This study was carried out to investigate the morphological changes of gonadotropes in pituitary gland and spermatogenic cells in testis, obtained from 150 of 3-year-old immature and mature male rainbow trout (Oncorhynchus mykiss) during the reproductive cycles from March to February in the following year. In the maturation cycle of the pituitary gonadotropes of cultured rainbow trout, three periods can be distinguished i.e. a period of resting(March-August), a period of full spermatogenesis (September-November), and a period of breeding (December-February). The ultrastructures of the gonadotropes largely parallel the cyclical changes in the tests. The seminiferous tubules contain all spermatogenetic stages and sperm cells in a period of early maturation. At first, the size of the nucleus and cytoplasm decrease gradually at every stages from spermatogonia to spermatids. In the secondary spermatocytes, the small mitochondria are located over the outer cytoplam. In spermatids, the cytoplasmic masses move toward the posterior part of the nucleus. In spermatids, the two large mitochondria are located over the cytoplasm. In spermatids, the cytoplasmic masses move towark the posterior part of the nucleus. In spermatids, the two large mitochondria are located over the cytoplasm and begin to elongate. In spermatozoa, the surface of the nucleus devreases in volume. Examination by TEM shows that the nuclear envelope and plasma membrane are slightlywrinkled and closely adhered to the nucleus of spermatozoa. Two oval mitochondria are quite separated and the flagellum is inserted into the base of the spermatozoa head.The axoneme in this fish has the typical pattern such as nine peripheral doublets and a central doublet(9+2). there are remarkable individual differences in the size and morphology of spermatozoa head as observed by transmission and scanning electron microscopy.
The maturity and spawning of Korean flounder Glyptocephalus stelleri were investigated using samples randomly collected in the East Sea of Korea from May 2005 to September 2008. Gonadosomatic index began to increase in December and reached a maximum between March and May. After spawning it began to decrease from June. The spawning period was March to June, and the main spawning period was April to May. Annual reproductive cycles of this species can be divided into four successive stages; immature stage (July~November), maturing stage (December~February), mature stage (March~April) and spent stage (May~June). The fecundity ranged from 15,146 eggs at 24.1 cm (TL) to 101,491 eggs at 38.1 cm (TL). The relationship between total length (TL) and fecundity (F) can be expressed as $F=0.0004TL^{3.449}$ ($R^2=0.663$), with F increasing with TL. The TL at 50% group maturity ($TL_{50%}$) was estimated to be 25.6 cm.
Seasonal changes in biochemical composition of muscle, gonad-viceral, mass and whole body of the cultured ark shell, Scapharca broughtonii in the Gamag bay of Yeosu city were studied from December 2008 to November 2009 in relation to environmental condition and reproductive cycles. Average monthly water temperature in the winter was in the range of $7-12^{\circ}C$ and $20-25^{\circ}C$ in the summer, while the salinity fluctuated in the range of 30.1%-33.8‰ on the average. Seasonal fluctuation of the concentration of nutrient salt was the highest in September ($13.04{\mu}g/L$) with average annual concentration of $4.6{\mu}g/L$. The main spawning season of the ark shell was during the months of July and August, and the gonads were in inactive stage during the winter. The gonad-visceral mass contained lower amounts of proteins than the other body parts. The most marked changes in body composition were lipids and carbohydrates within the gonad-visceral mass, and protein for each of the organs was relatively consistent throughout the year. All the parts in the visceral sac displayed the highest changes during the gametogenic cycle while the contents of moisture and lipid within the visceral act displayed somewhat inverse relations with each other. Moisture content was the lowest during the inactive stage during which the lipid content is the highest. The lipid content was the lowest immediately following spawning with increase in the moisture content as the lipid is being consumed. Protein mass within the visceral sac was low in comparison to the muscle mass. It is deemed that carbohydrates, lipids and proteins in the visceral sac play the major role as the source of energy during the development process of the gonads, and used for maintenance of base metabolism when available food is scarce.
Implantation itself is governed by an array of endocrine, paracrine and autocrine modulators, of embryonic and maternal origin. Window of implantation is the unique temporal and spatial expression of factors allows the embryo to implant via signaling, appositioning, attachment, and invasion in a specific time frame of $2{\sim}4$ days. When the embryo has arrived in the uterine cavity, a preprogrammed sequence of events occurs, which involves the production and secretion of a multitude of biochemical factors such as cytokines, growth factors, and adhesion molecules by the endometrium and the embryo, thus leading to the formation of a receptive endometrium. Cytokines such as LIF, CSF-1, and IL-1 have all been shown to play important roles in the cascade of events that leads to implantation. Integrin, L-selectin ligands, glycodelin, mucin-1, HB-EGF and pinopodes are involved in appositioning and attachment. The embryo also produces cytokines and growth factors (ILs, VEGF) and receptors for endometrial signals such as LIF, CSF-1, IGF and HB-EGF. The immune system and angiogenesis play an important role. The usefulness of these factors to assess endometrial receptivity and to estimate the prognosis for pregnancy in natural and artificial cycles remains to be proven. Integrins, pinopodes, glycodelin and LIF (from biopsies) are promising candidates; from uterine flushings, glycodelin and LIF are also candidates. The ideal serum marker is not available, but VEGF, glycodelin and CSF have some clinical implications. Further evaluation that includes larger groups of infertile women and fertile controls are needed to elucidate whether their presence in plasma, flushing fluid, or endometrial samples can be used as some kind of a screening tool to assess endometrial function and prognosis for pregnancy before and after artificial reproductive therapy. A better understanding of their function in human implantation may lead to therapeutic intervention, thereby improving the success rate in reproduction treatment. New molecular techniques are becoming available for measuring both embryonic and endometrial changes prior to and during implantation. The use of predictive sets of markers may prove to be more reliable than a single marker. Ultimately, the aim is to use these tools to increase implantation in artificial cycles and consequently improve live-birth rates.
Objective: This study was performed to evaluate the effect of oxytocin antagonist on the outcome of IVF/ICSI cycles in infertile patients with repeated failure of IVF/ICSI treatment. Method: Forty patients who had experienced two or more failures of IVF/ICSI treatment without low ovarian reserve, were recruited for this prospective randomized study. All patients received controlled ovarian stimulation (COS) using GnRH antagonist multidose protocol (MDP). For the intervention group, intravenous administration of atosiban (mixed vasopressin $V_{1A}$/oxytocin antagonist) started with a bolus dose 6.75 mg one hour before embryo transfer (ET) and continued at an infusion rate of 18 mg/hour. After ET, administered atosiban was reduced to 6 mg/hour and continued for 2 hours. The main efficacy endpoints were clinical pregnancy rate and implantation rate. Results: Patients' characteristics were comparable in the intervention and control groups. COS parameters and IVF results were also similar. The number of uterine contractions for 3 minutes measured just before ET was significantly lower in the intervention group than control group ($3.5{\pm}1.4$ vs $8.7{\pm}2.2$, p<0.001). While there was no statistically significant difference in the clinical pregnancy rate between control group and intervention group (20.0% and 40.0%, p=0.168), the implantation rate was significantly higher in the intervention group, with 16.9% (11/65) compared with 6.0% (4/67) in the control group (p=0.047). There were no differences in ectopic pregnancy rate and miscarriage rate between the two groups. Conclusion: This study demonstrates that administration of oxytocin antagonist during ET can improve the implantation rate probably by decreasing the frequency of uterine contractions in infertile patients undergoing IVF/ICSI treatment.
Objective : To identify the factors affecting the complete fetal loss following multifetal pregnancy reduction (MFPR). Design: Retrospective clinical study. Methods : A total of 256 consecutive treatments of MFPR in IVF-ET cycles performed between 1992 through 2000 in Samsung Cheil hospital were analyzed. MFPR was done around 8 weeks of gestation by transvaginal ultrasono-guided aspiration in multiple pregnancies and reduced to singleton or twins. Stepwise logistic regression was performed to identify the factors affecting the final outcome of pregnancy after MFPR. Dependent variable was complete fetal loss and the independent variables were maternal age, paternal age, initial number of gestational sac (iGSNO), initial number of fetal heart beat, the number of remaining live fetus after MFPR, and chorionicity. Results: The total survival rate was 87.9%, and total fetal loss rate after MFPR was 12.1%. Total fetal loss occurred within four weeks from MFPR procedure was 1.95%. Total loss occurred after four weeks of procedure and before 24 gestational weeks was 8.2%. Seventy nine percent (202/256) of pregnancies delivered after 34 weeks of gestation. The survival rate of pregnancies reduced to singleton was significantly higher than that of pregnancies reduced to twins (93.5% vs. 86.7%, p<0.05). The mean ($\pm$SEM) gestational age at delivery was $36.2{\pm}1.0$ and $34.1{\pm}0.5$ weeks for pregnancies reduced to singletons and twins, respectively (p=0.065). Logistic regression analysis revealed that the maternal age, the number of initial gestational sac (iGSNO), and the number of remaining live fetus after MFPR significantly affected the rate of total fetal loss (Z = 0.174'age + 0.596'iGSNO + 1.324'remaining fetuses -12.07), (p<0.05). Conclusions: MFPR seems to be a relatively safe and efficient method to improve the obstetric outcome in high order multiple pregnancy. Because the maternal age, the number of initial gestational sac and the remaining live fetuses after MFPR affect the total fetal loss rate, restriction of the number of transferred embryos according to the age and MFPR to singleton fetus could be considered for the better obstetric outcome in IVF pregnancy.
Estradiol-17${\beta}$($E_2$) levels in the blood were estimated according to varying the time and amount of the administration of $Clomid^{(R)}$. $Clomid^{(R)}$ were administered on the 2nd, 3rd and 4th menstrual day corresponding to the recruitment period and on the 5th menstrual day corresponding to the selection period of the ovarian follicles, respectively. And $Clomid^{(R)}$ were administered 50 mg, 100 mg and 150 mg/day, repectively. The effects of the sequential HMG to $E_2$ levels in the blood were also estimated. The results were as following : 1. Blood $E_2$ levels according to the day and amount of administration of $Clomid^{(R)}$ were the highest in the group 3(D $2{\sim}6$, 150 mg/day, with HMG) and the lowest in the group 6(D $5{\sim]9$, 50 mg/day, without HMG). $E_2$ levels showed increasing tendency to 0 day. 2. In the cases of the administration of $Clomid^{(R)}$ during the $2nd{\sim}6th$ menstrual day, $E_2$ levels according to the amount were similar among groups and showed increasing tendency daily. 3. In the cases of administration of $Clomid^{(R)}$ during the $2nd{\sim}6th$ menstrual day, $E_2$ levels according to the sequential HMG independent of the amount of $Clomid^{(R)}$ were higher in the with HMG group than without HMG groups. 4. In the case of the administration of $Clomid^{(R)}$ during the $5th{\sim}9th$ menstrual day, $E_2$ levels according to the amount were the highest in the 100 mg/day group and the lowest in the 50mg/day group. 5. In the cases of administration of $Clomid^{(R)}$ independent of the amount during the 5th${\sim}$9th menstrual day, $E_2$ levels according to the sequential HMG were higher in the with HMG group than without HMG group. 6. $E_2$ levels according to the amount independent of the day of the administration of $Clomid^{(R)}$ were the highest in the 100 mg/day group and 150 mg/day, 50 mg/day group in low sequence. 7. $E_2$ levels according to the sequential HMG independent of the day and amount of the administration of $Clomid^{(R)}$ were higher in the with HMG group than the without HMG group. 8. $E_2$ levels according to the day of the administration of $Clomid^{(R)}$ independent of the amount of $Clomid^{(R)}$ and sequential HMG were the highest in the group D 2${\sim}$6 and the lowest in the group D 5${\sim}$9. According to the above results, there were higher $E_2$ levels in the group with sequential HMG than without HMG. Therefore, the hypothesis, postulated initially by the author, was not verified that sequential HMG would not affect the $E_2$ levels which were related to the process of the selection of the ovarian follicle in the connection with 'FSH window'. Because it may be the stimulation after the selection of later predominant follicle. And the highest level of $E_2$ was estimated in the $Clomid^{(R)}$ 150 mg/day group with sequential HMG on the 2nd${\sim}$6th day, and the higher levels were estimated in the 2nd${\sim}$6th day, 3rd${\sim}$7th day and 4th${\sim}$8th day groups than the 5th${\sim}$9th day group. The lower levels were estimated in the $Clomid^{(R)}$ 50 mg/day group without HMG than 100 mg/day and 150 mg/day on the 5th${\sim}$9th day. Therefore, further study will be needed that combines analyses of the E2 levels in the blood according to the various administration of $Clomid^{(R)}$ with or without sequential HMG and determination of the numbers and size of the ovarian follicles by ultrasonogram.
Objective: This study was conducted to investigate the effect of vitrification on the implantation and the pregnancy of human blastocysts. Method: The transfer of the frozen-thawed blastocysts by the slow freezing or vitrification was performed between January 1998 and July 1999. The zygotes derives from IVF were cocultured with cumulus cells in YS medium containing 20% hFF for 5days. Two or three of the best balstocysts produced on day 5 were transferred into the uterus, and then supernumerary blastocysts were randomly divided into two groups. One was frozen by slow freezing and the other was frozen by vitrification method. The slow freezing procedure was performed in two steps (5% glycerol and 9% glycerol + 0.2 M sucrose for 10 min, respectively) using programmed freezer ($-2^{\circ}C$/min to $-7^{\circ}C$, manual seeding at $-7^{\circ}C$, $-0.3^{\circ}C$/min to $-38^{\circ}C$ and plunged into $LN_{2}$). The blastocysts frozen by slow freezing were thawed at $36^{\circ}C$ then removed glycerol in 7 steps. The vitrification procedure was performed in three steps (10% glycerol for 5 min, 10% glycerol + 20% ethylene glycol for 5 min, 25% glycerol + 25% ethylene glycol and directly $LN_{2}$ within 1 min). The blastocysts frozen by vitrification were thawed at $20^{\circ}C$ water then removed cryoprotectant in 3 steps. In each group, thawed blastocysts were cocultured with cumulus cells in YS medium containing 20% hFF for 18h and transferred into the uterus. The implantation rate was evaluated per transferred blastocysts and the pregnancy rate was evaluated per transfers. Results: The survival rate of vitrified group (74.5%) was higher than slow freezing group (68.0%), but not significant. When 98 thawed blastocysts of vitrification were transferred in 40 cycles, 19 pregnancies (clinical pregnancy rate; 47.5%) were established. One miscarriage occurred in the eighth week of pregnancy (ongoing pregnancy rate; 45.0%). 7 pregnancies were ongoing, 11 pregnancies went to term, and 16 healthy infants were born. The Implantation rate was 31.6%. These results were higher than those obtained by the slow freezing (clinical pregnancy rate; 40.3%, ongoing pregnancy rate; 32.5% and implantation rate; 25.3%), but not significant. Conclusion: Vitrification is a simple, quick and economical method when compared to slow freezing. It will be chosen as a good method of human embryo freezing in IVF-ET programs.
Objective: Birth sex ratio (BSR) with human IVF-ET program is an interesting subject of social and scientific issue but very little information is available in Korea. This study was performed to assess the BSR with IVF-ET and to suggest the effective factors on the BSR. Methods: The national data from 1991~2008 were obtained from governmental Statistics Korea and the delivery data of human IVF-ET program on 2007 and 2008 were provided from the Ministry for Health, Welfare and Family Affairs. The BSR were statistically analyzed according to methods of IVF and to transferred embryos from fresh or frozen-thawed cycles. Results: The BSRs of Korean populations were over 1.10 up to 2002, and then it declined and maintained to 1.06 as balance BSR on 2007 and 2008. In human IVF-ET program, the BSRs were 1.07 on 2007 and 1.06 on 2008, respectively. Conventional IVF on 2008 showed the highest BSR as 1.10, and ICSI the lowest on 2008 as 1.01. There was no significant difference of BSRs related to the methods of in vitro fertilization and the feature of transferred embryos. Conclusion: The BSR of Korea showed 1.06~1.07 as normal and balanced state on 2007 and 2008 both general populations and human IVF-ET program. To the best of our knowledge, this is the first study to evaluate the BSR of human IVF-ET in Korea. There is a need to expand the further studies for national statistics and influencing factors on the BSR with IVF-ET.
Objective: To evaluate the outcomes of in vitro fertilization and embryo transfer (IVF-ET) in women over 40 years of age. Methods: A total of 170 patients (271 cycles) over 40 years of age who underwent IVF-ET at Seoul Women's Hospital (Incheon, Korea) were analyzed in this study retrospectively. The patients were grouped into the women <44 years old group and the women $\geq$44 years old group. Statistical analysis was performed using Student's t-test and Fisher's exact test as appropriate. Results: An overall clinical pregnancy rate per retrieval was 11% (30/271). Of these, clinical miscarriage rate were 33% (10/30) and the overall delivery rate was 7.4% (20/271) per retrieval, respectively. The women $\geq$44 years old group had significantly higher cancellation rate (13% vs. 25%), lower number of retrieved oocytes (6.17$\times$4.62 vs. 4.13$\times$4.07), decreased number of 2PN (4.83$\times$3.61 vs. 3.46$\times$3.12), and reduced embryos for transfer (3.52$\times$1.72 vs. 2.81$\times$1.83) than the women <44 years old group. We found significantly lower clinical pregnancy rate (13.0% vs. 2.1%) and live birth rate (9.0% vs. 0.0%) in the women $\geq$44 years old group than the women <44 years old group. Conclusion: The present study has shown that IVF outcome is seriously impaired in the women $\geq$44 years old.
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