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Peran Transforming Growth Factor? terhadap Tingkat Kematangan dan Kejadian Apoptosis Oosit Sapi pada kultur In Vitro Widjiati -; Rimayanti -; Arief boediono; Agus Setiadi
Jurnal Veteriner Vol 11 No 2 (2010)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

Low productivity of in vitro embryo production at blastosis level appears to be as a result of imperfectoocyte maturation which causes imperfect oocyte growth, and in turn will affect the embryo growth. Inaddition to hormonal factor, growth factor plays significant role in maturation process of oocytes. Asgrowth factor might has a significant role during maturation process, a study was conducted to determinewhether transforming growth factor ? (TGF ?) isolated from oocytes of cumulus complex is required for coculture and in vitro embryo production.. Oocytes were collected from follicles with the diameter of 3-5 mmand > 5 mm. Then the oocytes were cultured for 22 hours at 38.5o with 5% CO2 atmosphere in tissueculture medium (TCM) 199 supplemented with 5 ?g/mg luteonizing hormone (LH), 3% bovine serumalbumin (BSA) 50 ?g/ml gentamycin sulfat and three different levels of TGF ? (12,85 pg/ml, 25,7 pg/mland 38,55 pg/ml). The oocyte maturation and number of apoptosis cells were examined. The result showedthat oocyte maturation in medium supplemented TGF ? at the dose of 38,55 pg/ml was better than in thatat dose of 12,85pg/ml or 25,7 pg/ml. The best maturation was observed at metaphase II stage of oocytedevelopment. No apoptosis was observed during maturation of oocytes. Supplementation of TGF ? at thedose of 38,55 pg/ml in culture medium increased the oocyte maturation without causing a significantapoptosis in vitro.
Aktivasi Oosit Menggunakan Strontium Klorida setelah Injeksi dengan Spermatozoa Domba Hasil Pengeringbekuan (OOCYTE ACTIVATION USING STRONTIUM CHLORIDE FOLLOWING INJECTION OF FREEZE-DRIED RAM SPERMATOZOA) Takdir Saili; Ita Djuwita; Mohamad Agus Setiadi; Srihadi Agungpriyono; Arief Boediono
Jurnal Veteriner Vol 13 No 3 (2012)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

One of the factors that inhibit the formation of male pronuclei following injection of freeze-dried ramspermatozoa was the absence of artificial activation during oocyte incubation after the injection. Therefore,in this experiment the ability of strontium chloride (SrCl2) to improve oocyte activation followingintracytoplasmic sperm injection (ICSI) was evaluated. Aceto lacmoid staining was used to assessdecondensation and pronucleus formation following ICSI. Results of this experiment revealed that freezedriedspermatozoa had the ability to decondense and to form 1PN following injection into oocytes evenwithout artificial activation, but failed to form 2PN. However, 40% of 2PN oocytes were obtained when theinjected oocytes was first incubated for 20 minutes in medium containing 50 mM strontium chloride thensubsequently incubated for 10 hours in medium without strontium. On the contrary, the 2PN oocytes werenot observed either in injected oocyte neither without artificial activation nor in non-injected oocytes withartificial activation. In conclusion, freeze-dried ram spermatozoa were able to decondense and to support2PN formation following ICSI and artificial activation using strontium.
THE DEVELOPMENT OF FOLLICLES AND OOCYTES VIABILITY FROM EWE OVARIUM POST-INTRAUTERINE TRANSPLANTATION TO PSEUDOPREGNANT RABBIT Ramadhan Sumarmin; Adi Winarto; Tutty Laswardi Yusuf; Arief Boediono
Jurnal Veteriner Vol 9 No 3 (2008)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

The objective of this study was to evaluate the development of ewe follicles and oocytes viabilityfollowing an intrauterine transplantation of the ovaries into pseudopregnant rabbit. The experiment wasconducted in day 1 of pseudopregnant rabbit. Following transplantation for 5, 7, and 9 days, the ovarieswere recollected. The development of follicles determined by counting the number of follicles in paraffinembedded ovaries after staining with haematoxylin-eosin (HE). The viability of oocytes was determined byslicing the ovaries. Oocytes were incubated in CO2 incubator with 5% CO2, 38°C for 24 hours. After maturation,the oocytes were stained with 2% aceto-orcein to determine the nuclear oocytes status. The result showedthat follicles were detected in all stages of their development (primordial, primary, preantral, and antralfollicle stages), but their number decreased significantly (P<0.05) 5, 7 or 9 days after transplantation,except for those at primordial stage which at day 5 post-transplantation (634.7±56.88) were not significantlydifferent to the control (683.7±61.55). After maturation, the oocytes that were able to reach the M-II phaseat day 5 and day 7 post-transplantation were 35.05% and 35.24% respectively. They were significantly(P<0.05) lower than the control (56.65%). In conclusion, the development of follicles and oocytes viability inthe ewe ovaries in pseudopregnant rabbits was still preserved during intrauterine transplantation.
Mice Oocytes Respond after Vitrification Followed by Artifical Activation Using a Various Concentration of Strontium Chloride and Cytochalasin B Muhammad Rosyid Ridlo; Rini Widyastuti; Alkaustariyah Lubis; Mokhamad Fahrudin; Arief Boediono
Jurnal Veteriner Vol 19 No 3 (2018)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

Oocyte vitrification is the important part of gamete preservation for further purpose. The objective of this study was to evaluate the response or development of vitrified-mice oocyte following activation using various concentrations of Strontium Chloride (Sr Cl2). Oocytes were collected from superovulation-induced female mice. Oocytes vitrification was then performed using a gradual equilibration of 2 M Ethylene Glycol in 0.25 M sucrose and 7 M Ethylene Glycol on 0.5 sucrose. Subsequently, the vitrified oocytes were thawed and activated using various Strontium Chloride concentration in each group. Control 1 is unvitrified oocyte and without Sr Cl2. Control 2 is unvitrified oocyte then activated by 20 mM Sr Cl2. Zero (0) mM Sr Cl2 is vitrified oocyte without Sr Cl2. Group Ten (10) mM is vitrified oocyte then activated by 10 mM Sr Cl2. Group Twenty (20) mM is vitrified oocyte then activated by 20 mM Sr Cl2. The viability of vitrified-thawed oocytes was observed based on ooplasm integrity. Whereas the oocytes respond to artificial activation was observed based on pronucleus formation after 10 hours of activation. The result showed that 39% of oocyte degenerated following vitrification. The respond of vitrified-thawed oocytes following artificial activation using Strontium Chloride was significantly lower compared to fresh oocytes (p<0,05). Interestingly the highest percentage of activated oocytes (36.36%) was present in a group achieved 20 mM Strontium Chloride. As conclusion is Strontium Chloride 20mM has a best result (36,36%) to activate vitrified oocyte than 0 mM and 10 mM of Strontium Chloride.
Kebuntingan Hasil Transfer Blastosis Mencit yang Dibekukan dengan Metode Vitrifikasi Kriolup I Wayan Batan; I Ketut Suatha; Ita Djuwita; Nining Handhayani; Wahono Esti Prasetyaningtyas; Ketut Adnyane Mudite; Bibiana W Lay; Supar -; Arief Boediono
Jurnal Veteriner Vol 12, No 3 (2011)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

The aim of the study was to assess the viability of vitrified embryo using cryoloop as a carrier ofembryo. The blastocyst stage embryos were collected from superovulated mice. Embryos were frozenusing vitrification method and vitrified embryos were loaded on copper filament cryoloop before dipped inliquid nitrogen. The viability of vitrified embryos was assess in vitro by medium cultered and in vivo bytransfered them to recipient mice. The result shows the viability of vitrified embryos was 85,7% after 24hours cultured and the embryos were born from two pregnant recipient mice out of nine (22%) or fouroffspring out of 63 trasfered embryos (6%). In conclusion, vitrified blatocyst stage embryos using cryoloopas a carrier could keep the viability of the embryos and they could be transfered to the recipient mice andwere born normally.
DEVELOPMENT OF MICE AND HAMSTER EMBRYOS IN KSOMAA AND HECM-6 MEDIUM Bayu Rosadi; M Agus Setiadi; Dondin Sajuthi; Arief Boediono
Jurnal Veteriner Vol 9 No 4 (2008)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

The purpose of the present study was to investigate the viability of mice and hamster embryos developed in Kalium Simplex Optimized Medium amino acid (KSOMaa) and Hamster Embryo Culture Medium-6 (HECM-6) medium. Female DDY mice were superovulated by injection i.p. of 5 IU Pregnant Mare Serum Gonadotropine (PMSG) and 5 IU Human Chorionic Gonadotropine (hCG) in 48 h interval, hamster (Phodopus campbelli) injected by 2.5 IU PMSG and 2.5 IU hCG 48 h later. Then females were mated with fertile males. Eight-cell embryos were recovered at day 3 after natural mating. The mice embryos were cultured in KSOMaa+5% NBCS (New Born Calf Serum) (T1) and HECM-6+5% NBCS (T2), the hamster embryos were cultured in KSOMaa+5% NBCS (T3) and HECM-6 + 5% NBCS (T4) for further development at 37oC in a humidified atmosphere of 5% CO2 in air for 48 h. The examinations were replicated five times. The T1 embryos developed to compact morulla and early blastocyst 100% (140/140), 92.1% (129/140) to blastocyst and expanded blastocyst, and 22.9% (32/140) became hatching/hatched. The T3 reached 100% (60/60) to compact morulla and early blastocyst, 85.0% (51/60) blastocyst, and 48.3% (29/60) expanded blastocyst, no embryo observed hatching/hatced. The T2 embryos had more expanded blastocyst than T3 (P<0.05), hatching/hatched rate higher than T1 and T3 but lower than T4 (P<0.05). Shortly, KSOMaa enable to support 8-cell stage mice and hamster embryo, but the hamster embryo developed lower at expanded blastocyst stage. HECM-6 is more appropriate than KSOMaa to support 8-cell mice embryos development and suitable to develop 8-cell stage hamster embryos.
Sel Kumulus Sebagai Feeder Layer pada Kultur Stem Cells Embrionic Mencit (CUMULUS CELLS AS FEEDER LAYER IN CULTURE OF MOUSE EMBRYONIC STEM CELLS) Thomas Mata Hine; Arief Boediono; Iman Supriatna; Dondin Sajuthi
Jurnal Veteriner Vol 13 No 2 (2012)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

The purpose of this study was to verify the effectiveness of cumulus cells as a feeder layer in supportingthe growth of mouse embryonic stem cells. Embryos at blastocyst stage were exposed in pronase solution,and then incubated in rabbit anti-mouse antibody and guinea pig complement to lyse and separate thetrophoblast cells from the inner cell mass. Inner cell mass subsequently cultured in a petri dish containinga cumulus feeder layer, mouse embryonic fibroblasts, or without a feeder layer, in stem cells medium. Theresulting stem cells colony passaged in trypsin solution, pipetted repeatedly to produce subcolonies orsingle cells, and cultured as before in some new petri dishes. Characterization of stem cells was identifiedby using alkaline phosphatase staining. The results showed the effectiveness of cumulus cells as feederlayer for culture of mouse embryonic stem cells was comparable with mouse embryonic fibroblasts, andboth of them were better than without a feeder layer method. The number of primary colony, cell lines, andcolony growth rate increased 41.30, 8.70, and 54.20%, respectively, while doubling time was shorter 10:21hours compared to the growth without feeder layer method. These results prove that the cumulus feederlayer effectively supports the growth of mouse embryonic stem cells.
Seleksi Spermatozoa Domba Garut dengan Metode Sentrifugasi Gradien Densitas Percoll Heri Sujoko; Mohamad Agus Setiadi; Arief Boediono
Jurnal Veteriner Vol 10 No 3 (2009)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

Preparation of a good quality of sperm is required in order to increase the efficacy of in vitro fertilization.One method of a good sperm preparation is the use of Percoll density gradient centrifugation. This studywas aimed to determine the optimal combination of speed and time of centrifugation for the spermpreparation with the lowest mortality rate. Fresh semen collected from Garut sheep with the concentrationof 2 million cells per ml was used in this study. The experimental design used was a completely randomizedfactorial design with one control group and sixteen combination of speed and time of centrifugation. Theywere 1),(1) 200 xg for 5 min; (2) 200 xg for 10 min; (3) 200 xg for 15 min; (4) 200 xg for 20 min; (5) 300 xg for5 min; (6) 300 xg for 10 min; (7) 300 xg for 15 min; (8) 300 xg for 20 min; (9) 400 xg for 5 min; (10) 400 xg for10 min; (11) 400 xg for 15 min; (12) 400 xg for 20 min; (13) 500 xg for 5 min; (14) 500 xg for 10 min; (15) 500xg for 15 min; and (16) 500 xg for 20 min. All of control and treatments combination were replicated forthree times. Each control and combined treatment were replicated for three times. The variables observedwere the concentration of sperm, the percentage of live sperms, the percentage of progressively motilesperms and the percentage of normal sperm. The data obtained were analyzed by two way analysis ofvariance (ANOVA) and the differences between treatment groups were subjected for Duncan’s MultipleRange Test (DMRT). The best result was obtained using 400 xg for 15 minutes which showed the spermconcentration of 66.66 ± 12.22 millions per ml, the living sperms of 74.82± 1.53%, the percentage ofprogressively motile sperm of 57.56±8.42% and the percentage of normal sperm of 86±1.73%).
PERUBAHAN VIABILITAS DAN STRUKTUR SUBSELULER SPERMATOZOA DOMBA SETELAH PENGERINGBEKUAN Takdir Saili; I Ketut Mudite Adnyana; Ronny Rachman Noor; Mohamad Agus Setiadi; Srihadi Agungpriyono; Arief Boediono
Jurnal Veteriner Vol 10 No 4 (2009)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

Several methods i.e. cooling, freezing, and freeze-drying have been widely used to preserve spermatozoa with various degree of success. Freeze-drying appears to provide a method to preserve spermatozoa in a dry state without requiring liquid nitrogen for storing frozen spermatozoa. Freeze-drying procedures can have a detrimental effect on plasma membrane and acrosomal cap of the spermatozoa. In this experiment study, the viability and subcellular changes of freeze-dried ram spermatozoa were evaluated using staining method and scanning electron microscopy. The results revealed that all freeze-dried spermatozoa were dead following evaluation using eosin staining and Hoechst-propidium iodide staining methods. Morover, plasma membrane and acrosomal cap of freeze-dried ram spermatozoa was disrupted observed using scanning electron microscope.
Perpanjangan Usia Tikus Tua Setelah Xenotransplantasi Human Umbilical Cord Mesenchymal Stem Cells (hUC-MSCs) Elpita Tarigan; Adisti Dwijayanti; Frans Dhyanagiri Suyatna; Indra Bachtiar; Sandy Qlintang; I Ketut Mudite Adnyane; Arief Boediono
Jurnal Veteriner Vol 21 No 4 (2020)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (218.261 KB) | DOI: 10.19087/jveteriner.2020.21.4.539

Abstract

Currently, mesenchymal stem cells (MSCs) for implementing regenerative medicine in aging become interest in medical research science, especially in degerative disease and other aging problems. This research was aimed to determine the effectiveness of hUC-MSCs on inhibiting the aging process through the lifetime of the rat and the effect of intravenous administration of hUC-MSCs in phisiologycally aging female rat on the blood analysis. This study was used 40 aged female rats with 29-30 months of age divided into four groups with 10 rats each. The control rat group was given physiological NaCl (0.9%) 0.5 mL, and the treated rat group was given hUC-MSCs 1x107 cells/kg body weight in 0.5 mL NaCl 0.9%, wasinjected intravenously in caudo lateralis tail vein with stratified frequency; one time injection (SC1), three times injections (SC3) and five times injections (SC5). Perifer blood was collected from retro-ortbital sinus vein 30 days before and after injection of hUC-MSCs for hematology and blood chemistry analysis. Based on the results were obtained, it indicated that hUC-MSCs increased the survival of aging rat werein treatment group, life span of rats was extended up to 40 months compared to the average life of control rat aged up to 34±2 months. The injection of hUC-MSCs 1x107 cells/kg of body weight with one, three andfive times injection were affected to blood profiles and blood chemistry with correlation were low. The conclusions are hUC-MSCs extend the lifespan of aging rat and were affect the blood in general but in normal range of aging rat, affect in ALT and creatinin as tissue repair and tolerated by aging rat.
Co-Authors . AULANI’AM . Herdis . Yulnawati A.S. Satyaningtijas Abinawanto Abinawanto Abinawanto Adi Winarto Adisti Dwijayanti Adkhilni Utami Adkhilni Utami Adrian Situmeang Adrian Situmeang Adrian Situmeang Adrien Jems Akiles Unitly Adrien Jems Akiles Unitly, Adrien Agus Harsoyo Agus Harsoyo Agus Oman Sudrajat Agus Setiadi Ahmed, Ifty Al Azhar Al Mukhlas Fikri AL-AZHAR AL-AZHAR Alfred O. M. Dima Alif Iman Fitrianto Alif Iman Fitrianto Alimuddin Alimuddin Alkaustariyah Lubis Amrozi Anak Agung Gede Sugianthara Anak Agung Istri Sri Wiadnyani Andri Maruli Tua Lubis Andri Maruli Tua Lubis Andriyanto A Andriyanto Andriyanto ANOM BOWOLAKSONO Arie Adrianus Polim AS Aku, AS Aucky Hinting Aulia Miftakhur Rahman Ayu Mulia Sundari Bambang Kiranadi Batara Sirait Bayu Rosadi Berry Juliandi BIBIANA W LAY Bibiana W Lay Boenjamin Setiawan Boenjamin Setiawan Budiariati, Vista Cahayadi, Sigit Daru Cece Sumantri Chairun Nisa Citra Noviana Cutnya’ Shaliran Nazlie (Alm) Dedy D. Solihin Diah Nugrahani Pristihadi Dian Anggraini Djaswadi Dasuki Djoko Walujo Dody Dharmawan Trijuno, Dody Dharmawan Dondin Sajuthi Dwi Budiono Dwiranti, Astari Elpita Tarigan Eni Kusrini EVY AYU ARIDA Farid A. Moeloek Ferry Sandra Frans Dhyanagiri Suyatna Frans Dhyanagiri Suyatna Frans Dhyanagiri Suyatna Funahashi, Hiroaki Hadi, Restu S. Handina Rakhmawati Handina Rakhmawati Harry Murti Harry Murti Harry Murti Harry Murti Harry Murti Hartanti Kusumaningtyas HERA MAHESHWARI HERDIS Herdis . herdis herdis Heri Sujoko Heru Setijanto I Ketut Mudite Adnyana I Ketut Suatha I Wayan Batan Ichsan Ichsan Iis Diatin Iman Supriatna Indra Bachtiar Indra Kusuma Irma H Suparto Irma Herawati Suparto Irma Suryani ITA DJUWITA Ita Djuwita Ita Djuwita Ita Fauzia Hanoum, Ita Fauzia Ivan Sini Karisma Mardatillah Karisma Mardatillah KARTINI ERIANI Kartini Eriani KARTINI ERIANI Kartiwa, R. Angga Kelvin Yaprianto Kelvin Yaprianto Kelvin Yaprianto Ketut Adnyane Mudite Krido Brahmo Putro Kusdiantoro Mohamad Kusumaningtyas, Hartanti Latifah Kosim Darusman Lea Tarliyah Lindiawati, Riris Lubis, Andri Maruli Tua M Agus Setiadi M. Haviz Madihah Madihah Madihah Madihah, Madihah Maman Surachman Mas Rizky A.A. Syamsunarno Maula, Yogi Nikmatul Miraprahesti, Retti N. Mohamad Fakhrudin Mohammad Ghozali Mohammad Ghozali, Mohammad Mokhamad Fahrudin MOZES R. TOELIHERE MUHAMMAD AGUS SUPRAYUDI Muhammad Gunawan Muhammad Gunawan Muhammad Gunawan Muhammad Rizal Muhammad Rizal MUHAMMAD RIZAL MUHAMMAD RIZAL' Muhammad Rosyid Ridlo Muhammad Zairin JR Muhammad Zairin Jr. MULYOTO PANGESTU MULYOTO PANGESTU Muslim Muslim NASTITI KUSUMORINI Nastiti Kusumorini Nastiti Kusumorini Nining Handayani Nining Handayani Nining Handhayani Noer Muhammad Dliyaul Haq Noer Muhammad Dliyaul Haq, Noer Muhammad Dliyaul Nurhayati, Retno Wahyu Nurhidayat - Nurhidayat Nurhidayat Nurhidayat Nurhidayat Nuril Farizah Nursanti, Risa Nuzulia, Nur Aisyah Prakoso, Nurul Muhammad Prasetyaningtyas, Wahono Puspitasari, Riris Rachmat Herman Rahmaniyah, Wiwit Ridhani Rahminiwati, Min Rakhmawati, Handina Ramadhan Sumarmin Rangga Setiawan Rangga Setiawan Ratih Rinendyaputri Ratih Rinendyaputri Ratih Rinendyaputri Resti Rahma Dianti Ridi Arif Rimayanti - Rini Widyastuti Riris L. Puspitasari Riris L. Puspitasari Ronny Rachman Noor Salsabila, Cyntia Bella Sandy Qlintang SATRIYAS ILYAS Satya Gunawan Shofwal Widad Sigit Prastowo Siti Darodjah Rasad Situmeang, Adrian Sony H. Sumarsono Sony Heru Sumarsono SONY HERU SUMARSONO SONY HERU SUMARSONO Sri Catur Setyawatiningsih Srihadi Agungpriyono Subangkit, Mawar Sulistiono, Sumarsono, Sony H. Sumarsono, Sony Heru Sundari, Ayu Mulia Supar - Supar . Sutarya Enus Sutiman Bambang Sumitro TAKDIR SAILI TARUNI SRI PRAWAST MIEN KAOMINI ANY ARYANI DEDY DURYADI SOLIHIN Taufik Jamaan Thomas Mata Hine Trevino A. Pakasi Tri Aprilliana Wulandari TRINIL SUSILOWATI Tutik Wrediati Tutik Wresdiyati Tutty Laswardi Yusuf Tutty Laswardi Yusuf Tuty Laswardi Yusuf Uswatun Hasanah Vincentia Maria Wahono Esthi Prasetyaningtyas Wahono Esti Prasetyaningtyas Wahono Esti PrasetyoningtyaserB Wahyudin Wasmen Manalu Watanabe, Seiichi Widjiati w Wildan Mubarok Wining Astini Wining Astini Wiwit Ridhani Rahmaniyah Wulandari, Tri Aprilliana Yessie Widya Sari Yoga Yuniadi Yuhara Sukra Yuhara Sukra Yulnawati . YULNAWATI YULNAWATI Yundari, Yundari Yushinta Fujaya