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Cartoons are children's favorite programs, but as parents, it is very important to monitor what kind of fun programs they are watching because not all of them are recommendable for your child. Among them is the one series that is called the Happy Tree Friends. Although it looks like an innocent and harmless cartoon, it leaves a lot to be desired. It is full of violent scenes and the main theme seems to be death.

Cartoons should also be a way through which your child learns speech, surprisingly, Happy Tree Friends cartoon has some character who neither speak nor make sensible dialogue but instead they mumble some incomprehensible word. This is the best way to kill your child's speech confidence ad if you want the best for your child, keep him or her away from this cartoon.

This program is available on the Internet and it is therefore crucial that you monitor what your children have access to from the Internet. It went online back in 2000, when it was represented at film festivals. It gained popularity the world over in countries like Canada, Brazil, Italy and Russia. From here, it was transferred to the television channels.

All the characters in this cartoon series are personified animals who share some common characteristics and this is how you are able to identify them. They have two front buck teeth and heart shaped noses. The episodes all start in a peaceful way but transgresses into violence. The creators of the Happy Tree fiends have released four DVDs with these animal shows and it has been a great success.

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Materials and Methods Used for Studying Standard Freezing and Thawing Protocols for Cryopreservation of Biopsied Embryos at the Blastocyst Stage

Experimental design

Test group: between February 2005 and October 2007,148 human IVF combined PGD cycles were performed at our center. A total 173 blastocysts were available for freezing. Of those, 44 were euploid embryos and 129 were aneuploid embryos.

Control group: during the study period, 54 frozen embryo transfer cycles were performed using intact frozen-thawed blastocyst including 185 non-biopsied embryos.

Embryo biopsy

Embryo biopsy was performed with three micropipettes at 68-70 hours after egg retrieval. Biopsy medium contains a HEPES buffered HTF lacking Ca2+ and Mg2+ ions with EDTA( 0.5mn) and HSA (5mg/ml)(Irine Scientific, USA). Zona drilling was performed by using acidified Tryode’s Solution (PH 2.1-2.5). A single blastomere was removed per embryo. After biopsy, embryos were cultured for 24 to 48 hours before transfer. Following fresh embryo transfer, per informed consent, remaining blastocysts graded A or B were frozen in separate vials based on PGD outcome (euploid or aneuploid).

Embryo freezing and thawing

Freezing protocol: Embryos were sequentially equilibrated in F1 and F2 blastocyst freeze media (Irvine Scientific,USA) for 10 minutes respectively at room temperature. Then, they were loaded into vials with 0.5ml F2 medium. Cooling was conducted in freezing machine (Freeze Control CL-8800, Biogenics, USA) as followings: cooling rate was 2°C/minute from 20°C to -6.5°C, held embryos for 5 minutes at -6.5°C, and then manual seeding was done and held embryos for another10minutes at -6.5°C. Cooling rate was -0.3°C/minute from -6.5°C to -38°C. As the temperature reached to -38°C, held embryos for 5 minutes and plunged into LN2. All blastocysts were kept frozen for a minimum of one month before thawing.

Thawing protocol: Vials were removed from storage tank, held at room temperature for 1 minute and immersed in 37°C water bath; embryos were taken out of vials and transferred to T1 and T2 blastocyst thaw media (Irvine Scientific, USA) sequentially for 10 minutes each at room temperature .After embryos were equilibrated in T2 medium , they were cultured in blastocyst culture medium ( Global +10% SSS, LifeGlobal).

Embryo evaluation and frozen embryo transfer

Embryo survivability was determined one hour after thawing .Any blastocyst with >50% or >90% viable cells was labeled as survival embryo or better survival embryo respectively. Embryo with total lysed cells was recorded separately. The formation of blastocyst cavity was evaluated after thawing 8 hours. Normal embryos with relative good quality were selected for transfer in each cycle. Aneuploid embryos were discarded after evaluation.

Clinical outcome

Clinical pregnancy was diagnosed by ultrasound with heart beat after embryo transfer 4 weeks. Implantation rate was obtained by the percentage of gestational sacs with fetal heart beat divided by the total number of embryos transferred.

Statistical analysis

Data was analyzed by ?2-test.

About the Author

Myself webmaster of http://www.vermesh.com/ , a service provider of assured fertility solutions, specializes in Tubal Reversal Treatment, Tubal Ligation Reversal, Fertility Clinics, Egg Donation, Embryo Transfer, Fertility Doctor, Gestational Surrogacy, and In vitro fertilization

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