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Evaluation the Efficacy of HBA Sperm- Hyaluronan Binding Assay Method in Infertility Centers 2 2 Introduction: The novel development of the (HBA) sperm- hyaluronan binding assay has now been routinely used in some laboratories worldwide to predict sperm maturity and functionality. The purpose of this study was to evaluate the correlation between HBA score and histone deficiency, protamine content and DNA fragmentation. Methods: During April- March 2012 patients who were enrolled at Shiraz- Human Assisted Reproductive Center were included in this study. Semen sample from 100 men were assessed during this study. Patients were divided into two groups, group A (%15≤HBA≤%55) n = 50 and group B (HBA≥ %55) n = 50. Sperm analysis carried out according to World Health Organization criteria. Histon efficiency, protamine content, and DNA integrity were assessed by Aniline blue, Chromomycine A3 staining and sperm chromatin dispersion test. Results: The results show positive correlation between sperm concentration and motility with percentage HA-bounded sperms while a negative correlation with percentage abnormal morphology. Histone deficiency assessed using Aniline blue staining show significant difference in group A 18.22±1.06 compared with group B 32.43± 2.90. Furthermore, the percentage HAbounded sperm showed a significant negative correlation with protamine deficiency assessed by CMA3 staining spermatozoa (15.06± 0.88 in group A versus. 75.17± 2.37 in group B). The results also show a positive correlation between intact DNA with HA bound ability. Conclusion: The results indicating that HBA had a high degree of selectivity for sperm with histone efficiency, protamine quality, and high DNA integrity. These findings are important from the points of view of human sperm DNA integrity, sperm function, and the potential efficacy of HA-mediated sperm selection for human assisted reproduction techniques. 3 8 2013/07/15 1392/4/24 2013/10/10 1392/7/18 Forouzan Absalan Forouzan Absalan Department of anatomy Iran forouzan_absalan@yahoo.com Sirous Rostami Sirous Rostami Iran s_rostami@yahoo.com Abdollah Zabihi Abdollah Zabihi Iran a.zabihi@yahoo.com (HBA) Sperm- Hyaluronan Binding Assay Aniline Blue Staining Sperm Chromatin Dispersion Test Chromomycine A 3 Staining 1. Sperm nucleus decondensation, hyaluronic acid binding and##oocyte activation capacity: different markers of sperm immaturity?##Case reports. J Assist Reprod Genet. 2012; 29: 353-355.##2. Wyrobek AJ, Eskenazi B, Young S, Arnheim N, Tiemann-##Boege I, Jabs BW, Glaser RL, Pearson FS, Evenson D. Advancing##age has differential effects on DNA damage, chromatin##integrity, gene mutations, and aneuploidies. Proc Natl##Acad Sci USA. 2006; 103: 9601- 9606.##3. Cayli S, Jakab A, Ovari L, Delpiano E, Celik-Ozenci C, Sakkas##D, Ward D and Huszar G. Biochemical markers of sperm##function: male fertility and sperm selection for ICSI. Reprod##Biomed Online. 2003; 7: 462–468.##4. Cayli S, Sakkas D, Vigue L, Demir R, and Huszar G. Cellular##maturity and apoptosis in human sperm: creatine kinase,##caspase-3 and Bcl-xl levels in mature and diminished maturity##sperm. Mol Hum Reprod. 2004; 10: 365-372.##5. Ye H, Huang G, Gao Y, Liu D Y. Relationship between human##sperm- hyaluronan binding assay and fertilization rate##in conventional in vitro fertilization. Hum Reprod. 2006;##21(6): 1545-1550.##6. Jakab A, Sakkas D, Delpiano E, Cayli S, Kovanci E, Ward##D, Revelli A, Huszar G: Intracytoplasmic sperm injection: a##novel selection method for sperm with normal frequency of##chromosomal aneuploidies. Fertil Steril. 2005; 84: 1665-1673.##7. Huszar G, Ozenci CC, Cayli S, Zavaczki Z, Hansch E, Vigue##L: Hyaluronic acid binding by human sperm indicates cellular##maturity, viability, and unreacted acrosomal status. Fertil##Steril. 2003; 79(Suppl 3): 1616-1624.##8. Huszar G, Jakab A, Sakkas D, Ozenci CC, Caily S, Ozkavukcu##S: Fertility testing and ISCI sperm selection by hyaluronic##acid binding: clinical and genetic aspects. Reprod Biomed##Online. 2007; 14: 650-663.##9. Huszar G, Ozenci CC, Cayli S et al. Hyaluronic acid binding##by human sperm indicates cellular maturity, viability and##unreacted acrosomal status. Fertile Steril. 2003; 79 (supplementary##3): 1616-24.##10. Midatlantic Diagnostics, Inc. HBA Sperm Hyaluronan##Binding Assay Information sheet. http:// www.midatlanticdiagnostics.##com/products/sperm/hba.htm##11. World Health Organization. Laboratory manual for the##examination of human semen and sperm cervical mucus interaction.##4th ed. Cambridge: Cambridge University Press;##1999.##12. Kruger TF, Menkveld R, Stander FS, Lombard CJ, der##Merwe JP Wan, van Zyl JA. Sperm morphologic features as a##prognostic factor in in vitro fertilization. Fertil Steril. 1986; 46.##8##February 2014, Volume 11, Number 1##13. Fernandes JL, Muriel L, Rivero MT, Goyanes V, Vasquez##R, Alvarez JG. The sperm chromatin dispersion test: a simple##method for the determination of sperm DNA fragmentation.##J Androl.2003; 24: 59–66.##14. Nasr-Esfahani MH, Razavi S, Vahdati AA, Fathi F, Tavalaee##M: Evaluation of sperm selection procedure based on hyaluronic##acid binding ability on ICSI outcome. J Assist Reprod##Genet. 2008; 25: 197-203.##15. Parmegiani L, Cognigni GE, Bernardi S, Troilo E, Ciampaglia##W, Filicori M: "Physiologic ICSI": hyaluronic acid (HA)##favors selection of spermatozoa without DNA fragmentation##and with normal nucleus, resulting in improvement of embryo##quality. Fertil Steril. 2010, 93: 598-604.##16. Simon L, Brunborg G, Stevenson M, Lutton D, Mcmanus##J, Lewis SE. Clinical significance of sperm DNA damage in##assisted reproduction outcome. Hum Reprod. 2010; 25: 1594-##1608.##17. Sakkas D, Mariethoz E, St. John JC. Abnormal sperm parameters##in humans are indicative of an abortive apoptotic##mechanism linked to the Fas-mediated pathway. Exp Cell##Res. 1999; 251: 350-355##18. Huszar G, Ozenci CC, Jakab A, Vigue L. A double chember##device for advanced sperm testing: semen analysis and an##assay for sperm maturity by hialuronic acid (HA) binding. J##Androl (Suppl.) 2002; 23(S1): 55.##19. Celik-Ozenci C, Jakab A, Kovacs T, Catalanotti J, Demir R,##Bray-Ward P, and Ward D, Huszae G. Sperm selection for##ICSI: shape properties do not predict the absence or presence##of numerical chromosomal aberrations. Hum Reprod 2004;##9: 2052-2059.##20.Yagci A, Nurk W, Stronk J, Huszar G. Spermatozoa bound##to solid state hyaluronic acid show chromatin structure with##high DNA chain integrity: an acridine orange fluorescence##study. J Androl. 2010; 31: 566-572.##21. Worrilow KC, Eid S, Woodhouse D, Perloe M, Smith S, Witmyer##J, Ivani K, Khoury C, Ball GD, Elliot T, Lieberman J. Use##of hyaluronan in the selection of sperm for intracytoplasmic##sperm injection (ICSI); significant improvement in clinical##outcomes-multicenter, double blinded and randomized trial.##Hum Reprod. 2012; 28: 306–314.##22. Alkhayal A, San Gabriel M, Ziedan K, Alrabeeah K, Noel##D, Zini A. Sperm DNA and chromatin integrity in semen##samples used for intrauterine insemination. J Assist Reprod##Jenet. 2013; 30(11): 1519-24.##1. Sperm nucleus decondensation, hyaluronic acid binding and##oocyte activation capacity: different markers of sperm immaturity?##Case reports. J Assist Reprod Genet. 2012; 29: 353-355.##2. Wyrobek AJ, Eskenazi B, Young S, Arnheim N, Tiemann-##Boege I, Jabs BW, Glaser RL, Pearson FS, Evenson D. Advancing##age has differential effects on DNA damage, chromatin##integrity, gene mutations, and aneuploidies. Proc Natl##Acad Sci USA. 2006; 103: 9601- 9606.##3. Cayli S, Jakab A, Ovari L, Delpiano E, Celik-Ozenci C, Sakkas##D, Ward D and Huszar G. Biochemical markers of sperm##function: male fertility and sperm selection for ICSI. Reprod##Biomed Online. 2003; 7: 462–468.##4. Cayli S, Sakkas D, Vigue L, Demir R, and Huszar G. Cellular##maturity and apoptosis in human sperm: creatine kinase,##caspase-3 and Bcl-xl levels in mature and diminished maturity##sperm. Mol Hum Reprod. 2004; 10: 365-372.##5. Ye H, Huang G, Gao Y, Liu D Y. Relationship between human##sperm- hyaluronan binding assay and fertilization rate##in conventional in vitro fertilization. Hum Reprod. 2006;##21(6): 1545-1550.##6. Jakab A, Sakkas D, Delpiano E, Cayli S, Kovanci E, Ward##D, Revelli A, Huszar G: Intracytoplasmic sperm injection: a##novel selection method for sperm with normal frequency of##chromosomal aneuploidies. Fertil Steril. 2005; 84: 1665-1673.##7. Huszar G, Ozenci CC, Cayli S, Zavaczki Z, Hansch E, Vigue##L: Hyaluronic acid binding by human sperm indicates cellular##maturity, viability, and unreacted acrosomal status. Fertil##Steril. 2003; 79(Suppl 3): 1616-1624.##8. Huszar G, Jakab A, Sakkas D, Ozenci CC, Caily S, Ozkavukcu##S: Fertility testing and ISCI sperm selection by hyaluronic##acid binding: clinical and genetic aspects. Reprod Biomed##Online. 2007; 14: 650-663.##9. Huszar G, Ozenci CC, Cayli S et al. Hyaluronic acid binding##by human sperm indicates cellular maturity, viability and##unreacted acrosomal status. Fertile Steril. 2003; 79 (supplementary##3): 1616-24.##10. Midatlantic Diagnostics, Inc. HBA Sperm Hyaluronan##Binding Assay Information sheet. http:// www.midatlanticdiagnostics.##com/products/sperm/hba.htm##11. World Health Organization. Laboratory manual for the##examination of human semen and sperm cervical mucus interaction.##4th ed. Cambridge: Cambridge University Press;##1999.##12. Kruger TF, Menkveld R, Stander FS, Lombard CJ, der##Merwe JP Wan, van Zyl JA. Sperm morphologic features as a##prognostic factor in in vitro fertilization. Fertil Steril. 1986; 46.##8##February 2014, Volume 11, Number 1##13. Fernandes JL, Muriel L, Rivero MT, Goyanes V, Vasquez##R, Alvarez JG. The sperm chromatin dispersion test: a simple##method for the determination of sperm DNA fragmentation.##J Androl.2003; 24: 59–66.##14. Nasr-Esfahani MH, Razavi S, Vahdati AA, Fathi F, Tavalaee##M: Evaluation of sperm selection procedure based on hyaluronic##acid binding ability on ICSI outcome. J Assist Reprod##Genet. 2008; 25: 197-203.##15. Parmegiani L, Cognigni GE, Bernardi S, Troilo E, Ciampaglia##W, Filicori M: "Physiologic ICSI": hyaluronic acid (HA)##favors selection of spermatozoa without DNA fragmentation##and with normal nucleus, resulting in improvement of embryo##quality. Fertil Steril. 2010, 93: 598-604.##16. Simon L, Brunborg G, Stevenson M, Lutton D, Mcmanus##J, Lewis SE. Clinical significance of sperm DNA damage in##assisted reproduction outcome. Hum Reprod. 2010; 25: 1594-##1608.##17. Sakkas D, Mariethoz E, St. John JC. Abnormal sperm parameters##in humans are indicative of an abortive apoptotic##mechanism linked to the Fas-mediated pathway. Exp Cell##Res. 1999; 251: 350-355##18. Huszar G, Ozenci CC, Jakab A, Vigue L. A double chember##device for advanced sperm testing: semen analysis and an##assay for sperm maturity by hialuronic acid (HA) binding. J##Androl (Suppl.) 2002; 23(S1): 55.##19. Celik-Ozenci C, Jakab A, Kovacs T, Catalanotti J, Demir R,##Bray-Ward P, and Ward D, Huszae G. Sperm selection for##ICSI: shape properties do not predict the absence or presence##of numerical chromosomal aberrations. Hum Reprod 2004;##9: 2052-2059.##20.Yagci A, Nurk W, Stronk J, Huszar G. Spermatozoa bound##to solid state hyaluronic acid show chromatin structure with##high DNA chain integrity: an acridine orange fluorescence##study. J Androl. 2010; 31: 566-572.##21. Worrilow KC, Eid S, Woodhouse D, Perloe M, Smith S, Witmyer##J, Ivani K, Khoury C, Ball GD, Elliot T, Lieberman J. Use##of hyaluronan in the selection of sperm for intracytoplasmic##sperm injection (ICSI); significant improvement in clinical##outcomes-multicenter, double blinded and randomized trial.##Hum Reprod. 2012; 28: 306–314.##22. Alkhayal A, San Gabriel M, Ziedan K, Alrabeeah K, Noel##D, Zini A. Sperm DNA and chromatin integrity in semen##samples used for intrauterine insemination. J Assist Reprod##Jenet. 2013; 30(11): 1519-24. ##

Adipose Stem Cells as a Feeder Layer Reduce Apoptosis and p53 Gene Expression of Human Expanded Hematopoietic Stem Cells Derived from Cord Blood 2 2 Introduction: Human hematopoietic stem cells (hHSCs) have been used for transplantation in hematologic failures. Because the number of hHSCs per cord blood unit is limited, the expansion of these cells is important for clinical application. It has been reported that cytokines and feeder layer provide a perspective to in vitro expansion of hHSCs. In this regard, cord blood CD34+ cells expanded after co-culture with adipose-derived stem cells (ADSCs) feeder layer and cytokine conditions were analyzed in terms of apoptosis and the expression level of p53, a tumor suppressor gene. Methods: Three cultures of cord blood CD34+ cells were prepared ex-vivo for 7 days, including cytokines stem cell factor (SCF), thrombopoietin (TPO), and fetal liver tyrosine kinase 3 ligand (Flt3L) with ADSCs feeder layer, cytokines without feeder layer, and a culture on micro porous membrane with cytokines. Apoptosis rate and the expression level of P53 gene in all groups were analyzed by real-time PCR. Results: The data showed that p53 expression and apoptosis rate in CD34+ fresh cells were higher than other culture systems. P53 gene expression in the expanded cells at both cytokine cultures with and without ADSCs feeder layer decreased, but it was lower than other culture systems at co-culture system with cytokines (P < 0.05). Apoptosis rate in expanded cord blood CD34+ with feeder layer was significantly lower than the group without ADSCs feeder layer. Conclusion: It could be concluded that direct culture of CD34+on feeder layer ADSCs affects the expansion of HSCs. 9 14 2013/07/152013/05/23 1392/3/2 2013/10/102013/11/17 1392/8/26 Tahereh Foroutan Tahereh Foroutan Kharazmi University, Tehran, Iran Iran foroutan@khu.ac.ir Aysan Farhadi Aysan Farhadi Iran Bahram Mohammad Soltani Bahram Mohammad Soltani Iran Hematopoietic Stem Cell Adipose Feeder Layer p53 1. Chotinantakul K, Leeanansaksiri W. Hematopoietic stem cell##Development, Niches and signaling pathways. Bone Marrow##Research 2012; 6 (3):1-16.##2. Glettig D, Kaplan D. Extending human hematopoietic Stem##Cell Survival in vitro with adipocyte. Bioresearch open access##2013; 2(3):179-185.##3. Patricia D, Conrad G, Stephen G. Ex vivo expansion of hematopoietic##cells from umbilical cord blood for clinical transplantation.##Leukocyte Biology 1998; 64(8):147-155.##4. Pant V, Cardama A, Guillermina L. The p53 pathway in hematopoiesis:##lessons from mouse models, implications for##humans. Blood 2012; 120 (26): 5118-5127.##5. Jones SN, Hancock AR, Vogel H, Donehower LA, Bradley A.##Overexpression of Mdm2 in mice reveals a p53-independent##role for Mdm2 in tumorigenesis. Proc Natl Acad Sci USA##1998; 95(26): 15608-15612.##6. Tung SS, Parmar S, Robinson SN, et al. Ex vivo expansion of##umbilical cord blood for transplantation. Best Pract Res Clin##Haematol 2010; 23: 245-57.##7. Jang YK, Jung DH, Jung MH. Mesenchymal stem cells feeder##layer from human umbilical cord blood for ex vivo expanded##growth and proliferation of hematopoietic progenitor cells.##Ann Hematol 2006; 85: 212-25.##8. Alakel N, Jing D, Muller K, Bornhauser M. Ehninger G. Ordemann##R. Direct contact with mesenchymal stromal cells##affects migratory behavior and gene expression profile of##CD133+ hematopoietic stem cells during ex vivo expansion.##Exp Hemato 2009; 37(4): 504-13##9. Oodi A, Noruzinia M, Habibi M, Nikougoftar M, Soltanpour##MS, Khorshidfar M, Amirizadeh N. Expression of p16 Cell##Cycle inhibitor in human Cord blood CD34+expanded cells##following co-culture with bone marrow-derived mesenchymal##stem cells. Hematology 2012; 17(6): 334-40.##10. Amirizadeh N, Oodi A, Nikougoftar M, Soltanpour MS.##Expression and promoter methylation changes of the##P15INK4b during ex vivo cord blood CD34+ cell expansion##following co-culture with mesenchymal stromal cells. Hematology##2013; 18(5): 260-8.##1. Chotinantakul K, Leeanansaksiri W. Hematopoietic stem cell##Development, Niches and signaling pathways. Bone Marrow##Research 2012; 6 (3):1-16.##2. Glettig D, Kaplan D. Extending human hematopoietic Stem##Cell Survival in vitro with adipocyte. Bioresearch open access##2013; 2(3):179-185.##3. Patricia D, Conrad G, Stephen G. Ex vivo expansion of hematopoietic##cells from umbilical cord blood for clinical transplantation.##Leukocyte Biology 1998; 64(8):147-155.##4. Pant V, Cardama A, Guillermina L. The p53 pathway in hematopoiesis:##lessons from mouse models, implications for##humans. Blood 2012; 120 (26): 5118-5127.##5. Jones SN, Hancock AR, Vogel H, Donehower LA, Bradley A.##Overexpression of Mdm2 in mice reveals a p53-independent##role for Mdm2 in tumorigenesis. Proc Natl Acad Sci USA##1998; 95(26): 15608-15612.##6. Tung SS, Parmar S, Robinson SN, et al. Ex vivo expansion of##umbilical cord blood for transplantation. Best Pract Res Clin##Haematol 2010; 23: 245-57.##7. Jang YK, Jung DH, Jung MH. Mesenchymal stem cells feeder##layer from human umbilical cord blood for ex vivo expanded##growth and proliferation of hematopoietic progenitor cells.##Ann Hematol 2006; 85: 212-25.##8. Alakel N, Jing D, Muller K, Bornhauser M. Ehninger G. Ordemann##R. Direct contact with mesenchymal stromal cells##affects migratory behavior and gene expression profile of##CD133+ hematopoietic stem cells during ex vivo expansion.##Exp Hemato 2009; 37(4): 504-13##9. Oodi A, Noruzinia M, Habibi M, Nikougoftar M, Soltanpour##MS, Khorshidfar M, Amirizadeh N. Expression of p16 Cell##Cycle inhibitor in human Cord blood CD34+expanded cells##following co-culture with bone marrow-derived mesenchymal##stem cells. Hematology 2012; 17(6): 334-40.##10. Amirizadeh N, Oodi A, Nikougoftar M, Soltanpour MS.##Expression and promoter methylation changes of the##P15INK4b during ex vivo cord blood CD34+ cell expansion##following co-culture with mesenchymal stromal cells. Hematology##2013; 18(5): 260-8. ##

Histomorphometry of Golden Hamster Ovaries 2 2 Introduction: Genital system is important herd in animal's creation. It is necessary for animal reproduction. Mammal has two sex in genital system such as male and female in separated animals. Animal reproduction destroyed by disorder of genital system. In this case, animals cannot produce an animal similar to them. Finally it cause overthrow a species of animals. Methods: Histomorphologic characteristic of hamster ovary studied in this research for a human model. 5 right and 5 left ovary of puberty hamster is studied. Studied biometric parameters were length, wildness and thickness. Results: This research results showed which right hamster ovary had 5/08±1/6mm in length, 3/05 ± 0/56 mm in wide ,and 2/88 ± 0/327 mm in diameter and left ovary had 4/68 ± 1/253 mm in length, 2/2 ± 0/703 mm in wide and 2/4 ± 0/609 mm in diameter. Countering of different type of hamster ovarian follicles were showed which number of primordial follicles were 1628/33± 305/6049, number of primary follicles were 307/33 ± 56/76, number of secondary follicles were 140 ± 60/63, number of tertiary follicles were 9± 0 and number of graafian follicles were 8 ± 3/6. Total count of hamster ovarian follicles was 2192/66± 178/46. Histological studies showed which covering epithelium, parenchyma of hamster ovary is similar to another mammal, and zuna plucida is forming in primary follicles when two cuboidal cell layers are observed around oocytes. Conclusion: Finally, because hamsters has short living period in comparing to human, therefore they have low ovarian follicles and oocytes but hamster ovarian follicles and oocyte were similar to human follicles and oocyte in histologic characteristics. 15 24 2013/07/152013/05/232013/08/7 1392/5/16 2013/10/102013/11/172013/12/16 1392/9/25 Hamid Karimi Hamid Karimi Department of Basic Science, Iran karimi@tabrizu.ac.ir Mohsen Abbasi Mohsen Abbasi Iran Fatemeh Balazadehkocheh Fatemeh Balazadehkocheh Iran Belal Hasanzadeh Belal Hasanzadeh Iran Embryology Female Genital System Hamster Histology 1. Aharoni I.. Ecerpt from memories of a Hebrew zoologist.##In: Yerganian G (1972) Histology and cytogenetics of human.##Progress Experimental Tumor Research. 1942; 16:##2-41.##2. Besselsen, D.G. Biology of laboratory Rodents. Research##Animal Methods. 2002; 433-453.##3. Cushman R.A., Allan M.F., Kuehn L.A., Snelling W.M.,##Cupp, A.S. Evaluation of antral follicle count ovarian morphology##in crossbred beef cows: Investigation of influence##of the estrous cycle, age and birth weight; 2009.##4. Eurell J. A. C., Frappier, B.L. Dellmann\'s Textbook of Veterinary##Histology, John Wiley & Sons. California; 2006.##5. Gatterman r., Friotzscche, P., Winandy R., Neumann k.##Comparative studies bopdu mass, body measurement and##organ weight of wild-derived and laboratory golden hamsters##(Mesocricetus auratus); 2002.##6. Gattermann, R. 70 Jahre Goldhamster in menschiher obhut-##wie gro sind die Unterschied zu seinen wildlebenden##Verwandten? Tierlaboratorium. 2000; 23: 86-99.##7. Junqueira L.C. Carneiro J. Basic histology, 10th Ed. Long##Medical Book, London: McGraw-Hill; 2003. P. 449-453.##8. Loft B, Murton RK. (1973). Reproduction in birds, In: Avian##Biology. ED. Farmer DS and King JR Academic press##Inc, NY. 3:5-40.##9. Murphy MR. History the capture and domestication of##the syrian golden hamster (Mesocricetus auratus) Waterhouse.##IN: the hamster. Reproduction and biology (Siegel##HI, ed). New York: Planum Press; 1985.##10. Noakes, DE. Parkinson, TJ. and England, G.C.W. Arthur\'s##Veterinary Reproduction and Obstetrics, 8th Ed,##London; 2001.##11. Ozdemir D., Aydin, A., Yilmaz, S., Dine G., and Atalar,##O. Observation on morphology of the porcupine (Hystrix##cristata m). Vetyrinarki Archive. 2005; 75(2): 129-135.##24##February 2014, Volume 11, Number 1##12. Sadrkhanloo, R Abassi, M. Evaluation of number of different##of ovarian follicles in buffalo. Journal of the Faculty##of Veterinary Medicine, University of Tehran, 1999; 54 (4):##35-39.##13. Shay JW, Porter KR, and Prescott MD. The surface morphology##of CHO (Chinese hamster Ovary) cells following##enucleation. Proc. Nat. Aca. Sci. USA. 1979; 71 (8): 3059-##3063.##14. Telfer EE. The development of methods for isolation and##culture of preantral follicles from bovine and porcine ovaries,##Theriogenology. 1996; 45(1): 101-110.##15. Totay SM, Singh G, Tanja M, Pawshe CH, and Talwar##GP. In vitro Maturation, Fertilization and development of##follicular oocytes from buffalo (Bubalius bubalis) J.Reprod.##Frt, 1992; (95): 597-609.##16. Williams Carmen J, Erickson Gregory F. Morphology and##Physiology of the Ovary. 2012; Chapter 1. Available from:##http://health.com/Textbook%20of%20Endocrinology/##OVARY.##1. Aharoni I.. Ecerpt from memories of a Hebrew zoologist.##In: Yerganian G (1972) Histology and cytogenetics of human.##Progress Experimental Tumor Research. 1942; 16:##2-41.##2. Besselsen, D.G. Biology of laboratory Rodents. Research##Animal Methods. 2002; 433-453.##3. Cushman R.A., Allan M.F., Kuehn L.A., Snelling W.M.,##Cupp, A.S. Evaluation of antral follicle count ovarian morphology##in crossbred beef cows: Investigation of influence##of the estrous cycle, age and birth weight; 2009.##4. Eurell J. A. C., Frappier, B.L. Dellmann\'s Textbook of Veterinary##Histology, John Wiley & Sons. California; 2006.##5. Gatterman r., Friotzscche, P., Winandy R., Neumann k.##Comparative studies bopdu mass, body measurement and##organ weight of wild-derived and laboratory golden hamsters##(Mesocricetus auratus); 2002.##6. Gattermann, R. 70 Jahre Goldhamster in menschiher obhut-##wie gro sind die Unterschied zu seinen wildlebenden##Verwandten? Tierlaboratorium. 2000; 23: 86-99.##7. Junqueira L.C. Carneiro J. Basic histology, 10th Ed. Long##Medical Book, London: McGraw-Hill; 2003. P. 449-453.##8. Loft B, Murton RK. (1973). Reproduction in birds, In: Avian##Biology. ED. Farmer DS and King JR Academic press##Inc, NY. 3:5-40.##9. Murphy MR. History the capture and domestication of##the syrian golden hamster (Mesocricetus auratus) Waterhouse.##IN: the hamster. Reproduction and biology (Siegel##HI, ed). New York: Planum Press; 1985.##10. Noakes, DE. Parkinson, TJ. and England, G.C.W. Arthur\'s##Veterinary Reproduction and Obstetrics, 8th Ed,##London; 2001.##11. Ozdemir D., Aydin, A., Yilmaz, S., Dine G., and Atalar,##O. Observation on morphology of the porcupine (Hystrix##cristata m). Vetyrinarki Archive. 2005; 75(2): 129-135.##24##February 2014, Volume 11, Number 1##12. Sadrkhanloo, R Abassi, M. Evaluation of number of different##of ovarian follicles in buffalo. Journal of the Faculty##of Veterinary Medicine, University of Tehran, 1999; 54 (4):##35-39.##13. Shay JW, Porter KR, and Prescott MD. The surface morphology##of CHO (Chinese hamster Ovary) cells following##enucleation. Proc. Nat. Aca. Sci. USA. 1979; 71 (8): 3059-##3063.##14. Telfer EE. The development of methods for isolation and##culture of preantral follicles from bovine and porcine ovaries,##Theriogenology. 1996; 45(1): 101-110.##15. Totay SM, Singh G, Tanja M, Pawshe CH, and Talwar##GP. In vitro Maturation, Fertilization and development of##follicular oocytes from buffalo (Bubalius bubalis) J.Reprod.##Frt, 1992; (95): 597-609.##16. Williams Carmen J, Erickson Gregory F. Morphology and##Physiology of the Ovary. 2012; Chapter 1. Available from:##http://health.com/Textbook%20of%20Endocrinology/##OVARY. ##

Could Melissa Officinalis Extract Restore Streptozotocin-Induced Spatial Memory Impairment in Rats? 2 2 Introduction: Alzheimer disease is a progressive and irreversible neuropsychiatric disorder. Melissa Officinalis improves anxiety and clinical dementia symptoms caused by AD therefore, the purpose of the present study was to evaluate the effect of Mellisa extract on spatial memory deficit induced by STZ in MWM examination of male wistar rats. Methods: In the present study, 112 male wistar rats (220-270g) were used. Spatial learning deficit was induced by bilateral ICV injection of STZ (3mg/kg) via cannula then, the Melissa extract in different doses administrated by gavage and Morris Water Maze for measurement of spatial learning parameters was performed. Results: The results of this study demonstrated that gavage of the 200mg/kg of Melissa officinalis in combination with STZ, followed by significant reduction in two parameters: Time and Distance. Conclusion: Totally, the data indicate possible therapeutic value of M. officinalis extract on spatial learning improvement. 25 32 2013/07/152013/05/232013/08/72013/04/21 1392/2/1 2013/10/102013/11/172013/12/162013/09/29 1392/7/7 Fatemeh Sabbaghziarani Fatemeh Sabbaghziarani Tehran University of Medical Sciences Iran fateme_2008z@yahoo.com Sharareh Rasapour Sharareh Rasapour Khwarizmi University Iran shararerasapoor@yahoo.com Keywan Mortezaee Keywan Mortezaee Tehran University of Medical Sciences Iran keywan987@yahoo.com Shahrbano Oryan Shahrbano Oryan Khwarizmi University Iran sh_oryan@yahoo.com Mohammad Sharifzadeh Mohammad Sharifzadeh Tehran University of Medical Sciences Iran msharifzadeh@sina.tums.ac.ir Mahnaz Khanavi Mahnaz Khanavi Tehran University of Medical Sciences Iran khanavim@tums.ac.ir Gholamreza Hassanzadeh Gholamreza Hassanzadeh Tehran University of Medical Sciences Iran hassanzadeh@tums.ac.ir STZ Melissa Officinalis Spatial Memory 1. Lopez V, Martın S, Gomez-Serranillos MP, Carretero ME,##K. Jager A, and Calvo MI. Neuroprotective and Neurological##Properties of Melissa officinalis. Neurochem Res. 2009;##34:1955-1961.##2. Khalili M, and Hamzeh F. Effects of Active Constituents of##Crocus sativus L., Crocin on Streptozocin-Induced Model of##Sporadic Alzheimer\'s Disease in Male Rats. Iran. Biomed. J.##2010; 14 (1 & 2): 59-65.##3. Ondrejovic M, Kraic F, Benkovicova H, and, Silhar S. Optimisation##of Antioxidant Extraction from Lemon Balm (Melissa##officinalis). Czech J. Food Sci.2012; 30 (4): 385-393.##4. Fazli D, Malekirad AA, Pilevarian AA, Salehi H, Zerratpishe##A, Rahzani K, and Abdollahi M. Effects of Melissa officinalis##L. on Oxidative Status and Biochemical Parameters in Occupationally##Exposed Workers to Aluminum: A Before after##Clinical Trial. Int. J. Pharmacol. 2012; 8(2): 455-458.##5. Soodi M, Naghdi N, Hajimehdipoor H, Choopani S, and##Sahraei E. Memory-improving activity of Melissa officinalis##extract in naïve and scopolamine-treated rats. RPS. 2014; 9(2):##107-114.##6. Bayat M, Azami Tameh A, Ghahremani MH, Akbari M,##Ejtemaei Mehr Sh, Khanavi M and Hassanzadeh G. Neuroprotective##properties of Melissa officinalis after hypoxicischemic##injury both in vitro and in vivo. DARU Journal of##Pharmaceutical Sciences. 2012; 20: 42.##7. Hassanzadeh G, Pasbakhsh P, Akbari M, Shokri S, Ghahremani##MH, Amin Gh, Kashani I, and Azami Tameh A. Neuroprotective##Properties of Melissa Officinalis L. Extract Against##Ecstasy-Induced Neurotoxicity. Cell Journal (Yakhteh). 2011;##13(1): 25-30.##8. Hajiaghaee R, and Akhondzadeh S. Herbal medicine in##the treatment of Alzheimer’s disease. Journal of Medicinal##Plants. 2012; 11(41).##9. Kita T, Wagner G C, and Nakashima T. Current Research on##Methamphetamine-Induced Neurotoxicity: Animal Models##of Monoamine Disruption. J Pharmacol Sci. 2003. 92: 178 –##195.##10. Blokland A, Jolles J. Behavioral and biochemical effects of##an ICV injection of streptozotocin in old lewis rats. Pharmacol##Biochem Behav. 1994; 47(4): 833-7.##11. Lannert H, Hoyer S. Intracerebroventricular administration##of streptozotocin causes long-term diminutions in learning##and memory abilities and in cerebral energy metabolism in##adult rats. Behav Neurosci. 1998; 112(5): 1199-208.##12. Perry EK, Pikering AT, Wang WW, Houghton PJ, and Perry##NS. Medicinal plants and Alzheimer’s disease: integrating##ethnobotanical and contemporary scientific evidence. J Altern##Complement Med. 1998; 4: 419-28.##13. Perry EK, Pikering AT, Wang WW, Houghton PJ, and Perry##NS. Medicinal plants and Alzheimer’s disease: from etnobotany##to phytotherapy. J Pharm Pharmacol. 1999; 51: 527-34.##14. Wake G, Court J, Pickering A, Lewis R, Wilkins R, Perry E.##CNS acetylcholine receptor activity in European medicinal##plants traditionally used to improve failing memory. J Ethnopharmacol.##2000; 69: 105-14.##32##February 2014, Volume 11, Number 1##15. Ferreira A, Proneca C, Serralheiro MLM, Araujo MEM. The##in vitro screening for acetylcholinesterase inhibition and antioxidant##activity of medicinal plants from Portugal. Journal##of Ethnopharmacology. 2006; 108: 31-37.##16. Behl C. Alzheimer’s disease and oxidative stress: implications##for novel therapeutic approaches. Progress in Neurobiology.##1999; 57: 301-323.##17. Ryan MR, and Byrne O. Plant-insect coevolution and inhibition##of acetylcholinesterase. Journal of Chemical Ecology.##1988; 14: 1965-1975.##18. Akhondzadeh S, Noroozian M, Mohammadi M, Ohadinia##S, Jamshidi AH, and Khani M. Melissa officinalis extract in##the treatment of patients with mild to moderate Alzheimer’s##disease. A double blind, randomized and placebo-controlled##trial. J Neurol Neurosurg Psychiatr. 2003; 74: 863-6.##19. Iwasaki K, Satoh-Nakagawa T, Maruyama M, Monma Y,##Nemoto M, and Tomita N. A randomized, observer-blind,##Controlled trial of the traditional Chinese medicine Yi-Gan##San for improvement of behavioral and psychological symptoms##and activities of daily living in dementia patients. J Clin##Psychiatr. 2005; 66:248-52.##20. Sadraei H, Ghannadi A, and Malekshahi K. Relaxant effect##of essential oil of Melissa officinalis and citral on rat ileum##contractions. Fitoterapia. 2003; 74: 445-52.##21. Kennedy DO, Little W, Scholey AB. Attenuation of laboratory-##induced stress in humans after acute administration##of Melissa officinalis (lemon balm). Psychosom Med. 2004;##66: 607-13.##22. Kennedy DO, Scholey AB, Tildesley NTJ, Perry EK, Wesnes##KA. Modolation of mood and cognitive performance following##acute administration of Melissa officinalis (lemon balm).##Pharmacol Biochem Behav. 2002; 72: 953-64.##23. Kennedy DO, Wake G, Savelev S, Tildesley NT, Perry EK,##Wesnes KA, et al. Modolation of mood and cognitive performance##following administration of single doses of Melissa##officinalis (lemon balm) with human CNS nicotinic and muscarinic##receptor binding properties. Neuropsychopharmacology.##2003; 28: 1871-81.##24. Ballard C, O’Brien J, Reichelt K, Perry E. Aromatherapy as##a safe and effective treatment for the management of agitation##in severe dementia: the results of a double blind, placebo##Controlled trial. J Clin Psychiatr. 2002; 63: 553-8.##25. Rogers SL, Friedholf LT. Long term efficacy and safety of##donepezil in the treatment of Alzheimer’s disease: an interim##analysis of the results of a US multicenter open-label extension##study. Eur. Neuropsychopharmacol. 1998; 8: 657.##26. Le Bars PL, Katz MM, Berman N. Randomized trial of an##extract of Ginkgo biloba for dementia. J. Am. Med. Assoc.##1997; 278: 1327-32.##27. Holscher C. Possible causes of Alzheimer’s disease: amyloid##fragment, free radicals, and calcium homeostasis. Neurobiology##of Disease. 1998; 5: 129-141.##1. Lopez V, Martın S, Gomez-Serranillos MP, Carretero ME,##K. Jager A, and Calvo MI. Neuroprotective and Neurological##Properties of Melissa officinalis. Neurochem Res. 2009;##34:1955-1961.##2. Khalili M, and Hamzeh F. Effects of Active Constituents of##Crocus sativus L., Crocin on Streptozocin-Induced Model of##Sporadic Alzheimer\'s Disease in Male Rats. Iran. Biomed. J.##2010; 14 (1 & 2): 59-65.##3. Ondrejovic M, Kraic F, Benkovicova H, and, Silhar S. Optimisation##of Antioxidant Extraction from Lemon Balm (Melissa##officinalis). Czech J. Food Sci.2012; 30 (4): 385-393.##4. Fazli D, Malekirad AA, Pilevarian AA, Salehi H, Zerratpishe##A, Rahzani K, and Abdollahi M. Effects of Melissa officinalis##L. on Oxidative Status and Biochemical Parameters in Occupationally##Exposed Workers to Aluminum: A Before after##Clinical Trial. Int. J. Pharmacol. 2012; 8(2): 455-458.##5. Soodi M, Naghdi N, Hajimehdipoor H, Choopani S, and##Sahraei E. Memory-improving activity of Melissa officinalis##extract in naïve and scopolamine-treated rats. RPS. 2014; 9(2):##107-114.##6. Bayat M, Azami Tameh A, Ghahremani MH, Akbari M,##Ejtemaei Mehr Sh, Khanavi M and Hassanzadeh G. Neuroprotective##properties of Melissa officinalis after hypoxicischemic##injury both in vitro and in vivo. DARU Journal of##Pharmaceutical Sciences. 2012; 20: 42.##7. Hassanzadeh G, Pasbakhsh P, Akbari M, Shokri S, Ghahremani##MH, Amin Gh, Kashani I, and Azami Tameh A. Neuroprotective##Properties of Melissa Officinalis L. Extract Against##Ecstasy-Induced Neurotoxicity. Cell Journal (Yakhteh). 2011;##13(1): 25-30.##8. Hajiaghaee R, and Akhondzadeh S. Herbal medicine in##the treatment of Alzheimer’s disease. Journal of Medicinal##Plants. 2012; 11(41).##9. Kita T, Wagner G C, and Nakashima T. Current Research on##Methamphetamine-Induced Neurotoxicity: Animal Models##of Monoamine Disruption. J Pharmacol Sci. 2003. 92: 178 –##195.##10. Blokland A, Jolles J. Behavioral and biochemical effects of##an ICV injection of streptozotocin in old lewis rats. Pharmacol##Biochem Behav. 1994; 47(4): 833-7.##11. Lannert H, Hoyer S. Intracerebroventricular administration##of streptozotocin causes long-term diminutions in learning##and memory abilities and in cerebral energy metabolism in##adult rats. Behav Neurosci. 1998; 112(5): 1199-208.##12. Perry EK, Pikering AT, Wang WW, Houghton PJ, and Perry##NS. Medicinal plants and Alzheimer’s disease: integrating##ethnobotanical and contemporary scientific evidence. J Altern##Complement Med. 1998; 4: 419-28.##13. Perry EK, Pikering AT, Wang WW, Houghton PJ, and Perry##NS. Medicinal plants and Alzheimer’s disease: from etnobotany##to phytotherapy. J Pharm Pharmacol. 1999; 51: 527-34.##14. Wake G, Court J, Pickering A, Lewis R, Wilkins R, Perry E.##CNS acetylcholine receptor activity in European medicinal##plants traditionally used to improve failing memory. J Ethnopharmacol.##2000; 69: 105-14.##32##February 2014, Volume 11, Number 1##15. Ferreira A, Proneca C, Serralheiro MLM, Araujo MEM. The##in vitro screening for acetylcholinesterase inhibition and antioxidant##activity of medicinal plants from Portugal. Journal##of Ethnopharmacology. 2006; 108: 31-37.##16. Behl C. Alzheimer’s disease and oxidative stress: implications##for novel therapeutic approaches. Progress in Neurobiology.##1999; 57: 301-323.##17. Ryan MR, and Byrne O. Plant-insect coevolution and inhibition##of acetylcholinesterase. Journal of Chemical Ecology.##1988; 14: 1965-1975.##18. Akhondzadeh S, Noroozian M, Mohammadi M, Ohadinia##S, Jamshidi AH, and Khani M. Melissa officinalis extract in##the treatment of patients with mild to moderate Alzheimer’s##disease. A double blind, randomized and placebo-controlled##trial. J Neurol Neurosurg Psychiatr. 2003; 74: 863-6.##19. Iwasaki K, Satoh-Nakagawa T, Maruyama M, Monma Y,##Nemoto M, and Tomita N. A randomized, observer-blind,##Controlled trial of the traditional Chinese medicine Yi-Gan##San for improvement of behavioral and psychological symptoms##and activities of daily living in dementia patients. J Clin##Psychiatr. 2005; 66:248-52.##20. Sadraei H, Ghannadi A, and Malekshahi K. Relaxant effect##of essential oil of Melissa officinalis and citral on rat ileum##contractions. Fitoterapia. 2003; 74: 445-52.##21. Kennedy DO, Little W, Scholey AB. Attenuation of laboratory-##induced stress in humans after acute administration##of Melissa officinalis (lemon balm). Psychosom Med. 2004;##66: 607-13.##22. Kennedy DO, Scholey AB, Tildesley NTJ, Perry EK, Wesnes##KA. Modolation of mood and cognitive performance following##acute administration of Melissa officinalis (lemon balm).##Pharmacol Biochem Behav. 2002; 72: 953-64.##23. Kennedy DO, Wake G, Savelev S, Tildesley NT, Perry EK,##Wesnes KA, et al. Modolation of mood and cognitive performance##following administration of single doses of Melissa##officinalis (lemon balm) with human CNS nicotinic and muscarinic##receptor binding properties. Neuropsychopharmacology.##2003; 28: 1871-81.##24. Ballard C, O’Brien J, Reichelt K, Perry E. Aromatherapy as##a safe and effective treatment for the management of agitation##in severe dementia: the results of a double blind, placebo##Controlled trial. J Clin Psychiatr. 2002; 63: 553-8.##25. Rogers SL, Friedholf LT. Long term efficacy and safety of##donepezil in the treatment of Alzheimer’s disease: an interim##analysis of the results of a US multicenter open-label extension##study. Eur. Neuropsychopharmacol. 1998; 8: 657.##26. Le Bars PL, Katz MM, Berman N. Randomized trial of an##extract of Ginkgo biloba for dementia. J. Am. Med. Assoc.##1997; 278: 1327-32.##27. Holscher C. Possible causes of Alzheimer’s disease: amyloid##fragment, free radicals, and calcium homeostasis. Neurobiology##of Disease. 1998; 5: 129-141. ##

Combined Effect of Ginger and Pumpkin Seed Extracts on Rat Testis and Serum Biochemical Parameters after Cyclophosphamide Treatment 2 2 Introduction: Cyclophosphamide is a chemotherapy drug with several side effects on various organs such as male reproductive system that can cause infertility. In this study, we assessed combined effect of ginger and pumpkin extractson rat testis after CP injection. Methods: Forty adult male rats were randomly divided into 4 groups: The control group received intraperitoneal injection of isotonic saline solution. The cyclophosphamide (CP) group received a single dose of cyclophosphamide (100 mg kg_1BW) intraperitoneally. Combined extracts (ginger+ pumpkin) group received orally 300 mg combined extracts and combined extracts(ginger+ pumpkin) +CP groups received orally 300 mg combined extracts for a period of 6 weeks after CP injection. Results: Our results showed that although ginger extract could not change testis weight, testosterone, malondialdehyde (MDA) and ROS, antioxidant level in serum was increased significantly. Epithelium thickness and tube diameter were decreased in combined groups with or without CP in comparison to control group. The combined extract could improve histological changes in both combined extract and combined extract+ CP compared to CP group, which could be attributed to the higher serum level of antioxidants. Conclusion: The administration of combined extracts 33 40 2013/07/152013/05/232013/08/72013/04/212013/08/14 1392/5/23 2013/10/102013/11/172013/12/162013/09/292013/12/24 1392/10/3 Forouzan Mohamadi Forouzan Mohamadi Kashan University of Medical Sciences Iran forouzan.mohamadi@gmail.com Hossein Nikzad Hossein Nikzad Kashan University of Medical Sciences Iran nikzad_h@kaums.ac.ir Aliakbar Taherian Aliakbar Taherian Kashan University of Medical Sciences Iran Aliakbar Taherian Mohsen Taghizadeh Mohsen Taghizadeh Kashan University of Medical Sciences Iran mohsenta44@yahoo.com Abolfazl Azami-Tameh Abolfazl Azami-Tameh Kashan University of Medical Sciences Iran aazami@kaums.ac.ir Homayon Naderian Homayon Naderian Kashan University of Medical Sciences Iran naderian_h@kaums.ac.ir Mohammad Ali Atlasi Mohammad Ali Atlasi Kashan University of Medical Sciences Iran atlasi-m@kaums.ac.ir Testis Ginger Pumpkin Cyclophosphamide Biochemical parameters 1. Mohammadi F,Nikzad H,Taghizadeh M, Azami-Tameh A,##Hosseini M. Protective effect of Zingiber officinale extract##on rat testis after cyclophosphamide treatment. Andrologia.##2014. 46: 680-686.##2. Kashani HH, Malekzadeh Shiravani S,Hoshmand F.The##effect of aqueous extract of Salep prepared from root-tubers##of Dactylorhiza maculate (Orchidaceae) on the testes##and sexual hormones of immature male mice. Journal of##Medicinal Plants Research. 2012. 6(24): 4102-4106.##3. Lamfon HA. Effect of fenugreek seed extract on carbendazim-##inhibited spermatogenesis in albino rats. Journal of##Applied Pharmaceutical Science. 2012. 2(04): 09-13.##4. Aghaei S, Nikzad H, Taghizadeh M, Azami Tameh A,##Taherian A. Protective effect of Pumpkin seed extract##on sperm characteristics, biochemical parameters and##epididymal histology in adult male rats treated with Cyclophosphamide.##Andrologia. 2014. 46: 927-935.##5. Rezvanfar M,Sadrkhanlou RA. Protection of cyclophosphamide-##induced toxicity in reproductive tract histology,##sperm characteristics, and DNA damage by an herbal##source; evidence for role of free-radical toxic stress. Hum##Exp Toxicol. 2008. 27(12): 901-910.##6. Masala A, Faedda R, Alagna S, Satta A, Chiarelli G, Rovasio##PP. Use of testosterone to prevent cyclophosphamideinduced##azoospermia. Ann Intern Med. 1997. 126: 292-295.##7. Kirkland R, Bongiovanni AM, Cornfeld D, McCormick JB,##Parks JS.Gonadotropin responses to luteinizing releasing##factor in boys treated with cyclophosphamide for nephrotic##syndrome. J Pediatr. 1976. 89: 941-944.##8. Hoorweg-Nijman JJ.Cyclophosphamide-induced disturbance##of gonadotropin secretion manifesting testicular##damage. Acta endocrinologica. 1992. 126(2): 143-148.##9. Mohammadi F, Nikzad H,Taherian A,Amini Mahabadi J,##Salehi M. Effects of Herbal Medicine on Male Infertility.##Anatomical Sciences. 2013. 10(4): 3-16.##10. Erasmus N, Solomon M,Fortuin K, Henkel R.Effect of##Eurycoma longifolia Jack (Tongkat Ali) extract on human##spermatozoa in vitro. Andrologia. 2012. 44: 308-314##11. Khaki A, Fathiazad F, Nouri M,Afshin A, Hamadeh##DVM.The effects of Ginger on spermatogenesis and sperm##parameters of rat. Iranian Journal of Reproductive Medicine.##2009. 7(1): 7-12.##12. Morakinyo OA, Achema PU, Adegoke OA.Effect of Zingiber##Officinale (Ginger) on Sodium Arsenite Induced Reproductive##Toxicity in Male Rats. African Journal of Biomedical##Research. 2010. 13(1): 39-45.##13. Morakinyo A, Adeniyi OS, Arikawe AP.Effects of Zingiber##officinale on reproductive functions in the male rat.##African Journal of Biomedical Research. 2008. 11(3): 329-##334.##14. Amara S, Abdelmelek H, Garrel C, Guiraud P, Douki##T.Preventive effect of zinc against cadmium-induced oxidative##stress in the rat testis. J Reprod Dev. 2008. 54(2):##129-134.##15. Nkang A,Omokaro D,Egbe A, Amanke G.Variations in##fatty acid proportions during desiccation of Telfairia occidentalis##seeds harvested at physiological and agronomic##maturity. African Journal of Biotechnology. 2004. 2(2): 33-##39.##16. Abd El-Ghany M, Dalia A, Hafez Soha M. Biological##study on the effect of pumpkin seeds and zinc on reproductive##potential of male rats. Faculty of Specific Education##Mansoura University,Egypt April. 2010: 14-15.##17. Gundidza G, Mmbengwa VM,Magwa ML , Ramalivhana##NJ, Mukwevho NT.Aphrodisiac properties of some Zimbabwean##medicinal plants formulations. African Journal of##Biotechnology. 2009. 8(22): 6402-6407.##18. Oyeyemi, M., S. Olukole, and O. Esan Sperm morphological##studies of West African Dwarf Bucks treated with##pumpkin plant (Cucurbita pepo). Int J Morphol, 2008. 26:##121-126.##19. Ilbey YO, Ozbek E, Simsek A, Otunctemur A, Cekmen##M,Somay A.Potential chemoprotective effect of melatonin##in cyclophosphamide-and cisplatin-induced testicular##damage in rats. Fertil Steril. 2009. 92(3): 1124-1132.##20. Tsantili E, Shin Y, Nock J, Watkins C.Antioxidant concentrations##during chilling injury development in peaches.##Postharvest Biol Technol. 2010. 57(1): 27-34.##21. Djeridane A, Yousfi M, Nadjemi B, Boutassouna D,##Stocker P.Antioxidant activity of some Algerian medicinal##plants extracts containing phenolic compounds. Food##Chem. 2006. 97(4): 654-660.##22. George j.The handbook of experimental animals, The##laboratory rat. 2000, San Diego, Calif. London: Academic.##145-152.##23. Mohammadi F, Nikzad H, Taghizadeh M, Moravveji##SA.Effect of pumpkin extract regimen on testicular structure##and serum biochemical parameters in cyclophosphamide-##treated adult rats. KAUMS Journal (FEYZ). 2013.##17(5): 438-446.##24. Saalu L, Kpela T, Benebo AS, Oyewopo AO, Anifowope##EO, Oguntola JA.The Dose-Dependent Testiculoprotective##and Testiculotoxic Potentials of Telfairia occidentalis Hook##f. Leaves Extract in Rat. International Journal of Applied##Research in Natural Products. 2010. 3(3): 27-38.##25. Selvakumar E, Prahalathan C,Sudharsan P, Varalakshmi##P.Chemoprotective effect of lipoic acid against cyclophosphamide-##induced changes in the rat sperm. Toxicol. 2006.##217(1): 71-78.##26. Das UB, Mallick M, Debnath J, Ghosh D.Protective effect##of ascorbic acid on cyclophosphamide-induced testicular##gametogenic and androgenic disorders in male rats. Asian##J Androl. 2002. 4(3): 201-208.##27. Elangovan N, Chiou T, Tzeng W.Cyclophosphamide##treatment causes impairment of sperm and its fertilizing##ability in mice. Toxicol. 2006. 222(1): 60-70.##28. Nwanna EE, Oboh G.Antioxidant and Hepatoprotective##Properties of Polyphenol Extracts from Telfairia occidentaux##(Fluted Pumpkin) Leaves on Acetaminophen Induced##Liver Damage. Pakistan Journal of Biological Sciences.##2007. 10(16): 2682-2687.##40##February 2014, Volume 11, Number 1##29. Gossell-Williams M, Davis A, O\'Connor N.Inhibition##of testosterone-induced hyperplasia of the prostate of##sprague-dawley rats by pumpkin seed oil. Journal of Medicinal##Food. 2006. 9(2): 284-286.##30. Tsai YS, Tong Y, Cheng J, Lee C.Pumpkin seed oil and##phytosterol-F can block testosterone/prazosin-induced##prostate growth in rats. Urologia Internationalis. 2006.##77(3): 269-274.##31. Bourre JM, Dumont O, Durand G. Dose-effect of dietary##oleic acid: oleic acid is conditionally essential for some organs.##Reproduction Nutrition Development. 2004. 44(4):##371-380.##32. Khaki A, Iraj S, Bazi P,Imani SAM.Comparative Study of##Aminoglycosides(Gentamicin & Streptomycin) and Fluoroquinolone##(Ofloxacin) Antibiotics on Testis Tissue in##Rats: Light and Transmission Electron Microscopic Study.##Pak. J. Med. Sci. 2009. 25(4): 624-629.##33. Grzanna R, Lindmark L, Frondoza CG. Ginger-an herbal##medicinal product with broad anti-inflammatory actions.##Journal of medicinal food. 2005. 8(2): 125-132.##1. Mohammadi F,Nikzad H,Taghizadeh M, Azami-Tameh A,##Hosseini M. Protective effect of Zingiber officinale extract##on rat testis after cyclophosphamide treatment. Andrologia.##2014. 46: 680-686.##2. Kashani HH, Malekzadeh Shiravani S,Hoshmand F.The##effect of aqueous extract of Salep prepared from root-tubers##of Dactylorhiza maculate (Orchidaceae) on the testes##and sexual hormones of immature male mice. Journal of##Medicinal Plants Research. 2012. 6(24): 4102-4106.##3. Lamfon HA. Effect of fenugreek seed extract on carbendazim-##inhibited spermatogenesis in albino rats. Journal of##Applied Pharmaceutical Science. 2012. 2(04): 09-13.##4. Aghaei S, Nikzad H, Taghizadeh M, Azami Tameh A,##Taherian A. Protective effect of Pumpkin seed extract##on sperm characteristics, biochemical parameters and##epididymal histology in adult male rats treated with Cyclophosphamide.##Andrologia. 2014. 46: 927-935.##5. Rezvanfar M,Sadrkhanlou RA. Protection of cyclophosphamide-##induced toxicity in reproductive tract histology,##sperm characteristics, and DNA damage by an herbal##source; evidence for role of free-radical toxic stress. Hum##Exp Toxicol. 2008. 27(12): 901-910.##6. Masala A, Faedda R, Alagna S, Satta A, Chiarelli G, Rovasio##PP. Use of testosterone to prevent cyclophosphamideinduced##azoospermia. Ann Intern Med. 1997. 126: 292-295.##7. Kirkland R, Bongiovanni AM, Cornfeld D, McCormick JB,##Parks JS.Gonadotropin responses to luteinizing releasing##factor in boys treated with cyclophosphamide for nephrotic##syndrome. J Pediatr. 1976. 89: 941-944.##8. Hoorweg-Nijman JJ.Cyclophosphamide-induced disturbance##of gonadotropin secretion manifesting testicular##damage. Acta endocrinologica. 1992. 126(2): 143-148.##9. Mohammadi F, Nikzad H,Taherian A,Amini Mahabadi J,##Salehi M. Effects of Herbal Medicine on Male Infertility.##Anatomical Sciences. 2013. 10(4): 3-16.##10. Erasmus N, Solomon M,Fortuin K, Henkel R.Effect of##Eurycoma longifolia Jack (Tongkat Ali) extract on human##spermatozoa in vitro. Andrologia. 2012. 44: 308-314##11. Khaki A, Fathiazad F, Nouri M,Afshin A, Hamadeh##DVM.The effects of Ginger on spermatogenesis and sperm##parameters of rat. Iranian Journal of Reproductive Medicine.##2009. 7(1): 7-12.##12. Morakinyo OA, Achema PU, Adegoke OA.Effect of Zingiber##Officinale (Ginger) on Sodium Arsenite Induced Reproductive##Toxicity in Male Rats. African Journal of Biomedical##Research. 2010. 13(1): 39-45.##13. Morakinyo A, Adeniyi OS, Arikawe AP.Effects of Zingiber##officinale on reproductive functions in the male rat.##African Journal of Biomedical Research. 2008. 11(3): 329-##334.##14. Amara S, Abdelmelek H, Garrel C, Guiraud P, Douki##T.Preventive effect of zinc against cadmium-induced oxidative##stress in the rat testis. J Reprod Dev. 2008. 54(2):##129-134.##15. Nkang A,Omokaro D,Egbe A, Amanke G.Variations in##fatty acid proportions during desiccation of Telfairia occidentalis##seeds harvested at physiological and agronomic##maturity. African Journal of Biotechnology. 2004. 2(2): 33-##39.##16. Abd El-Ghany M, Dalia A, Hafez Soha M. Biological##study on the effect of pumpkin seeds and zinc on reproductive##potential of male rats. Faculty of Specific Education##Mansoura University,Egypt April. 2010: 14-15.##17. Gundidza G, Mmbengwa VM,Magwa ML , Ramalivhana##NJ, Mukwevho NT.Aphrodisiac properties of some Zimbabwean##medicinal plants formulations. African Journal of##Biotechnology. 2009. 8(22): 6402-6407.##18. Oyeyemi, M., S. Olukole, and O. Esan Sperm morphological##studies of West African Dwarf Bucks treated with##pumpkin plant (Cucurbita pepo). Int J Morphol, 2008. 26:##121-126.##19. Ilbey YO, Ozbek E, Simsek A, Otunctemur A, Cekmen##M,Somay A.Potential chemoprotective effect of melatonin##in cyclophosphamide-and cisplatin-induced testicular##damage in rats. Fertil Steril. 2009. 92(3): 1124-1132.##20. Tsantili E, Shin Y, Nock J, Watkins C.Antioxidant concentrations##during chilling injury development in peaches.##Postharvest Biol Technol. 2010. 57(1): 27-34.##21. Djeridane A, Yousfi M, Nadjemi B, Boutassouna D,##Stocker P.Antioxidant activity of some Algerian medicinal##plants extracts containing phenolic compounds. Food##Chem. 2006. 97(4): 654-660.##22. George j.The handbook of experimental animals, The##laboratory rat. 2000, San Diego, Calif. London: Academic.##145-152.##23. Mohammadi F, Nikzad H, Taghizadeh M, Moravveji##SA.Effect of pumpkin extract regimen on testicular structure##and serum biochemical parameters in cyclophosphamide-##treated adult rats. KAUMS Journal (FEYZ). 2013.##17(5): 438-446.##24. Saalu L, Kpela T, Benebo AS, Oyewopo AO, Anifowope##EO, Oguntola JA.The Dose-Dependent Testiculoprotective##and Testiculotoxic Potentials of Telfairia occidentalis Hook##f. Leaves Extract in Rat. International Journal of Applied##Research in Natural Products. 2010. 3(3): 27-38.##25. Selvakumar E, Prahalathan C,Sudharsan P, Varalakshmi##P.Chemoprotective effect of lipoic acid against cyclophosphamide-##induced changes in the rat sperm. 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Tsai YS, Tong Y, Cheng J, Lee C.Pumpkin seed oil and##phytosterol-F can block testosterone/prazosin-induced##prostate growth in rats. Urologia Internationalis. 2006.##77(3): 269-274.##31. Bourre JM, Dumont O, Durand G. Dose-effect of dietary##oleic acid: oleic acid is conditionally essential for some organs.##Reproduction Nutrition Development. 2004. 44(4):##371-380.##32. Khaki A, Iraj S, Bazi P,Imani SAM.Comparative Study of##Aminoglycosides(Gentamicin & Streptomycin) and Fluoroquinolone##(Ofloxacin) Antibiotics on Testis Tissue in##Rats: Light and Transmission Electron Microscopic Study.##Pak. J. Med. Sci. 2009. 25(4): 624-629.##33. Grzanna R, Lindmark L, Frondoza CG. Ginger-an herbal##medicinal product with broad anti-inflammatory actions.##Journal of medicinal food. 2005. 8(2): 125-132. ##

Maternal Exposure to Silver Nanoparticles in Mice: Effects on Dams’ Reproductive Performance and Pups’ Neurobehavioral Ontogeny 2 2 Introduction: - Significant increase in usage of silver nanoparticles (AgNPs) in consumer products, has increased its exposures to human and animals. Although there are some reports regarding the effects of AgNPs on complex organisms, no report was found as to its effects on neurobehavioral ontogeny. Methods: To investigate the effects of maternal exposure to AgNPs on development of neurobehavioral reflexes as well as physical indexes of growthand development during preweaning period, virgin female NMRI mice were treated with zero, 5 and 50 μg/mouse AgNPs. AgNPs were injected subcutaneously (S.C) to female mice, immediately following male exposure and at once every three days until parturition. Reproductive performance of dams were assessed and home cage activity and developmental landmarks of all pups were observed daily. Gross necropsy was performed on the 28th day. Results: No significant differences were observed in dams' and pups' weight. Survival rate was decreased (p<0.05) in NP5 group and a clear hyperactivity was observed in NP50 group. Prenatal exposure to AgNPs delayed the development of the some pups' neurobehavioral reflexes which were supposed to take place during the first four days following birth. The weight of spleen was decreased in AgNPs treated mice and the weight of liver was increased in male offspring who was exposed to AgNPs before birth. Conclusion: This study revealed that prenatal exposure to AgNPs delayed neurobehavioral development during the early stages of the pre-weaning period. Furthermore, more attention should be paid to alteration of lymphoid organ and liver weights of offspring. 41 52 2013/07/152013/05/232013/08/72013/04/212013/08/142013/06/10 1392/3/20 2013/10/102013/11/172013/12/162013/09/292013/12/242013/10/26 1392/8/4 Masome Rashno Masome Rashno Shahid Chamran University of Ahvaz, Ahvaz, Iran. Iran masome.rashno@yahoo.com Seyed Reza Fatemi Tabatabaei Seyed Reza Fatemi Tabatabaei Shahid Chamran University of Ahvaz, Ahvaz, Iran. Iran Fatemi_r@scu.ac.ir Mahmood Khaksary Mahabady Mahmood Khaksary Mahabady Shahid Chamran University of Ahvaz, Ahvaz, Iran. Iran mkhaksarymahabady@yahoo.com Shahab Ghaderi Shahab Ghaderi Shahid Chamran University of Ahvaz, Ahvaz, Iran. Iran shahabghaderi33@yahoo.com Silver Nanoparticles Neonatal Period Neurobehavioral Ontogeny 1. Benn TM, Westerhoff P. Nanoparticle silver released into##water from commercially available sock fabrics. Environ Sci##Technol 2008; 42: 4133-4139.##2. Nel A, Xia T, Madler L, Li N. Toxic potential of materials at##the Nano level. Science 2006; 311: 622-627.##3. Chaloupka K, Malam Y, Seifalian AM. Nanosilver as a##new generation of nanoproduct in biomedical applications.##Trends Biotechnol 2010; 28: 580-588.##4. Wong KKY, Liu X. Silver nanoparticles—the real ‘‘silver bullet’’##in clinical medicine? Med Chem Commun 2010; 1: 125-##131.##5. Wilkinson LJ, White RJ, Chipman JK. Silver and nanoparticles##of silver in wound dressings: a review of efficacy and##safety. J Wound Care 2011; 20: 543-549.##6. Hansen SF, Maynard A, Baun A, Tickner JA. \'Late lessons##from early warnings for nanotechnology\'. Nat Nanotechnol##2008; 3/8: 444-447.##7. Hong JS, Kim S, Lee SH, Jo E, Lee B, Yoon J, et al. Combined##repeated-dose toxicity study of silver nanoparticles with the##reproduction/developmental toxicity screening test. Nanotoxicology##2014; 8: 349-362.##8. Tang J, Xiong L, Wang S, Wang J, Liu L, Li J, et al. Distribution,##translocation and accumulation of silver nanoparticles##in rats. J Nanosci Nanotechnol 2009; 9: 4924-4932.##9. Rahman MF, Wang J, Patterson TA, Saini UT, Robinson BL,##Newport GD, et al. Expression of genes related to oxidative##stress in the mouse brain after exposure to silver-25 nanoparticles.##Toxicol Lett 2009; 187: 15-21.##10. Park EJ, Yi J, Kim Y, Choi K, Park K. Silver nanoparticles induce##cytotoxicity by a Trojan-horse type mechanism. Toxicol##in Vitro 2010; 24: 872-878.##11. Wang Z, Qu G, Su L, Wang L, Yang Z, Jiang J, et al. Evaluation##of the biological fate and the transport through biological##barriers of nanosilver in mice. Curr Pharm Des 2013; 19:##6691-6697.##12. Lee Y, Choi J, Kim P, Choi k, Kim S, Shon W, et al. A transfer##of silver nanoparticles from pregnant rat to offspring. Toxicol##Res 2012; 28: 139-141.##13. Tang J, Xiong L, Wang S, Wang J, Liu L, Li H, et al. Influence##of silver nanoparticles on neurons and blood-brain barrier##via subcutaneous injection in rats. Appl Surf Sci 2008; 225:##502-504.##14. Fatemi M, Roodbari NH, Ghaedi K, Naderi G. The effects##of prenatal exposure to silver nanoparticles on the developing##brain in neonatal rats. J Biol Res-Thessalon 2013; 20:##233-242.##15. Philbrook NA, Winn LM, Afrooz AR, Saleh NB, Walker##VK. The effect of TiO (2) and Ag nanoparticles on reproduction##and development of Drosophila melanogaster and CD-1##mice. Toxicol Appl Pharmacol 2011; 257: 429-436.##16. Kim YS, Kim JS, Cho HS, Rha DS, Kim JM, Park JD, et al.##Twenty-eight-day oral toxicity, genotoxicity, and genderrelated##tissue distribution of silver nanoparticles in Sprague-##Dawley rats. Inhal Toxicol 2008; 20: 575-583.##51##February 2014, Volume 11, Number 1##17. Kim JS, Song KS, Sung JH, Ryu HR, Choi BG, Cho HS,##et al. Genotoxicity, acute oral and dermal toxicity, eye##and dermal irritation and corrosion and skin sensitization##evaluation of silver nanoparticles. Nanotoxicology 2013; 7:##953-960.##18. Yu WJ, Son JM, Lee J, Kim SH, Lee IC, Baek HS, et al. Effects##of silver nanoparticles on pregnant dams and embryo-fetal##development in rats. Nanotoxicology 2013; [Epub ahead of##print].##19. Khaksary Mahabady M. The evaluation of teratogenicity##of nanosilver on skeletal system and placenta of rat##fetuses in prenatal period. Afr J Pharm Pharmaco 2012;##6: 419-424.##20. Fox WM. Reflex-ontogeny and behavioral development of##the mouse. Animal Behavior 1965; 13: 234-241.##21. Tsujii H, Hoshishima K. The effect of the administration of##trace amounts of metals to pregnant mice upon the behavior##and learning of their offspring. J Fac Agric Shinshu Univ##1979; 16: 13-27.##22. Tamashiro KL, Wakayama T, Blanchard RJ, Blanchard DC,##Yanagimachi R. Postnatal growth and behavioral development##of mice cloned from adult cumulus cells. Biol Reprod##2000; 63: 328-334.##23. Austin CA, Umbreit TH, Brown KM, Barber DS, Dair BJ,##Francke-Carroll S, et al. Distribution of silver nanoparticles##in pregnant mice and developing embryos. Nanotoxicology##2012; 6: 912-922.##24. Powers CM, Slotkin TA, Seidler FJ, Badireddy AR, Padilla S.##Silver nanoparticles alter zebrafish development and larval##behavior: distinct roles for particle size, coating and composition.##Neurotoxicol Teratol 2011; 33: 708-714.##25. Ratnasooriya WD, Jayakody JR, Premakumara GA. Adverse##pregnancy outcome in rats following exposure to a##Salacia reticulata (Celastraceae) root extract. Braz J Med Biol##Res 2003; 36: 931-935.##26. Ivani S, Karimi I, Tabatabaei SR. Biosafety of multiwalled##carbon nanotube in mice: a behavioral toxicological approach.##J Toxicol Sci 2012; 37: 1191-1205.##27. Li Y, Zhang Y, Yan B. Nanotoxicity overview: nano-threat##to susceptible populations. Int. J Mol Sci 2014; 15: 3671-3697.##28. Johnston HJ, Hutchison G, Christensen FM, Peters S,##Hankin S, Stone V. A review of the in vivo and in vitro toxicity##of silver and gold particulates: particle attributes and##biological mechanisms responsible for the observed toxicity.##Crit Rev Toxicol 2010; 40: 328-346.##29. Liu JY, Hurt RH. Ion release kinetics and particle persistence##in aqueous nano-silver colloids. Environ Sci Technol##2010; 44: 2169-2175.##30. Liu Y, Guan W, Ren G, Yang Z. The possible mechanism of##silver nanoparticle impact on hippocampal synaptic plasticity##and spatial cognition in rats. Toxicol Lett 2012; 209: 227-##231.##31. Ataei ML, Ebrahimzadeh-bideskan AR. The effects of nanosilver##and garlic administration during pregnancy on neuron##apoptosis in rat offspring hippocampus. Iran. J Basic Med Sci##2014; 17: 411-418.##32. Crawley JN, Paylor R. A proposed test battery and constellations##of specific behavioral paradigms to investigate the behavioral##phenotypes of transgenic and knockout mice. Horm##Behav 1997; 31: 197-211.##33. Shoults-Wilson WA, Reinsch BC, Tsyusko OV, Bertsch PM,##Lowry GV, Unrine JM. Effect of silver nanoparticle surface##coating on bioaccumulation and reproductive toxicity in##earthworms (Eisenia fetida). Nanotoxicology 2011; 5: 432-##444.##34. Asharani PV, Wu Y, Gong Z, Valiyaveettil S. Toxicity of silver##nanoparticles in zebrafish models. Nanotechnology 2008;##19: 255102.##35. Bar-Ilan O, Albrecht RM, Fako VE, Furgeson DY. Toxicity##assessments of multisized gold and silver nanoparticles in##zebrafish embryos. Small 2009; 5: 1897-1920.##36. Griffitt RJ, Luo J, Gao J, Bonzongo JC, Barber DS. Effects of##particle composition and species on toxicity of metallic nanomaterials##in aquatic organisms. Environ Toxicol Chem 2008;##27: 1972-1978.##37. Cui Y, Chen X, Zhou Z, Lei Y, Ma M, Cao R, et al. Prenatal##exposure to nanoparticulate titanium dioxide enhances##depressive-like behaviors in adult rats. Chemosphere 2014;##96: 99-104.##38. Mohammadipour A, Fazel A, Haghir H, Motejaded F,##Rafatpanah H, Zabihi H, et al. Maternal exposure to titanium##dioxide nanoparticles during pregnancy; impaired##memory and decreased hippocampal cell proliferation in##rat offspring. Environ Toxicol Pharmacol 2014; 37: 617-625.##39. Jackson P, Hougaard KS, Boisen AMZ, Jacobsen NR, Jensen##KA, Møller P, et al. Pulmonary exposure to carbon black by##inhalation or instillation in pregnant mice: Effects on liver##DNA strand breaks in dams and offspring. Nanotoxicology##2011; 6: 486-500.##40. Lim JH, Kim SH, Lee IC, Moon C, Kim SH, Shin DH, et##al. Evaluation of maternal toxicity in rats exposed to Multiwalled##carbon nanotubes during pregnancy. Environ Health##Toxicol 2011; 26: e2011006.##41. Biederman J. Attention-deficit/hyperactivity disorder: a selective##overview. Biol Psychiatry 2005; 57: 1215-1220.##42. Hussain SM, Javorina AK, Schrand AM, Duhart HM, Ali##SF, Schlager JJ. The interaction of manganese nanoparticles##with PC-12 cells induces dopamine depletion. Toxicol Sc##2006; 92: 456-463.##43. Wang J, Rahman MF, Duhart HM, Newport GD, Patterson##TA, Murdock RC, et al. Expression changes of dopaminergic##system-related genes in PC12 cells induced by manganese,##silver, or copper nanoparticles. Neurotoxicology 2009; 30:##926-933.##44. Powers CM, Wrench N, Ryde IT, Smith AM, Seidler FJ, Slotkin##TA. Silver impairs neurodevelopment: studies in PC12##cells. Environ Health Perspect 2010; 118: 73-9.##45. Powers CM, Badireddy AR, Ryde IT, Seidler FJ, Slotkin##TA. Silver nanoparticles compromise neurodevelopment##in PC12 cells: critical contributions of silver ion, particle##size, coating, and composition. Environ Health Perspect##2011; 119: 37-44.##52##February 2014, Volume 11, Number 1##46. Powers CM, Levin ED, Seidler FJ, Slotkin TA. Silver exposure##in developing zebrafish produces persistent synaptic##and behavioral changes. Neurotoxicol Teratol 2011; 33: 329-##32.##47. Hadrup N, Loeschner K, Mortensen A, Sharma AK, Qvortrup##K, Larsen EH, et al. The similar neurotoxic effects of##nanoparticulate and ionic silver in vivo and in vitro. Neurotoxicology##2012; 33: 416-423.##48. Laviola G, Adriani W, Gaudino C, Marino R, Keller F. Paradoxical##effects of prenatal acetylcholinesterase blockade on##neuro-behavioral development and drug-induced stereotypies##in reeler mutant mice. Psychopharmacology 2006; 187:##331-344.##49. Kim S, Ryu DY. Silver nanoparticle-induced oxidative##stress, genotoxicity and apoptosis in cultured cells and animal##tissues. J Appl Toxicol 2012; 33: 78-89.##50. Lee Y, Kim P, Yoon J, Lee B, Choi K, Kil KH, et al. Serum##kinetics, distribution and excretion of silver in rabbits following##28 days after a single intravenous injection of silver nanoparticles.##Nanotoxicology 2013; 7: 1120-1130.##51. Kim YS, Song MY, Park JD, Song KS, Ryu HR, Chung YH, et##al. Subchronic oral toxicity of silver nanoparticles. Part Fibre##Toxicol 2010; 7: 20.##1. Benn TM, Westerhoff P. Nanoparticle silver released into##water from commercially available sock fabrics. Environ Sci##Technol 2008; 42: 4133-4139.##2. Nel A, Xia T, Madler L, Li N. Toxic potential of materials at##the Nano level. Science 2006; 311: 622-627.##3. Chaloupka K, Malam Y, Seifalian AM. Nanosilver as a##new generation of nanoproduct in biomedical applications.##Trends Biotechnol 2010; 28: 580-588.##4. Wong KKY, Liu X. Silver nanoparticles—the real ‘‘silver bullet’’##in clinical medicine? Med Chem Commun 2010; 1: 125-##131.##5. Wilkinson LJ, White RJ, Chipman JK. Silver and nanoparticles##of silver in wound dressings: a review of efficacy and##safety. J Wound Care 2011; 20: 543-549.##6. Hansen SF, Maynard A, Baun A, Tickner JA. \'Late lessons##from early warnings for nanotechnology\'. Nat Nanotechnol##2008; 3/8: 444-447.##7. Hong JS, Kim S, Lee SH, Jo E, Lee B, Yoon J, et al. Combined##repeated-dose toxicity study of silver nanoparticles with the##reproduction/developmental toxicity screening test. Nanotoxicology##2014; 8: 349-362.##8. Tang J, Xiong L, Wang S, Wang J, Liu L, Li J, et al. Distribution,##translocation and accumulation of silver nanoparticles##in rats. J Nanosci Nanotechnol 2009; 9: 4924-4932.##9. Rahman MF, Wang J, Patterson TA, Saini UT, Robinson BL,##Newport GD, et al. Expression of genes related to oxidative##stress in the mouse brain after exposure to silver-25 nanoparticles.##Toxicol Lett 2009; 187: 15-21.##10. Park EJ, Yi J, Kim Y, Choi K, Park K. Silver nanoparticles induce##cytotoxicity by a Trojan-horse type mechanism. Toxicol##in Vitro 2010; 24: 872-878.##11. Wang Z, Qu G, Su L, Wang L, Yang Z, Jiang J, et al. Evaluation##of the biological fate and the transport through biological##barriers of nanosilver in mice. Curr Pharm Des 2013; 19:##6691-6697.##12. Lee Y, Choi J, Kim P, Choi k, Kim S, Shon W, et al. A transfer##of silver nanoparticles from pregnant rat to offspring. Toxicol##Res 2012; 28: 139-141.##13. Tang J, Xiong L, Wang S, Wang J, Liu L, Li H, et al. Influence##of silver nanoparticles on neurons and blood-brain barrier##via subcutaneous injection in rats. Appl Surf Sci 2008; 225:##502-504.##14. Fatemi M, Roodbari NH, Ghaedi K, Naderi G. The effects##of prenatal exposure to silver nanoparticles on the developing##brain in neonatal rats. J Biol Res-Thessalon 2013; 20:##233-242.##15. Philbrook NA, Winn LM, Afrooz AR, Saleh NB, Walker##VK. The effect of TiO (2) and Ag nanoparticles on reproduction##and development of Drosophila melanogaster and CD-1##mice. Toxicol Appl Pharmacol 2011; 257: 429-436.##16. Kim YS, Kim JS, Cho HS, Rha DS, Kim JM, Park JD, et al.##Twenty-eight-day oral toxicity, genotoxicity, and genderrelated##tissue distribution of silver nanoparticles in Sprague-##Dawley rats. Inhal Toxicol 2008; 20: 575-583.##51##February 2014, Volume 11, Number 1##17. Kim JS, Song KS, Sung JH, Ryu HR, Choi BG, Cho HS,##et al. Genotoxicity, acute oral and dermal toxicity, eye##and dermal irritation and corrosion and skin sensitization##evaluation of silver nanoparticles. Nanotoxicology 2013; 7:##953-960.##18. Yu WJ, Son JM, Lee J, Kim SH, Lee IC, Baek HS, et al. Effects##of silver nanoparticles on pregnant dams and embryo-fetal##development in rats. Nanotoxicology 2013; [Epub ahead of##print].##19. Khaksary Mahabady M. The evaluation of teratogenicity##of nanosilver on skeletal system and placenta of rat##fetuses in prenatal period. Afr J Pharm Pharmaco 2012;##6: 419-424.##20. Fox WM. Reflex-ontogeny and behavioral development of##the mouse. Animal Behavior 1965; 13: 234-241.##21. Tsujii H, Hoshishima K. The effect of the administration of##trace amounts of metals to pregnant mice upon the behavior##and learning of their offspring. J Fac Agric Shinshu Univ##1979; 16: 13-27.##22. Tamashiro KL, Wakayama T, Blanchard RJ, Blanchard DC,##Yanagimachi R. Postnatal growth and behavioral development##of mice cloned from adult cumulus cells. Biol Reprod##2000; 63: 328-334.##23. Austin CA, Umbreit TH, Brown KM, Barber DS, Dair BJ,##Francke-Carroll S, et al. Distribution of silver nanoparticles##in pregnant mice and developing embryos. Nanotoxicology##2012; 6: 912-922.##24. Powers CM, Slotkin TA, Seidler FJ, Badireddy AR, Padilla S.##Silver nanoparticles alter zebrafish development and larval##behavior: distinct roles for particle size, coating and composition.##Neurotoxicol Teratol 2011; 33: 708-714.##25. Ratnasooriya WD, Jayakody JR, Premakumara GA. Adverse##pregnancy outcome in rats following exposure to a##Salacia reticulata (Celastraceae) root extract. Braz J Med Biol##Res 2003; 36: 931-935.##26. Ivani S, Karimi I, Tabatabaei SR. Biosafety of multiwalled##carbon nanotube in mice: a behavioral toxicological approach.##J Toxicol Sci 2012; 37: 1191-1205.##27. Li Y, Zhang Y, Yan B. Nanotoxicity overview: nano-threat##to susceptible populations. Int. J Mol Sci 2014; 15: 3671-3697.##28. Johnston HJ, Hutchison G, Christensen FM, Peters S,##Hankin S, Stone V. A review of the in vivo and in vitro toxicity##of silver and gold particulates: particle attributes and##biological mechanisms responsible for the observed toxicity.##Crit Rev Toxicol 2010; 40: 328-346.##29. Liu JY, Hurt RH. Ion release kinetics and particle persistence##in aqueous nano-silver colloids. Environ Sci Technol##2010; 44: 2169-2175.##30. Liu Y, Guan W, Ren G, Yang Z. The possible mechanism of##silver nanoparticle impact on hippocampal synaptic plasticity##and spatial cognition in rats. Toxicol Lett 2012; 209: 227-##231.##31. Ataei ML, Ebrahimzadeh-bideskan AR. The effects of nanosilver##and garlic administration during pregnancy on neuron##apoptosis in rat offspring hippocampus. Iran. J Basic Med Sci##2014; 17: 411-418.##32. Crawley JN, Paylor R. A proposed test battery and constellations##of specific behavioral paradigms to investigate the behavioral##phenotypes of transgenic and knockout mice. Horm##Behav 1997; 31: 197-211.##33. Shoults-Wilson WA, Reinsch BC, Tsyusko OV, Bertsch PM,##Lowry GV, Unrine JM. Effect of silver nanoparticle surface##coating on bioaccumulation and reproductive toxicity in##earthworms (Eisenia fetida). Nanotoxicology 2011; 5: 432-##444.##34. Asharani PV, Wu Y, Gong Z, Valiyaveettil S. Toxicity of silver##nanoparticles in zebrafish models. Nanotechnology 2008;##19: 255102.##35. Bar-Ilan O, Albrecht RM, Fako VE, Furgeson DY. Toxicity##assessments of multisized gold and silver nanoparticles in##zebrafish embryos. Small 2009; 5: 1897-1920.##36. Griffitt RJ, Luo J, Gao J, Bonzongo JC, Barber DS. Effects of##particle composition and species on toxicity of metallic nanomaterials##in aquatic organisms. Environ Toxicol Chem 2008;##27: 1972-1978.##37. Cui Y, Chen X, Zhou Z, Lei Y, Ma M, Cao R, et al. Prenatal##exposure to nanoparticulate titanium dioxide enhances##depressive-like behaviors in adult rats. Chemosphere 2014;##96: 99-104.##38. Mohammadipour A, Fazel A, Haghir H, Motejaded F,##Rafatpanah H, Zabihi H, et al. Maternal exposure to titanium##dioxide nanoparticles during pregnancy; impaired##memory and decreased hippocampal cell proliferation in##rat offspring. Environ Toxicol Pharmacol 2014; 37: 617-625.##39. Jackson P, Hougaard KS, Boisen AMZ, Jacobsen NR, Jensen##KA, Møller P, et al. Pulmonary exposure to carbon black by##inhalation or instillation in pregnant mice: Effects on liver##DNA strand breaks in dams and offspring. Nanotoxicology##2011; 6: 486-500.##40. Lim JH, Kim SH, Lee IC, Moon C, Kim SH, Shin DH, et##al. Evaluation of maternal toxicity in rats exposed to Multiwalled##carbon nanotubes during pregnancy. Environ Health##Toxicol 2011; 26: e2011006.##41. Biederman J. Attention-deficit/hyperactivity disorder: a selective##overview. Biol Psychiatry 2005; 57: 1215-1220.##42. Hussain SM, Javorina AK, Schrand AM, Duhart HM, Ali##SF, Schlager JJ. The interaction of manganese nanoparticles##with PC-12 cells induces dopamine depletion. Toxicol Sc##2006; 92: 456-463.##43. Wang J, Rahman MF, Duhart HM, Newport GD, Patterson##TA, Murdock RC, et al. Expression changes of dopaminergic##system-related genes in PC12 cells induced by manganese,##silver, or copper nanoparticles. Neurotoxicology 2009; 30:##926-933.##44. Powers CM, Wrench N, Ryde IT, Smith AM, Seidler FJ, Slotkin##TA. Silver impairs neurodevelopment: studies in PC12##cells. Environ Health Perspect 2010; 118: 73-9.##45. Powers CM, Badireddy AR, Ryde IT, Seidler FJ, Slotkin##TA. Silver nanoparticles compromise neurodevelopment##in PC12 cells: critical contributions of silver ion, particle##size, coating, and composition. Environ Health Perspect##2011; 119: 37-44.##52##February 2014, Volume 11, Number 1##46. Powers CM, Levin ED, Seidler FJ, Slotkin TA. Silver exposure##in developing zebrafish produces persistent synaptic##and behavioral changes. Neurotoxicol Teratol 2011; 33: 329-##32.##47. Hadrup N, Loeschner K, Mortensen A, Sharma AK, Qvortrup##K, Larsen EH, et al. The similar neurotoxic effects of##nanoparticulate and ionic silver in vivo and in vitro. Neurotoxicology##2012; 33: 416-423.##48. Laviola G, Adriani W, Gaudino C, Marino R, Keller F. Paradoxical##effects of prenatal acetylcholinesterase blockade on##neuro-behavioral development and drug-induced stereotypies##in reeler mutant mice. Psychopharmacology 2006; 187:##331-344.##49. Kim S, Ryu DY. Silver nanoparticle-induced oxidative##stress, genotoxicity and apoptosis in cultured cells and animal##tissues. J Appl Toxicol 2012; 33: 78-89.##50. Lee Y, Kim P, Yoon J, Lee B, Choi K, Kil KH, et al. Serum##kinetics, distribution and excretion of silver in rabbits following##28 days after a single intravenous injection of silver nanoparticles.##Nanotoxicology 2013; 7: 1120-1130.##51. Kim YS, Song MY, Park JD, Song KS, Ryu HR, Chung YH, et##al. Subchronic oral toxicity of silver nanoparticles. Part Fibre##Toxicol 2010; 7: 20. ##

Evaluation of Different Genotypes at Codons 11, 72 and 248 of p53 Gene in Samples of Endometriosis 2 2 Introduction: Endometriosis is a prevalent gynecological disorder among women which is diagnosed by the growth of endometrial tissue outside of uterus and is mainly accompanied by severe pelvic pain and infertility. P53 also known as cellular tumor antigen P53 inside codons 11, 72 and 248 are contained with single nucleotide changes in which tends to be nearly rampant.This will probably be increasing the chances of endometriosis infection to some great extent. Our aim was to evaluate the connection between endometriosis and polymorphism inside the codons 11, 72 and 248 of P53 gene. Methods: In this study, single nucleotide changes in codons 11, 72 and 284 of TP53 gene among 44 persons infected with endometriosis and the same studying population for noninfected group have been examined. After primer design and amplification of polymorphic sequences by PCR, the polymorphisms of related codons have been evaluated by the digestion method (RFLP). Results: In this study on the codon 72, there were seen differences in the distribution of genotype frequencies of normal polymorphic subjects and control subjects with endometriosis. At codons 11 and 248, there were observed no significant correlations between polymorphic and normal genotypes of endometriosis and non-endometriosis groups. Conclusion: According to the results, we can say that probably polymorphism 53 58 2013/07/152013/05/232013/08/72013/04/212013/08/142013/06/102013/07/19 1392/4/28 2013/10/102013/11/172013/12/162013/09/292013/12/242013/10/262013/12/18 1392/9/27 Hajar Dahim Hajar Dahim department of biochemistry Iran h.dahim Kahin Shahani Kahin Shahani department of biochemistry Iran shahani@IAUFALA.ac.ir Ahmad Sabanizadeh Ahmad Sabanizadeh department of anatomy Iran shebani54@yahoo.com mohsen mohsen Iran taghavi164@yahoo.com Polymorphisms Gene P53 Endometriosis ## ##

Development of an Anaglyph Anatomical Atlas of Rabbit as a New Generation of Printable 3D Anatomical Atlases 2 2 Introduction: To improve anatomical atlases, we tried to create a printable 3D atlas of rabbit's anatomy based on the Anaglyph. Methods: In this study, twenty white rabbits euthanized and dissected. All 3D images were reconstructed by means of "stereo photo maker (SPM®)" software. Results: By this atlas, users can easily understand nomenclature, the topography and, certainly, the transposition of the anatomical structures in each image even if they are not at the dissecting room. Conclusion: The novelty of this atlas may encourage students to spend more time and concentrate on the material more as opposed to the classical atlases, thereby, leading to better understanding of anatomy. It is worth mentioning that this atlas has been prepared as an example for other works. 59 65 2013/07/152013/05/232013/08/72013/04/212013/08/142013/06/102013/07/192013/04/21 1392/2/1 2013/10/102013/11/172013/12/162013/09/292013/12/242013/10/262013/12/182013/08/10 1392/5/19 سید مسعود ذوالحواریه Seyed Masoud Zolhavarieh Department of Clinical Sciences, Faculty of Para veterinary Science, Bu-Ali Sina University, Hamedan, Iran. Iran mzolhavarieh@basu.ac.ir مهران وطنچیان Mehran Vatanchian Department of Anatomy and Cell biology, School of Medicine, North Khorasan University of Medical Sciences, Bojnord, Iran. Iran vatanchian@nkums.ac.ir Anatomy Rabbit Education Anaglyph Three- dimensional Image 1. Salazar I. Coming changes in veterinary anatomy: what is or##should be expected? J vet med educ. 2002; 29(3): 126-30.##2. Aziz MA, Mckenzie JC, Wilson JS, Cowie RJ, Ayeni SA,##Dunn BK. The human cadaver in the age of biomedical informatics.##Anat Rec. 2002; 269(1): 20-32.##3. Cahill D, Leonard R, Marks S. A comment on recent teaching##of human anatomy in the United States. Surg Radiol Anat.##2000; 22(2): 69-71.##4. Gregory SR, Cole TR. The changing role of dissection in medical##education. JAMA. 2002; 287(9): 1180-1.##5. Evans HE, DeLahunta A. Guide to the dissection of the dog:##WB Saunders; 2004.##6. Older J. Anatomy: A must for teaching the next generation.##The Surgeon. 2004; 2(2): 79-90.##7. Marks SC. The role of three-dimensional information in##health care and medical education: The implications for##anatomy and dissection. Clin Anat. 2000; 13(6): 448-52.##8. Gatesy SM, Shubin NH, Jenkins Jr FA. Anaglyph stereo imaging##of dinosaur track morphology and microtopography.##Palaentol Electron. 2005; 8(1): 814.##9. Ackerman MJ. The visible human project: a resource for education.##Acad Med. 1999; 74(6): 667-70.##10. Spitzer VM, Whitlock DG. The visible human dataset: the##anatomical platform for human simulation. Anat Rec. 1998;##253(2): 49-57.##11. Bacro T, Gilbertson B, Coultas J. Web-delivery of anatomy##video clips using a CD-ROM. Anat Rec. 2000; 261(2): 78-82.##12. Gatti A. The use of ADAM software in podiatry. Clin Podiatr##Med Surg. 1993; 10(3): 563.##13. Inwood MJ, Ahmad J. Development of instructional, interactive,##multimedia anatomy dissection software: A studentled##initiative. Clin Anat. 2005; 18(8): 613-7.##14. Gehrmann S, Höhne K, Linhart W, Pflesser B, Pommert##A, Riemer M, et al. A novel interactive anatomic atlas of the##hand. Clin Anat. 2006; 19(3): 258-66.##15. Bryson-Richardson RJ, Berger S, Schilling TF, Hall TE, Cole##NJ, Gibson AJ, et al. FishNet: an online database of zebrafish##anatomy. BMC Biol. 2007; 5(1): 34.##16. Nicholson DT, Chalk C, Funnell WRJ, Daniel SJ. Can virtual##reality improve anatomy education? A randomised##controlled study of a computer-generated three-dimensional##anatomical ear model. Med Educ. 2006; 40(11): 1081-7.##65##February 2014, Volume 11, Number 1##17. Yelnik J, Bardinet E, Dormont D, Malandain G, Ourselin S,##Tandé D, et al. A three-dimensional, histological and deformable##atlas of the human basal ganglia. I. Atlas construction##based on immunohistochemical and MRI data. Neuroimage.##2007; 34(2): 618-38.##18. Chang AJ, Kim HJ, Choi JW, Yu KY. Ghosting reduction##method for color anaglyphs. Proceedings of SPIE Stereoscopic##Displays and Applications XIX. 2008; 6803: 68031G.##19. Hristov H, Kostov D, Vladova D. Topographical anatomy##of some abdominal organs in rabbits. TJS. 2006; 4(3): 7-10.##20. Zotti A, Banzato T, Cozzi B. Cross-sectional anatomy of the##rabbit neck and trunk: comparison of computed tomography##and cadaver anatomy. Res Vet Sci. 2009; 87(2): 171-6.##21. Zolhavarieh SM, Nourian A, Sadeghi-Nasab A. A new##method for on-farm euthanasia with animal welfare considerations.##IJVS. 2011; 6(1/2): 55-63.##22. Zraqou J, Alkhadour W, Qahwaji R, Ipson S, Ugail H. Enhanced##3D perception using Super-resolution and saturation##control techniques for solar images. UbiCC. 2009; 4(4): 68-90.##23. Popesko P, Rajtová V, Horák J, Timčáková I. A colour atlas##of the anatomy of small laboratory animals: Wolfe; 1992.##1. Salazar I. Coming changes in veterinary anatomy: what is or##should be expected? J vet med educ. 2002; 29(3): 126-30.##2. Aziz MA, Mckenzie JC, Wilson JS, Cowie RJ, Ayeni SA,##Dunn BK. The human cadaver in the age of biomedical informatics.##Anat Rec. 2002; 269(1): 20-32.##3. Cahill D, Leonard R, Marks S. A comment on recent teaching##of human anatomy in the United States. Surg Radiol Anat.##2000; 22(2): 69-71.##4. Gregory SR, Cole TR. The changing role of dissection in medical##education. JAMA. 2002; 287(9): 1180-1.##5. Evans HE, DeLahunta A. Guide to the dissection of the dog:##WB Saunders; 2004.##6. Older J. Anatomy: A must for teaching the next generation.##The Surgeon. 2004; 2(2): 79-90.##7. Marks SC. The role of three-dimensional information in##health care and medical education: The implications for##anatomy and dissection. Clin Anat. 2000; 13(6): 448-52.##8. Gatesy SM, Shubin NH, Jenkins Jr FA. Anaglyph stereo imaging##of dinosaur track morphology and microtopography.##Palaentol Electron. 2005; 8(1): 814.##9. Ackerman MJ. The visible human project: a resource for education.##Acad Med. 1999; 74(6): 667-70.##10. Spitzer VM, Whitlock DG. The visible human dataset: the##anatomical platform for human simulation. Anat Rec. 1998;##253(2): 49-57.##11. Bacro T, Gilbertson B, Coultas J. Web-delivery of anatomy##video clips using a CD-ROM. Anat Rec. 2000; 261(2): 78-82.##12. Gatti A. The use of ADAM software in podiatry. Clin Podiatr##Med Surg. 1993; 10(3): 563.##13. Inwood MJ, Ahmad J. Development of instructional, interactive,##multimedia anatomy dissection software: A studentled##initiative. Clin Anat. 2005; 18(8): 613-7.##14. Gehrmann S, Höhne K, Linhart W, Pflesser B, Pommert##A, Riemer M, et al. A novel interactive anatomic atlas of the##hand. Clin Anat. 2006; 19(3): 258-66.##15. Bryson-Richardson RJ, Berger S, Schilling TF, Hall TE, Cole##NJ, Gibson AJ, et al. FishNet: an online database of zebrafish##anatomy. BMC Biol. 2007; 5(1): 34.##16. Nicholson DT, Chalk C, Funnell WRJ, Daniel SJ. Can virtual##reality improve anatomy education? A randomised##controlled study of a computer-generated three-dimensional##anatomical ear model. Med Educ. 2006; 40(11): 1081-7.##65##February 2014, Volume 11, Number 1##17. Yelnik J, Bardinet E, Dormont D, Malandain G, Ourselin S,##Tandé D, et al. A three-dimensional, histological and deformable##atlas of the human basal ganglia. I. Atlas construction##based on immunohistochemical and MRI data. Neuroimage.##2007; 34(2): 618-38.##18. Chang AJ, Kim HJ, Choi JW, Yu KY. Ghosting reduction##method for color anaglyphs. Proceedings of SPIE Stereoscopic##Displays and Applications XIX. 2008; 6803: 68031G.##19. Hristov H, Kostov D, Vladova D. Topographical anatomy##of some abdominal organs in rabbits. TJS. 2006; 4(3): 7-10.##20. Zotti A, Banzato T, Cozzi B. Cross-sectional anatomy of the##rabbit neck and trunk: comparison of computed tomography##and cadaver anatomy. Res Vet Sci. 2009; 87(2): 171-6.##21. Zolhavarieh SM, Nourian A, Sadeghi-Nasab A. A new##method for on-farm euthanasia with animal welfare considerations.##IJVS. 2011; 6(1/2): 55-63.##22. Zraqou J, Alkhadour W, Qahwaji R, Ipson S, Ugail H. Enhanced##3D perception using Super-resolution and saturation##control techniques for solar images. UbiCC. 2009; 4(4): 68-90.##23. Popesko P, Rajtová V, Horák J, Timčáková I. A colour atlas##of the anatomy of small laboratory animals: Wolfe; 1992. ##