Determination of Hepatitis B Virus Genotypes in Yazd, Central Province of Iran 2 2 Background and Aims: Hepatitis B virus (HBV) infects the liver and causes acute and chronic hepatitis, hepatocellular carcinoma and cirrhosis. HBV has been divided to eight genotypes (A–H) and subgenotypes of A, B, C and F. For the first time, we determined HBV genotypes in infected samples by INNO-LiPA method in Yazd, central province of Iran. Materials and Methods: This study was performed on samples suspected of HBV infection. The sera of fifteen out of ninety-five samples that had shown positive results by RT-PCR were used for HBV genotyping by using INNO-LiPA HBV genotyping assay. Results and Conclusions: Seven (46.7%) out of fifteen samples were female. The mean age of the patients was 37.8±14.3 years. The average number copy of HBV in infected samples was 1.04×106±4.74×105 Copies/ml. All fifteen infected samples had genotype D. Our results showed that HBV genotype D was the only detectable genotype in Yazd, central province of Iran. A further study with a larger sample size in different provinces of Iran is needed to identify HBV genotypes in Iran. 81 86 2015/08/18 1394/5/27 2015/08/18 1394/5/27 Hossein Hadinedoushan Hossein Hadinedoushan Immunology Department, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. Immunology Department, Reproductive Immunology Research Center, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. Iran hhadi_2000@yahoo.com Mahdi Dehghan-Manshadi Mahdi Dehghan-Manshadi Immunology Department, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran Iran Fateme Zare Fateme Zare Immunology Department, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran Iran HBV Genotyping Hepatitis B Virus INNO-LIPA [1]. Lee WM. Hepatitis B virus infection. New England journal of medicine 1997;337(24):1733-45.##[2]. Shepard CW, Simard EP, Finelli L, Fiore AE, Bell BP. Hepatitis B virus infection:epidemiology and vaccination. Epidemiologic reviews 2006; 28(1):112-25.##[3]. Kumar A. Innate immune responses in hepatitis B virus (HBV) infection. Virologyjournal 2014; 11(1):22.##[4]. Osiowy C, Giles E. Evaluation of the INNO-LiPA HBV genotyping assay fordetermination of hepatitis B virus genotype. Journal of clinical microbiology 2003; 41(12):5473-477.##[5]. Kramvis A, Kew M, François G. Hepatitis B virus genotypes. Vaccine 2005; 23(19):2409-423.##[6]. Attaullah S, Rehman Su, Khan S, Ali I, Ali S, Khan SN. Prevalence of hepatitis B virusgenotypes in HBsAg positive individuals of Afghanistan. Virol J. 2011; 8:281.##[7]. Ali M, Idrees M, Ali L, Hussain A, Ur Rehman I, Saleem S et al. Hepatitis B virusin Pakistan: A systematic review of prevalence, risk factors, awareness status and genotypes. Virol J 2011; 8: 102.##[8]. Atalay MA, Gokahmetoglu S, Aygen B. Genotypes of hepatitis B virus in CentralAnatolia, Kayseri, Turkey. Saudi Med J. 2011; 32(4):360-63.##[9]. Khaled IAA, Mahmoud OM, Saleh AF, Baioumi EA. Prevalence of HBVGenotypes in Egypt among Hepatitis Patients. Journal of American Science 2010; 6(11):185-90.##[10]. Zhu CT, Dong CL. Characteristics of general distribution of hepatitis B virusgenotypes in China. Hepatobiliary Pancreat Dis Int 2009; 8(4):397-401.##[11]. Norder H, Couroucé A-M, Coursaget P, Echevarria J, M e, Lee S-D et al. Geneticdiversity of hepatitis B virus strains derived worldwide: genotypes, subgenotypes, and HBsAg subtypes. Intervirology 2004; 47(6):289-309.##[12]. Hussain M, Chu C-J, Sablon E, Lok AS. Rapid and sensitive assays for determination of hepatitis B virus (HBV) genotypes and detection of HBV precore and core promoter variants. Journal of clinical microbiology 2003; 41(8): 3699-705.## ##

Determining the Prevalence and Detection of the Most Prevalent Virulence Genes in Acinetobacter Baumannii Isolated From Hospital Infections 2 2 Background and Aims: Acinetobacter baumannii is mostly a cause of septicaemia, pneumonia and urinary tract infections following hospitalization of patients with more severe illnesses. The aim of this study was to determine the prevalence and detection of the most prevalent virulence genes in A. baumannii isolated from hospital infections of two largest hospitals of Tehran, Iran. Materials and Methods: In this cross-sectional study, 500 clinical specimens were obtained from various types of hospital infections over a period of 6 month, consisting of blood (98 samples), phlegm (141), urine (92), pus (134) and cerebrospinal fluid (35) from patients admitted to the Payambaran and Baqiyatallah Hospitals in Tehran. The isolated organisms were identified based on colony morphology, microscopic characteristics and various biochemical tests according to some standard laboratory methods. Conventional polymerase chain reaction (PCR) was employed to confirm the identity of the isolates. Results: A. baumannii was isolated from 121 (24.2%) of the 500 cultured samples. The highest isolation of A. baumannii was observed in blood samples while cerebrospinal fluid had the least. The isolation rate recorded for blood samples in this study was 43.87%. fimH gene was the most virulence gene detected in 74% of samples, sfa/focDE and afa/draBC genes were next highly detected genes, respectively. The lowest isolations were observed in PapG III, papC and PapG II genes. Conclusions: High prevalence of A. baumannii and their virulence genes showed hospital prevalence of these bacteria, thereby causing many problems for infections control and treatment. Therefore, determining this bacterium by molecular methods and designing conservation programs for the control of different infections in parts of the hospital seems to be urgently needed. 87 97 2015/08/182015/08/18 1394/5/27 2015/08/182015/08/18 1394/5/27 Hassan Momtaz Hassan Momtaz Department of Microbiology, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran Iran Seyed Morteza Seifati Seyed Morteza Seifati Department of Microbiology, Ashkezar Branch, Islamic Azad University, Ashkezar, Iran. Iran Marziyeh Tavakol Marziyeh Tavakol Department of Microbiology, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran Iran marziyeh.tavakol@yahoo.com Acinetobacter baumannii Hospital infections Virulence genes 1. Abdel-El-Haleem D. Acinetobacter: environmental and biotechnological applications. Afr J Biotechnol. 2003; 2(4):71-75.##2. Pantophlet R, Brade L, Brade H. Use of a murine O-antigen-specific monoclonal antibody to identify Acinetobacter strains of unnamed genomicspecies 13 Sensu Tjernberg and Ursing. J Clin Microbiol.1999; 37(6):1693-698.##3. Espinal P, Roca I, Vila J. Clinical impact and molecular basis of antimicrobial resistance in non-baumannii Acinetobacter. Future Microbiol. 2011; 6(5):495-511.##4. Nemec A, Krizova L, Maixnerova M, van der Reijden TJ, Deschaght P, Passet V et al. Genotypic and phenotypic characterization of the Acinetobacter calcoaceticus-Acinetobacter baumannii complexwith the proposal of Acinetobacter pittii sp. nov. (FormerlyAcinetobacter genomic species3) andAcinetobacternosocomialissp. nov. (Formerly Acinetobacter genomic species 13TU). Res Microbiol. 2011; 162(4):393-404.##5. Gerner-Smidt P, Tjernberg I, Ursing J.Reliability of phenotypic tests for identification of Acinetobacter species. J Clin Microbiol. 1991; 29(2):277-282.##6. 6- [6]. Pendleton JN, Gorman SP, Gilmore BF. Clinical relevance of the ESKAPE pathogens. Expert Rev Anti Infect Ther. 2013; 11(3):297-308.##7. 7- [7].Ong CW, Lye DC, Khoo KL, Chua GS, Yeoh SF, Leo YS, et al. Severe community-acquired Acinetobacter baumannii pneumonia: an emerging highly lethal infectious diseasein the Asia-Pacific. Respirology 2009; 8- 14(8):1200-205.##8. Leung WS, Chu CM, Tsang KY, Lo FH, Lo KF, Ho PL. Fulminant community-acquired Acinetobacter baumannii pneumonia as a distinct clinical syndrome. Chest 2006;129(1):102-109.##9. Anstey NM, Currie BJ, Hassell M, Palmer D, Dwyer B, Seifert H. Community acquired bacteremic Acinetobacter pneumonia in tropical Australia is caused by diverse strains of Acinetobacter baumannii, with carriage in the throat in at-risk groups. J Clin Microbiol. 2002; 40(2):685-86.##10. Jones RN. Microbial etiologies of hospital-acquired bacterial pneumonia andventilator-associated bacterial pneumonia. Clin Infect Dis. 2010; 51:81-87.##11. Vincent JL, Rello J, Marshall J, Silva E, Anzueto A, Martin CD, et al. International 15- study of the prevalence and outcomes of infection in intensive care units. JAMA 2009; 302(21):2323-329.##12. Dorsey CW, Beglin MS, Actis LA. Detection and analysis of iron uptakecomponents expressed by Acinetobacter baumannii clinical isolates. J Clin Microbiol. 2003; 41(9):4188-193.##13. Schreckenberger PC, Graevenitz A. Nonfermentative gramnegative rods. In PRMurray, Manual of Clinical Microbiology, ASM, Washington 1999:539-543.##14. Yun HC, Branstetter JG, Murray CK. Osteomyelitis in military personnel woundedin Iraq and Afghanistan. J Trauma 2008; 64(2):163-68.##15. Thom KA, Johnson JK, Lee MS, Harris AD. Environmental contamination because ofmultidrug-resistant Acinetobacter baumannii surrounding colonized orinfected patients. Am J Infect Control 2011; 39(9):711-15.##16. Weber DJ, Rutala WA, Miller MB, Huslage K, Sickbert-Bennett E. Role ofhospital surfaces in the transmission of emerging health care-associated pathogens: norovirus, Clostridium difficile, and Acinetobacter species. Am J Infect Control 2010; 38(5):25-33.##17. Peleg AY, Seifert H, Paterson DL. Acinetobacter baumannii: emergence of asuccessful pathogen. Clin Microbiol Rev. 2008; 21(3):538-82.##18. Bergogne-Bérézin E, Towner KJ. Acinetobacter spp. As nosocomial pathogens: microbiological, clinical, and epidemiological features. Clin Microbiol Rev. 1996; 9(2):148-65.##19. Morgan DJ, Liang SY, Smith CL, Johnson JK, Harris AD, Furuno JP. et al. Frequentmultidrug-resistant Acinetobacter baumannii contamination of gloves, gowns, and hands of healthcare workers. Infect Control Hosp Epidemiol. 2010; 31(7):716-21.##20. Thom KA, Harris AD, Johnson JA, Furuno JP. Low prevalence of Acinetobacterbaumannii colonization on hospital admission. Am J Infect Control.2010; 38(4):329-31.##21. Roca I, Espinal P, Vila-Farrés X, Vila J. The Acinetobacter baumannii Oxymoron:Commensal Hospital Dweller Turned Pan-Drug-Resistant Menace. Front Microbiol. 2012; 3:148.##22. Higgins PG, Dammhayn C, Hackel M, Seifert H. Global spread of carbapenem-resistant Acinetobacter baumannii.J Antimicrob Chemother. 2010; 65(2):233-38.##23. Doughari HJ.,Ndakidemi PA, Human IS,Benade S. Virulence, resistance genes and transformation amongst environmental isolates of Escherichia coli and Acinetobacter spp. J Microbiol. Biotechnol. 2012; 22(1): 25-33.##24. Connell I, Agace W, Klemm P, Schembri M, Marild S, Svanborg C. Type 1 fimbrialexpression enhances Escherichia coli virulence for the urinary tract. Proc. Natl. Acad. Sci. USA 1996; 93(18): 9827-832.##25. Barnhart, M.M. and M.R. Chapman, 2006.Curli biogenesis and function. Annu. Rev. Microbiol; 60: 131-147.##26. Johnson JR, Stell AL. Extended virulence genotypes of Escherichia coli strains frompatients with urosepsis in relation to phylogenic and host compromise. J Infect Dis. 2000; 181:261-72.##27. Le Bouguenec CL, Archambaud M, Labigne A. A rapid and specific detection of 32- the pap, afa and sfa adhesin-encoding operons in uropathogenic Escherichia coli strains by polymerase chain reaction. J Clin Microbiol. 1992; 30:1189-193.##28. Gophna U, Barlev M, Seijffers R, Oelschlager TA, Hacker J, Ron EZ. Curli 34- fibers mediate internalization of Escherichia coli by eukariotic cells. Infect Immun. 2001; 69: 2659- 665.##29. Sarén A, Virkola R, Hacker J, Korhonen TK. The cellular form of human fibronectin as 36- an adhesion target for the S fimbriae of meningitis-associated Escherichia coli. Infect Immun. 1999; 67(5): 2671-676.##30. Blanco J, Alonso MP, Gonzalez EA, Blanco M, Garabal JI. Virulence factors o f 38- bacteraemic Escherichia coli with particular reference to production of cytotoxic necrotizing factor (CNF) by P-fimbriate strains. J Med Microbiol. 1990; 31: 175-83.##31. Montenegro MA, Bitter-Suermann D, Timmis JK, Agüero ME, Cabello FC, Sanyal SC, Timmis KN. Tra T gene sequences, serum resistance and pathogenicity-related factors in clinical isolates of Escherichia coli and other gramnegative bacteria. J Gen Microbiol. 1985; 131: 1511-521.##32. Rosenberg M, Perry A, Bayer EA, Gutnick DL, Rosenberg E, Ofek I. Adherence ofAcinetobacter calcoaceticus RAG-1 to human epithelial cells and to hexadecane. Inect Immun. 1981; 33(1):29-33.##33. Rosenberg M, Bayer EA, Delarea J, Rosenberg E. Role of Thin Fimbriae inAdherence and Growth of Acinetobacter calcoaceticus RAG-1 on Hexadecane. Appl Environ Microbiol 1982; 44(4):929-37.##34. Kaplan N, Rosenberg E, Jann B, Jann K.Structural studies of the capsular polysaccharide of Acinetobacter calcoaceticus BD4. Eur J Biochem. 1985; 152(2):453-58. [35]. Actis LA, Tolmasky ME, Crosa LM,Crosa JH. Effect of iron-limiting conditions 45- on growth of clinical isolates of Acinetobacter baumannii. J Clin Microbiol 1993; 31(10):2812-815.##35. Russo TA, Luke NR, Beanan JM, Olson R, Sauberan SL, MacDonald U, et al. The K1capsular polysaccharide of Acinetobacter baumannii strain 307-0294 is a major virulence factor. Infect Immun.2010; 78(9):3993-4000.##36. Donlan RM. Biofilms: microbial life on surfaces. Emerg Infect Dis. 2002; 8(9):881-90.##37. Wroblewska MM, Sawicka-Grzelak A, Marchel H, Luczak M, Sivan A. Biofilmproduction by clinical strains of Acinetobacter baumannii isolated from patients hospitalized in twotertiary care hospitals. FEMS Immunol Med Microbiol. 2008; 53(1):140-44.##38. de Breij A, Gaddy J, van der Meer J, Koning R, Koster A, van den Broek P, et al. CsuA/BABCDE-dependent pili are not involved in the adherence of Acinetobacter baumannii ATCC19606(T) tohuman airway epithelial cells and their inflammatory response. Res Microbiol.2009; 160(3):213-18. [40]. Gaddy JA, Actis LA. Regulation ofAcinetobacter baumannii biofilm formation. Future Microbiol. 2009; 4(3):273-78.##39. Antunes LC, Imperi F, Carattoli A, ViscaP. Deciphering the multifactorial nature of Acinetobacter baumannii pathogenicity. PLoS One 2011; 6(8):22674. ##40. [42]. Cerqueira GM, Peleg AY. Insights intoAcinetobactebaumannii pathogenicity. IUBMB Life 2011; 63(12):1055-1060.##41. Braun G, Vidotto MC. Evaluation ofadherence, hemagglutination and presence of genes codifying for virulence factors of Acinetobacter baumannii causing urinary tract infection. Mem Inst Oswaldo Cruz. 2004; 99(8): 839-44.##42. Cappuccino JG, Sherman N. Microbiology Lab Manual by James G. Cappuccino andNatalie Sherman 4th edition, Benjamin-Cummings Publishing Company 1996; ISBN: 9780805305739.##43. Chiang MC, Kuo SC, Chen YC, Lee YT, Chen TL, Fung CP. Polymerase chain reactionassay for the detection of Acinetobacter baumannii in endotracheal aspirates from patients in the intensive care unit. J Microbiol Immunol Infect. 2011; 44(2):106-10.##44. Yamamoto S, Terai A, Yuri K, KurazonoH, Takeda Y, Yoshida O. Detection of urovirulence factors in Escherichia coli by multiplex polimerase chain reaction. FEMS Immunol Med Microbiol. 1995; 12:85-90.##45. Blanco M, Blanco JE, Blanco J.Polymerase chain reaction for detection of Escherichia coli strains producing cytotoxic necrotizing factor type 1 and 2 (CNF1 and CNF2). J Microbiol Meth. 1996; 26:95-101.##46. Schubert S, Rakin A, Karch H, Carniel E,Hessemann J. Prevalence of the “high-pathogenicity island” of Yersinia species among Escherichia coli strains that are pathogenic to humans. Infect Immun.1998; 66:480-85.##47. Huang SH, Wass C, Fu Q, Prasadarao NV, Stins M, Kim KS. Escherichia coli invasion ofbrain microvascular endothelial cells in vitro and in vivo: molecular cloning and characterization of invasion gene ibe10. Infect Immun. 1995; 63:4470-475.##48. 59- [50]. Johnson JR, Brown JJ. Colonization with and acquisition of uropathogenic Escherichiacoli strains as revealed by polymerase chain reaction-based detection. J Infect Dis.1998; 177:1120-124.##49. Marklund BI, Tennent JM, Garcia E: Horizontal gene transfer of the Escherichiacoli pap and prs pili operons as a mechanism for the development of tissue-specific adhesive properties. Mol Microbiol 1992; 6:2225-242.##50. Jaggi N, Sissodia P, Sharma L. Acinetobacter baumannii isolates in a tertiarycare hospital: Antimicrobial resistance and clinical significance. J Microbiol Infect Dis. 2012; 2(2):57-63.##51. Siau H, Yuen KY, Wong SS, Ho PL, Luk WK: The epidemiology of acinetobacterinfections in Hong Kong. J Med Microbiol 1996; 44(5):340-7.##52. Shanthi M, Sekar U. Multi-drug resistant Pseudomonas aeruginosa and Acinetobacterbaumannii infections among hospitalized patients: risk factors and outcomes. J Assoc Physicians India 2009;57:636, 638-40,645.##53. Beavers SF, Blossom DB, Wiemken TL,##54. 65- Kawaoka KY, Wong A, Goss L,et al. Comparison of risk factors for recovery of Acinetobacter baumannii during outbreaks at two Kentucky hospitals, 2006. Public Health Rep. 2009; 124(6):868-74.##55. del Mar Tomas M, Cartelle M, Pertega S, Beceiro A, Llinares P, Canle D,et al. Hospitaloutbreak caused by a carbapenem-resistant strain of Acinetobacter baumannii: patient prognosis andrisk-factors for colonization and infection. Clin Microbiol Infect.2005; 11(7):540-##56. Rahbar M, Hajia M. Detection and quantitation of the etiologic agents ofventilator-associated pneumonia in endotracheal tubeaspirates from patients in Iran. Infect Control Hosp Epidemiol.2006; 27(8):884-85.##57. Rahbar M, Mehrgan H, Aliakbari NH. Prevalence of antibiotic-resistant Acinetobacterbaumannii in a 1000-bed tertiary care hospital in Tehran, Iran. Indian J Pathol Microbiol. 2010; 53(2):290-93.##58. Lautenbach E, Synnestvedt M, Weiner MG, Bilker WB, Vo L, Schein J, Kim M.##59. Epidemiology and impact of imipenem resistance in Acinetobacter baumannii. Infect Control Hosp Epidemiol. 2009; 30(12):1186-192.##60. Marchaim D, Navon-Venezia S, Schwartz D, Tarabeia J, Fefer I, Schwaber MJ, CarmeliY. Surveillance cultures and duration of carriage of multidrug-resistant Acinetobacter baumannii. J Clin Microbiol 2007; 45(5):1551-555.##61. Maragakis LL, Cosgrove SE, Song X, Kim D, Rosenbaum P, Ciesla N, et al. Anoutbreak of multidrug-resistant Acinetobacter baumannii associated with pulsatile lavage wound treatment. JAMA 2004; 292(24):3006-3011.##62. Villegas MV, Hartstein AI. Acinetobacter outbreaks, 1977-2000. Infect Control HospEpidemiol. 2003; 24(4):284-95.##63. Hornick DB, Allen BL, Horn MA, Clegg S. Fimbrial types among respiratory isolatesbelonging to the family Enterobacteriaceae. J Clin Microbiol 1991; 29: 1795-800.##64. Köljelg S, Vuopio-Varkila JV, Lyytikäinen O, Mikelsaar M, Wadström T. Cell surfaceproperties of Acinetobacter baumannii. APMIS 1996; 104: 659-66. ##65. Mohajeri P, Rezaei Z, Sharbati S, Rasi H, Rostami Z, Farahani A, Khodarahmi R.Frequency of Adhesive Virulence Factors in Carbapenemase-producing Acinetobacter baumannii Isolated from Clinical Samples in West of Iran. Asian J Bio Sci. 2014; 7:158-64. ##66. McConnell MJ, Actis L, Pachon J.Acinetobacter baumannii: human infections, factors contributing to pathogenesis and animal models. FEMS Microbiol Rev.2013; 37: 130-55.##67. Ofek, I, Doyle RJ. Bacterial Adhesion to Cells and Tissues. 1st Edn. Champion andHall, London, UK, 1994: 513-61.##68. Sokurenko EV, Hasty DL, and D.E. Dykhuizen DE. Pathoadaptive mutations:Gene loss and variation in bacterial pathogens. Trends Microbiol. 1999;7(5):191-95.##69. Joly-Guillou ML. Clinical impact and pathogenicity of Acinetobacter. Clin MicrobiolInfect. 2005; 11(11): 868-73.## ##

The Effect of Nonpolar Fraction of Carum Copticum Essence on Acetylcholine Induced Contraction in the Ileum of Isolated Rats 2 2 Background and Aims: Our previous study has demonstrated that essence of Carum copticum reduces contraction activity of ileum in rat. The present study was designed to find out the effects of nonpolar fraction of Carum copticum essence (NFCCE) on mechanical activity of the isolated ileum in rat. Materials and Methods: For evaluation of spasmolythic property of fraction, different doses of the solution were added to organ bath after acetylcholine with concentration of 10-4 molar (M), and for assessment of antispasmodic effect of fraction, different doses of the solution were added to the organ bath before acetylcholine with concentration of 10-9 up to 10-2 M. Then isotonic contractions of ileum were recorded through an isolated tissue chamber in an organ bath using oscillographic device. Results: Our findings showed that the spasmolytic effect of NFCCE in concentrations of 25, 50, and 100 ng/ml significantly reduces acetylcholine (10-4M)-induced contractions (p< 0.05). Also antispasmodic effect of NFCCE on logarithmic concentrations of acetylcholine (10-9 up to 10-2M) indicates that in the presence of 10-3 M acetylcholine, the maximum (100%) and minimum (84.9%), inhibition of contraction is induced by concentrations of 100 ng/ml and 25 ng/ml, respectively (p< 0.05). Conclusions: The spasmolytic effect of this study is probably resulted by anti-cholinergic response of NFCCE on isolated ileum of rats. 98 104 2015/08/182015/08/182015/08/18 1394/5/27 2015/08/182015/08/182015/08/18 1394/5/27 Seyed Hassan Hejazian Seyed Hassan Hejazian Department of Physiology, Herbal Medicine Research Center, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran Iran Sara Sadegh Zade Sara Sadegh Zade Department of Physiology, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran Iran Mahila Lotfi Mahila Lotfi Department of Physiology, International Campus, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. Iran Fatemeh Safari Fatemeh Safari Department of Physiology, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran Iran fa.cardio@gmail.com Acetylcholine Carum copticum Ileum Spasmolythic effect [1]. Khajeh M, Yamini Y, Sefidkon F, Bahramifar N. Comparison of essential oil of Carum Copticum obtained by supercritical carbon dioxide extraction and hydrodistillation methods. Food Chem. 2004; 86: 587–91. ##[2]. Avesina, Law in Medicine vol. 2, Soroush Press, Tehran, p. 187, 985. ##[3]. Devasankaraiah G, Hanin I, Haranath Ps, Ramanamurthy PS. Cholinomimetic effects of aqueous extracts from Carum Copticum seeds. Br J Pharmacol. 1974; 52(4): 613–14. ##[4]. Dashti-Rahmatabadi MH, Hejazian SH, Morshedi A, Rafati A. The analgesic effect of Carum Copticum extract and morphine on phasic pain in mice. J Ethnopharmacol. 2007; 109: 226-28. ##[5]. Boskabady MH, Alizadeh M, Jahanbin B. Bronchodilatory effect of Carum Copticum in airways of asthmatic patients. Therapie. 2007; 62(1): 23-29. ##[6]. Boskabady MH, Jandaghi P, Kiani S, Hasanzadeh L. Antitussive effect of Carum Copticum in guinea pigs. J Ethnopharmacol. 2005; 97:79-82. ##[7]. Hejazian SH, Sepehri H, Dashti MH, Mahdavi SM. Does essential oil of Carum copticum affect acetylcholine-induced contraction in isolated rat's Ileum? African Journal of Pharmacy and Pharmacology. 2011; 5: 1432-435. ##[8]. Hejazian SH, Morowatisharifabad M, Mahdavi SM. Relaxant effect of Carum Copticum on intestinal motility in ileum of rat, World Journal of Zoology 2007; 2:15-18. ##[9]. Hejazian SH, Dashti-Rahmatabadi MH, Mahdavi SM. The effect of Carum Copticum extract on acetylcholine induced contraction in isolated rat's ileum. J Acupuncture Meridian Stud. 2009; 2:275-78. ##[10]. Hejazian SH, Bagheri SM, Safari F. Spasmolytic and Antispasmodic action of Trachyspermum ammi Essence on Rat's Ileum Contraction. North american medical sciense 2014; 6:643-47. ##[11]. Unno T, Matsuyama H, Izumi Y, Yamada M, Wess J, Komori S. Roles of M2 and M3 muscarinic receptors in cholinergic nerve-induced contractions in mouse ileum studied with receptor knockout mice. Br J Pharmacol. 2006; 149: 1022-30. ##[12]. Sá-Nunes A, Corrado AP, Baruffi MD, Faccioli LH, Disodium cromoglycate prevents ileum hyperreactivity to histamine in Toxocara canis-infected guinea pigs. Pharmacol Res. 2003; 48(5): 451-55. ##[13]. Roberts SJ, Papaioannou M, Evans BA ,Summers RJ. Characterization of beta-adrenoceptor mediated smooth muscle relaxation and the detection of mRNA for beta1-, beta-2- and beta3-adrenoceptors in rat ileum. Brit. J. Pharmacol. 1999; 127(4): 949–61. ##[14]. Van Crombruggen K, Van Nassauw L, Timmermans JP, Lefebvre RA. Inhibitory purinergic P2 receptor characterisation in rat distal colon. Neuropharmacology. 2007; 53:257-71. ##[15]. Zizzo MG, Mulè F, Serio R. Functional evidence for GABA as modulator of the contractility of the longitudinal muscle in mouse duodenum: role of GABA (A) and GABA(C) receptors. Neuropharmacology 2007; 52:1685-90. ##[16]. Kito Y, Suzuki H. Effects of Dai-kenchu-To on spontaneous activity in the mouse small intestine. J Smooth Muscle Res. 2006; 42: 189-201. ##[17]. Boskabady MH, Moetamedshariati V. Bronchodilatory and anti-cholinergic effects of Carum Copticum on isolated guinea-pig tracheal chain. Eur Respir J. 1996; 23, 28s. ##[18]. Boskabady MH, Shaikhi J. Inhibitory effect of Carum Copticum on histamine (H1) receptors of isolated guinea-pig tracheal chain. J. Ethnopharmacol. 2000; 69(3): 217-27. ##[19]. Gilani A.H, Jabeen Q, Ghayur MN, Janbaz KH, Akhtar MS. Studies on the antihypertensive,antispasmodic,bronchodilator and hepatoprotective activities of the Carum Copticum seed extract. J. Ethnopharmacol. 2005; 98:127-135. ##[20]. Hejazian SH, Bagheri SM, Fattahi A. Role of thymol in inhibition of acetylcholine induced contraction of isolated Rat's Ileum. Physiology and pharmacology 2013;17: 216-23. ##[21]. Ceccatto VM, Coelho-de-Souza AN, Gomes MD, Lahlou S, Leal-Cardoso JH, Lima FC, et al. Vasorelaxant effect of the monoterpenic phenol isomers, carvacrol and thymol, on rat isolated aorta. Fundam Clin Pharmacol. 2009; 24: 341-50. ##[22]. Szentandrássy N, Szigeti G, Szegedi C, Sárközi S, Magyar J, Bányász T, et al. Effect of thymol on calcium handling in mammalian ventricular Myocardium. Life Sci. 2004; 74(7): 909-21. ##[23]. Boskabady MH, Ramazni M, Tabai T. Relaxant effects of different fractions of essential oil from Carum Copticum on gunea pig tracheal chains. Phytother Res. 2003; 17(10): 1145-149. ## ##

Molecular Detection of Chicken Anemia Virus from Native Larry-breed Chickens in Chaharmahal-va-Bakhtiyari Province, Iran 2 2 Background and Aims: Finding prevalence of chicken anemia virus (CAV) infection in native chickens is necessary to avoid transmission of infection to commercial flocks. In this paper we attempt to describe molecular detection of chicken anemia virus in native Larry-breed chickens in Chaharmahal-va-Bakhtiyari province in Iran for the first time. Materials and Methods: Blood samples were collected from 100 native Larry-breed chickens (5-8 months old) in Shahrekord, Lordegan, Brogen and Koohrang, i.e.four cities of Chaharmahal-va-Bakhtiyari province. To detect CAV, Polymerase chain reaction (PCR) was undertaken on isolated DNA from blood samples using a pair of CAV specific primers that produced a 374 base pair fragment. Results: PCR analysis detected CAV in 12 of 100 (12%) tested blood samples. Conclusions: The results revealed that native Larry chicks were not free from CAV infection in Chaharmahal-va-Bakhtiyari province and vaccination against CAV should be taken into account in native farms. 105 111 2015/08/182015/08/182015/08/182015/08/18 1394/5/27 2015/08/182015/08/182015/08/182015/08/18 1394/5/27 Neda Eskandarzade Neda Eskandarzade Department of Biochemistry, School of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman, Iran. Iran Alireza Heydarnezhad Alireza Heydarnezhad Research Committee Student, Ilam University of Medical Sciences, Ilam, Iran. Iran Yousef Valizadeh Yousef Valizadeh Research Committee Student, Ilam University of Medical Sciences, Ilam, Iran. Iran Mona Shokouhi Mona Shokouhi Research Committee Student, Ilam University of Medical Sciences, Ilam, Iran. Iran Alireza Shafizadeh Kholenjani Alireza Shafizadeh Kholenjani Research Committee Student, Ilam University of Medical Sciences, Ilam, Iran. Iran Chicken Anemia Virus Larry-breed PCR [1]. Von Bulow V, Schat KA. Chicken Infectious Anemia. In: Diseases of Poultry. Chapter 2. Iowa State University Press: Ames IA; 2013: 249. ##[2]. Yuasa N, Taniguchi T, Yoshida I. Isolation and some characteristics of an agent inducing anemia in chicks. Avian Dis. 1979; 23:366-85. ##[3]. Miller MM, Schat KA. Chicken infectious anaemia virus: an example of the ultimate host-parasite relationship. Avian Pathol. 2004; 48:734-45. ##[4]. Rosenberger JK, Cloud SS. The effects of age, route of exposure and coinfections with infectious bursal disease virus on the pathogenicity and transmissibility of chicken anemia agent (CAA). Avian Dis. 1989; 33:753-59. ##[5]. Cloud SS, Rosenberger JK, Lillehoj HS. Immune dysfunction following infection with chicken anemia agent and infectious bursal disease virus. II. Alterations of in vitro lymphoproliferation and in vivo. Vet Immunol Immunopathol. 1992; 34:353-66. ##[6]. Rosenberger JK, Cloud SS. Chicken anaemia virus. Poult Sci. 1998; 77:1190-192. ##[7]. Yuasa N, Imai K. Pathogenicity and antigenicity of eleven isolates of chicken anemia agent (CAA). Avian Pathol. 1986; 15:639-45. ##[8]. Yuasa N, Noguchi T, Furuta K, Yoshida I. Maternal antibody and its effect on the susceptibility of chicks to chicken anemia agent. Avian Dis. 1980; 24:197-202. ##[9]. Chettle NJ, Eddy RK, Wyeth PJ, Lister SA. An outbreak of disease due to chicken anemia agent in England. Vet Rec. 1989; 124:211-15. ##[10]. McIlroy SG, McNulty S, Bruce DW, Smyth JA, Goodall EA, Alcorn MJ. Economic effects of clinical chicken anemia agent infection on profitable broiler production. Avian Dis. 1992; 36:566-74. ##[11]. McNulty MS, McIlroy SG, Bruce DW, Todd D. Economic effects of subclinical chicken anemia agent infection in broiler chickens. Avian Dis. 1991; 35: 263-68. ##[12]. Von Bulow V, Fuchs B, Bertram M. In vitro studies of the causative agent infectious anemia (CAA) in chickens: multiplication, titration, serum neutralization test and the indirect immunfluorescent test. Zentralbl Veterinar med. 1985; 32:679-93. ##[13]. Schat KA, Van Santen V. Chicken infectious anemia. In: Diseases of Poultry, 12th ed. Saif YM, Fadly AM, Glisson JR, McDougald LR, Nolan LK, Swayne DE, eds. Wiley-Blackwell: Ames IA; 2008: 211-235. ##[14]. Todd D, Creelan JL, Mackie DP, Rixon F, McNulty MS. Purification and biochemical characterization of chicken anemia agent. J Gen Virol. 1990; 71:19-23. ##[15]. Dren CN, Koch G, Kant A, Verschueren CAJ, Van der Eb AJ, Noteborn MHN. A hot start PCR for the laboratory diagnosis of CAV. In: International symposium on infectious bursal disease and chicken infectious anemia. Germany: Rauischholzhausen. 1994:413-420. ##[16]. Noteborn MHM, Verschueren CAJ, Van Roozelaar DJ, Veldkamp S, Van der Eb AJ, De Boer GF. Detection of chicken anaemia virus by DNA hybridisation and polymerase chain reaction. Avian Pathol. 1992; 21:107-18. ##[17]. Soine C, Watson SK, Rybicki E, Lucio B, Nordgren RM, Parrish CR. Schat KA. Determination of the detection limit of the polymerase chain reaction for chicken infectious anemia virus. Avian Dis. 1993; 37:467-76. ##[18]. Tham KM. Polymerase chain reaction analysis of chicken anemia virus DNA in fresh and paraffin-embedded tissues. In: PCR: protocols for diagnosis of human and animal virus diseases. Becker Y and Darai G, eds. Berlin: Springer-Verlag; 1995: 555-63. ##[19]. Todd D, Mawhinney KA, McNulty MS. Detection and differentiation of chicken anemia virus isolates by using the polymerase chain reaction. J Clin Microbiol. 1992; 30:1661-66. ##[20]. Tham KM, Stanislawek WL. Detection of chicken anaemia agent DNA sequences by the polymerase chain reaction. Arch Virol. 1992; 127:245–55. ##[21]. Toroghi R, Shoushtari AH, Charkhkar S, Neyazi MH. The first report of Chicken Infectious Anaemia occurrence among Iranian broiler flocks, Proceedings of 13th Iranian Veterinary Congress, Iran Vet Association 2003: 240. ##[22]. Mahzounieh M, Karimi I, Zahraei Salehi T. Serologic evidence of chicken infectious anemia in commercial chicken flocks in Shahrekord, Iran. Int Poult Sci. 2005; 4:500-503. ##[23]. Gholami-Ahangaran M, Zia-Jahromi N. Serological and molecular identification of subclinical chicken anaemia virus infection in broiler chickens in Iran. African Journal of Microbiol Res. 2012; 6:4471-474. ##[24]. Iwata N, Fujino M, Tuchiya AK, Iwata A, Otaki Y, Ueda S. Development of an enzyme-linked immunosorbent assay using recombinant chicken anaemia virus proteins expressed in a baculovirus vector system. J Vet Med Sci. 1998; 60:175-80. ##[25]. Cardona C, Lucio B, Oconnell P, Jagne J, Schat K. Humeral immune responses to chicken infectious anemia virus in three strains of chickens in a closed flock. Avian Dis. 2000; 44:661-67. ##[26]. Ballal A, Elhussein AM, Abdelrahim ISA. Serological survey of chicken infectious anemia in commercial chicken flocks in Khartoum state, Sudan. J Anim Vet Adv. 2005; 4:666-67. ##[27]. Farkas T, Maeda K, Sugiura H, Kal K, Hirai K, Otsuki K, Hayashi TA. Serological survey of chickens, Japanese quail, pigeons, ducks and crows for antibodies to chicken anemia virus in Japan. Avian Pathol. 1998; 27:316-20. ##[28]. Hadimli HH, Erganis O, Guler L, Uan US. Investigation of chicken infectious anemia virus infection by PCR and ELISA in chicken flocks. Turk J Vet Anim Sci. 2008; 32:79-84. ##[29]. Roussan DA. Serological survey on the prevalence of CIVA in commercial broiler chicken flock in Northern Jordan. Int Poult Sci. 2006; 5:544-46. ##[30]. Oluwayelu DO, Todd D. Rapid identification of chicken anemia virus in Nigerian backyard chickens by polymerase chain reaction combined with restriction ##endonuclease analysis. African J Biotech. 2008; 7:271-75. ##[31]. Hernandez-Divers SM, Villegas P, Prieto F, Unda JC, Stedman N, Ritchie B, et al. A survey of selected avian pathogens of backyard poultry in northwestern Ecuador. J Avian Med Surg. 2006; 20:147-58. ##[32]. Barrios PR, Marin SY, Resende M, Rios RL, Resende JS, Horta RS, et al. Occurrence of chicken anemia virus in backyard chickens of the Metropolitan region of Belo Horizonte, Minas Gerais. Brazilian J of Poult Sci. 2009; 11:135-38. ##[33]. Von Bulow V, Schat KA. Chicken infectious anemia: In: Diseases of Poultry, 10th ed. Calnek BW, Barnes JH, Beard CW, McDougald LR, Saif YM, eds. Iowa: Iowa State University Press Ames; 1997: 739-56. ##[34]. Markowski-Grimsrud CJ, Schat KA. Infection with chicken anemia virus impairs the generation of antigenspecific ##cytotoxic T lymphocytes. Immunol. 2003; 109:283–94. ##[35]. McConnell CD, Adair BM, McNulty MS. Effects of chicken anemia virus on cell-mediated immune function in chickens exposed to the virus by a natural route. Avian Dis. 1993; 37:366–74. ##[36]. Shoushtari AH, Ezzi A, Bozorgmehri MH Gudarzi H. Coinfection of broilers with Influeza, chicken infectious anemia and Marek viruses. 3rd Iranian Congress of Virology 2006. ##[37]. Farhoodi M, Toroghi R, Bassami MR, Kianizadeh M. Chicken infectious anemia virus mong broiler flocks in Iran. Archive of Razi Institute 2007; 62: 1-6. ##[38]. Ezzi A, Shoushtari A, Marjanmehr H, Toroghi R, Tavasoly A, Bahmani-nejad MA. Experimental studies of pathogenecity of chicken infectious anaemia virus (3 isolates) in Iran. Archives of Razi Institute 2012; 67: 13-19.## ##

Assessment of Alpha-1 Antitrypsin Deficiency in Patients with Severe Chronic Obstructive Pulmonary Disease 2 2 Background and Aims: Chronic obstructive pulmonary disease (COPD) is a kind of pulmonary diseases characterized by chronic obstruction of lung that is in the form of a diffuse narrowing of airways resulting in air flow resistance. Alpha-1 antitrypsin (AAT) deficiency is genetically relatively common risk factor in patients with COPD throughout the world and the exact cause of its prevalence is unknown. We therefore performed a study to determine the frequency of AAT deficiency in patients with severe COPD compared to the healthy controls. Materials and Methods: In this cross-sectional case control study, AAT serum level in 60 patients with severe COPD for whom the history and spirometry test with FEV1<50% had been confirmed based on gold criteria as well as 60 healthy controls, were tested using commercial kit and nephelometry method. Results: The lowest serum levels of AAT measured in patients was <0.349 g/l and the highest was 3.099 g/l. These were obtained in healthy subjects as 1.180 g/l and 4.195 g/l respectively. Out of 60 patients, 4 (6.7%) had partial deficiency of AAT (AAT<1 g/l) and 6 (10%) had definite shortage of AAT (AAT<0.5 g/l). In healthy subjects, we did not find any definite and relative lack of AAT. The comparison of results obtained from these two groups indicated a significant difference between frequency of AAT (P=0.001). Conclusions: Our findings revealed the frequency of AAT deficiency, as a factor involved in COPD disease, to be 10% and can be the reason for the high prevalence and severity of COPD in Zahedan city. 112 120 2015/08/182015/08/182015/08/182015/08/182015/08/18 1394/5/27 2015/08/182015/08/182015/08/182015/08/182015/08/18 1394/5/27 Nezarali Moulaei Nezarali Moulaei Internal Medicine Department, Research Centre for Tropical and Infectious Diseases, Zahedan University of Medical Sciences, Zahedan, Iran. Iran Hossein Ali Khazaei Hossein Ali Khazaei Immunology and Hematology Department, Clinical Immunology Research Center, Zahedan University of Medical Sciences, Zahedan, Iran. Iran Vida Pejman Vida Pejman Zahedan University of Medical Sciences, Zahedan, Iran. Iran Alph-1-antitrypsin COPD Zahedan [1]. Molloy K, Hersh CP, Morris VB, Carroll TP, O'Connor CA, Lasky-Su JA, et al. Clarification of the risk of chronic obstructive pulmonary disease in α1-antitrypsin deficiency PiMZ heterozygotes. Am J Respir Crit Care Med. 2014; 15; 189(4): 419-27. ##[2]. Dewar M, Curry RW Jr. Chronic obstructive pulmonary disease: diagnostic considerations. Am Fam Physician. 2006; 73(4):669-76. ##[3]. Zutler M, Quinlan PJ, Blanc PD. Alpha-1-antitrypsin deficient man presenting with lung function decline associated with dust exposure: a case report. J Med Case Reports 2011; 5:154. ##[4]. Barbu C, Iordache M, Man MG. Inflammation in COPD: pathogenesis, local and systemic effects. Rom J Morphol Embryol. 2011; 52(1): 21-7. ##[5]. Celli BR, Halbert RJ, Nordyke RJ, Schan B. Airway obstruction in never smokers: result from the Third Natinoal Health and Nutrition Examination Survey. Am J Med. 2009; 118:1364-72. ##[6]. Khazaei HA, Moulaei NA, Nasiraldin Tabatabei SM, Shahverdi H. Seroepidemiology characteristic of respiratory illnesses in elderly patients. Annals of Biological Research ( In press). ##[7]. Sant Anna CA, Stelmach R, Zanetti Feltrin MI, Filho WJ, Chiba T, Cukier A. Evaluation of health-related quality of life in low-income patients with COPD receiving long term oxygen therapy. Chest 2003; 123(1):136-41.##[8]. Murray CJL, Lopez AD. In: The global burden of disease: a comprehensive assessment of mortality and disability from diseases, injuries and risk factors in 1990 and projected to 2020. Cambridge, MA: Harvard University Press: 2005. ##[9]. Murray CJ, Lopez AD. Alternative projections of mortality and disability by cause 1990-2020: Global burden of disease study. Lancet 2000; 349(9064):1498-504. ##[10]. Clancy J, Turner C. Smoking and COPD: the impact of nature-nurture interactions. Br J Nurs. 2013 Jul 25-Aug 7; 22(14); 820: 822-26. ##[11]. Leuenberger P, Schwartz J, Ackermann-Liebrich U, Blaser K, Bolognini G, Bongard JP, et al. Passive smoking exposure in adults and chronic respiratory symptoms (SAPALDIA study).Swiss Study on Air Pollution and lung Disease in Adults, SAPALDIA Team. Am J Respir Crit Care Med. 2005; 150(5 Pt 1):1222-228 ##[12]. Eisner MD, Klein J, Hammond SK, Koren G, Lactao G, Iribarren C. Directly measured second hand smoke exposure and asthma health outcomes. Thorax 2005;60(10): 814-21. ##[13]. Smith CA, Harrison DJ. Association between polymorphism in gene for microsomal epoxide hydrolase and susceptibility to emphysema. Lancet 2009; 350(9078):630-33. ##[14]. Hakim IA, Harris R, Garland L, Cordova CA, Mikhael DM, Sherry Chow HH. Gender difference in systemic oxidative stress and antioxidant capacity in current and former heavy smokers. Cancer Epidemiol Biomarkers Prev. 2012; 21(12):2193-200. ##[15]. Thomson NC, Chaudhuri R, Livingston E. Asthma and cigarette smoking. Eur Respir J. 2004; 24(5):822-33. ##[16]. American Thoracic Society / European Respiratory Society Statement Standards for the diagnosis and management of individuals with Alpha-1antitrypsin deficiency. American Journal of Respiratory and Critical Care Medicine 2003; 168; 818-900. ##[17]. Stockley RA. α1-antitrypsin deficiency: what has it ever done for us? Chest 2013; 144(6):1923-29. ##[18]. Jonigk D, Al-Omari M, Maegel L, Müller M, Izykowski N, Hong J, et al. Anti-inflammatory and immunomodulatory properties of α1-antitrypsin without inhibition of elastase. Proc Natl Acad Sci U S A. 2013; 110(37): 15007–5012. ##[19]. Bergin DA, Reeves EP, Hurley K, Wolfe R, Jameel R, Fitzgerald S, et al. The circulating proteinase inhibitor α-1 antitrypsin regulates neutrophil degranulation and autoimmunity.Sci Transl Med. 2014; 1;6(217):217ra1 .##[20]. Tobin MJ, Cook PJL, Hutchison DCS. Alpha 1-antitrypsin deficiency: the clinical and physiological feature of pulmonary emphysema in subjects homozygous for Pi type Z. Br JDS Chest 1990; 77:14-27. ##[21]. Stockley RA. Alpha1-antitrypsin review. Clin Chest Med. 2014; 35(1):39-50. ##[22]. Valiulis A, Utkus A, Stukas R, Valiulis A, Siderius L. Introducing standards of the best medical practice for patients with inherited alpha-1-antitrypsin deficiency in central Eastern Europe. Prilozi. 2014; 35(1):106-13. ##[23]. Nadi E, Hajelouei M, Abduli R. Study the frequency of alpha-1 antitrypsin in patients with chronic obstructive pulmonary disease admitted to Ekbatan hospital of Hamadan city. Journal of Ghom universuty of medical sciences 2011;5(1):7-11(In Persian). ##[24]. Spínola C, Bruges-Armas J, Pereira C. Alpha-1-antitrypsin deficiency in Madeira (Portugal): the highest prevalence in the world. Respir Med. 2009; 103(10):1498-502. ##[25]. Eriksson S, Carison J, Velez R. Risks for Cirrhosis and Primary Liver Cancer in Alpha-1 Antitrypsin Deficiency. N Engl J Med. 1986; 314:736-39. ##[26]. Siri D, Farah H, Hogarth DK. Distinguishing alpha1-antitrypsin deficiency from asthma. Ann Allergy Asthma Immunol. 2013; 111(6):458-64. ##[27]. Kulpati DD. Prevalence of alpha-1-antitrypsin deficiency in C.O.P.D. and some other respiratory illnesses. J Assoc Physicians India. 1977; 25(7):443-49. ##[28]. Browne RJ, Mannino DM, Khoury MJ. Alpha 1-antitrypsin deficiency deaths in the United States from 1979-1991.An analysis using multiple-cause mortality data. Chest 1996; 110(1):78-83. ##[29]. Lieberman J, Winter B, Sastre A. Alpha 1-antitrypsin Pi-types in 965 COPD patients. Chest 1986; 89(3):370-73. ##[30]. Hersh CP, Dahl M, Ly NP, Berkey CS, Nordestgaard BG, Silverman EK. Chronic obstructive pulmonary disease in α1-antitrypsin PI MZ heterozygotes: a meta-analysis. Thorax 2004; 59:843-49. ##[31]. Dahl M, Tybjaerg-Hansen A, Lange P, Vestbo J, Nordestgaard BG. Change in lung function and morbidity from chronic obstructive pulmonary disease in alpha1-antitrypsin MZ heterozygotes: A longitudinal study of the general population. Ann intern mad. 2005; 19:270-79. ##[32]. Rodriguez F, Jardí R, Costa X, Cotrina M, Galimany R, Vidal R, et al. Rapid screening for alpha1-antitrypsin deficiency in patients with chronic obstructive pulmonary disease using dried blood specimens. Am J Respir Crit Care Med. 2002; 15; 166(6):814-17.## ##

Investigation of ELISA and PCR for Diagnosis of Infectious Mononucleosis 2 2 Background and Aims: Infectious mononucleosis (IM)is the clinical manifestation of primary infection with Epstein-Barr virus (EBV). Humans are the only known reservoir of EBV. Regarding the problems in diagnosis of the disease, the purpose of this study was to assess Enzyme-linked immunosorbent assay (ELISA) and Nested polymerase chain reaction (PCR) as a diagnostic tool for this disease. Materials and Methods: 50 samples were collected from the suspicious patients with EBV and 50 samples from the healthy individuals as the control and both techniques were applied for them. Results: The results showed that 76% of the patients and 14% of the control samples had EBV DNA in serum with PCR. Statistical analysis showed significant difference between the patient and the control samples for infection with EBV (P < 0.0001). Samples were classified into three groups according to the ELISA that were acute phase (20%), recent infection or convalescence phase (14%) and past infection (66%), respectively. Conclusions: Comparing the two methods, the results of the ELISA test indicated that ELISA would be the best method to be used for the diagnosis of IM. Our results suggest that serology may be more sensitive and could be performed as the initial screening test for acute EBV infection. Although, the PCR test is routinely used as an accurate method for detection of the pathogens with a higher specificity and sensitivity comparing the immunoassay, in IM, ELISA seems to be the best method for detecting antibodies against EBV. 121 127 2015/08/182015/08/182015/08/182015/08/182015/08/182015/08/18 1394/5/27 2015/08/182015/08/182015/08/182015/08/182015/08/182015/08/18 1394/5/27 Shirin Taleifard Shirin Taleifard Biology Department, Faculty of Sciences, University of Isfahan, Isfahan, Iran. Iran Rasoul Roghanian Rasoul Roghanian Biology Department, Faculty of Sciences, University of Isfahan, Isfahan, Iran. Iran Majid Buzari Majid Buzari Biology Department, Faculty of Sciences, University of Isfahan, Isfahan, Iran. Iran Hasan Salehi Hasan Salehi Infectious Diseases Department, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran. Iran ELISA Epstein-Barr virus Infectious mononucleosis PCR [1]. Kim M, Wadke M. Comparative evaluation of two test methods (Enzyme Immunoassay and latex fixation) for the detection of heterophil antibodies in Infectious Mononucleosis. J Clin Microbiol. 1990; 28(11): 2511- 513. ##[2]. Dolcetti R, Carbone A. Epstein- Barr virus infection and chronic lymphocytic leukemia:a possible progression factor? Infect Agents Cancer 2010; 22(5):22. ##[3]. Cook L, Midgett J, Willis D, Clinton B, Folds D. Evaluation of latex based heterophil antibody assay for diagnosis of acute Infectious Mononucleosis. J Clin Microbiol. 1987; 25(12): 2391- 394. ##[4]. Ohga S, Nomura A, Takada H, Hara T. Immunological aspects of Epstein- /Barr virus infection. CRIT REV ONCOL HEMAT. 2002; 44: 203-15. ##[5]. Paschal M, Clerici P. Serological diagnosis of Epstein- Barr virus infections: problems and solutions. World J Virol. 2012; 1(1): 31- 43. ##[6]. Maurmann S, Fricke L, Wagner HJ, Schlenke P, Hennig H, Steinhoff J, et al. Molecular parameters for precise diagnosis of asymptomatic Epstein-Barr virus reactivation in healthy carriers. J Clin Microbiol. 2003; 41(1):5419- 428. ##[7]. Bhaduri-McIntosh S, Landry ML, Nikiforow S, Rotenberg M, El-Guindy A, Miller G.Serum IgA Antibodies to Epstein- Barr virus (EBV) Early Lytic Antigens Are Present in Primary EBV Infection. J Infec. Dis. 2007; 195:483- 92. ##[8]. Chan KH, NG MH, Seto WH, Peiris JS. Epstein- Barr virus (EBV) DNA in sera of patients with primary EBV infection. J Clin Microbiol. 2001; 30(11):4152-154. ##[9]. Ohga S, Nomura A, Takada H, Hara T. Immunological aspects of Epstein- /Barr virus infection. Critical Reviews in Oncology/Hematology 2002; 44:203-15. ##[10]. Coskun O, Sener K, Kilic S, Erdem H, Yaman H, Besirbellioglu A, et al. Stress-related Epstein- Barr virus reactivation. Clin Exp Med. 2010; 10:15-20. ##[11]. Hakim H, Gibson C, Pan J, Strivastava K, Gu Z, Bankowsk MJ, Hayden RT. Comparison of various blood compartments and reporting units for the detection and quantification of Epstein- Barr virus in peripheral blood. J Clin Microbiol. 2007; 45(7):2151-155. ##[12]. Hofer M, Weber A, Haffner K, Berlis A, Haffner K, Brelis A, et al. Acute hemorrhagic Leukoencephalitis (Hurst΄s disease) linked to Epstein-Barr virus infection. Acta Neuropathol. 2005; 109: 226-30. ##[13]. Gopalkrishna V, Aggarval N, Malhotra VL, Koranne RV, Mohan VP, Mittal A, Das B.C. Chlamidia trochomatis and human Papilomavirus infection in Indian with sexually transmitted disease and cervical precancerous and cancerous lesions. Clin Microbiol Infec. 2000; 6:88-93. ##[14]. Telenti A, Marshall W F, Smith FT. Detection of Epestein- Barr virus by polymerase chain reaction. J Clin Microbiol. 1990; 28(10): 2187-190. ##[15]. Okay TS, Del Negro MB, Yamamato L, Raiz Junior R. Detection of EBV DNA in serum samples of an immunosupressed child during a three years follow-up: association of clinical and PCR data with active infection. Rev Inst Med Trop São Paulo. 2005; 47(2): 99- 102. ##[16]. Sitki D, Edwards R, Covington M, Traub N. Biology of Epstein- Barr virus during Infectious Mononucleosis. J Infec Dis. 2004; 189:483- 92. ##[17]. She R, Stevenson J, Phansalkar AR, Hillyard DR, Litwin CM, Petti CA. Limitations of polymerase chain reaction testing for diagnosing acute Epstein-Barr virus infections. Diagn Microbiol Infect Dis. 2007; 58:333- 35. ##[18]. Berger C, Day P, Meier G, Zingg W, Bossart W, Nadal D. Dynamics of Epstein- Barr virus DNA Levels in Serum During EBV-Associated Disease. J Med Virol. 2001; 64:505- 12. ##[19]. Kozic S, Vince A, Bes JI, Rode OD, Lepej SZ, Poljak M, et al. Evaluation of a commercial real-time PCR assay for quantitation of Epstein- Barr virus DNA in different groups of patients. J Virol Methods. 2006; 135:263- 68. ##[20]. Svahn A, Magnusson M, Jagdahl L, Schloss L, Kahlmeter G, Linde A. Evaluation of three commercial Enzyme-Linked Immunosorbent Assays and two latex agglutination assays for diagnosis of primary Epstein-Barr Virus infection. J Clin Microbiol. 1997; 35(15): 2728-732.## ##

Pattern of Human Hydatic Cyst in the Province of Yazd, Iran (2006-2011) 2 2 Background and Aims: Hydatidosis is a major health problem caused by larva stage of cestodes belonging to the genus Echinococcus granulosus with a cosmopolitan distribution. Zoonoses have a mountain side distribution in Iran. Echinococcus granulosus takes places in two models sylvatic and domestic. Both life-cycles are present in Iran and human is considered as an afferent host. The prevalence rate of hydatidosis is different, ranging from 1-220 cases in 100000 individuals and mortality rate of hydatidosis up to 2-4%. The aim of the present study was to explore the frequency of human hydatic cyst in Yazd province, Iran from 2006 to2011. Materials and Methods: This is a retrospective study applied through census for five years which investigated all of the profiles concerned with surgeries. Results: A total of 26911 surgeries were performed on individuals for five years 12 cases (0.045%) were infected with hydatic cyst out of which 9 cases (75%) were affected through liver, 1 case (33.34%) through pulmonary, 1 case (33.33%) through intestine and 1 case (33.33%) through cerebellum. 50% of the infected individuals were females. Morbidity age was 8-69 years, 7(66.67%) cases who were infected with Echinococcus granulosus were natives and 5 cases (33.33%) were non-natives. Among non-native individuals 2 cases were foreigners. Conclusions: Yazd province is a dry area and the desert cycle of Echinococcus granulosus is present in it. Jackals and wolves act as final hosts and goats and camels are the intermediated hosts. Based on the findings compared with those in other areas, the prevalence rate in these areas are similar requiring more control in using water, beverages, fruits and vegetables. 128 133 2015/08/182015/08/182015/08/182015/08/182015/08/182015/08/182015/08/18 1394/5/27 2015/08/182015/08/182015/08/182015/08/182015/08/182015/08/182015/08/18 1394/5/27 Ali Fattahi Bafghi Ali Fattahi Bafghi Department of Medical Parasitology & Mycology, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. Iran Mahin Ghafourzadeh Mahin Ghafourzadeh Department of Laboratory Sciences, School of Paramedicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. Iran Echinococcus granulosus Human hydatidosis Yazd Zoonoses [1]. WHO, Fact sheet N 377, Updated March 2014. ##[2]. Anvari MH, Moubedi I, Masoud J, Mansourian A. Cameles camelu sdromed arius as intermediate host of Echinococcus granulosus in the central region of Iran, Journal of Shahid Sadoughi University of medical sciences 2001; 8(4):74-79. ##[3]. Peter MS. Progress in diagnosis, treatment and elimination of echinococcosis and cysticercoids’. Parasitol Int. 2006; 55:7-13. ##[4]. Stamatelos MC, Sargedi CH, Stefanaki C, Stefanaki Ch, Safioleas C, Matthaiopoulou I, Safioleas M. Anthelminthic treatment: An adjuvant therapeutic strategy against Echinococcus granulosus. Parasitol Int. 2009; 58(2):115-20. ##[5]. Yang YR, Rosenzvit MC, Zhang LH, Zhang JZ, McManus DP. Molecular study of Echinococcus in west-central china. Parasitology 2005; 131:547-55. ##[6]. Yasuhito S, Minoru N, Kazuhiro N, HiroshiY, Akira I. Recombinant antigens for serodiagnosis of cysticercoids’ and echinococcosis. Parasitol Int. 2006; 55: 69-73. ##[7]. Arene EOI. Prevalence of hydatidosis in domestic livestock in the Niger. Delta. Trop Anim Health Prod. 1985; 17:3-4. ##[8]. Ekert J, Gemmell MA, Matays Z, Soulsby JL. Directive pour la surveillance et la prévention de l’echnococcose/ hydatidosis et la lutte contre ces maladies.Genève: OMS ; 1984. ##[9]. David H, Wen Y, Tiaoying L, Yongfu X, Xingwang CH, Yan H, et al. Control of hydatidosis. Parasitol Int. 2006; 55: 247-52. ##[10]. Torgerson PR, Heath DD. Transmission dynamics and control options for Echinococcus granulosus. Parasitology 2003; 127:143-58. ##[11]. Hadighi R, Mirhadi F, Rokni M. Evaluation of a dot-ELISA for the serodiagnosis of human hydatid disease. Pak J Med Sci. 2003; 19(4):268-71. ##[12]. Shiryazdi SM, Mirshamsi MH, Hosseini B, Ebadi M. Cases of the hydatid cyst that were operated upon in Yazd. Journal of Shahid Sadoughi University of Medical Sciences and Health Services 2000; 1(8):25-30. ##[13]. Larrieu EJ, Frider B. Human cystic echinococcosis: contributions to the natural history of the disease. Ann Trop Med Parasitol. 2001; 9597):679-87. ##[14]. Pezeshki A, Kia EB, Gholizadeh A, Koohzare A. An analysis of hydatid cyst surgeries in Tehran Milad Hospital, Iran, during 2001-2004. Pakistan Journal of Medical Sciences 2007; 23(1): 138-40. ##[15]. Ahmadi NA, Hamidi MA. Retrospective analysis of human cystic echinococcosis in Hamedan province, an endemic region of Iran. Ann Trop Med Parasitol. 2008; 102:603-609. ##[16]. Rokni MB. Echinococcosis /hydatidosis in Iran, Iranian Journal of Parasitology 2009; 4(2):1-16 ##[17]. Rostaminejad M, Nazemalhosseini Mojarad E, Norouzina M, Fasihi Harandi M. Echinococcosis: based on molecular studies in Iran, Gastroenterology and Herpetology From Bed to Bench 2010; 3(4):169-76. ##[18]. World Health Organization. International classification of ultrasound images in cystic echinococcosis for application in clinical and field epidemiological settings. In: PAIR, an option for the treatment of cystic echinococcosis. WHO, Geneva, 2001. ##[19]. Malik AA, ul Bari S, Younis M, Wani KA, Rather AA. Primary splenic hydatidosis. Indian J Gastroenterol.2011; 30(4):175-77. ##[20]. Jabbari Nooghabi A, Raoufian K, Motie MR. Concomitant Splenic and Hepatic Hydatidosis: Report of Two Cases and Review of the Literature. Acta Med Iran. 2015; 53( 1):74-77.## ##

The Homing of Spermatogonial Cells in the Cavities of a Novel Nanoscaffold 2 2 Background and Aims: In this lab trial, the effect of scaffold based on human serum albumin (HSA) and hydroxyapatite nanoparticles (HA NPs) on mouse spermatogonial cell line (SCL) was investigated. Materials and Methods: To synthesize HA NPs, calcium nitrate and diammonium phosphate at pH 13 were gently added and heated at 100 ºC for 24 hours. Then serial concentrations of HA NPs was separately added to 500 mg/mL of HSA, and immediately placed in the 100 ºC water bath. Then, all scaffolds were cut, and incubated with SCL for 6h, 12h, and 24h at 37 ºC. Finally, the cell count was read, and homing of the cells was examined by optical microscopy. Results: It was found that the quantity of cells did not change by increase in concentration of HA NPs. On the other hand, increased incubation time led to decrease in cell count. Light microscopic observation of scaffold cavities after incubation showed the homing of spermatogonial cells. Conclusions: This promising scaffold must be more investigated in vitro and in vivo, and may be suitable for making artificial testis. 134 142 2015/08/182015/08/182015/08/182015/08/182015/08/182015/08/182015/08/182015/08/19 1394/5/28 2015/08/182015/08/182015/08/182015/08/182015/08/182015/08/182015/08/182015/08/19 1394/5/28 Saeide Nezami Saridar Saeide Nezami Saridar Department of Biology, Islamic Azad University, Ashkezar Branch, Ashkezar, Yazd, Iran. Iran Seyedhossein Hekmatimoghaddam Seyedhossein Hekmatimoghaddam Department of Laboratory Sciences, School of Paramedicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. Iran Mona Yadegar Mona Yadegar Department of Biology, Islamic Azad University, East Tehran Branch, Tehran, Iran. Iran Human serum albumin Hydroxyapatite Scaffold Spermatogonia Testis [1]. Grimaldi P, Di Giacomo D, Geremia R. The endocannabinoid system and spermatogenesis. Frontiers endocrinol. 2013; 4:192. ##[2]. Valli H, Phillips BT, Shetty G, Byrne JA, Clark AT, Meistrich ML, et al. Germline stem cells: toward the regeneration of spermatogenesis. Fertil Steril. 2014; 101:3-13. ##[3]. Dores C, Alpaugh W, Dobrinski I. From in vitro culture to in vivo models to study testis development and spermatogenesis. Cell Tissue Res. 2012; 349:691-702. ##[4]. Jahnukainen K, Stukenborg JB. Clinical review: Present and future prospects of male fertility preservation for children and adolescents. J Clin Endocrinol Metabol. 2012; 97:4341-51. ##[5]. Brinster RL. Male germline stem cells: from mice to men. Science 2007; 316:404-405. ##[6]. Aslam I, Fishel S, Moore H, Dowell K, Thornton S. Fertility preservation of boys undergoing anti-cancer therapy: a review of the existing situation and prospects for the future. Hum Reprod. 2000; 15:2154-159. ##[7]. Hovatta O, Foudila T, Siegberg R, Johansson K, von Smitten K, Reima I. Pregnancy resulting from intracytoplasmic injection of spermatozoa from a frozen-thawed testicular biopsy specimen. Hum Reprod. 1996; 11:2472-473. ##[8]. Jiang X, Cao HQ, Shi LY, Ng SY, Stanton LW, Chew SY. Nanofiber topography and sustained biochemical signaling enhance human mesenchymal stem cell neural commitment. Acta Biomaterialia. 2012; 8:1290-302. ##[9]. Delgado-Rivera R, Harris SL, Ahmed I. Increased FGF-2 secretion and ability to support neurite outgrowth by astrocytes cultured on polyamide nanofibrillar matrices. Matrix Biol. 2009; 28:137-47. ##[10]. Vacanti JP, Langer R. Tissue engineering: the design and fabrication of living replacement devices for surgical reconstruction and transplantation. Lancet 1999; 354:SI32–SI4. ##[11]. Nur EKA, Ahmed I, Kamal J, Schindler M, Meiners S. Three-dimensional nanofibrillar surfaces promote self-renewal in mouse embryonic stem cells. Stem Cells 2006; 24:426–33. ##[12]. Nur EKA, Ahmed I, Kamal J, Schindler M, Meiners S. Three-dimensional nanofibrillar surfaces induce activation of Rac. Biochem Biophys Res Commun. 2005; 331:428-34. ##[13]. Schindler M, Ahmed I, Kamal J. A synthetic nanofibrillar matrix promotes in vivo-like organization and morphogenesis for cells in culture. Biomaterials 2005; 26:5624-631. ##[14]. Srouji S, Kizhner T, Suss-Tobi E, Livne E, Zussman E. 3-D Nanofibrous electrospun multilayered construct is an alternative ECM mimicking scaffold. J Mater Sci Mater Med. 2008; 19:1249-55. ##[15]. Hou Y, Fei R, Burkes JC, Lee SD, Munoz-Pinto D, Hahn MS, et al. Thermoresponsive nanocomposite hydrogels: Transparency, rapid deswelling and cell release. J Biomater Tiss Eng. 2011; 1. ##[16]. Junka R, Valmikinathan CM, Kalyon DM, Yu X. Laminin functionalized biomimetic nanofibers for nerve tissue engineering. J Biomater Tiss Eng. 2013; 3:494-502. ##[17]. Sheikh FA, Kanjwal MA, Macossay J, Muhammad MA, Cantu T, Barakat NA, et al. Fabrication of mineralized collagen from bovine waste materials by hydrothermal method as promised biomaterials. J Biomater Tiss Eng. 2011; 1. ##[18]. Rafeeqi T, Kaul G. Carbon nanotubes as a scaffold for spermatogonial cell maintenance. J Biomed Nanotech 2010; 6:710-17. ##[19]. Eslahi N, Hadjighassem MR, Joghataei MT, Mirzapour T, Bakhtiyari M, Shakeri M, et al. The effects of poly L-lactic acid nanofiber scaffold on mouse spermatogonial stem cell culture. Int J Nanomed. 2013; 8:4563-76. ##[20]. Yoshioka T, Yamamoto K, Kobashi H, Tomita M, Tsuji T. Receptor-mediated endocytosis of chemically modified albumins by sinusoidal endothelial cells and Kupffer cells in rat and human liver. Liver 1994; 14:129-37. ##[21]. Tripathi G, Basu B. A porous hydroxyapatite scaffold for bone tissue engineering: Physico-mechanical and biological evaluations. Ceramics International 2012; 38:341-49. ##[22]. Francis GL. Albumin and mammalian cell culture: implications for biotechnology applications. Cytotechnology 2010; 62:1-16. ##[23]. Schnitzer JE, Oh P. Albondin-mediated capillary permeability to albumin. Differential role of receptors in endothelial transcytosis and endocytosis of native and modified albumins. J Biol Chem. 1994; 269:6072-82. ##[24]. Kuchar M, Vankova L, Petrokova H, Cerny J, Osicka R, Pelak O, et al. Human interleukin-23 receptor antagonists derived from an albumin-binding domain scaffold inhibit IL- 23-dependent ex vivo expansion of IL-17-producing T-cells. Proteins 2013. ##[25]. Ferrero-Gutierrez A, Menendez-Menendez Y, Alvarez-Viejo M, Meana A, Otero J. New serum-derived albumin scaffold seeded with adipose-derived stem cells and olfactory ensheathing cells used to treat spinal cord injured rats. Histol Histopathol. 2013; 28:89-100. ##[26]. Gallego L, Junquera L, Meana A, Garcia E, Garcia V. Three-dimensional culture of mandibular human osteoblasts on a novel albumin scaffold: growth, proliferation, and differentiation potential in vitro. Int J Oral Max Impl. 2010; 25:699-705. ##[27]. Gallego L, Junquera L, Meana A, Alvarez-Viejo M, Fresno M. Ectopic bone formation from mandibular osteoblasts cultured in a novel human serum-derived albumin scaffold. J Biomater Appl. 2010; 25:367-81. ##[28]. Weszl M, Skaliczki G, Cselenyak A, Kiss L, Major T, Schandl K, et al. Freeze-dried human serum albumin improves the adherence and proliferation of mesenchymal stem cells on mineralized human bone allografts. J Orthop Res. 2012; 30:489-96. ##[29]. Nseir N, Regev O, Kaully T, Blumenthal J, Levenberg S, Zussman E. Biodegradable scaffold fabricated of electrospun albumin fibers: mechanical and biological characterization. Tiss Eng Part C, Methods 2013; 19:257-64. ##[30]. Luisi I, Pavan S, Fontanive G, Tossi A, Benedetti F, Savoini A, et al. An albumin-derived peptide scaffold capable of binding and catalysis. Plos One 2013; 8:e56469. ##[31]. Wu B, Braun A, Ediger S, Huang W, Lilischkis, Schäfer K-H. Surface quality and biocompatibility of porous hydroxyapatite scaffolds for bone tissue engineering. Physica Status Solidi (a) 2013; 210:957-63. ##[32]. Dey S, Pal S. Evaluation of collagen-hydroxyapatite scaffold for bone tissue engineering. 13th International Conference on Biomedical Engineering (IFMBE Proceedings) 2009; 23:1267-270. ##[33]. Leukers B, Gulkan H, Irsen SH, Milz S, Tille C, Schieker M, et al. Hydroxyapatite scaffolds for bone tissue engineering made by 3D printing. J Mater Sci Mater Med. 2005; 16:1121-124. ##[34]. Kwon BJ, Kim J, Kim YH, Lee MH, Baek HS, Lee DH, et al. Biological advantages of porous hydroxyapatite scaffold made by solid freeform fabrication for bone tissue regeneration. Artif Organs 2013; 37:663-70.## ##