Judul: Produksi Embrio Kloning Mencit dengan Pengembangan Teknik Transfer Inti Sel Somatis
Abstrak: Embryonic stem cells can be obtained from an embryo generated through fertilization, recently it has been reported that an embryo can also be generated asexually through Somatic Cell Nuclear Transfer (SCNT). This procedure will overcome the ethical issue regarding the usage of embryo that was initially generated for reproductive purposes. The low efficiency of reproductive cloning is one of the reasons to explore the use of SCNT for biomedical purpose, specifically through nuclear transfer Embryonic Stem Cell (ntESC) production. Cloned embryos undergo DNA hypermethylation, which is thought to be one of the causes of sub-optimal growth. This study has five main objective that include the following: (1) to obtain an optimized oocyte superovulation, activation method, and enucleation using mice oocytes as a model; (2) to produce mouse cloned embryo, parthenogenetic embryo, and in vivo fertilized embryo; (3) to study the stages of pre-implantation development in vitro culture; (4) to determined the cumulus cell cycle using flow cytometry; (5) to study the development of SCNT technique by adding Scriptaid as an inhibitory enzyme HDAC (histone deacetylase). This study showed that using doses of 7.5 IU PMSG and hCG hormones can increase the oocyte cell production optimization through superovulation treatment. The most optimal method of activation is medium CZB & CB & SrCl2 (fresh medium) for six hours (method C) to activate oocyte cells until it reaches the level of activation of 97.21%. The addition of 3% sucrose able to increase the enucleation rate. The result of the research indicated that SCNT application is able to produce cloned embryos which are capable to develop to blastocyst stage (3,2%). In addition artificial activation of oocytes could produce parthenogenetic embryos which are capable to develop up to the blastocyst stage (8,6%). The results show that cumulus cells used as nuclei donors are in the G0/G1 cell cycle phase. The cumulus cells measure at 5-7 μm in diameter and accounts for 70,84% of the total isolated cumulus cell population. The enucleation process could achieve an efficiency rate of 49%, while the nuclear transfer achieves a 40,8% efficiency rate. The addition of Scriptaid successfully increase the embryonic cloning at the blastocyst stage’s success rate to 10,8%. This efficiency rate is three times higher compared to the control, which only shows a 3,2% success rate. Optimization of the superovulation treatment, enucleation, and the activation method could improve the efficiency of SCNT application to produce cloned mice embryos. Cumulus cells were used as donor nuclei has been confirmed at G0 or G1 stage of the cell cycle. Embryos cloned mice could be generated through the SCNT application. Preimplantation development of fertilized and parthenogenetic embryos still better than the cloned embryos. The addition of Scriptaid and TSA could increase the level of efficiency of up to threefold compared with controls.
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Judul: Produksi Embrio pada Berbagai Fase Estrus Mencit setelah Penyuntikan Kombinasi PMSG-hCG dan r-hFSH – r-hCG.
Abstrak: Perkembangan teknologi rekayasa genetika menghasilkan suatu inovasi sediaan hormonal gonadotropin rekombinan yang dapat diaplikasikan untuk meningkatkan efisiensi reproduksi yang bermanfaat di masa mendatang. Penelitian ini bertujuan mempelajari dan mengidentifikasi efektivitas respons stimulasi superovulasi pemberian sediaan hormon gonadotropin eksogen (PMSG/Folligon® dan hCG/Chorulon®) dengan hormon rekombinan (r-FSH:r-LH/Pergoveris® dan r-hCG/Ovidrel®) terhadap tahap perkembangan embrio di fase siklus estrus yang optimal pada produksi embrio secara in vivo pada mencit (Mus musculus). Hewan coba yang digunakan dalam penelitian ini adalah mencit betina dan jantan yang telah diaklimatisasi selama 2 minggu. Penelitian ini dilakukan pada dua kelompok perlakuan. Pada kelompok pertama diberikan sediaan kombinasi Folligon® dan Chorulon® dan kelompok kedua diberikan sediaan kombinasi Pergoveris® dan Ovidrel®. Untuk mengetahui fase estrus pada mencit, masing-masing kelompok perlakuan dilakukan pemeriksaan ulas vagina. Seluruh kelompok perlakuan diinjeksi Folligon®-Chorulon® dan Pergoveris®-Ovidrel® secara intraperitoneal. Perkawinan mencit dilakukan setelah penyuntikan Chorulon® atau Ovidrel®. Koleksi embrio dilakukan pada hari ke-3 setelah dikawinkan. Hasil penelitian menunjukkan rata-rata jumlah ovulasi embrio mencit pada setiap fase siklus estrus tidak berpengaruh nyata. Respon stimulasi superovulasi yang optimal cenderung terdapat pada mencit yang distimulasi menggunakan hormon kombinasi Pergoveris® dan Ovidrel® pada fase estrus., The development of genetic engineering technology has resulted in an innovation of recombinant gonadotropin hormonal preparations that can be applied to increase reproductive efficiency which is beneficial in the future. This study aims to study and identify the effectiveness of the superovulation stimulation response by administering exogenous gonadotropin hormones (PMSG/Folligon® and hCG/Chorulon®) with recombinant hormones (r-FSH:r-LH/Pergoveris® and r-hCG/Ovidrel®) on the stages of embryo development in the optimal estrus cycle in embryo production in vivo in mice (Mus musculus). The experimental animals used in this study were female and male mice that had been acclimatized for 2 weeks. This research was conducted in two treatment groups. The first group was given a combination of Folligon® and Chorulon® and the second group was given a combination of Pergoveris® and Ovidrel®. To determine the estrus phase in mice, each treatment group was examined by vaginal smear. All treatment groups were injected intraperitoneally with Folligon®-Chorulon® and Pergoveris®-Ovidrel®. The mating of mice was carried out after injection of Chorulon® or Ovidrel®. Embryo collection was carried out on the 3rd day after the mating. The results showed that the average number of ovulations of mouse embryos in each phase of the estrous cycle had no significant effect. Optimal superovulation stimulation responses tend to occur in mice stimulated using a combination of Pergoveris® and Ovidrel® hormones during the estrous phase.
Keyword: mice, Folligon®-Chorulon®, Pergoveris®-Ovidrel®, superovulation

Judul: Iklim Pulau Bali
Abstrak: Curah hujan rata-rata bulanan di Bali berkisar antara 47 mm sampai 327 mm, dengan curah hujan maksimum jatuh pada bulan Januari dan minimum pada bulan Agustus. Total hujan setahun rata-rata di Bali 1901 mm. Pada muim angin barat, curah hujan di utara pegunungan relatif sama dengan di selatan pegunungan, sedangkan pada musim angin timur curah hujan di selatan pegunungan lebih tinggi daripada di utara pegunungan. Isohyet tiap bulan di Bali menunjukkan pengaruh ого- grafis terjadi di bagian timur P. Bali yang mengakibatkan di selatan pegunungan lebih basah daripada di di utara pegunungan selama musim kemarau (monsoon timur). Dari isohyet tersebut dapat juga dilihat bahwa potensi air tertinggi ada di bagian timur P. Bali ini khususnya di bawah kaki G. Agung. Musim angin barat di Bali terjadi pada bulan Desember sampai dengan bulan Maret, dan pada bulan April sampai bulan September merupakan musim angin timur. Musim panca roba terjadi pada bulan Oktober sampai bulan November. ..
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