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Mehdi Hajmohammadi,1 Seyed Davar Siadat,2 Masoud Ghorbani,3,4,* Mehdi Shafiee Ardestani,5,* Shahram Teimourian,6 Vahid Asgari,3 Reza Ahangari Cohan,3 Mostafa Hajmohammadi,5 Akram Hajmohammadi,7 Ramezan Behzadi,8 Saied Rajab Nezhad,9 Nabiollah Namvar Asl10 1Department of Research and Biotechnology, 2Department of Microbiology, 3Department of Virology, Pasteur Institute of Iran, Tehran Iran; 4Department of Virology, University of Ottawa, Ottawa, ON, Canada; 5Department of Radiopharmacy, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran; 6Department of Medical Genetics, Iran University of Medical Sciences, Tehran, 7Faculty of Pharmacy, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, 8Laboratory Animal Management of North Research Center, Pasteur Institute of Iran, 9Department of Research and Development, Alhavi Pharmaceutical, Tehran, 10Pasteur Institute of Iran, Department of Animal Sciences, Karaj, Iran *These authors contributed equally tothis work Abstract: In recent years, many experiments have been conducted for the production and evaluation of anticancer glycoconjugated vaccines in developed countries and many achievements have been accomplished with Globo H derivatives. In the current experiment, a new chemically designed triplicate version of (Globo H)3–diethylenetriamine pentaacetic acid (DTPA)–KLH antigen was synthesized and characterized. Immunization with (Globo H)3-DTPA-KLH, a hexasaccharide that is a member of a family of antigenic carbohydrates that are highly expressed in various types of cancers conjugated with DTPA and KLH protein, induced a high level of antibody titer along with an elevated level of IL-4 in mice. Treatment of tumors with the collected sera from immunized mice decreased the tumor size in nude mice as well. None of the immunized mice illustrated any sign of tumor growth after injection of MCF-7 cells compared to the control animals. These findings, based on the newly presented structure of the Globo H antigen, lend exciting and promising evidence for clinical advancement in the development of a therapeutic vaccine in the future. Keywords: (Globo H)3-DTPA-KLH, glycoconjugate vaccines, breast cancer

Objective: This study introduced a simple method for bovine blastocyst vitrification.Materials and Methods: Bovine blastocysts were produced in vitro by means of a wholeco-culture system with vero cells. The blastocysts were randomly divided 1:3 into either vitrification(100 blastocysts) or control (43 blastocysts) groups. For vitrification,expanded - blastocystswere incubated first in equilibration medium for 8 minutes and then in the vitrificationsolution for 1 minute. The blastocysts were then loaded in the tip of a handmade cryotip forimmediate - deep freezing in liquid nitrogen. Frozen embryos were then warmed by directlyimmersing the tips in sequential warming solutions . warmed embryos were cultured for afurther period of 48 hours when the ratios of re-expansion, hatching and degeneration werecompared with the control group.Results: After warming, in the vitrified and control groups the ratios of re-expansion were78.5% ± 0.067 and 81.6% ± 0.072, the ratios of hatching were 43.7% ± 0.083 and 49.8%± 0.089 and the ratios of degeneration were 36% ± 0.082 and 22.3% ± 0.087, respectively,which were not significantly different between the two groups.Conclusion: Post - warming survival of the vitrified and non - vitrified embryos were notsignificantly different, handmade cryotips can be used as an efficient and feasible device forbovine blastocysts vitrification.

In vitro embryo production and embryo vitrification of genetically superior cows that culled inevitably due to health problems can accelerate genetic progress. This study was carried out to investigate whether maternal age and health status effects of high genetic merit cows affect cryosurvival and developmental competence of IVP embryos. In this sense, the effects of ageing and four common culling causes of dairy cows [repeat breeding (RPB), udder problems (UPM), chronic endometritis (CRE), and lameness (LAM)] on in vitro embryo development, and in vivo developmental competence after embryo vitrification were evaluated. The mean number of oocytes obtained per cow did not vary significantly between donors indifferent groups. Cleavage rates in RPB (86.0+/-4.2%), SEN (81.3+/-2.5%) and CRE (77.6+/-6.3%) cows which were comparable to control (95.9+/-1.5%) but were significantly higher than the related rate of UPM donors (50.6+/-2.6%). Importantly, there was no significant difference between the blastocyst rates of different groups. Mean overall survival rate was not different between the groups and was not affected by the blastocyst production rate. There was no significant difference between pregnancy rates of different groups. The results of the present study indicated that in cattle, neither ageing, nor these four diseases affect ovarian potential in terms of the yield and quality of in vitro embryo development.