes induction of genes which encode CC-chemokines and their receptors. In this study, a recombinant vector containing Tax-1 gene was made and tested for its ability to induce CCR5 (CC chemokine receptor 5) expression in K562 cell line. Methods: In order to perform this research, two blood samples of HTLV-1 positive were obtained from Urmia blood transfusion center. After DNA extraction, a complete sequence of Tax-1 gene was amplified by specific primers. Recombinant vectors carrying Tax-1 gene were synthesized and transformed into Escherichia coli (E. coli). After bacteria transformation, bacteria containing recombinant plasmid were selected and purified. Then, the recombinant shuttle vectors, pCDNA3.1-TAX, were transfected into the cell culture (K562 cell line). Expression of CCR5 was measured after 72 hr by Syber Green Real-Time PCR method compared to control cell culture. Normalization was done with GAPDH as a standard gene. Results: Cloning of Tax-1 gene in the vector, pCDNA3.1 was confirmed by colony PCR, restriction digestion, and sequencing methods. Expression of Tax-1 and CCR5 genes were confirmed by real time PCR and also, expression of CCR5 gene showed an 8-fold increase in comparison to mock-treated controls (p<0.05). Conclusion: Our data suggested that recombinant Tax-1 may have the enhancing effect on CCR5 expression rate at mRNA levels in K562 cell line. Further studies are necessary to evaluate the effect of pCDNA3.1-TAX on cell surface CCR5 expression. 67 71 Nasrin Haghnazari Sadaghiani Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine Iran Lila Pirayeshfard Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine Iran Afsaneh Aghaie Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine Iran Zohreh Sharifi Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine Iran CCR5Human T-cell Leukemia VirusTax-1 protein 10354.pdf Gallo RC. The discovery of the first human retrovirus: HTLV-1 and HTLV-2. Retrovirology 2005;2:17.##Gessain A, Cassar O. Epidemiological aspects and world distribution of HTLV-1 infection. Front Microbiol 2012;3:388.##Giam CZ, Jeang KT. HTLV-1 Tax and adult T-cell leukemia. Front Biosci 2007;12:1496-1507.##Clark J, Saxinger C, Gibbs WN, Lofters W, Lagranade L, Deceulaer K, et al. Seroepidemiologic studies of human T-cell leukemia/lymphoma virus type I in Jamaica. Int J Cancer 1985;36(1):37-41.##Yasunaga J, Matsuoka M. Molecular mechanisms of HTLV-1 infection and pathogenesis. Int J Hematol 2011;94(5):435-442.##Verdonck K, Gonzalez E, Van Dooren S, Vandamme AM, Vanham G, Gotuzzo E. Human T-lymphotropic virus 1: recent knowledge about an ancient infection. Lancet Infect Dis 2007;7(4):266-281.##Pilotti E, Bianchi MV, De Maria A, Bozzano F, Romanelli MG, Bertazzoni U, et al. HTLV-1/-2 and HIV-1 co-infections: retroviral interference on host immune status. Front Microbiol 2013;4:372.##Casoli C, Pilotti E, Bertazzoni U. Molecular and cellular interactions of HIV-1/HTLV coinfection and impact on AIDS progression. AIDS Rev 2007;9(3):140-149.##Grassmann R, Aboud M, Jeang K-T. Molecular mechanisms of cellular transformation by HTLV-1 Tax. Oncogene 2005;24(39):5976-5985.##Beilke MA. Retroviral coinfections: HIV and HTLV: taking stock of more than a quarter century of research. AIDS Res Hum Retroviruses 2012;28(2):139-147.##Barrios CS, Abuerreish M, Lairmore MD, Castillo L, Giam CZ, Beilke MA. Recombinant human T-cell leukemia virus types 1 and 2 Tax proteins induce high levels of CC-chemokines and downregulate CCR5 in human peripheral blood mononuclear cells. Viral Immunol 2011;24(6):429-439.##Sharma V, Lorey SL. Autocrine role of macrophage inflammatory protein-1 beta in human T-cell lymphotropic virus type-I tax-transfected Jurkat T-cells. Biochem Biophys Res Commun 2001;287(4):910-913.##Woitas R P, Ahlenstiel G, Iwan A, Rockstroh JK, Brackmann HH, Kupfer B, et al. Frequency of the HIV-protective CC chemokine receptor 5-delta32/delta32 genotype is increased in hepatitis C. Gastroenterology 2002;122(7):1721-1728.##Pfaffl MW. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 2001;29(9):e45.##Brites C, Alencar R, Gusmao R, Pedroso C, Netto EM, Pedral-Sampaio D, et al. Co-infection with HTLV-1 is associated with a shorter survival time for HIV-1-infected patients in Bahia, Brazil. AIDS 2001;15(15):2053-2055.##Pedroso C, Netto EM, Weyll N, Brites C. Coinfection by HIV-1 and human lymphotropic virus type 1 in Brazilian children is strongly associated with a shorter survival time. J Acquir Immune Defic Syndr 2011;57 Suppl 3:S208-211.##Tulius Silva M, de Melo Espindola O, Bezerra Leite AC, Araujo A. Neurological aspects of HIV/human T lymphotropic virus coinfection. AIDS Rev 2009;11(2):71-78.##Beilke MA, Theall KP, O'Brien M, Clayton JL, Benjamin SM, Winsor EL, et al. Clinical outcomes and disease progression among patients coinfected with HIV and human T lymphotropic virus types 1 and 2. Clin Infect Dis 2004;39(2):256-263.##Chavance M, Neisson-Vernant C, Quist D, Monplaisir N, Armengaud B, Chout R. HIV/HTLV-I coinfection and clinical grade at diagnosis. J Acquir Immune Defic Syndr Hum Retrovirol 1995;8(1):91-95.##Shibata D, Brynes RK, Rabinowitz A, Hanson CA, Slovak ML, Spira TJ, et al. Human T-cell lymphotropic virus type I (HTLV-I)-associated adult T-cell leukemia-lymphoma in a patient infected with human immunodeficiency virus type 1 (HIV-1). Ann Intern Med 1989;111(11):871-875.##Lasky LA, Nakamura G, Smith DH, Fennie C, Shimasaki C, Patzer E, et al. Delineation of a region of the human immunodeficiency virus type 1 gp120 glycoprotein critical for interaction with the CD4 receptor. Cell 1987;50(6):975-985.##de Castro-Amarante MF, Pise-Masison CA, McKinnon K, Washington Parks R, Galli V, Omsland M, et al. Human T cell leukemia virus type 1 infection of the three monocyte subsets contributes to viral burden in humans. J Virol 2016;90(5):2195-2207.##Moriuchi M, Moriuchi H, Fauci AS. HTLV type I Tax activation of the CXCR4 promoter by association with nuclear respiratory factor 1. AIDS Res Hum Retroviruses 1999;15(9):821-827.##Twizere JC, Springael JY, Boxus M, Burny A, Dequiedt F, Dewulf JF, et al. Human T-cell leukemia virus type-1 Tax oncoprotein regulates G-protein signaling. Blood 2007;109(3):1051-1060.##Barmania F, Pepper MS. C-C chemokine receptor type five (CCR5): an emerging target for the control of HIV infection. Appl Transl Genom 2013;2:3-16.##

Characterization of Monoclonal and Polyclonal Antibodies Recognizing Prostate Specific Antigen: Implication for Design of a Sandwich ELISA 2 2 Background: Prostate cancer is the second most common cancer in men. Prostate-Specific Antigen (PSA) is a tumor-associated glycoprotein with enzymatic activity which is secreted by the prostate gland. Following entry to the blood, 70-90% of PSA forms complexes with protease inhibitors and its enzymatic activity is inhibited. The serum level of PSA is increased and the rate of free PSA (fPSA) to total PSA is decreased in prostate cancer patients. Therefore, measurement of PSA and fPSA in serum is very valuable for diagnosis and prognosis of prostate cancer. Methods: In the present study, five anti PSA monoclonal Antibodies (mAb) were characterized by Enzyme-Linked Immunosorbent Assay (ELISA) and immunoblotting. For designing a sandwich ELISA, epitope specificity of these antibodies was studied by a competition ELISA. Free PSA was purified by electroelution technique from seminal plasma and used to produce polyclonal anti-fPSA antibody in rabbit. Purified polyclonal antibody (pAb) and mAbs were conjugated with HRP enzyme and Biotin (Bio) to set up the sandwich ELISA. Results: Three of the mAbs were found to recognize PSA similarly. One of these mAbs (2G3) was paired with anti-fPSA pAb to detect fPSA in serum. Eventually, serum fPSA concentration of 356 subjects was measured and compared by our designed ELISA and a commercial ELISA kit. Our results demonstrated a significant correlation (r=0.68; p<0.001) between the two assays. Sensitivity and specificity of our designed ELISA was 72.4 and 82.8%, respectively. Conclusion: These results imply suitability of our designed ELISA for detection of fPSA in patients with prostate cancer. 72 79 Sahar Raoofi Mohseni Department of Immunology, Faculty of Public Health, Tehran University of Medical Sciences Department of Immunology, Faculty of Public Health, Tehran University of Medical Sciences Iran Forough Golsaz-Shirazi Department of Immunology, Faculty of Public Health, Tehran University of Medical Sciences Department of Immunology, Faculty of Public Health, Tehran University of Medical Sciences Iran Mostafa Hosseini Department of Epidemiology and Biostatistics, Faculty of Public Health, Tehran University of Medical Sciences Department of Epidemiology and Biostatistics, Faculty of Public Health, Tehran University of Medical Sciences Iran Jalal Khoshnoodi Department of Immunology, Faculty of Public Health, Tehran University of Medical Sciences Department of Immunology, Faculty of Public Health, Tehran University of Medical Sciences Iran Tannaz Bahadori Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Mohammad Ali Judaki Department of Immunology, Faculty of Public Health, Tehran University of Medical Sciences Department of Immunology, Faculty of Public Health, Tehran University of Medical Sciences Iran Mahmood Jeddi-Tehrani Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR Iran Fazel Shokri Department of Immunology, Faculty of Public Health, Tehran University of Medical SciencesMonoclonal Antibody Research Center, Avicenna Research Institute, ACECR Department of Immunology, Faculty of Public Health, Tehran University of Medical SciencesMonoclonal Antibody Research Center, Avicenna Research Institute, ACECR IranIran ELISAMonoclonal antibodiesProstate cancerProstate specific antigen 10351.pdf Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D. Global cancer statistics. CA Cancer J Clin 2011;61(2):69-90.##Yousef GM, Diamandis EP. The new human tissue kallikrein gene family: structure, function, and association to disease. Endocr Rev 2001;22(2):184-204.##Stenman UH, Abrahamsson PA, Aus G, Lilja H, Bangma C, Hamdy FC, et al. Prognostic value of serum markers for prostate cancer. Scand J Urol Nephrol Suppl 2005; (216):64-81.##Shariat SF, Semjonow A, Lilja H, Savage C, Vickers AJ, Bjartell A. Tumor markers in prostate cancer I: blood-based markers. Acta Oncol 2011;50 Suppl 1:61-75.##Adhyam M, Gupta AK. A review on the clinical utility of PSA in cancer prostate. Indian J Surg Oncol 2012;3(2):120-129.##Lilja H, Oldbring J, Rannevik G, Laurell C. Seminal vesicle-secreted proteins and their reactions during gelation and liquefaction of human semen. J Clin Invest 1987;80(2):281-285.##Mattsson JM, Ravela S, Hekim C, Jonsson M, Malm J, Närvänen A, et al. Proteolytic activity of prostate-specific antigen (PSA) towards protein substrates and effect of peptides stimulating PSA activity. PloS One 2014;9(9):e107819.##Jung K, Brux B, Lein M, Rudolph B, Kristiansen G, Hauptmann S, et al. Molecular forms of prostate-specific antigen in malignant and benign prostatic tissue: biochemical and diagnostic implications. Clin Chem 2000;46(1):47-54.##Catalona WJ, Smith DS, Ratliff TL, Dodds KM, Coplen DE, Yuan JJ, et al. Measurement of prostate-specific antigen in serum as a screening test for prostate cancer. N Engl J Med 1991;324(17):1156-1161.##Thompson IM, Pauler DK, Goodman PJ, Tangen CM, Lucia MS, Parnes HL, et al. Prevalence of prostate cancer among men with a prostate-specific antigen level≤ 4.0 ng per milliliter. N Engl J Med 2004;350(22):2239-2246.##Balk SP, Ko YJ, Bubley GJ. Biology of prostate-specific antigen. J Clin Oncol 2003;21(2):383-391.##Sp I, Ramona I, Sukesh. The efficiency of the serum prostate specific antigen levels in diagnosing prostatic enlargements. J Clin Diagn Res 2013;7(1):82-84.##Stenman UH, Leinonen J, Zhang WM, Finne P. Prostate-specific antigen. Semin Cancer Biol 1999;9(2):83-93.##Ulmert D, O'brien MF, Bjartell AS, Lilja H. Prostate kallikrein markers in diagnosis, risk stratification and prognosis. Nat Rev Urol 2009;6(7):384-391.##de Vries S. Active surveillance and prostate cancer screening. 1st ed. Rotterdam: Erasmus University Rotterdam; 2009. 151 p.##Bayat AA, Ghods R, Shabani M, Mahmoudi AR, Yeganeh O, Hassannia H, et al. Production and characterization of monoclonal antibodies against human prostate specific antigen. Avicenna J Med Biotechnol 2015;7(1):2-7.##Black MH, Grass CL, Leinonen J, Stenman UH, Diamandis EP. Characterization of monoclonal antibodies for prostate-specific antigen and development of highly sensitive free prostate-specific antigen assays. Clin Chem 1999;45(3):347-354.##Matsumoto K, Konishi N, Hiasa Y, Kimura E, Takahashi Y, Shinohara K, et al. A highly sensitive enzyme-linked immunoassay for serum free prostate specific antigen (f-PSA). Clin Chim Acta 1999;281(1):57-69.##matsumoto K, Konishi N, Samori T, Kimura E, Doi M, Kato S, et al. ELISA for a complexed antigen with a monoclonal antibody blocking reaction with the free antigen-assay-specific for complexed prostate-specific antigen. J Immunol Methods 2000;234(1-2):99-106.##Zhu L. Development of novel assays for measuring different molecular forms of prostate specific antigen [dissertation]. [Finland]: University of Helsinki Helsinki; 2009. 74 p.##Ito K, Yamamoto T, Ohi M, Kurokawa K, Suzuki K, Yamanaka H. Free/total PSA ratio is a powerful predictor of future prostate cancer morbidity in men with initial PSA levels of 4.1 to 10.0 ng/mL. Urology 2003;61(4):760-764.##Tessmer U, Quack T, Donn F, Leuner A, Dernick R. Biological activity of prostate‐specific antigen isolated by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and electroelution. Electrophoresis 1995; 16(1):793-799.##Sá-Pereira P, Duarte J, Costa-Ferreira M. Electroelution as a simple and fast protein purification method: isolation of an extracellular xylanase from Bacillus sp. CCMI 966. Enzyme Microb Technol 2000;27(1-2):95-99.##Lei Z, Anand A, Mysore KS, Sumner LW. Electroelution of intact proteins from SDS-PAGE gels and their subsequent MALDI-TOF MS analysis. Methods Mol Biol 2007;355:353-363.##Frenette G, Deperthes D, Tremblay RR, Lazure C, Dubé JY. Purification of enzymatically active kallikrein hK2 from human seminal plasma. Biochim Biophys Acta Gen Subj 1997;1334(1):109-115.##

Development and Comparison of In-house Line Probe Assay (LiPA) and SYBR Green Real-time PCR Regarding the Detection of Periodontal Pathogens 2 2 Background: Periodontal disease, which can become a chronic condition, is an inflammatory disease that upsets the soft and hard structures supporting the teeth. The aim of the present study was to design and develop an in-house Line Probe Assay (LiPA), to detect putative periodontitis-related bacterial pathogens, and compare it with SYBR Green Real-time PCR. Methods: The LiPA method was launched using biotinylated 16s rRNA universal primers and specific probes for each of the five bacteria including Aggregatibacter actinomycetemcomitans, Prevotella intermedia, Tannerella forsythia, Porphyromonas gingivalis and Treponema denticola. For this, optimized quantities of the primers and specific probes were dotted onto nylon membrane stripes in a defined pattern. Hybridization was performed between the probes and the single-stranded biotinylated PCR products. The stripes were developed via biotin-streptavidin reaction. Ultimately, the analytical and diagnostic sensitivity and specificity of the in-house LiPA was evaluated and compared with SYBR Green Real-time PCR. Results: The detection limit of the LiPA was 2760 copies of targeted genes. In testing analytical specificity, only signals corresponding to the specific biotinylated products were produced. The calculated diagnostic sensitivity of the LiPA for the five bacterial targets ranged from 96.4 to 100%, whereas the diagnostic specificity was between 90.9 and 100%. Comparing the results, no noticeable difference (p=0.4795) was observed between the two methods. Conclusion: To screen periodontal pathogens, a simple, inexpensive and accurate method is desirable. The in-house LiPA, having advantages such as high specificity and sensitivity, and the ability to detect five major periodontal pathogens, offers the option of evaluating samples without the need for a post-PCR platform. 80 87 Mohammad Soleimani Department of Microbiology, Qom Branch, Islamic Azad University Department of Microbiology, Qom Branch, Islamic Azad University Iran Mohammad Reza Zolfaghari Department of Microbiology, Qom Branch, Islamic Azad University Department of Microbiology, Qom Branch, Islamic Azad University Iran PeriodontitisReal-time PCRSensitivity and specificity 10353.pdf Hyvärinen K, Laitinen S, Paju S, Hakala A, Suominen-Taipale L, Skurnik M, et al. Detection and quantification of five major periodontal pathogens by single copy gene-based real-time PCR. Innate Immun 2009;15(4):195-204.##Sheiham A, Netuveli GS. Periodontal diseases in Europe. Periodontol 2000 2002;29(1):104-121.##Brown LJ, Löe H. Prevalence, extent, severity and progression of periodontal disease. Periodontol 2000 1993;2:57-71.##Chala S, Abouqal R, Abdallaoui F. Prevalence of apical periodontitis and factors associated with the periradicular status. Acta odontol Scand 2011;69(6):355-359.##Slots J. The predominant cultivable microflora of advanced periodontitis. Scand J Dent Res 1977;85(2):114-121.##Haffajee AD, Cugini MA, Tanner A, Pollack RP, Smith C, Kent RL Jr, et al. Subgingival microbiota in healthy, well-maintained elder and periodontitis subjects. J Clin Periodontol 1998;25(5):346-353.##Soleimani M, Zolfaghari MR, Morovvati A. Development and comparison of conventional PCR and SYBR green real time PCR for detection of aggregatibacter actinomycetemcomitans and Tannerella for sythensis. Jundishapur J Microbiol 2013;6(8):e6757.##Slots J, Ting M. Actinobacillus actinomycetemcomitans and porphyromonas gingivalis in human periodontal disease: occurrence and treatment. Periodontol 2000 1999;20(1):82-121.##Kozarov EV, Dorn BR, Shelburne CE, Dunn WA Jr, Progulske-Fox A. Human atherosclerotic plaque contains viable invasive actinobacillus actinomycetemcomitans and porphyromonas gingivalis. Arterioscler Thromb Vasc Biol 2005;25(3):e17-e8.##Suzuki N, Yoshida A, Saito T, Kawada M, Nakano Y. Quantitative microbiological study of subgingival plaque by real-time PCR shows correlation between levels of tannerella forsythensis and fusobacterium spp. J Clin Microbiol 2004;42(5):2255-2257.##Ashimoto A, Chen C, Bakker I, Slots J. Polymerase chain reaction detection of 8 putative periodontal pathogens in subgingival plaque of gingivitis and advanced periodontitis lesions. Oral Microbiol Immunol 1996;11(4):266-273.##Maeda H, Fujimoto C, Haruki Y, Maeda T, Kokeguchi S, Petelin M, et al. Quantitative real‐time PCR using TaqMan and SYBR Green for actinobacillus actinomycetemcomitans, porphyromonas gingivalis, prevotella intermedia, tetQ gene and total bacteria. FEMS Immunol Med Microbiol 2003;39(1):81-86.##Coffey J, Choudhry M, Shlossman M, Makin IRS, Singh VK. Multiplex real‐time PCR detection and relative quantification of periodontal pathogens. Clin Exp Dent Res 2016;2(3):185-192.##Wen D, Zhang C. Universal multiplex PCR: a novel method of simultaneous amplification of multiple DNA fragments. Plant Methods 2012;8(1):32.##Wong ML, Medrano JF. Real-time PCR for mRNA quantitation. Biotechniques 2005;39(1):75-85.##Ling DI, Zwerling AA, Pai M. GenoType MTBDR assays for the diagnosis of multidrug-resistant tuberculosis: a meta-analysis. Eur Respir J 2008;32(5):1165-1174.##Abe C, Ogata H, Kawata K, Hiraga T, Takashima T, Suetake T. [Detection of rifampin-resistant Mycobacterium tuberculosis by line probe assay (LiPA)]. Kekkaku 2000;75(10):575-581. Japanese.##Naidoo P, du Toit E, Dunbar R, Lombard C, Caldwell J, Detjen A, et al. A comparison of multidrug-resistant tuberculosis treatment commencement times in MDRTBPlus line probe assay and Xpert(R) MTB/RIF-based algorithms in a routine operational setting in Cape Town. PLoS One 2014;9(7):e103328.##Ritter C, Lucke K, Sirgel FA, Warren RW, van Helden PD, Bottger EC, et al. Evaluation of the AID TB resistance line probe assay for rapid detection of genetic alterations associated with drug resistance in Mycobacterium tuberculosis strains. J Clin Microbiol 2014;52(3):940-946.##Menton JF, Kearney K, Morgan JG. Development of a real-time RT-PCR and reverse line probe hybridisation assay for the routine detection and genotyping of noroviruses in Ireland. Virol J 2007;4:86.##Wang J, Bakkers J, Melchers WJ. [Genotyping of human papillomavirus genotyes in cervical scrapes by line probe assay]. Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za zZhi 1999;13(1):9-12. Chinese.##Noppornpanth S, Sablon E, De Nys K, Lien TX, Brouwer J, Van Brussel M, et al. Genotyping hepatitis C viruses from Southeast Asia by a novel line probe assay that simultaneously detects core and 5′ untranslated regions. J Clin Microbiol 2006;44(11):3969-3974.##Andonov A, Chaudhary RK. Subtyping of hepatitis C virus isolates by a line probe assay using hybridization. J Clin Microbiol 1995;33(1):254-256.##Avó AP, Agua-Doce I, Andrade A, Pádua E. Hepatitis C virus subtyping based on sequencing of the C/E1 and NS5B genomic regions in comparison to a commercially available line probe assay. J Med Virol 2013;85(5):815-822.##Cai Q, Zhao Z, Liu Y, Shao X, Gao Z. Comparison of three different HCV genotyping methods: core, NS5B sequence analysis and line probe assay. Int J Mol Med 2013;31(2):347-352.##Comanor L, Elkin C, Leung K, Krajden M, Kronquist K, Nicolas K, et al. Successful HCV genotyping of previously failed and low viral load specimens using an HCV RNA qualitative assay based on transcription-mediated amplification in conjunction with the line probe assay. J Clin Virol 2003;28(1):14-26.##Liu CH, Liang CC, Liu CJ, Lin CL, Su TH, Yang HC, et al. Comparison of abbott realTime HCV genotype II with versant line probe assay 2.0 for Hepatitis C virus genotyping. J Clin Microbiol 2015;53(5):1754-1757.##Nadkarni MA, Martin FE, Jacques NA, Hunter N. Determination of bacterial load by real-time PCR using a broad-range (universal) probe and primers set. Microbiology 2002;148(Pt 1):257-266.##Baehni P, Guggenheim B. Potential of diagnostic microbiology for treatment and prognosis of dental caries and periodontal diseases. Crit Rev Oral Biol Med 1996;7(3):259-277.##Frank JA, Reich CI, Sharma S, Weisbaum JS, Wilson BA, Olsen GJ. Critical evaluation of two primers commonly used for amplification of bacterial 16S rRNA genes. Appl Environ Microbiol 2008;74(8):2461-2470.##Geevarghese A, Baskaradoss JK, Al Dosari A. Application of 16s RRNA in identifying oral microflora-a review of literature. World Appl Sci J 2012;17(10): 1303-1307.##Hutter G, Schlagenhauf U, Valenza G, Horn M, Burgemeister S, Claus H, et al. Molecular analysis of bacteria in periodontitis: evaluation of clone libraries, novel phylotypes and putative pathogens. Microbiology 2003;149(Pt 1):67-75.##Majidzadeh-A K, Morovvati A, Soleimani M, Langeroudi AG, Merat S, Jabbari H. Development and application of an In-house line probe assay for Hepatitis C virus genotyping. Hepat Mon 2013;13(5):e6767.##Aberle SW, Kletzmayr J, Watschinger B, Schmied B, Vetter N, Puchhammer-Stöckl E. Comparison of sequence analysis and the INNO-LiPA HBV DR line probe assay for detection of lamivudine-resistant hepatitis B virus strains in patients under various clinical conditions. J Clin Microbiol 2001;39(5):1972-1974.##Albert H, Bwanga F, Mukkada S, Nyesiga B, Ademun JP, Lukyamuzi G, et al. Rapid screening of MDR-TB using molecular line probe assay is feasible in Uganda. BMC Infect Dis 2010;10:41.##Barnard M, Gey van Pittius NC, van Helden PD, Bosman M, Coetzee G, Warren RM. The diagnostic performance of the genoType MTBDRplus version 2 line probe assay is equivalent to that of the Xpert MTB/RIF assay. J Clin Microbiol 2012;50(11):3712-3716.##Melchers WJ, Bakkers JM, Wang J, de Wilde PC, Boonstra H, Quint WG, et al. Short fragment polymerase chain reaction reverse hybridization line probe assay to detect and genotype a broad spectrum of human papillomavirus types. Clinical evaluation and follow-up. Am J Pathol 1999;155(5):1473-1478.##

Expression and Activity of Platelet Endothelial Nitric Oxide Synthase are Decreased in Patients with Coronary Thrombosis and Stenosis 2 2 Background: Nitric Oxide (NO) which is synthesized by endothelial Nitric Oxide Synthase (eNOS) in both vascular tissues and platelets plays an important role as a protective mediator in the cardiovascular system. It modulates blood pressure, vasodilation and thrombosis. In this regard, eNOS activity and gene expression in platelets and NO levels in patients’ plasmas with Coronary Thrombosis (CT) and stenosis diseases were determined.   Methods: Blood samples were collected from 60 subjects that where divided into three equal groups [without coronary artery disease, with CT and less than 70% Coronary Stenosis (CS)]. NO concentration in plasma was measured by the Griess reagent system. The eNOS activity was assessed based on a fluorimetric detection system in platelets and the gene expression was quantified by the real time-reverse transcription-polymerase chain reaction.  Results: There was a significant decrease in the amount of NO concentration in the plasma of subjects with CT (0.53±0.09 µM, p<0.01) and CS (1.31±0.11 µM, p<0.01) compared to the control group (2.6±0.10 µM). The activity levels of eNOS enzyme were significantly lower in patients’ platelets with CT (0.68±0.013 UF/mn, p<0.01) and CS (0.85±0.017 UF/mn, p<0.01) than the control cases (1.29±0.019 UF/mn). These data were consistent with the reduction of the expression levels of eNOS in patients with CT (75 folds) and CS (4 folds) lower than the control cases. Conclusion: Patients with CT and CS possessed reduced eNOS activity and gene expression in their platelets. Decreased plasma concentration of NO in these patients confirmed the potential significance of the diagnostic and prognostic value of NO in the subjects’ plasma with vascular disease risk. 88 93 Zahra Emami Department of Biochemistry, Tarbiat Modares UniversityDepartment of Biochemistry, Faculty of Medicine, Shiraz University of Medical Sciences Department of Biochemistry, Tarbiat Modares UniversityDepartment of Biochemistry, Faculty of Medicine, Shiraz University of Medical Sciences IranIran Alireza Mesbah Namin Department of Biochemistry, Tarbiat Modaress University Department of Biochemistry, Tarbiat Modaress University Iran Javad Kojuri Department of Cardiology, Faculty of Medicine, Shiraz University of Medical Sciences Department of Cardiology, Faculty of Medicine, Shiraz University of Medical Sciences Iran Farideh Mashayekhi Jalali Department of Biochemistry, Faculty of Medicine, Arak University of Medical Sciences Department of Biochemistry, Faculty of Medicine, Arak University of Medical Sciences Iran Mozhgan Rasti Department of Biochemistry, Faculty of Medicine, Shiraz University of Medical Sciences Department of Biochemistry, Faculty of Medicine, Shiraz University of Medical Sciences Iran Cardiovascular diseaseseNOSNitric oxidePlateletsReal time PCRThrombosis 10349.pdf Liu VW, Huang PL. Cardiovascular roles of nitric oxide: a review of insights from nitric oxide synthase gene disrupted mice. Cardiovasc Res 2008;77(1):19-29.##Ciftçi C, Melil S, Cebi Y, Ersoz M, Cagatay P, Kılıcgedik M, et al. Association of endothelial nitric oxide synthase promoter region (T-786C) gene polymorphism with acute coronary syndrome and coronary heart disease. Lipids Health Dis 2008;7:5.##Fleming I, Busse R. Molecular mechanisms involved in the regulation of the endothelial nitric oxide synthase. Am J Physiol Regul Integr Comp Physiol 2003;284(1):R1-12.##Papapetropoulos A, Rudic RD, Sessa WC. Molecular control of nitric oxide synthases in the cardiovascular system. Cardiovasc Res 1999;43(3):509-520.##Shimasaki Y, Saito Y, Yoshimura M, Kamitani S, Miyamoto Y, Masuda I, et al. The effects of long-term smoking on endothelial nitric oxide synthase mRNA expression in human platelets as detected with real-time quantitative RT-PCR. Clin Appl Thromb Hemost 2007;13(1):43-51.##Shah AM. Inducible nitric oxide synthase and cardiovascular disease. Cardiovasc Res 2000;45(1):148-155.##Dashwood MR, Loesch A. Inducible nitric oxide synthase and vein graft performance in patients undergoing coronary artery bypass surgery: physiological or pathophysiological role? Curr Vasc Pharmacol 2014; 12(1):144-151.##Marjanovic JA, Stojanovic A, Brovkovych VM, Skidgel RA, Du X. Signaling-mediated functional activation of inducible nitric-oxide synthase and its role in stimulating platelet activation. J Biol Chem 2008;283(43):28827-28834.##Kumar P, Arora S, Sagar TS, Das N, Bhattacharjee J. Evaluation of endothelial and platelet functions with intergenotypic variation of eNOS Glu 298 Asp gene polymorphism in relation to postmenopausal women. Int J Biol Med Res 2010;4:272-276.##Lauer T, Preik M, Rassaf T, Strauer BE, Deussen A, Feelisch M, et al. Plasma nitrite rather than nitrate reflects regional endothelial nitric oxide synthase activity but lacks intrinsic vasodilator action. Proc Natl Acad Sci US 2001;98(22):12814-12819.##Li Z, Delaney MK, O'Brien KA, Du X. Signaling during platelet adhesion and activation. Arterioscler Thromb Vasc Biol 2010;30(12):2341-2349.##Dosenko VE, Zagoriy VY, Haytovich NV, Gordok OA, Moibenko AA. Allelic polymorphism of endothelial NO-synthase gene and its functional manifestations. Acta Biochim Pol 2006;53(2):299-302.##Jayachandran M, Miller VM. Ovariectomy upregulates expression of estrogen receptors, NOS, and HSPs in porcine platelets. Am J Physiol Heart Circ Physiol 2002;283(1):H220-226.##Randriamboavonjy V, Schrader J, Busse R, Fleming I. Insulin induces the release of vasodilator compounds from platelets by a nitric oxide-G kinase-VAMP-3-dependent pathway. J Exp Med 2004;199(3):347-356.##Teo IA, Choi JW, Morlese J, Taylor G, Shaunak S. LightCycler qPCR optimisation for low copy number target DNA. J Immunol Methods 2002;270(1):119-133.##Kleninbongard P, Dejam A, Lauer T, Rassaf T, Schindler A, Picker O, et al. Plasma nitrite reflects constitutive nitric oxide synthase activity in mammals. Free Radic Biol Med 2003;35(7):790-796.##Schulz R, Rassaf T, Massion PB, Kelm M, Balligand JL. Recent advances in the understanding of the role of nitric oxide in cardiovascular homeostasis. Pharmacolo Ther 2005;108(3):225-256.##Ferreiro CR, Chagas AC, Carvalho MH, Dantas AP, Scavone C, Souza LC, et al. Expression of inducible nitric oxide synthase is increased in patients with heart failure due to ischemic disease. Braz J Med Biol Res 2004;37(9):1313-1320.##Gkaliagkousi E, Ritter J, Ferro A. Platelet-derived nitric oxide signaling and regulation. Circ Res 2007;101(7):654-662.##Alonso D, Radomski MW. Nitric oxide, platelet function, myocardial infarction and reperfusion therapies. Heart Fail Rev 2003;8(1):47-54.##Ikeda H, Takajo Y, Murohara T, Ichiki K, Adachi H, Haramaki N, et al. Platelet-derived nitric oxide and coronary risk factors. Hypertension 2000;35(4):904-907.##Chrapko WE, Juracz P, Radomski MW, Lara N, Archer S, Le Melledo JM. Decreased platelet nitric oxide synthase activity and plasma nitric oxide metabolites in major depressive disorder. Biol Psychiatry 2004;56(2):129-134.##Chu Y, Heistad DD, Knudtson KL, Lamping KG, Faraci FM. Quantification of mRNA for endothelial NO synthase in mouse blood vessels by real-time polymerase chain reaction. Arterioscler Thromb Vasc Biol 2002;22(4):611-616.##Förstermann U, Boissel JP, Kleinert H. Expressional control of the “constitutive” isoforms of nitric oxide synthase (NOS I and NOS III). FASEB J 1998;12(10):773-790.##Uematsu M, Ohara Y, Navas JP, Nishida K, Murphy TJ, Alexander RW, et al. Regulation of endothelial cell nitric oxide synthase mRNA expression by shear stress. Am J Physiol 1995;269(6 Pt):C1371-1378.##Kleinert H, Wallerath T, Euchenhofer C, Ihrig-Biedert I, Li H, Forstermann U. Estrogens increase transcription of the human endothelial NO synthase gene: analysis of the transcription factors involved. Hypertension 1998;31(2):582-588.##Sessa WC, Pritchard K, Seyedi N, Wang J, Hintze TH. Chronic exercise in dogs increases coronary vascular nitric oxide production and endothelial cell nitric oxide synthase gene expression. Circ Res 1994;74(2):349-539.##Moncada S, Higgs A. The L-arginine-nitric oxide pathway. N Engl J Med 1993;329(27):2002-2012.##Kelm M, Rath J. Endothelial dysfunction in human coronary circulation: relevance of the l-arginine-NO-pathway. Basic Res Cardiol 2001;96(2):107-127.##

Identification of Potential Lead Molecules for Zika Envelope Protein from in Silico Perspective 2 2 Background: Zika virus is the family member of flavivirus with no reported clinically approved drugs or vaccines in the market till date. This virus is spread by Aedes mosquitoes, and can also be transmitted through sexual contact or blood transfusions. There are reported medical conditions like microcephaly among new-borns delivered by infected pregnant women. The envelope protein of Zika virus is associated with virulence, tropism, mediation of receptor binding and membrane fusion. ED1-EDII domain (K1 loop pocket) is an integral part of the envelope protein and a potential drug target. In the present study, the purpose was to identify the potential lead molecules to dock against K1 loop which could be later considered as flavivirus entry inhibitors. Methods: Multiple sequence alignment method was considered for the analysis of indels in envelope protein. Phylogenetic tree was constructed based on the alignment. Aliphatic index, GRAVY scores and hydropathy plot of the envelope proteins were calculated for the flavivirus family members. Zika envelope protein was homology modeled and considered for protein-ligand docking analysis with chemical compounds of known functions. Results: As per in silico based analysis, the envelope protein of Zika virus is highly hydrophilic with the least number of amino acid deletions compared to rest of the family members. During docking studies, it was observed that compounds like NITD, compound 6, P02, Doxytetracycline and Rolitetracycline show better binding affinity with Zika envelope protein compared to dengue virus. Conclusion: These better binding compounds could be the promising lead molecules for Zika envelope protein which could better block the viral entry. 94 103 Selvaa Kumar C Faculty of Biotechnology and Bioinformatics, D.Y. Patil Deemed to be University, CBD Belapur Faculty of Biotechnology and Bioinformatics, D.Y. Patil Deemed to be University, CBD Belapur India Shine Devarajan Faculty of Biotechnology and Bioinformatics, D.Y. Patil Deemed to be University, CBD Belapur Faculty of Biotechnology and Bioinformatics, D.Y. Patil Deemed to be University, CBD Belapur India AedesDengue virusEnvelope proteinFlavivirusMicrocephalyZika virus 10340.pdf Dick GW, Kitchen SF, Haddow AJ Zika virus. I. Isolations and serological specificity. Trans R Soc Trop Med Hyg 1952;46(5):509-520.##Dick GW. Zika virus. II. Pathogenicity and physical properties. Trans R Soc Trop Med Hyg 1952;46(5):521-534.##de Oliveira WK, Carmo EH, Henriques CM, Coelho G, Vazquez E, Cortez-Escalante J, et al. Zika virus infection and associated neuroloic disorders in Brazil. N Engl J Med 2017;376(16):1591-1593.##Azari-Hamidian S. Checklist of Iranian mosquitoes (Diptera: Culicidae). J Vector Ecol 2007;32(2):235-242.##Mardani M. Update on Zika virus infections. Arch Clin Infect Dis 2016;11(2):69-71.##Payne S. Family Flaviviridae. Viruses from understanding to investigation. Academic Press; 2017,129-139.##Beltrán-Silva SL,Chacón-Hernández SS, Moreno-Palacios E, Pereyra-Molina JA. Clinical and differential diagnosis: Dengue, chikungunya and Zika. Rev Med Hosp Gen Méx 2018;81(3):146-153.##Malone RW, Homan J, Callahan MV, Glasspool-Malone J, Damodaran L, Schneider Ade B, et al. Zika virus: medical countermeasure development challenges. PLoS Negl Trop Dis 2016;10(3):e0004530.##Sikka V, Chattu VK, Popli RK, Galwankar SC, Kelkar D, Sawicki S G, et al. The emergence of Zika virus as a global health security threat: a review and a consensus statement of the INDUSEM joint working group. J Glob Infect Dis 2016;8(1):3-15.##Oster AM, Brooks JT, Stryker JE, Kachur RE, Mead P, Pesik NT, et al. Interim guidelines for prevention of sexual transmission of Zika virus-United States, 2016. MMWR Morb Mortal Wkly Rep 2016;65(5):120-121.##Schuler-Faccini L, Ribeiro EM, Feitosa IM, Horovitz D D, Cavalcanti DP, Pessoa A, et al. Possible association between Zika virus infection and microcephaly-Brazil. MMWR Morb Mortal Wkly Rep 2015;65(3):59-62.##Logan IS. ZIKA-How fast does this virus mutate? Dongwuxue Yanjiu 2016;37(2):110-115.##Kuno G, Chang GJ. Full-length sequencing and genomic characterization of Bagaza, Kedougou, and Zika viruses. Arch Virol 2007;152(4):687-696.##Enfissi A, Codrington J, Roosblad J, Kazanji M, Rousset D. Zika virus genome from the Americas. Lancet 2016;387(10015):227-228.##Baronti C, Piorkowski G, Charrel RN, Boubis L, Leparc-Goffart I, de Lamballerie X. Complete coding sequence of Zika virus from a French polynesia outbreak in 2013. Genome Announc 2014;2(3):pii:e00500-514.##Bollati M, Alvarez K, Assenberg R, Baronti C, Canard B, Cook S, et al. Structure and functionality in flavivirus NS-proteins: perspectives for drug design. Antiviral Res 2010;87(2):125-148.##Lindenbach BD, Thiel HJ, Rice CM. Flaviviridae: The viruses and their replication. 5th rev.ed.Knipe DM, Howley PM. Fields virology. Philadelphia: Lippincott Williams & Wilkins; 2007. 1101 p.##Stadler K, Allison SL, Schalich J, Heinz FX. Proteolytic activation of tick-borne encephalitis virus by furin. J Virol 1997;71(11):8475-8481.##Speight G, Coia G, Parker MD, Westaway EG. Gene mapping and positive identification of the non-structural proteins NS2A, NS2B, NS3, NS4B and NS5 of the flavivirus Kunjin and their cleavage sites. J Gen Virol 1998;69(Pt 1):23-34.##Zhang X, Ge P, Yu X, Brannan JM, Bi G, Zhang Q, et al. EM structure of the mature dengue virus at 3.5-Å resolution. Nat Struct Mol Biol 2012;20(1):105-110.##Kudelko M, Brault JB, Kwok K, Li MY, Pardigon N, Peiris JS, et al. Class II ADP-ribosylation factors are required for efficient secretion of dengue viruses. J Biol Chem 2012;287(1):767-777.##McMinn PC. The molecular basis of virulence of the encephalogenic flaviviruses. J Gen Virol 1997;78(Pt 11):2711-2722.##Rey FA, Heinz FX, Mandl C, Kunz C, Harrison SC. The envelope glycoprotein from tick-borne encephalitis virus at 2 Å resolution. Nature 1995;375(6529):291-298.##Modis Y, Ogata S, Clements D, Harriso SC. A ligand binding pocket in the dengue virus envelope glycoprotein. Proc Natl Acad Sci USA 2003;100(12):6986-6991.##Zhang Y, Zhang W, Ogata S, Clements D, Strauss JH, Baker TS, et al. Conformational changes of the flavivirus E glycoprotein. Structure 2004;12(9):1607-1618.##Lee E, Lobigs M. Substitutions at the putative receptor-binding site of an encephalitic flavivirus alter virulence and host cell tropism and reveal a role for glycolsaminoglycans in entry. J Virol 2000;74(19):8867-8875.##Seligman SJ, Bucher DJ. The importance of being outer: consequences of the distinction between the outer and inner surfaces of flavivirus glycoprotein. Trends Microbiol 2003;11(3):108-110.##Modis Y, Ogata S, Clements D, Harrison SC. Variable surface epitopes in the crystal structure of dengue virus type 3 envelope glycoprotein. J Virol 2005;79(2):1223-1231.##Modis Y, Ogata S, Clements D, Harrison SC. Structure of the dengue virus envelope protein after membrane fusion. Nature 2004;427(6972):313-319.##Bressanelli S, Stiasny K, Allison SL, Stura EA, Duquerroy S, Lescar J, et al. Structure of a flavivirus envelope glycoprotein in its low-pH induced membrane fusion conformation. EMBO J 2004;23(4):728-738.##Sirohi D, Chen Z, Sun L, Klose T, Pierson TC, Rossmann MG, et al. The 3.8 Å resolution cyro-EM structure of Zika virus. Science 2016;352(6284):467-470.##Stiasny K, Allison SL, Marchler-Bauer A, Kunz C, Heinz FX. Structural requirements for low-pH-induced rearrangements in the envelope glycoprotein of tick-borne encephalitis virus. J Virol 1996;70(11):8142-8147.##Allison SL, Stiasny K, Stadler K, Mandl CW, Heinz FX. Mapping of functional elements in the stem-anchor region of tick-borne encephalitis virus envelope protein E. J Virol 1999;73(7):5605-5612.##Orlinger KK, Hoenninger VM, Kofler RM, Mandl CW. Constructon and mutagenesis of an artificial bicistronic tick-borne encephalitis virus genome reveals an essential function of the second transmembrane region of protein E in flavivirus assembly. J Virol 2006;80(24):12197-12208.##Boutet E, Lieberherr D, Tognolli M, Schneider M, Bansal P, Bridge AJ, et al. UniProtKB/Swiss-Prot, the manually annotated section of the UniProt Knowledge Base: how to use the entry view. Methods Mol Biol 2016;1374:23-54.##Gasteiger E, Hoogland C, Gattiker A, Hoogland C, Gattiker A, Duvaud S, et al. Protein identification and analysis tools on the ExPASy server. In: Walker JM, editor. The proteomics protocols handbook. UK: Humana Press; 2005. p. 571-607.##Argos P, Rossman MG, Grau UM, Zuber H, Frank G, Tratschin JD. Thermal stability and protein structure. Biochemistry 1979;18(25):5698-5703.##Ikai A. Thermostability and aliphatic index of globular proteins. J Biochem 1980;88(6):1895-1898.##Kyte J, Doolittle RF. A simple method for displaying the hydropathic character of a protein. J Mol Biol 1982;157(1):105-132.##Sievers F, Wilm A, Dineen D, Gibson TJ, Karplus K, Li W, et al. Fast, scalable generation of high‐quality protein multiple sequence alignments using Clustal Omega. Mol Syst Biol 2011;7:539.##Dai L, Song J, Lu X, Deng YQ, Musyoki AM, Cheng H, et al. Structures of the Zika virus envelope protein and its complex with a Flavivirus broadly protective antibody. Cell Host Microbe 2016;19(5):696-704.##Zhao H, Fernandez E, Dowd KA, Speer SD, Platt DJ, Gorman MJ, et al. Structural basis of Zika virus-specific antibody protection. Cell 2016;166(4):1016-1027.##Robbiani DF, Bozzacco L, Keeffe JR, Khouri R, Olsen PC, Gazumyan A, et al. Recurrent potent human neutralizing antibodies to Zika virus in Brazil and Mexico. Cell 2017;169(4):597-609.e11.##Wang Q, Yang H, Liu X, Dai L, Ma T, Qi J, et al. Molecular determinants of human neutralizing antibodies isolated from a patient infected with Zika virus. Sci Transl Med 2016;8(369):369ra179.##Guex N, Peitsch MC, Schwede T. Automated comparative protein structure modeling with SWISS-MODEL and Swiss-PdbViewer: a historical perspective. Electrophoresis 2009;30 Suppl 1:S162-173.##Lovell SC, Davis IW, Arendall WB 3rd, de Bakker PI, Word JM, Prisant MG, et al. Structure validation by Calpha geometry: phi,psi and Cbeta deviation. Proteins 2003;50(3):437-450.##Wiederstein M, Sippl MJ. ProSA-web: interactive web service for the recognition of errors in three-dimensional structures of proteins. Nucleic Acids Res 2007;35(web server issue):W407-410.##Maybridge Chemical Company Ltd. Available online: http://www.maybridge.com (accessed on 16 February 2011).##Kampmann T, Yennamalli R, Campbell P, Stoermer MJ, Fairlie DP, Kobe B, et al. In silico screening of small molecule libraries using the dengue virus envelope E protein has identified compounds with antiviral activity against multiple flaviviruses. Antiviral Res 2009;84(3):234-241.##Poh MK, Yip A, Zhang S, Smit JM, Wilschut J, Priestle JP, et al. A small molecule fusion inhibitor of dengue virus. Antiviral Res 2009;84(3):260-266.##Wang QY, Patel SJ, Vangrevelinghe E, Xu HY, Rao R, Jaber D, et al. A small-molecule dengue virus entry inhibitor. Antimicrob Agents Chemother 2009;53(5):1823-1831.##Zhou Z, Khaliq M, Suk JE, Patkar C, Li L, Kuhn RJ, et al. Antiviral compounds discovered by virtual screening of small-molecule libraries against dengue virus E protein. ACS Chem. Biol 2008;3(12):765-775.##Yang JM, Chen YF, Tu YY, Yen KR, Yang YL. Combinatorial computational approaches to identify tetracycline derivatives as flavivirus inhibitors. PLoS One 2007;2(5):e428.##Li Z, Khaliq M, Zhou Z, Post CB, Kuhn RJ, Cushman M. Design, synthesis, and biological 284 evaluation of antiviral agents targeting flavivirus envelope proteins. J Med Chem 2008;51(15):4660-4671.##Morris GM, Huey R, Lindstrom W, Sanner MF, Belew RK, Goodsell DS, et al. AutoDock4 and AutoDock Tools4: automated docking with selective receptor flexibility. J Comput Chem 2009;30(16):2785-2891.##

Prediction of RNA- and DNA-Binding Proteins Using Various Machine Learning Classifiers 2 2 Background: Nucleic acid-binding proteins play major roles in different biological processes, such as transcription, splicing and translation. Therefore, the nucleic acid-binding function prediction of proteins is a step toward full functional annotation of proteins. The aim of our research was the improvement of nucleic-acid binding function prediction. Methods: In the current study, nine machine-learning algorithms were used to predict RNA- and DNA-binding proteins and also to discriminate between RNA-binding proteins and DNA-binding proteins. The electrostatic features were utilized for prediction of each function in corresponding adapted protein datasets. The leave-one-out cross-validation process was used to measure the performance of employed classifiers. Results: Radial basis function classifier gave the best results in predicting RNA- and DNA-binding proteins in comparison with other classifiers applied. In discriminating between RNA- and DNA-binding proteins, multilayer perceptron classifier was the best one. Conclusion: Our findings show that the prediction of nucleic acid-binding function based on these simple electrostatic features can be improved by applied classifiers. Moreover, a reasonable progress to distinguish between RNA- and DNA-binding proteins has been achieved. 104 111 Mehdi Poursheikhali Asghari Department of Biophysics, Faculty of Biological Sciences, Tarbiat Modares University Department of Biophysics, Faculty of Biological Sciences, Tarbiat Modares University Iran Parviz Abdolmaleki Department of Biophysics, Faculty of Biological Sciences, Tarbiat Modares University Department of Biophysics, Faculty of Biological Sciences, Tarbiat Modares University Iran DNA-binding proteinsMachine-learning algorithmsRNA-binding proteins 10341.pdf Castello A, Fischer B, Frese CK, Horos R, Alleaume AM, Foehr S, et al. Comprehensive identification of RNA-binding domains in human cells. Mol Cell 2016;63(4):696-710.##Kloetgen A, Munch PC, Borkhardt A, Hoell JI, McHardy AC. Biochemical and bioinformatic methods for elucidating the role of RNA-protein interactions in posttranscriptional regulation. Brief Funct Genomics 2014;14(2):102-114.##Lunde BM, Moore C, Varani G. RNA-binding proteins: modular design for efficient function. Nat Rev Mol Cell Biol 2007;8(6):479-490.##Cai CZ, Han LY, Ji ZL, Chen X, Chen YZ. SVM-Prot: web-based support vector machine software for functional classification of a protein from its primary sequence. Nucleic Acids Res 2003;31(13):3692-3697.##Cai YD, Lin SL. Support vector machines for predicting rRNA-, RNA-, and DNA- binding proteins from amino acid sequence. Biochim Biophys Acta 2003;1648(1-2):127-133.##Han LY, Cai CZ, Lo SL, Chung MC, Chen YZ. Prediction of RNA-binding proteins from primary sequence by a support vector machine approach. RNA 2004;10(3):355-368.##Yu X, Cao J, Cai Y, Shi T, Li Y. Predicting rRNA-, RNA-, and DNA-binding proteins from primary structure with support vector machines. J Theor Biol 2006;240(2):175-184.##Shao X, Tian Y, Wu L, Wang Y, Jing L, Deng N. Predicting DNA- and RNA- binding proteins from sequences with kernel methods. J Theor Biol 2009;258(2):289-293.##Spriggs RV, Murakami Y, Nakamura H, Jones S. Protein function annotation from sequence: prediction of residues interacting with RNA. Bioinformatics 2009;25 (12):1492-1497.##Kumar M, Gromiha MM, Raghava GPS. SVM based prediction of RNA-binding proteins using binding residues and evolutionary information. J Mol Recognit 2011;24(2):303-313.##Peng CR, Liu L, Niu B, Lv YL, Li MJ, Yuan YL, et al. Prediction of RNA-binding proteins by voting systems. J Biomed Biotechnol 2011;2011:506205.##Agostini F, Zanzoni A, Klus P, Marchese D, Cirillo D, Targatlia GG. catRAPID omics: a web server for large-scale prediction of protein-RNA interactions. Bioinformatics 2013;29(22):2928-2930.##Klus P, Bolognesi B, Agostini F, Marchese D, Zanzoni A, Targatlia GG. The cleverSuite approach for protein characterization: predictions of structural properties, solubility, chaperon requirements and RNA-binding abilities. Bioinformatics 2014;30(11):1601-1608.##Livi CM, Klus P, Delli Ponti R, Targatlia GG. catRAPID signature: identification of ribonucleoproteins and RNA-binding regions. Bioinformatics 2016;32(5):773-775.##Ma X, Guo J, Sun X. Sequence-based prediction of RNA-binding proteins using random forest with minimum redundancy maximum relevance feature selection. Biomed Res Int 2015;2015:425810.##Zhang X, Liu S. RBPPred: predicting RNA-binding proteins from sequence using SVM. Bioinformatics 2017;33(6):854-862.##Sharan M, Forstner KU, Eulalio A, Vogel J. APRICOT: an integrated computational pipeline for the sequence-based identification and characterization of RNA-binding proteins. Nucleic Acids Res 2017;45(11):e96.##Shazman S, Mandel-Gutfreund Y. Classifying RNA-binding proteins based on electrostatic properties. PLoS Comput Biol 2008;4(8):e1000146.##Paz I, Kligun E, Bengud B, Mandel-Gutfreund Y. BindUP: a web server for non-homology-based prediction of DNA and RNA binding proteins. Nucleic Acids Res 2016;44(W1):W568-574.##Zhao H, Yang Y, Zhou Y. Structure-based prediction of RNA-binding domains and RNA-binding sites and application to structural genomics targets. Nucleic Acids Res 2011;39(8):3017-3025.##Zhao H, Yang Y, Zhou Y. Highly accurate and high-resolution function prediction of RNA binding proteins by fold recognition and binding affinity prediction. RNA Biol 2011;8(6):988-996.##Yang Y, Zhan J, Zhao H, Zhou Y. A new size-independent score for pairwise protein structure alignment and its application to structure classification and nucleic-acid binding prediction. Proteins 2012;80(8):2080-2088.##Yang Y, Zhao H, Wang J, Zhou Y. SPOT-Seq-RNA: predicting protein-RNA complex structure and RNA-binding function by fold recognition and binding affinity prediction. Methods Mol Biol 2014;1137:119-130.##Zhao H, Yang Y, Janga SC, Kao CC, Zhou Y. Prediction and validation of the unexplored RNA-binding protein atlas of the human proteome. Proteins 2014;82(4):640-647.##Puton T, Kozlowski L, Tuszynska I, Rother K, Bujnicki JM. Computational methods for prediction of protein-RNA interactions. J Struct Biol 2012;179(3):261-268.##Zhao H, Yang Y, Zhou Y. Prediction of RNA binding proteins comes of age from low resolution to high resolution. Mol BioSyst 2013;9(10):2417-2425.##Si J, Cui J, Cheng J, Wu R. Computational prediction of RNA-binding proteins and binding sites. Int J Mol Sci 2015;16(11):26303-26317.##Jones S, Barker JA, Nobeli I, Thornton JM. Using structural motif templates to identify proteins with DNA binding function. Nucleic Acids Res 2003;31(11):2811-2823.##Ahmad S, Sarai A. Moment-based prediction of DNA-binding proteins. J Mol Biol 2004;341(1):65-71.##Shanahan HP, Garcia MA, Jones S, Thornton JM. Identifying DNA-binding proteins using structural motifs and the electrostatic potential. Nucleic Acids Res 2004;32(16):4732-4741.##Bhardwaj N, Langolis RE, Zhao G, Lu H. Kernel-based machine learning protocol for predicting DNA-binding proteins. Nucleic Acids Res 2005;33(20):6486-6493.##Gao M, Skolnick J. A threading-based method for the prediction of DNA-binding proteins with application to the human genome. PLoS Comput Biol 2009;5(11):e1000567.##Nimrod G, Szilagyi A, Leslie C, Ben-tal N. Identification of DNA-binding proteins using structural, electrostatic and evolutionary features. J Mol Biol 2009;387(4):1040-1053.##Nimrod G, Schushan M, Szilagyi A, Leslie C, Ben-Tal N. iDBPs: a web server for the identification of DNA binding proteins. Bioinformatics 2010;26(5):692-693.##Zhao H, Yang Y, Zhou Y. Structure-based prediction of DNA-binding proteins by structural alignment and a volume-fraction corrected DFIRE-based energy function. Bioinformatics 2010;26(15):1857-1863.##Zhao H, Wang J, Zhou Y, Yang Y. Predicting DNA-binding proteins and binding residues by complex structure prediction and application to human proteome. PLoS One 2014;9(5):e96694.##Kumar M, Gromiha MM, Raghava GPS. Identification of DNA-binding proteins using support vector machines and evolutionary profiles. BMC Bioinformatics 2007;8:463.##Fang Y, Guo Y, Feng Y, Li M. Predicting DNA-binding proteins: approached from Chou s pseudo amino acid composition and other specific sequence features. Amino Acids 2008;34(1):103-109.##Nani L, Lumini A. Combining ontologies and dipeptide composition for predicting DNA-binding proteins. Amino Acids 2008;34(4):635-641.##Kumar KK, Pugalenthi G, Suganthan PN. DNA-Prot: Identification of DNA binding proteins from protein sequence information using random forest. J Biomol Struct Dyn 2009;26(6):679-686.##Nani L, Lumini A. An ensemble of reduced alphabets with protein encoding based on grouped weight for predicting DNA-binding proteins. Amino Acids 2009;36 (2):167-175.##Lin WZ, Fang JA, Xiao X, Chou KC. iDNA-Prot: identification of DNA binding proteins using random forest with grey model. PLoS One 2011;6(9):e24756.##Zou C, Gong J, Li H. An improved sequence based prediction protocol for DNA-binding proteins using SVM and comprehensive feature analysis. BMC Bioinformatics 2013;14:90.##Song L, Li D, Zeng X, Wu Y, Guo L, Zou Q. nDNA-prot: identification of DNA-binding proteins based on unbalanced classification. BMC Bioinformatics 2014;15:298.##Xu R, Zhou J, Liu B, Yao L, He Y, Zou Q, et al. enDNA-Prot: identification of DNA-binding proteins by applying ensemble learning. Biomed Res Int 2014;2014:294279.##Liu B, Xu J, Lan X, Xu R, Zhou J, Wang X, et al. iDNA-Protdis: identifying DNA-binding proteins by incorporating amino acid distance-pairs and reduced alphabet profile into the general pseudo amino acid composition. PLoS One 2014;9(9):e106691.##Lou W, Wang X, Chen F, Chen Y, Jiang B, Zhang H. Sequence based prediction of DNA-binding proteins based on hybrid feature selection using random forest and Gaussian Naïve Bayes. PLoS One 2014;9(1):e86703.##Niu XH, Hu XH, Shi F, Xia JB. Predicting DNA binding proteins using support vector machine with hybrid fractal features. J Theor Biol 2014;343:186-192.##Zhang Y, Xu J, Zheng W, Zhang C, Qiu X, Chen K, et al. newDNA-Prot: Prediction of DNA-binding proteins by employing support vector machine and a comprehensive sequence representation. Comput Biol Chem 2014;52:51-59.##Liu B, Xu J, Fan S, Xu R, Zhou J, Wang X. PseDNA-Pro: DNA-binding protein identification by combining Chou’s PseAAC and physicochemical distance transformation. Mol Inform 2015;34(1):8-17.##Xu R, Zhou J, Wang H, He Y, Wang X, Liu B. Identifying DNA-binding proteins by combining support vector machine and PSSM distance transformation. BMC Syst Biol 2015;9 Suppl 1:S10.##Zhang J, Gao B, Chai H, Ma Z, Yang G. Identification of DNA-binding proteins using multi-features fusion and binary firefly optimization algorithm. BMC Bioinformatics 2016;17(1):323.##Ahmad S, Sarai A. Analysis of electric moments of RNA-binding proteins: implications for mechanism and prediction. BMC Struct Biol 2011;11:8.##Kukic P, Nielsen JE. Electrostatics in proteins and protein-ligand complexes. Future Med Chem 2010;2(4):647-666.##Hall M, Frank E, Holmes G, Pfahringer B, Reutemann P, Witten IH. The WEKA data mining software: An update. SIGKDD Explor 2009;11(1):10-18.##Demir-Kavuk O, Kamada M, Akutsu T, Knapp EW. Prediction using step-wise L1, L2 regularization and feature selection for small data sets with large number of features. BMC Bioinformatics 2011;12:412.##Asadabadi EB, Abdolmaleki P. Predictions of protein-protein interfaces within membrane protein complexes. Avicenna J Med Biotechnol 2013;5(3):148-157.##Sing T, Sander O, Beerenwinkel N, Lengauer T. ROCR: visualizing classifier performance in R. Bioinformatics 2005;21(20):3940-3941.##R-package Development Core Team, R: a language and environment for statistical computing. R Foundation for Statistical Computing 2013, Vienna, Austria. URL http://www.R-project.org/.##

Enhancing Stability of Destabilized Green Fluorescent Protein Using Chimeric mRNA Containing Human Beta-Globin 5′ and 3′ Untranslated Regions 2 2 Background: In spite of recent progress in mRNA technologies and their potential applications for treatment of human diseases, problems such as the transient nature of mRNA limit the stability of gene up-regulation and, thus, potentially reduce mRNA efficiency for gene therapy. Using human β-globin 5′ and 3′ untranslated regions (UTRs), this study aimed to develop the different chimeric constructs of mRNAs to increase the stability of destabilized green fluorescent protein (EGFPd2) in HEK 293 cells.  Methods: Purified human β-globin (HBG) 5′-3′UTRs, and the coding sequence of destabilized green fluorescent protein (EGFPd2) were amplified separately and ligated to each other using SOEing PCR method in a different format. As controls, the original construct of EGFPd2 under the control of T7 promoter was used. Following in vitro transcription, HEK 293 cells were then transfected with several constructs and incubated at 37oC in a CO2 incubator. They were monitored under a fluorescence microscope every four hours for the first 24 hr, then every 12 hr afterwards. The resulting fluorescence was measured as a surrogate for translation efficiency and duration.  Results: By monitoring the HEK cells over 48 hr, cells transfected with mRNA with various HBG UTRs showed significantly different fluorescence intensity and stability in comparison with the pEGFPd2 prototype (control transcript) overtime. Overall, the images show that replacement of the 3′ UTR end of the prototype vector pGFPd2 with the 3′ end of β- globin mRNA increases the half-life of the chimeric mRNA for more than 32 hr.  Conclusion: This result indicates that β-globin 3′ UTR would definitely increase the half-life of mRNA and may help to decrease the mRNA therapeutic dosage in the treatment of diseases associated with mRNA therapy. 112 117 Setare Adibzadeh Diagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical SciencesDepartment of Medical Biotechnology, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences Diagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical SciencesDepartment of Medical Biotechnology, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences IranIran Majid Fardaei Department of Genetics, Faculty of Medicine, Shiraz University of Medical Sciences Department of Genetics, Faculty of Medicine, Shiraz University of Medical Sciences Iran Mohammad Ali Takhshid Diagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences Diagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences Iran Mohammad Reza Miri Diagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences Diagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences Iran Gholam Reza Rafiei Dehbidi Diagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences Diagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences Iran Ali Farhadi Diagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences Diagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences Iran Reza Ranjbaran Diagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences Diagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences Iran Parnian Alavi Diagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences Diagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences Iran Negin Nikouyan Diagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences Diagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences Iran Noorossadat Seyyedi Diagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences Diagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences Iran Samaneh Naderi Diagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences Diagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences Iran Alireaz Eskandari Diagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences Diagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences Iran Abbas Behzad-Behbahani Diagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences Diagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences Iran Beta-globinsGenetic therapyGreen flourescent proteinsHalf-lifemRNA 335.pdf Ginn SL, Alexander IE, Edelstein ML, Abedi MR, Wixon J. Gene therapy clinical trials worldwide to 2012–an update. J Gene Med 2013;15(2):65-77.##Mehta A, Haber JE. Sources of DNA double-strand breaks and models of recombinational DNA repair. Cold Spring Harbor Perspect Biol 2014;6(9):a016428.##Rabinovich PM, Komarovskaya ME, Ye ZJ, Imai C, Campana D, Bahceci E, et al. Synthetic messenger RNA as a tool for gene therapy. Hum Gene Ther 2006;17(10):1027-1035.##Matsui A, Uchida S, Ishii T, Itaka K, Kataoka K. Messenger RNA-based therapeutics for the treatment of apoptosis-associated diseases. Sci Rep 2015;5:15810.##Haimovich G, Medina DA, Causse SZ, Garber M, Millán-Zambrano G, Barkai O, et al. Gene expression is circular: factors for mRNA degradation also foster mRNA synthesis. Cell 2013;153(5):1000-1011.##Carlevaro-Fita J, Rahim A, Guigó R, Vardy LA, Johnson R. Cytoplasmic long noncoding RNAs are frequently bound to and degraded at ribosomes in human cells. RNA 2016;22(6):867-882.##Selinger DW, Saxena RM, Cheung KJ, Church GM, Rosenow C. Global RNA half-life analysis in Escherichia coli reveals positional patterns of transcript degradation. Genome Res 2003;13(2):216-223.##Pedersen M, Nissen S, Mitarai N, Svenningsen SL, Sneppen K, Pedersen S. The functional half-life of an mRNA depends on the ribosome spacing in an early coding region. J Mol Biol 2011;407(1):35-44.##Sharova LV, Sharov AA, Nedorezov T, Piao Y, Shaik N, Ko MS. Database for mRNA half-life of 19 977 genes obtained by DNA microarray analysis of pluripotent and differentiating mouse embryonic stem cells. DNA Res 2009;16(1):45-58.##Walsh A, Tock M, Mallen M, Kaberdin V, Gabain Av, McDowall K. Cleavage of poly (A) tails on the 3′-end of RNA by ribonuclease E of Escherichia coli. Nucleic Acids Res 2001;29(9):1864-1871.##Dreyfus M, Régnier P. The poly (A) tail of mRNAs: bodyguard in eukaryotes, scavenger in bacteria. Cell 2002;111(5):611-613.##Ysla RM, Wilson GM, Brewer G. Assays of adenylate uridylate-rich element-mediated mRNA decay in cells. Methods Enzymol 2008;449:47-71.##Hitti E, Khabar K. Sequence variations affecting AU-rich element function and disease. Front Biosci (Landmark Ed) 2011;17:1846-1860.##Wu X, Bartel DP. Widespread influence of 3′-end structures on mammalian mRNA processing and stability. Cell 2017;169(5):905-917.e11.##Peixeiro I, Silva AL, Romão L. Control of human β-globin mRNA stability and its impact on beta-thalassemia phenotype. Haematologica 2011;96(6):905-913.##Chen CY, Ezzeddine N, Shyu AB. Messenger RNA half‐life measurements in mammalian cells. Methods Enzymol 2008;448:335-357.##Hocine S, Singer RH, Grünwald D. RNA processing and export. Cold Spring Harb Perspect Biol 2010;2(12):a000752.##Charlesworth A, Meijer HA, de Moor CH. Specificity factors in cytoplasmic polyadenylation. Wiley Interdiscip Rev RNA 2013;4(4):437-461.##Halbeisen R, Galgano A, Scherrer T, Gerber A. Post-transcriptional gene regulation: from genome-wide studies to principles. Cell Mol Life Sci 2008;65(5):798-813.##Li X, Zhao X, Fang Y, Jiang X, Duong T, Fan C, et al. Generation of destabilized green fluorescent protein as a transcription reporter. J Biol Chem 1998;273(52):34970-34975.##Matsuda T, Cepko CL. Controlled expression of transgenes introduced by in vivo electroporation. Proc Natl Acad Sci USA 2007;104(3):1027-1032.##Miri M, Behzad-Behbahani A, Fardaei M, Farhadi A, Talebkhan Y, Mohammadi M, et al. Construction of bacterial ghosts for transfer and expression of a chimeric hepatitis C virus gene in macrophages. J Microbiol Methods 2015;119:228-232.##Misra S. Human gene therapy: a brief overview of the genetic revolution. J Assoc Physicians India 2013;61(2):127-133.##Moore MJ. From birth to death: the complex lives of eukaryotic mRNAs. Science 2005;309(5740):1514-1518.##Russell J, Liebhaber SA. The stability of human beta-globin mRNA is dependent on structural determinants positioned within its 3'untranslated region. Blood 1996;87(12):5314-5323.##Yu J, Russell JE. Structural and functional analysis of an mRNP complex that mediates the high stability of human β-globin mRNA. Mol Cell Biol 2001;21(17):5879-5888.##Hino M, Ito H, Yamashiro Y, Hattori Y, Nitta T, Adhiyanto C. The+1,506 (A>C) mutation in the 3′ untranslated region affects β-globin expression. Hemoglobin 2012;36(4):399-406.##

Development of One-Step Tetra-primer ARMS-PCR for Simultaneous Detection of the Angiotensin Converting Enzyme (ACE) I/D and rs4343 Gene Polymorphisms and the Correlation with CAD Patients 2 2 Background: The Angiotensin Converting Enzyme (ACE) Insertion/Deletion and rs4343 gene polymorphisms could be associated with pathogenesis of essential hypertension and cardiovascular disorders and Coronary Artery Disease (CAD). In the present study, a fast and novel approach of multiplex Tetra-Primer Amplification Refractory Mutation System-PCR (T-ARMS-PCR) was developed for simultaneous detection of two SNPs including ACE I/D (rs4340) and 2350A>G (rs4343) of Angiotensin Converting Enzyme (ACE) gene.  Methods: The present research was performed using 148 blood samples taken from patients with CAD and 135 healthy individuals. One set of inner primers (for rs4343) and one set of outer primer pairs were designed for genotyping of Insertion/Deletion and rs4343 polymorphisms in single tube T-ARMS-PCR.  Results: Our results manifested that genotypes and alleles frequency of the ACE polymorphisms showed no statistically significant association between CAD patients and the control group. In addition, complete concordance was seen between sensitive Tetra-ARMS-PCR and sequencing method.  Conclusion: The technique is the first work for simultaneous detection of Insertion/Deletion polymorphism and rs4343 SNPs in ACE gene and the results were entirely according to those from an independent procedure. 118 123 Mohammad Mehdi Heidari Department of Biology, Faculty of Science, Yazd University Department of Biology, Faculty of Science, Yazd University Iran Mehdi Hadadzadeh Department of Cardiac Surgery, Afshar Hospital, Shahid Sadoughi University of Medical Sciences Department of Cardiac Surgery, Afshar Hospital, Shahid Sadoughi University of Medical Sciences Iran Hossein Fallahzadeh Research Center of Prevention and Epidemiology of Non-Communicable Disease, Shahid Sadoughi University of Medical Sciences Research Center of Prevention and Epidemiology of Non-Communicable Disease, Shahid Sadoughi University of Medical Sciences Iran Angiotensin converting enzyme (ACE) geneCoronary artery diseaseInsertion/ Deletion polymorphism 10365.pdf Ferrari R, Fox K. Heart rate reduction in coronary artery disease and heart failure. Nat Rev Cardiol 2016;13(8):493-501.##Ganesh SK, Arnett DK, Assimes TL, Basson CT, Chakravarti A, Ellinor PT, et al. Genetics and genomics for the prevention and treatment of cardiovascular disease: update: a scientific statement from the American Heart Association. Circulation 2013;128(25):2813-2851.##Heidari MM, Khatami M, Hadadzadeh M, Kazemi M, Mahamed S, Malekzadeh P, et al. Polymorphisms in NOS3, MTHFR, APOB and TNF-alpha genes and risk of coronary atherosclerotic lesions in Iranian patients. Res Cardiovasc Med 2016;5(1):e29134.##Khatami M, Heidari MM, Hadadzadeh M, Scheiber-Mojdehkar B, Bitaraf Sani M, Houshmand M. Simultaneous genotyping of the rs4762 and rs699 polymorphisms in angiotensinogen gene and correlation with Iranian CAD patients with novel Hexa-primer ARMS-PCR. Iran J Public Health 2017;46(6):811-819.##Zhu X, Bouzekri N, Southam L, Cooper RS, Adeyemo A, McKenzie CA, et al. Linkage and association analysis of angiotensin I-converting enzyme (ACE)-gene polymorphisms with ACE concentration and blood pressure. Am J Hum Genet 2001;68(5):1139-1148.##Wollinger LM, Dal Bosco SM, Rempe C, Almeida SE, Berlese DB, Castoldi RP, et al. Role of ACE and AGT gene polymorphisms in genetic susceptibility to diabetes mellitus type 2 in a Brazilian sample. Genet Mol Res 2015;14(4):19110-19116.##Firouzabadi N, Tajik N, Bahramali E, Bakhshandeh H, Maadani M, Shafiei M. Gender specificity of a genetic variant of angiotensin-converting enzyme and risk of coronary artery disease. Mol Biol Rep 2013;40(8):4959-4965.##Salonen R, Nyyssonen K, Porkkala E, Rummukainen J, Belder R, Park JS, et al. Kuopio atherosclerosis prevention study (KAPS). a population-based primary preventive trial of the effect of LDL lowering on atherosclerotic progression in carotid and femoral arteries. Circulation 1995;92(7):1758-1764.##Vargas-Alarcon G, Zamora J, Sanchez-Garcia S, Rodriguez-Perez JM, Cardoso G, Posadas-Romero C. Angiotensin-I-converting enzyme (ACE) insertion/ deletion polymorphism in Mexican patients with coronary artery disease. Association with the disease but not with lipid levels. Exp Mol Pathol 2006;81(2):131-135.##Bouzekri N, Zhu X, Jiang Y, McKenzie CA, Luke A, Forrester T, et al. Angiotensin I-converting enzyme polymorphisms, ACE level and blood pressure among Nigerians, Jamaicans and African-Americans. Eur J Hum Genet 2004;12(6):460-468.##Saddick SY. Angiotensin converting enzyme gene polymorphism studies: a case-control study. Saudi J Biol Sci 2015;22(3):327-331.##Ancelin ML, Carriere I, Scali J, Ritchie K, Chaudieu I, Ryan J. Angiotensin-converting enzyme gene variants are associated with both cortisol secretion and late-life depression. Transl Psychiatry 2013;3:e322.##Poorgholi L, Saffar H, Fathollahi MS, Davoodi G, Anvari MS, Goodarzynejad H, et al. Angiotensin- converting enzyme insertion/deletion polymorphism and its association with coronary artery disease in an Iranian population. J Tehran Heart Cent 2013;8(2):89-94.##Sameer AS, Syeed N, Tak SA, Bashird S, Nissar S, Siddiq MA. ACE I/D polymorphism in hypertensive patients of Kashmiri population. Cardiol Res 2010;1(1):1-7.##Rigat B, Hubert C, Corvol P, Soubrier F. PCR detection of the insertion/deletion polymorphism of the human angiotensin converting enzyme gene (DCP1) (dipeptidyl carboxypeptidase 1). Nucleic Acids Res 1992;20(6):1433.##Jia EZ, Xu ZX, Guo CY, Li L, Gu Y, Zhu TB, et al. Renin-angiotensin-aldosterone system gene polymorphisms and coronary artery disease: detection of gene-gene and gene-environment interactions. Cell Physiol Biochem 2012;29(3-4):443-452.##Abdollahi MR, Huang S, Rodriguez S, Guthrie PA, Smith GD, Ebrahim S, et al. Homogeneous assay of rs4343, an ACE I/D proxy, and an analysis in the British women's heart and health study (BWHHS). Dis Markers 2008;24(1):11-17.##Jiang MH, Su YM, Tang JZ, Shen YB, Deng XT, Yuan DS, et al. Angiotensin-converting enzyme gene 2350 G/A polymorphism and susceptibility to atrial fibrillation in Han Chinese patients with essential hypertension. Clinics (Sao Paulo) 2013;68(11):1428-1432.##

Triploidy and Routine Combined First Trimester Pregnancy Screening 2 2 Background: This report is about a pregnancy with a triploid fetus and underscores the potential of first trimester combined screening to detect this devastating chromosomal aberration earlier in pregnancy. This report is about a pregnancy with a triploid fetus identified from the first trimester combined screening and confirmed by amniocentesis.  Methods: A 28 year old, G5P2AB2 woman was referred to our clinic at 15 weeks of gestation due to a remarkable decrease of her first trimester double biochemical markers and therefore in the high-risk range for trisomy 13 and 18. The woman underwent amniocentesis which revealed a karyotype of 69,XXX. The parents opted for termination and in post mortem physical examination, a hydrocephalus fetus with marked Intra-Uterine Growth Retardation (IUGR) in addition to syndactyly of third and fourth digits, low set malformed ears, micrognathia and club foot, was seen.  Results: Our results and previous reports highlight the need to consider a somewhat consistent pattern of the first trimester combined screening in a pregnancy with triploidy and underscore the potential of this screening strategy to detect this chromosomal aberration earlier in pregnancy.  Conclusion: Early prenatal diagnosis of this syndrome would provide women an opportunity to terminate an affected pregnancy earlier. This is also important in preventing the risks of associated later induced abortion or obstetric complications. 124 126 Mitra Eftekhariyazdi Department of Obstetrics and Gynecology, Faculty of Medicine, Sabzevar University of Medical Sciences Department of Obstetrics and Gynecology, Faculty of Medicine, Sabzevar University of Medical Sciences Iran Ali Khaligh Sabzevar University of Medical Sciences Sabzevar University of Medical Sciences Iran Behnaz Suizi Department of Obstetrics and Gynecology, Faculty of Medicine, Sabzevar University of Medical Sciences Department of Obstetrics and Gynecology, Faculty of Medicine, Sabzevar University of Medical Sciences Iran Maryam Naghibi Nasab Department of Obstetrics and Gynecology, Faculty of Medicine, Sabzevar University of Medical Sciences Department of Obstetrics and Gynecology, Faculty of Medicine, Sabzevar University of Medical Sciences Iran Davood Zare-Abdollahi Genetics Research Center, University of Social Welfare and Rehabilitation Sciences Genetics Research Center, University of Social Welfare and Rehabilitation Sciences Iran Fetal growth retardationHydrocephalusNuchal translucency measurementSyndactylyTriploidy 10362.pdf Baer RJ, Flessel MC, Jelliffe-Pawlowski LL, Goldman S, Hudgins L, Hull AD, et al. Detection rates for aneuploidy by first-trimester and sequential screening. Obstet Gynecol 2015;126(4):753-759.##Engelbrechtsen L, Brøndum-Nielsen K, Ekelund C, Tabor A, Skibsted L; Danish Fetal Medicine Study Group. Detection of triploidy at 11-14 weeks' gestation: a cohort study of 198 000 pregnant women. Ultrasound Obstet Gynecol 2013;42(5):530-535.##Kazerouni NN, Currier RJ, Flessel M, Goldman S, Hennigan C, Hodgkinson C, et al. Detection rate of quadruple-marker screening determined by clinical follow-up and registry data in the statewide California program, July 2007 to February 2009. Prenat Diagn 2011;31(9):901-906.##Canick J. Prenatal screening for trisomy 21: recent advances and guidelines. Clin Chem Lab Med 2012;50(6):1003-1008.##Metcalfe A, Hippman C, Pastuck M, Johnson JA. Beyond trisomy 21: Additional chromosomal anomalies detected through routine aneuploidy screening. J Clin Med 2014;3(2):388-415.##Jauniaux E, Brown R, Snijders RJ, Noble P, Nicolaides KH. Early prenatal diagnosis of triploidy. Am J Obstet Gynecol 1997;176(3):550-554.##Daniel A, Wu Z, Bennetts B, Slater H, Osborn R, Jackson J, et al. Karyotype, phenotype and parental origin in 19 cases of triploidy. Prenat Diagn 2001;21(12):1034-1048.##Tørring N. First trimester combined screening - focus on early biochemistry. Scand J Clin Lab Invest 2016;76(6):435-447.##Jauniaux E, Brown R, Snijders RJ, Snijders RJ, Noble P, Nicolaides KH. Early prenatal diagnosis of triploidy. Am J Obstet Gynecol 1997;176(3):550-554.##Ferguson-Smith MA, Yates JR. Maternal age specific rates for chromosome aberrations and factors influencing them: report of a collaborative european study on 52965 amniocenteses. Prenat Diagn. 1984;4 Spec No:5-44.##Toufaily MH, Roberts DJ, Westgate MN, Holmes LB. Triploidy: Variation of phenotype. Am J Clin Pathol. 2016;145(1):86-95.##

Retraction: The Association of PON1 192 Q/R Polymorphism with the Risk of Idiopathic Male Infertility in Northern Iran 2 2 The Editorial Board of Avicenna Journal of Medical Biotechnology (AJMB) has decided to retract the original article entitled "The Association of PON1 192 Q/R Polymorphism with the Risk of Idiopathic Male Infertility in Northern Iran" published in the October-December 2018 issue due to a fact which is contrary to the scientific rules and regulation of AJMB. 127 127 Setareh Behrouzi Department of Biology, Faculty of Sciences, University of Guilan Department of Biology, Faculty of Sciences, University of Guilan Iran Farhad Mashayekhi Department of Biology, Faculty of Sciences, University of Guilan Department of Biology, Faculty of Sciences, University of Guilan Iran Mohammad Hadi Bahadori Department of Biology, Faculty of Sciences, University of Guilan Department of Biology, Faculty of Sciences, University of Guilan Iran Editorial 10388.pdf XXXX.XXXXC.NNNN.##