The purpose of this study was to evaluate the in vitro developmental promoting effect of human amniotic fluid (AF) on pronucleate 1-cell mouse embryos. The AF was obtained from five patients undergoing amniocentesis for the routine diagnosis of fetal abnormality. The supernatant was filtered ($0.22{\mu}m$) after centrifugation and stored at $-20^{\circ}C$. One-cell embryos were cultured in four study groups (10% AF, 0.4% BSA, 10% AF+0.4% BSA & no-supplement in Ham's F10) for 6 days (EXP. 1). Significantly more embryos hatched in 10% AF (P<0.0l), although no difference was found among other three groups. The embryos were also cultured in varous concentration of AF (0, 10, 50 & 100%) for 7 days (EXP. 2). The rate of hatched blastocysts was significantly higher in 10%- and 50% group than in 0% and 100%- one at day 6 (P<0.05) and day 7 (P<0.005), although no difference was found between 10% and 50%- group.
A teratogenic study on Original Woo-Whang-Chung-Sim-Won was carried out in SpragueDawley rats. Original Woo-Whang-Chung-Sim-Won suspended in distilled water was administered to pregnant dams by oral gavage during organogenesis period (from 7th to 17th day of gestation) at daily doses of 1/9, 1/3 and I pill/kg. About two-thirds of dams were sacrificed at 20th day of gestation to scrutinize the pregnant performances and fetal development, and the remaining dams were allowed to deliver. The growth, reflex, behaviour and reproductive function of F1 offsprings were examined. There was no treatment-related difference in body weight, food consumption and necropy findings of dams. No gross, skeletal and visceral abnormalities was observed in F1 fetuses from dams treated with Original Woo-Whang-Chung-Sim-Won. F1 offsprings did not show any treatment-related difference in growth, reflex, behaviour and reproductive peuformance. At caesarean section of F1 dams, no growth retardation and gross abnormality was observed in F2 fetuses. In conclusion, Original Woo-Whang-Chung-Sim-Won did not show any potential teratogenic activity in rats.
Objective: To report the successful delivery in a patient of recurrent spontaneous abortion caused by chromosomal abnormality. Material and Method: Case report. Results: Twelve oocytes were obtained by in vitro fertilization. Of eleven oocytes fertilized, two embryos turned out to be normal by using fluorescent in situ hybridization on blastomere biopsy. The patient succeeded in pregnancy and the result of amniocentesis was found to be normal. She delivered the healthy female baby by cesarean section. Conclusions: The successful delivery is possible in recurrent spontaneous abortion related with reciprocal translocation by using preimplantation genetic diagnosis.
Kim, Jin Yeong;Lim, Chun Kyu;Cha, Sun Hwa;Park, Soo Hyun;Yang, Kwang Moon;Song, In Ok;Jun, Jin Hyun;Park, So Yeon;Koong, Mi Kyoung;Kang, Inn Soo
Clinical and Experimental Reproductive Medicine
/
v.33
no.3
/
pp.179-187
/
2006
Objectives: The risk of aneuploidies of embryos increases in advanced maternal age or parental karyotype abnormality and it results in poor reproductive outcomes such as recurrent spontaneous abortion (RSA) or repeated implantation failure (RIF). Preimplantation genetic diagnosis for aneuploidy screening (PGD-AS) can be applied for better ART outcome by selecting chromosomally normal embryos. The aim of this study is to evaluate the clinical outcome of PGD-AS and which group can get much benefit from PGD-AS among the patients expected to have poor reproductive outcome. Methods: In 42 patients, 77 PGD cycles were performed for aneuploidy screening. Patients were allocated to 3 groups according to the indication of PGD-AS: group I-patients with old age (${\geq}37$) and RIF more than 3 times (n=11, mean age=42.2 yrs.), group II-patients with RSA (${\geq}3$ times) associated with aneuploid pregnancy (n=19, mean age=38.9 yrs.), group III-parental sex chromosome abnormality or mosaicism (n=18, mean age=29.6 yrs.) including Turner syndrome, Klinefelter syndrome and 47, XYY. PGD was performed by using FISH for chromosome 13, 16, 18, 21, X and Y in group I and II, and chromosome X, Y and 18 (or 17) in group III. Results: Blastomere biopsy was successful in 530 embryos and FISH efficiency was 92.3%. The proportions of transferable embryos in each group were $32.5{\pm}17.5%$, $23.0{\pm}21.7%$ and $52.6{\pm}29.2%$ (mean ${\pm}$ SD), respectively, showing higher normal rate in group III (group II vs. III, p<0.05). The numbers of transferred embryos in each group were $3.9{\pm}1.5$, $1.9{\pm}1.1$ and $3.1{\pm}1.4$ (mean ${\pm}$ SD), respectively. The clinical pregnancy rates per transfer was 0%, 30.0% and 20.0%, and it was significantly higher in group II (group I vs. group II, p<0.05). The overall pregnancy rate per transfer was 19.6% (10/51) and the spontaneous abortion rate was 20% (2/10) of which karyotypes were euploid. Nine healthy babies (one twin pregnancy) were born with normal karyotype confirmed on amniocentesis. Conclusion: Our data suggests that PGD-AS provides advantages in patients with RSA associated with aneuploidy or sex chromosome abnormality, decreasing abortion rate and increasing ongoing pregnancy rate. It is not likely to be beneficial in RIF group due to other detrimental factors involved in implantation.
Jo Qtae;Moon Hyo Bang;Cho Yong Chul;Kim Kwang Soo;Choy Eun Jung;Ko Sung Chung;Song Young Chae
Fisheries and Aquatic Sciences
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v.8
no.1
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pp.27-33
/
2005
This is a subsequent study to our previous finding that Pacific oysters, Crassostrea gigas, gained a so-called upper plateau concentration, around 30,000 ng/g dry weight digestive gland for benzo(a)pyrene, showed reproductive behavior but produced their ensuing reproductive outputs damaged. A serial dilution of sediment elutriates from Jinhae Bay, Korea, where pollution was progressive, were exposed to gametes of the Pacific oyster for 0, 5, 10, 20, 30, and 60 min to detail the pollutant effects on very young specimens. There was an apparent critical dilution over which adverse effects are evident. This was $10\%$ of the present sediment elutriate, corresponding to 0.3 ng/g on the basis of total polycyclic aromatic hydrocarbons (PAHs) for the oyster. Within the dilution the embryonic development was not influenced by the duration of exposure to its gamete stage. At higher dilutions over the critical dilution, occurrence of abnormality increased dependent on the pollutant dilution and the duration of exposure. Similar trends were also found in larval mortality. However, overall, the chemical toxicity was more significant to morphogenesis than to survival, suggesting a potential recruitment of the pollutants-induced abnormal larvae in the wild population to threaten the population integrity.
Park Dong-Heon;Jang Hyun-Yong;Kim Choung-Ik;Cheong Hee-Tae;Park Choon-Keun;Yang Boo-Keun
Toxicological Research
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v.21
no.2
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pp.167-173
/
2005
Bisphenol A (BPA) is a monomer used in the manufacture of a multitude of chemical products, including epoxy resins and polycarbonate. The objective of this study was to evaluate the effects of BPA administration on reproductive characteristics and blood hematological and chemical values in offspring of pregnant dams treated with BPA. BPA was administrated to pregnant mice by intraperitoneally injection with 0, 0.05, 0.5 and 5.0 mg/kg B.W. for 5 times at 3 days interval on gestation days 1-16. There were no treatment-related effects of BPA on reproductive organ weight in male offsprings at 45 days-of-age, but body weight was the lowest in 5.0mg BPA group when compared to other groups (P<0.05). No differences in semen characteristics (sperm concentration, viability, motility and abnormality) were observed between the control and BPA treatment groups. The WBC, HB, HT, MCV, MCH, MCHC, albumin, BUN and total protein of blood hematological and chemical values in male offsprings were not difference for any treatment groups, but RBC value in BPA groups was significantly increased comparing to the control group (P<0.05). The PLT value was slightly higher in 5.0mg BPA groups than in any other group, but not significantly difference among the experimental groups. In female offsprings, the effects of BPA didn't affect to the body and ovary weight, but the uterus weight in 5.0mg BPA group was slightly heavier than that of control group (P>0.05). No statically significant difference in blood hematological values in female offsprings were observed between the control group and BPA groups, but the concentration of albumin and BUN were significantly higher in 0.5mg BPA group when compared to control and other BPA treatment groups (P<0.05). The histological evaluation of testis and ovary in growing offspring at 45 days-of-age was not difference between the control group and BPA groups, but endometriosis of the uterus in female offspring was dramatically increased in 0.5 and 5.0mg BPA groups. These founding suggest that low concentration of BPA might not have a important role on reproductive ability or blood metabolite in offspring of pregnant dams treated with BPA.
This study was conducted to determine the relationship between elapsed time after semen preservation on the changes of bacteria and semen quality. Semen was diluted with BTS(Beltsville Thawing Solution) extender without antibiotic for 7 days and sperm parameter and fertility were measured. Sperm motility was measured by CASA and total bacteria number was counted after 22~24 hr incubation from counting agar plate in which sperm dilute to $10{\sim}10^6$ in 0.9% saline solution and inoculate to agar. Acrosomal integrity was measured by Chlortetracycline (CTC) staining. CTC patterns were uniform fluorescence over the whole head (pattern F), characteristic of incapacitated acrosome-intact spermatozoa; fluorescence-free band in the post-acrosomal region (pattern B), characteristic of capacitated acrosome-intact spermatozoa; and almost no fluorescence over the whole head except for a thin band in the equatorial segment (pattern AR), characteristic of acrosome reacted spermatozoa. Total number of bacteria was significantly increased (p<0.0001) 3 days after preservation. Sperm motility, viability, and morphological abnormality on elapsed time after preservation were lower from 5 ($77.24{\pm}6.47$, p<0.001) and 7 days ($77.24{\pm}6.47$, p<0.001) after preservation compared to 1 ($15.71{\pm}7.18$) and 3 days($18.39{\pm}7.22$) after preservation, respectively. Sperm viability was significantly lower ($53.25{\pm}35.03$, p<0.0001) at 7 days after preservation. Morphological abnormality of sperm was lower (p<0.001) at 1 ($15.71{\pm}7.18$) and 3 ($18.39{\pm}7.22$) days compared to 5 ($21.84{\pm}7.91$) and 7 ($22.59{\pm}9.93$) days after preservation. Acrosomal integrity and capacitation rate (pattern F) were significantly lower (p<0.001) from 5 days after preservation. Based on the data we obtained from this study suggested that semen preserved more than 5 days without antibiotic would not recommend use for artificial insemination.
The objective of this study was to evaluated the efficiency of sperm cryosurvival using each extenders Triladyl and LEY containing egg yolk to the cryopreservation of separated sperm by percoll in miniature pig. The ejaculated semen from miniature pig was separated by 65% percoll and un-separated sperm as a control before freezing. The freezing of diluted semen added with Triladyl containing egg yolk and LEY solution (solution I: 11% Lactose or Triladyl + egg yolk; solution II: solution I + glycerol + OEP). Analysis of sperm ability was estimated by viability, capacitation acrosome reaction using chlortetracycline (CIC) the morphologic abnormality and hypoosmotic swelling test(HOST). The groups were designed that as separated sperm by Percoll with Triladyl(ST) or LEY(SL) for cryopreservation. And unseparated sperm with Triladyl(UT) or LEY(UL). As a results, the viability was higher significantly(p<0.05) in ST, SL, UT than UL extender. The morphologic abnormality was measured significantly (p<0.05) lower in ST than other extenders. The AR-patterned in CTC analysis was measured significantly(p<0.05) lower in SL and UL than other extenders. In conclusion, using Triladyl extender resulted in viability and morphology of separated sperm by percoll that were effective than using LEY extender, but it resulted in capacitation acrosome reaction was lower than using LEY extender.
Jeong, Seon Young;Shin, Soon Ho;Jin, Young Guk;Ju, Sun Mi;Lee, Jung Sick
Korean Journal of Ichthyology
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v.18
no.2
/
pp.87-96
/
2006
Reproduction and organ system of the oily bitterling, Acheilognathus koreensis in Isa stream were investigated by means of histological methods. Sex ratio was observed 1 : 1 in female to male. The gonadosomatic index (GSI) of the female showed two peak in December and April. The ovipositor length index (OLI) represented irregular change. From the histological analysis of the organ system, epidermal atrophy and hyperplasia of pigment cell were observed in the skin. Epithelial layer lifting of the lamella and bifurcation of the filament were observed in the gill. Also histological changes as congestion, cytoplasmic degeneration of hepatic cell, glomerular dilatation, hyaline droplet degeneration of renal tubule epithelium and pycnosis of interstitial cell were identified in the liver and kidney, respectively.
As a fundamental study to increase the reproductive efficiency in cattle, highly motile sperm were separated and collected from raw semen, extended semen and frozen semen by different methods using various concentrations of bovine serum albumin or Tyrode's solution. Various characteristics and light microscopic and electron-microscopic morphology of sperm separated by different methods were compared. The results obtained were as follows; 1. The sperm separated from raw semen using bovine serum albumin showed significantly high value in motility, motile sperm count, percent of normal sperm and progressive motility, as compared with control sperm and revealed the highest sperm recovery rate when separated with 6% bovine serum albumin. 2. The sperm motility, percent of normal sperm and progressive motility of the highly motile sperm frozen after being separated from raw semen with bovine serum albumin, showed significantly high value than those of a control sperm and especially found the highest value when separated with 20% bovine serum albumin. 3. Light-microscopic percent of abnormality was significantly low in the prefrozen and postfrozen highly motile sperm separated with bovine serum albumin, as compared with control sperm. 4. Electron-microscopic finding of the highly motile sperm separated with bovine serum albumin showed low percent of deformity in the dilatation and vesiculation of cell membrane, in dilatation and density loss of acrosome than in those of control sperm. 5. It was impossible to separate the highly motile sperm from frozen semen with bovine serum albumin, but it was possible with Tyrode's solution. 6. Recovery rate of highly motile sperm from raw semen extended semen and frozen semen was the highest when the sperm pellet stood in Tyrde's solution for 80 minutes. 7. The highly motile sperm separated from raw semen, extended semen and frozen semen with Tyrode's solution showed significantly high value in motility, progressive motility and percent of normal sperm, as compared with control sperm. 8. Highly motile sperm, when separated from raw semen, extended semen and frozen semen with Tyrode's solution, showed significantly low percent of microscopic abnormality as compared with control sperm.
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