• Title/Summary/Keyword: Early Embryo

Search Result 590, Processing Time 0.045 seconds

Developmental Abnormality in Agricultural Region and Toxicity of the Fungicide Benomyl on Korea salamander, Hynobius leechii (한국산 도롱뇽(Hynobius leechii)의 농경지에서의 배 발생 이상과 살균제 Benomyl의 독성효과)

  • Choi, Yeoung-Ju;Yoon, Chun-Sik;Park, Joo-Hung;Jin, Jung-Hyo;Cheong, Seon-Woo
    • Korean Journal of Ecology and Environment
    • /
    • v.35 no.3 s.99
    • /
    • pp.198-212
    • /
    • 2002
  • A numerical variation and abnormalities were studied on egg bags and embryos of Korean salamander, Hynobius leechii from agricultural habitat. The teratogenic and toxic effects of fungicide benomyl were also investigated with early embryos from non-agricultural habitat. We collected 144 egg bags from agricultural region, and 3418 of early embryos were contained. The lengths of egg bags were varied from 10 to 23 cm and the most frequent length was 19 cm. The number of embryos was varied from 7 to 43, and the most frequent range was 22 to 26. Spontaneous abnormalities were occurred in 406 embryos among 116 egg bags, and 24 kinds of external abnormalities were found. Individuals showing severe external defect were histologically studied and they showed optic dyspalsia, thyroid carcinoma, somatic muscular dysplasia, partial biaxial structure, decrease of red blood cells in the heart, cephalic degeneration and intestinal dysplasia. 385 embryos from non-agricultural region were exposed to 200 nM${\sim}$ 1 ${\mu}$M of benomyl at blastula or gastrula for 12 days. All embryo were dead in the concentration of 1 ${\mu}$M (LD$_{100}$) and 75% of embryos were dead in 800nM of benomyl. Speciflc effect due to benomyl was acrania or cephalic dysplasia and this restult suggests that the benomyl inhibit stongly to the development of neural tissue. These abnormal developments may be caused by antimitotic action, inhibition of tubulin complex, destruction of microtubule, inhibitions of neurulation and closing of neural fold, and by the inhibition of the movement of neural crest cells.

Optimization of One-Step Dilution Method of Vitrified Bovine IVM/IVF/IVC Blastocysts (초자화 동결된 체외생산 소 배반포기배의 1 단계 융해 방법의 적정화)

  • Lee, K.S.;Kim, E.Y.;Nam, H.K.;Park, S.Y.;Park, E.M.;Yoon, S.H.;Park, S.P.;Chung, K.S.;Lim, J.H.
    • Korean Journal of Animal Reproduction
    • /
    • v.24 no.1
    • /
    • pp.89-95
    • /
    • 2000
  • This study was to establish an effective dilution technique in a vitrification of bovine blastocysts for the field trial. For vitrification, blastocysts were exposed in glycerol (G) and ethylene glycol (EG) mixture in m-DPBS supplemented with 10% FBS. Blastocysts were first exposed to 10% (v/v) G for 5 min, and subsequently were transferred to 10% G plus 20% EG (v/v) for 5 min. Finally, embryos were transferred to 25% G plus 25% EG (v/v) for 30 see and were placed in nitrogen vapor for 3 min, and then were plunged into L$N_2$. At thawing, the straw containing blastocysts was placed in air for 10 sec, and then plunged into a water bath at $25^{\circ}C$ until all ice had disappeared. They were placed in $25^{\circ}C$ and 36$^{\circ}C$ water according to treatment group for different time. Also, in vitro survival was assessed by the re-expansion and hatched rates at 24 hand 48 h postwarming, respectively. The results obtained in these experiments were summarized as follows; 1) In the survival rates of vitrified bovine blastocysts according to different dilution time at thawing, the data of 1 min group (86.6, 56.6%) were higher than those of other treatment groups (2 min; 93.5, 35.4%, 2.5 min; 76.9, 30.7%, 3 min; 88.8, 36.1% and 3.5 min; 83.7, 8.1%). 2) When the in vitro survival of vitrified groups according to different developmental stage was examined at 48 h after thawing using 1 min dilution method, the hatching rates of fast developed embryos (expanded blastocyst: 81.3%: early hatching blastocyst: 86.2%) were higher than that of delayed developed one (early blastocyst: 46.6%). 3) In addition, when the in vitro survival of vitrified groups according to different embryo age was compared, the hatched rates at 48 h after thawing of Day 7 (66.6%) and Day 8 embryos (60.0%) were significantly higher than that of Day 9 embryos (22.7%) (P<0.05). These results demonstrate that vitrified bovine IVM/IVF/IVC blastocysts can be successfully survived in vitro using one-step dilution (1 min) method.

  • PDF

Studies on Grain Filling and Quality Changes of Hard and Soft Wheat Grown under the Different Environmental Conditions (환경 변동에 따른 경ㆍ연질 소맥의 등숙 및 품질의 변화에 관한 연구)

  • Young-Soo Han
    • KOREAN JOURNAL OF CROP SCIENCE
    • /
    • v.17
    • /
    • pp.1-44
    • /
    • 1974
  • These studies were made at Suwon in 1972 and at Suwon, Iri, and Kwangju in 1973 to investigate grain filling process and variation of grain quality of NB 68513 and Caprock as hard red winter wheat, Suke #169 as soft red winter wheat variety and Yungkwang as semi-hard winter variety, grown under-three different fertilizer levels and seeding dates. Other experiments were conducted to find the effects of temperature, humidity and light intensity on the grain filling process and grain quality of Yungkwang and NB 68513 wheat varieties. These, experiments were conducted at Suwon in 1973 and 1974. 1. Grain filling process of wheat cultivars: 1) The frequency distribution of a grain weight shows that wider distribution of grain weight was associated with large grain groups rather than small grain group. In the large grain groups, the frequency was mostly concentrated near mean value, while the frequency was dispersed over the values in the small grain group. 2) The grain weight was more affected by the grain thickness and width than by grain length. 3) The grain weight during the ripening period was rapidly increased from 14 days after flowering to 35 days in Yungkwang and from 14 days after flowering to 28 days in NB 68513. The large grain group, Yungkwang was rather slowly increased and took a longer period in increase of endosperm ratio of grain than the small grain group, NB 68513. 4) In general, the 1, 000 grain weight was reduced under high temperature, low humidity, while it was increased under low temperature and high humidity condition, and under high temperature and humidity condition. The effect of shading on grain weight was greater in high temperature than in low temperature condition and no definite tendency was found in high humidity condition. 5) The effects of temperature, humidity and shading on 1, 000 grain weight were greater in large-grain group, Yungkwang than in small grain group, NB 68513. Highly significant positive correlation was found between 1, 000 grain weight and days to ripening. 6) The 1, 000 grain weight and test weight were increased more or less as the fertilizer levels applied were increased. However, the rate of increasing 1, 000 grain weight was low when fertilizer levels were increased from standard to double. The 1, 000 grain weight was high when planted early. Such tendency was greater in Suwon than in Kwangju or Iri area. 2. Milling quality: 7) The milling rate in a same group of varieties was higher under the condition of low temperature, high humidity and early maturing culture which were responsible for increasing 1, 000 grain weight. No definite relations were found along with locations. 8) In the varieties tested, the higher milling rate was found in large grain variety, Yungkwang, and the lowest milling rate was obtained from Suke # 169, the small grain variety. But the small grained hard wheat variety such as Caprock and NB 68513 showed higher milling rate compared with the soft wheat variety, Suke # 169. 9) There were no great differences of ash content due to location, fertilizer level and seeding date while remarkable differences due to variety were found. The ash content was high in the hard wheat varieties such as NB 68513, Caprock and low in soft wheat varieties such as Yungkwang and Suke # 169. 3. Protein content: 10) The protein content was increased under the condition of high temperature, low humidity and shading, which were responsible for reduction of 1, 000 grain weight. The varietal differences of protein content due to high temperature, low humidity and shading conditions were greater in Yungkwang than in NB 68513. 11) The high content of protein in grain within one to two weeks after flowering might be due to the high ratio of pericarp and embryo to endosperm. As grains ripen, the effects of embryo and pericarp on protein content were decreased, reducing protein content. However, the protein content was getting increased from three or four weeks after flowering, and maximized at seven weeks after flowering. The protein content of grain at three to four weeks after flowering increased as the increase of 1, 000 grain weight. But the protein content of matured grain appeared to be affected by daily temperature on calender rather than by duration of ripening period. 12) Highly significant positive correlation value was found between the grain protein content and flour protein content. 13) The protein content was increased under the high level of fertilizers and late seeding. The local differences of protein content were greater in Suwon than in Kwangju and Iri. 14) Protein content in the varieties tested were high in Yungkwang, NB 68513 and Caprock, and low in Suke # 169. However, variation in protein content due to the cultural methods was low in Suke # 169. 15) Protein yield per unit area was increased in accordance with increase of fertilizer levels and early maturing culture. However, nitrogen fertilizer was utilized rather effectively in early maturing culture and Yungkwang was the highest in protein yield per unit area. 4. Physio-chemical properties of wheat flour: 16) Sedimentation value was higher under the conditions of high temperature, low humidity and high levels of fertilizers than under the conditions of low temperature, high moisture and low levels of fertilizers. Such differences of sedimentation values were more apparent in NB 68513 and Caprock than Yungkwang and Suke # 169. The local difference of sedimentation value was greater in Suwon than in Kwangju and Iri. Even though the sedimentation value was highly correlated with protein content of grain, the high humidity was considered one of the factors affecting sedimentation value. 17) Changes of Pelshenke values due to the differences of cultural practices and locations were generally coincident with sedimentation values. 18) The mixing time required for mixogram was four to six minutes in NB 68513, five to seven minutes in Cap rock. The great variation of mixing time for Yungkwang and Suke # 169 due to location and planting conditions was found. The mixing height and area were high in hard wheat than in soft wheat. Variation of protein content due to cultural methods were inconsistent. However, the pattern of mixogram were very much same regardless the treatments applied. With this regard, it could be concluded that the mixogram is a kind of method expressing the specific character of the variety. 19) Even though the milling property of NB 68513 and Caprock was deteriorated under either high temperature and low humidity of high fertilizer levels and late seeding conditions, baking quality was better due to improved physio-chemical properties of flour. In contrast, early maturing culture deteriorated physio-chemical properties, milling property of grain and grain protein yield per unit area was increased. However, it might be concluded that the hard wheat production of NB 68513 and Caprock for baking purpose could be done better in Suwon than in Iri or Kwangju area. 5. Interrelationships between the physio-chemical characters of wheat flour: 20) Physio-chemical properties of flour didn't have direct relationship with milling rate and ash content. Low grain weight produced high protein content and better physio-chemical flour properties. 21) In hard wheat varieties like NB 68513 and Caprock, protein content was significantly correlated with sedimentation value, Pelshenke value and mixing height. However, gluten strength and baking quality were improved by the increased protein content. In Yungkwang and Suk # 169, protein content was correlated with sedimentation value, but no correlations were found with Pelshenke value and mixing height. Consequently, increase of protein content didn't improve the gluten strength in soft wheat. 22) The highly significant relationships between protein content and gluten strength and sedimentation . value, and between Pelshenke value, mixogram and gluten strength indicated that the determination of mixogram and Pelshenke value are useful for de terming soft and hard type of varieties. Determination of sedimentation value is considered useful method for quality evaluation of wheat grain under different cultural practices.

  • PDF

Effect of Cryoprotectant Kinds and Cell Stages on the Viability of Mouse Embryos Cryopreserved by OPP Vitrification (동결보호제의 종류 및 배발달단계가 OPP Vitrification 동결보존시 생쥐수정란의 생존성에 미치는 영향)

  • 공일근;조성균;조성근
    • Korean Journal of Animal Reproduction
    • /
    • v.23 no.1
    • /
    • pp.85-92
    • /
    • 1999
  • This study was designed to determine effect of cryoprotectant kinds and cell stages on OPP vitrification method in mouse embryos. The freezing speed, cryoprotectants and cell stage could affect of embryo viability following various vitrification methods. The vitrification solution used were consisting of 40% (v/v) ethylene glycol, 18% (w/v) Ficoll, 0.3 M sucrose solution in holding medium (D-PBS supplemented with 5% FCS: HM) (EFS) or 16.5% ethylene glycol , 16.5% dimethyl sulfoxide, 0.5 M sucrose in HM (EDS). The embryos were collected from oviduct at 18 h after hCG injection and then washed and cultured in mHTF medium until use. In experiment 1, the blastocysts were vitrified by OPP straw to determine the optimal vitrification solution of EFS or EDS. The post-thaw survival rates at re-expanded stage rates were significantly different between EFS and EDS (95.0 vs 100%), but at hatching stage was not different between EFS and EDS (90.0 vs 95.0%). respectively. In experiment 2, zygotes, 2-, 4-cell, morula and blastocysts were vitrified by OPP method to determine the acceptable of early stage embryos. The development rates to expanded blastocyst in zygote (70.0%) were significantly lower rather than those in 2-, 4- 8-cell, compacted morula or blastocyst (89.7, 90.0, 92.8, 97.6 or 97.5%), respectively. However, the cell number of post-thaw developed to expanded blastocyst in blastocyst and control blastocyst stage (39.6$\pm$2.81, 35.7$\pm$2.98) were significanty higher than those in zygote, 2-, 4-, 8-cell, compacted morula (29.8$\pm$3.21, 31.3$\pm$3.83, 29.3$\pm$3.58, 28.9$\pm$3.21 or 30.8$\pm$2.93). In experiment 3, the zygotes were exposed in VSl for 1, 2, and 3 min to the optimal exposed time. The cleavage rates (91.6, 88.5, 88.9%) and develop mental rates to blastocyst (83.3, 74.3 and 69.4%) depends on the exposed time in VSl were not significantly different among 1, 2, or 3 min, respectively. The cell number also were not significantly different among exposed time in VS1. respectively. These results indicate that OPP method could be useful for vitrification either EFS or EDS vitrification solution. The post-thaw survival rates at zygote were significantly lower than those at 2-, 4-, 8-cell, morula or blastocyst, respectively. The zygote stage were more sensitive rather than late stage embryos. The exposing time in VS1 for 1 min was better than that for 2 or 3 min, even it was not significantly different. The OPP vitrification method could be useful of mouse embryos either with EFS or EDS vitrification solution.

  • PDF

Studies on the Physiological Chemistry of Seed Development in Ginseng Seed (인삼식물의 종자발육 과정에 있어서의 생리화학적 연구)

  • Hee-Chun Yang
    • KOREAN JOURNAL OF CROP SCIENCE
    • /
    • v.17
    • /
    • pp.115-133
    • /
    • 1974
  • This study was done on the metabolism of chemical components during the seed development of ginseng. The changes of the chemical components were inspected in the following periods: from the early stage of flower organ formation to flowering time, from the early stage of fruiting to maturity, during the moisture stratification before sowing. From flower bud forming stage to meiosis stage, the changes in the fresh weight, dry weight, contents of carbohydrates, and contents of nitrogen compounds were slight while the content of TCA soluble phosphorus and especially the content of organic phosphorus increased markedly. From meiosis stage to microspore stage the fresh and dry weights increase greatly. Also, the total nitrogen content increases in this period. Insolub]e nitrogen was 62-70% of the total nitrogen content; the increase of insoluble nitrogen seems to have resulted form the synthesis of protein. The content of soluble sugar (reducing and non-reducing sugar) increases greatly but there was no observable increase in starch content. In this same period, TCA soluble phosphorus reached the maximum level of 85.4% of the total phosphorus. TCA insoluble phosphorus remained at the minimum content level of 14.6%. After the pollen maturation stage and during the flowering period the dry weight increased markedly and insolub]e nitrogen also increased to the level of 67% of the total nitrogen content. Also in this stage, the organic phosphorus content decreased and was found in lesser amounts than inorganic phosphorus. A rapid increase in the starch content was also observed at this stage. In the first three weeks after fruiting the ginseng fruit grows rapidly. Ninety percent of the fresh weight of ripened ginseng seed is obtained in this period. Also, total nitrogen content increased by seven times. As the fruits ripened, insoluble nitrogen increased from 65% of the total nitrogen to 80% while soluble nitrogen decreased from 35% to 20%. By the beginning of the red-ripening period, the total phosphoric acid content increased by eight times and was at its peak. In this same period, TCA soluble phosphorus was 90% of total phosphorus content and organic phosphorus had increased by 29 times. Lipid-phosphorus, nucleic acid-phosphorus and protein-phosphorus also increased during this stage. The rate of increase in carbohydrates was similar to the rate of increase in fresh weight and it was observed at its highest point three weeks after fruiting. Soluble sugar content was also highest at this time; it begins to decrease after the first three weeks. At the red-ripening stage, soluble sugar content increased again slightly, but never reached its previous level. The level of crude starch increased gradually reaching its height, 2.36% of total dry weight, a week before red-ripening, but compared with the content level of other soluble sugars crude starch content was always low. When the seeds ripened completely, more than 80% of the soluble sugar was non-reducing sugar, indicating that sucrose is the main reserve material of carbohydrates in ginseng seeds. Since endosperm of the ripened ginseng seeds contain more than 60% lipids, lipids can be said to be the most abundant reserve material in ginseng seeds; they are more abundant than carbohydrates, protein, or any other component. During the moisture stratification, ginseng seeds absorb quantities of water. Lipids, protein and starch stored in the seeds become soluble by hydrolysis and the contents of sugar, inorganic phosphorus, phospho-lipid, nucleic acid-phosphorus, protein phosphorus, and soluble nitrogen increase. By sowing time, the middle of November, embryo of the seeds grows to 4.2-4.7mm and the water content of the seeds amounts to 50-60% of the total seed weight. Also, by this time, much budding material has been accumulated. On the other hand, dry stored ginseng seeds undergo some changes. The water content of the seeds decreases to 5% and there is an observable change in the carbohydraes but the content of lipid and nitrogen compounds did not change as much as carbohydrates.

  • PDF

Effects of Y Chromosome Microdeletion on the Outcome of in vitro Fertilization (남성 불임 환자에서 Y 염색체 미세 결손이 체외 수정 결과에 미치는 영향)

  • Choi, Noh-Mi;Yang, Kwang-Moon;Kang, Inn-Soo;Seo, Ju-Tae;Song, In-Ok;Park, Chan-Woo;Lee, Hyoung-Song;Lee, Hyun-Joo;Ahn, Ka-Young;Hahn, Ho-Suap;Lee, Hee-Jung;Kim, Na-Young;Yu, Seung-Youn
    • Clinical and Experimental Reproductive Medicine
    • /
    • v.34 no.1
    • /
    • pp.41-48
    • /
    • 2007
  • Objective: To determine whether the presence of Y-chromosome microdeletion affects the outcome of in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) program. Methods: Fourteen couples with microdeletion in azoospermic factor (AZF)c region who attempted IVF/ICSI or cryopreserved and thawed embryo transfer cycles were enrolled. All of the men showed severe oligoasthenoteratoazoospermia (OATS) or azoospermia. As a control, 12 couples with OATS or azoospermia and having normal Y-chromosome were included. Both groups were divided into two subgroups by sperm source used in ICSI such as those who underwent testicular sperm extraction (TESE) and those used ejaculate sperm. We retrospectively analyzed our database in respect to the IVF outcomes. The outcome measures were mean number of good quality embryos, fertilization rates, implantation rates, $\beta$-hCG positive rates, early pregnancy loss and live birth rates. Results: Mean number of good quality embryos, implantation rates, $\beta$-hCG positive rates, early pregnancy loss rates and live birth rates were not significantly different between Y-chromosome microdeletion and control groups. But, fertilization rates in the Y-chromosome microdeletion group (61.1%) was significantly lower than that of control group (79.8%, p=0.003). Also, the subgroup underwent TESE and having AZFc microdeletion showed significantly lower fertilization rates (52.9%) than the subgroup underwent TESE and having normal Y-chromosome (79.5%, p=0.008). Otherwise, in the subgroups used ejaculate sperm, fertilization rates were showed tendency toward lower in couples having Y-chromosome microdeletion than couples with normal Y-chromosome. (65.5% versus 79.9%, p=0.082). But, there was no significance statistically. Conclusions: In IVF/ICSI cycles using TESE sperm, presence of V-chromosome microdeletion may adversely affect to fertilization ability of injected sperm. But, in cases of ejaculate sperm available for ICSI, IVF outcome was not affected by presence of Y-chromosome AZFc microdeletion. However, more larger scaled prospective study was needed to support our results.

The Effect of the Serum Progesterone and Estradiol Levels of hCG Administration Day on the Pregnancy and Fertilization Rate in IVF-ET Patients (체외수정 과배란 유도에서 hCG 주사 당일의 혈청 Progesterone과 Estradiol 농도가 수정율 및 임신율에 미치는 영향에 관한 연구)

  • Lee, Eun-Sook;Lee, Sang-Hoon;Bae, Do-Hwan
    • Clinical and Experimental Reproductive Medicine
    • /
    • v.23 no.1
    • /
    • pp.51-59
    • /
    • 1996
  • Controlled Ovarian hyperstimulation(COH) is generally used to obtain synchronous high quality oocytes in in vitro fertilization-embryo transfer(IVF-ET). Many investigators have studied the relationship between serum hormone levels and outcomes of IVF-ET because there is no accurate estimation method of oocyte quality. Early premature luteinization of follicles before oocyte retrieval is the most troublesome problem in COH for IVF-ET. Gonadotropin-releasing hormone agonists(GnRH-a) are used as adjuncts with gonadotropins for COH in patients undergoing in IVF. The possible benefits of GnRH-a pretreatment include improving oocyte quality, allowing a more synchronous cohort of follicles to be recruited, and preventing premature lueinization hormone surges. In COH of IVF cycles, we investigated whether an elevated progesterone(P4) level on the day of human chorionic gonadotropin(hCG) administration indicates premature luteinization and is associated with a lower fertilization rate. Many investigators have studied that the lower fertilization rates seen in patients with elevated P4 levels might result from an adverse effect of P4 on the oocytes. We hypothesizes that serum P4 levels around the day of hCG may be helpful prediction of out come in IVF-ET cycles. Success rates after COH of IVF-ET cycles are dependent upon many variable factors. Follicular factors including the number of follicles, follicular diameters and especially serum estradiol(E2) levels as an indirect measurement of follicular function and guality have been thought to influence the outcomes of IVF-ET. To assess whether serum P4 and E2 levels affect the fertilization and pregnancy rate, we reviewed the stimulation cycles of 113 patients (119 cycles) undergoing IVF-ET with short protocol with GnRH-a, from March 1993 to August 1994 retrospectively. The serum P4 and E2 levels were compared on the day of hCG in the pregnant group, 45 patients(47 cycles) and in the non-pregnant group, 68 patients (72 cycles) respectively. The serum E2 level in non-pregnant group was $1367{\pm}875.8$ pg/ml which was significantly lower than that of pregnant group, $1643{\pm}987.9$ pg/ml( p< 0.01 ). And the serum P4 level in non-pregnant group was $2.1{\pm}1.4$ ng/ml which was significantly higher than that of pregnant group, $1.0{\pm}0.7$ ng/ml( p< 0.001 ). The fertilization rate was $61.3{\pm}21.3%$ in pregnant group which was higher than that of non-pregnant group, $41.1{\pm}20.2%$ (p< 0.01). We suggest that the serum levels of P4 and E2 on the day of hCG administration are additional parameters that predict the outcomes of IVF-ET cycles.

  • PDF

A Study on the Histopathological Changes and Growth Inhibition of the Chick Embryos after Incubation with Radioactive Sulfur($^{35}S$) (방사성황산(放射性黃酸)($^{35}S$)이 부화계란(孵化鷄卵)의 발육(發育) 및 주요장기(主要臟器)의 병리조직상(病理組織像)에 미치는 영향(影響)에 관(關)한 연구(硏究))

  • Shin, Soo-Young
    • The Korean Journal of Nuclear Medicine
    • /
    • v.1 no.1
    • /
    • pp.37-54
    • /
    • 1967
  • The changes in histopathology of various organs and growth inhibition of the chick embryos incubated with radioactive sulfur ($^{35}S$) were experimentally studied. The various doses of $^{35}S$ were injected into the yolk sac at different intervals and the weight changes of the embryos were evaluated to determine the growth inhibition rates. The embryos sacrified on various incubation days were used for the study of histopathological changes in organs such as the bone, liver, kidney, gonad, and eye. Following were the results: 1) The weight changes of the $^{35}S$ treated groups were as follows: i. Embryos treated on the 5 th incubation day: No weight changes were noted on the 8th incubation day, however, the growth inhibition rate of 32.1% was noted in the group treated with $50{\mu}C$ and of 38.2% in the group treated with $150{\mu}C$ on the 12th incubation day. The rates were 9.1 and 12.1% on the 15th incubation day, and 6.5 and 10.6% on the 18th incubation day respectively. ii. Embryos treated on the 8th incubation day: The growth inhibition rates on the 12th, 15th and 18th incubation days in the groups treated with $50{\mu}C$ were 20.9, 25.9 and 18.8% and in those treated with $150{\mu}C$ were 20.0, 14.9 and 16.9% respectively. iii. Embryos treated on the 12th incubation day: The growth inhibition rates on the 15th and 18th in the groups treated with $50{\mu}C$ were 13.6 and 21.1% and in those treated with $150{\mu}C$ were 26.7 and 6.5% and in those treated with $250{\mu}C$ were 10.6 and 12.6% respectively. iv. Embryos treated on the 15th incubation day: The growth inhibition rates on the 18th in the groups treated with $50{\mu}C$ were 6.5% and in those treated with $150{\mu}C$ were 10.1% and in those treated with $250{\mu}C$ were 8.5% respectively. In summary, the longer the incubation days, the less the growth inhibition rates. II) The histopathological changes in the various organs were as follows: i. Bone: Hyperplasia and edematous changes of the bone cavity, irregular distribution of immature granular cells and increased number of the myeloblast, megakaryocyte and reticuloendothelial cells were noted. ii. Liver: The embryos treated with $150{\mu}C\;of\;^{35}S$ on the 8th incubation day showed necrosis and nucleolysis of the liver cell and abnormal enlargement of sinusoid on the 12th incubation day. The longer the incubation days, the more severe the changes such as the pyknotic artophy of the liver cells and heterochromatism. The embryos treated on the 5th incubation day with 50 and $150{\mu}C\;of\;^{35}S$ showed little changes, but sight enlargement and accumulation of serous fluid in the sinusoid on the 8th incubation day. iii. Kidney: No particular changes except atrophic changes of epithelium were noted in early stage, however, the infiltration of the granular cell and monocyte into the cortex and pyknotic changes of vascular glomeruli were noted in later stage. These changes were not closely related to the doses of $^{35}S$ given. iv. Gonad: The degenerative changes such as destruction of the immature germ cells, hyperplasia and vacuolization of the stroma were noted in testis and ovary. v. Eye: A slight distortion of the cornea and sclera was noted. The hypertrophy of inner layer and blood cell infiltration into the vascular layer of the choroid membrane were noted in embryo groups on the 12, 15 and 18th incubation days.

  • PDF

Acceleration of Early Embryonic Development by Induction of Acrosome Reaction in Intracytoplasmic Sperm Injection (세포질내 정자주입술 시행시 정자의 첨체반응이 수정란의 초기 발생과 임신율에 미치는 영향)

  • Lim, Y.J.;Lee, D.R.;Lee, J.E.;Kim, H.J.;Paik, H.R.;Yoon, H.S.;Shim, H.N.;Cho, J.H.;Roh, S.I.
    • Clinical and Experimental Reproductive Medicine
    • /
    • v.24 no.3
    • /
    • pp.311-318
    • /
    • 1997
  • Bypassing acrosome reaction and fusion process in intracytoplasmic sperm injection(ICSI), most of injected spermatozoa still contain intact acrosome contents and plasma membrane. It Is not known yet what acrosome contents and plasma membrane of spermatozoa have effect on the development of embryo. For further understanding of fertilization process after ICSI, we studied the time of pronucleus formation, disappearance and first cleavage in human zygote, and pregnancy rate in relation to acrosome reaction rate of spermatozoa after ICSI. Seventy cycles undergoing ICSI program were randomly selected. Sperm suspension from 38 cycles were treated 50% human follicular fluid(hFF) for 3 hours in order to induce acrosome reaction, others were not treated as control. Acrosome reaction in hFF treated and non-treated group was assessed by fluorescein isothiocyanate(FITC)-conjugated Arachis hypogea(PNA) and Pisum sativum agglutinin(PSA). Oocytes were classified into 'good' and 'poor' according to their morphology. After ICSI, fertilization of oocytes were assessed by detection of two pronuclei at 16 hours. The pronuclei disappearance and first cleavage of zygotes were observed at 24 hours, and then embryos were transferred to uterus after culture for 72 hours. The rate of acrosome reaction of spermatozoa in hFF treated group was significantly higher than that in control(p<0.01). Fertilization rates of good oocytes were not different both control and hFF treated group(81.3%(174/206) vs. 72.1%(102/130)). But, in poor oocytes, the fertilization rates in hFF treated group(72.1%(149/183)) were increased compared than those of control group (63.6%(98/140), p<0.01). In either good or poor oocytes, the rates of pronuclei disappearance in hFF treated-spermatozoa injected oocytes were higher than control (59.1%(103/174), 56.4%(84/149) vs. 32.4%(33/102), 37.8%(37/98), p<0.01). Also, the rates of thirst cleavage were increased in hFF treated group (31%(54/174), 24.1%(36/149)) compared than those of control group (10.8%(11/102), 13.2%(13/98), p<0.01). The pregnancy rates of hFF treated group (42.1%(16/38)) were slightly higher than control group (28.1%(9/32), p>0.05). But, the pregnancy rate of group which possessed more than one cleavaged zygote at 24 hours was higher than group which did not (45.2%(19/42) vs. 21.4%(6/28), p<0.05). From these results, the development of zygotes were faster in higher acrosome reacted sperm group than lower acrosome reacted sperm group after ICSI. Our results may be explained that acrosomal membrane and plasma membrane are easily detached from spermatozoa in acrosome reacted spermatozoa compared with acrosome intact sperm in the cytoplasm of oocyte during pronuclear formation. We conclude that the injection of acrosome reacted spermatozoa will increase the pregnancy rate as they can induce fast embryonic development in ICSI.

  • PDF

GnRH Antagonist Versus Agonist Flare-up Protocol in Ovarian Stimulation of Poor Responder Patients (저 반응군의 체외수정시술을 위한 과배란유도에 있어 GnRH Antagonist 요법과 GnRH Agonist Flare Up 요법의 효용성에 관한 연구)

  • Ahn, Young-Sun;Yeun, Myung-Jin;Cho, Yun-Jin;Kim, Min-Ji;Kang, Inn-Soo;Koong, Mi-Kyoung;Kim, Jin-Yeong;Yang, Kwang-Moon;Park, Chan-Woo;Kim, Hye-Ok;Cha, Sun-Hwa;Song, In-Ok
    • Clinical and Experimental Reproductive Medicine
    • /
    • v.34 no.2
    • /
    • pp.125-131
    • /
    • 2007
  • Objective: The aim of this study was to compare GnRH antagonist and agonist flare-up treatment in the management of poor responder patients. Methods: One hundred forty-four patients from Jan. 1, 2002 to Aug. 31, 2005 undergoing IVF/ICSI treatment who responded poorly to the previous cycle (No. of oocyte retrieved$\leq$5) and had high early follicular phase follicle stimulating hormone (FSH>12 mIU/ml were selected. Seventy-five patients received agonist flare-up protocol and 71 patients received antagonist protocol. We analyzed the number of oocytes retrieved, number of good embryos (GI, GI-1), total dose of hMG administered, implantation rate, cycle cancellation rate, pregnancy rate, live birth rate. Results: The cancellation rate was high in antagonist protocol (53.5% vs. 30.1%). The number of oocyte retrieved, the number of good embyos were high in agonist flare-up group. There was no statistical difference between GnRH agonist flare up protocol and GnRH antagonist protocol in implantation rate (14.5%, 10.1%), clinical pregnancy rate per transfer (29.4%, 21.2%) and live birth rate per transfer (21.6%, 18.2%). Although the result was not statistically significant, GnRH agonist flare up group showed a nearly doubled pregnancy rate and live birth rate per initial cycle than GnRH antagonist group. Conclusions: The agonist flare-up protocol appears to be slightly more effective than the GnRH antagonist protocol in implantation rate, pregnancy rate, live birth rate but shows statistically no significance. Agonist flare-up protocol improved the ovarian response in poor responders. However, based of the result of the study, we can expect improved ovarian response in poor responders by GnRH agonist flare up protocol.