Objective: An experiment was conducted to study the association between the single nucleotide polymorphisms (SNPs) in 5'-untranslated regions (5'-UTR) of equine chorionic gonadotropin (eCG) genes and the serum eCG levels. Methods: SNPs in 5'-UTR of eCG genes were screened across 10 horse breeds, including 7 Chinese indigenous breeds and 3 imported breeds using iPLEX chemistry, and the association between the serum eCG levels of 174 pregnant Da'an mares and their serum eCG levels (determined with ELISA) was analyzed. Results: Four SNPs were identified in the 5'-UTR of the $eCG{\alpha}$ gene, and one of them was unique in the indigenous breeds. There were 2 SNPs detected at the 5' end of the $eCG{\beta}$ subunit gene, and one of them was only found in the Chinese breeds. The SNP g.39948246T>C at the 5'-UTR of $eCG{\alpha}$ was associated significantly with eCG levels of 75-day pregnant mare serum (p<0.05) in Da'an mares. Prediction analysis on binding sites of transcription factors showed that the g.39948246T>C mutation causes appearance of the specific binding site of hepatocyte nuclear factor 3 forkhead homolog 2 (HFH-2), which is a transcriptional repressor belonging to the forkhead protein family of transcription factors. Conclusion: The SNP g.39948246T>C at the 5'-UTR of $eCG{\alpha}$ is associated with eCG levels of 75-day pregnant mare serum (p<0.05).
The embryonic development and hatchability of the artificially fertilized eggs of convict grouper (Hyporthodus septemfasciatus) ♀${\times}$giant grouper (Epinephelus lanceolatus) ♂ (CGGG) were compared to those of the maternal pure species (convict grouper ♀${\times}$♂, CG) to establish a novel grouper hybrid for aquaculture industry. The fertilized eggs were divided into nine 5-L beakers (3,000~5,000 eggs/beaker) filled with UV sterilized seawater and incubated at a temperature range of $23.5{\sim}24.8^{\circ}C$ (32.1~32.8 psu). The percentages of fertilization and hatching of CGGG were $69.4{\pm}1.5%$ and $59.0{\pm}5.1%$, respectively and were significantly lower than those of CG (p<0.05). The CGGG proceeded normal embryonic development similar to that of CG, but showed an irregular cleavage, immature embryonic body and spinal deformity in hatched larvae. The incubation time from fertilization to hatching of CGGG was 31 hrs, which was approximately 2 hrs slower than that of CG. Our study provided the possibility of mass production of grouper hybrid CGGG larvae.
A 10-week feeding trial was carried out to investigate the feasibility of using spent coffee ground (CG) as a potential feed ingredient for olive flounder. Growth, feed utilization, body composition and antioxidant enzyme activity were examined. A control diet was formulated and three other diets were prepared to contain 5, 10 or 15% CG (designated as Con, CG5, CG10 and CG15, respectively) by replacing for wheat flour. Two hundred forty fish ($104{\pm}0.7g/fish$) were allotted to 12 circular tanks of 400 L capacity at a density of 20 fish per tank and fed the experimental diets twice daily. At the end of the feeding trial, fish fed the CG5 diet exhibited significantly (P < 0.05) higher growth performance than those fed the control diet. Also, fish fed the CG10 diet had a comparable growth to that of the control group, but further increase of dietary CG inclusion level to 15% resulted in significant decrease of growth performance. Fish fed the CG15 diet showed significantly lower feed efficiency and protein efficiency ratio than other treatments. Significantly lower muscle protein content was observed in fish fed CG15 diet compared to the control. Significant reduction in plasma cholesterol concentration was found in fish fed CG15 diet compared to control. No significant changes were found in alkyle and superoxide radicals scavenging activities of plasma, muscle and liver among dietary treatments. Also, liver total protein, total antioxidant capacity, catalase and glutathione peroxidase activities were not significantly influenced by dietary inclusion of CG. According to these results, we concluded that CG can be included up to 10% in the diet for olive flounder without any adverse effects on growth, feed utilization and antioxidant enzyme activity.
Equine chorionic gonadotropin (eCG) consists of highly glycosylated ${\alpha}-$ and ${\beta}-subunits$ and is a unique member of the gonadotropin family, because it elicits the response characteristics of follicle stimulating hormone (FSH) and luteinizing hormone (LH) in species other than the horse. To directly assess the biological function of $rec-eCG{\beta}/{\alpha}$, we constructed mammalian expressing vectors of equine luteinizing hormone/chorionic gonadotropin receptors (eLH/CGR). The activity of $rec-eCG{\beta}/{\alpha}$ in vitro assayed in transient transfected CHO-K1 cells and in stably transfected PathHunter Parental cells with eLH/CGR was investigated. $rec-eCG{\beta}/{\alpha}$ was efficiently secreted in the CHO-K1 suspension cell media, and the quantity detected was about 200 mIU/ml from 1 to 7 days after transfection. In the western blot analysis, the $rec-eCG{\beta}/{\alpha}$ protein was broadly identified to be about 40~45 kDa molecular weight. The cAMP stimulation in CHO-K1 cells expressing eLH/CGR was determined to evaluate the activity of $rec-eCG{\beta}/{\alpha}$. The cAMP concentration increased in direct proportion to the concentration of the $rec-eCG{\beta}/{\alpha}$. The $EC_{50}$ value in the transient transfected CHO-K1 cells was $8.1{\pm}6.5ng$. The stable cell lines of eLH/CGR were established in the PathHunter Parental cells expressing ${\beta}-arrestin$. We found that $rec-eCG{\beta}/{\alpha}$ had full LH activity in the PathHunter Parental cells expressing eLH/CGR. The $EC_{50}$ value in transient and stable cells was $5.0{\pm}4.7ng/ml$ and $4.5{\pm}5.2ng/ml$, respectively. These results suggest that $rec-eCG{\beta}/{\alpha}$ has a biological activity in a cell expressing eLH/CGR. These stable cells expressed in PathHunter Parental cells could be useful for elucidating the functional mechanisms of deglycosylated $rec-eCG{\beta}/{\alpha}$ mutants.
This study was undertaken to test the hypothesis the hypothesis that treatment with hCG (5,000 IU) at the time of embryo transfer would enhance pregnancy rates in recipients, and the concentration of plasma urea nitrogen(PUN) in recipients was related to the effect of hCG on the reproductive performance. Blood samples were taken according to experimental condition for the assessment of the endogenous plasma progesterone concentration and plasma urea nitrogen. Concentrations of progesterone in plasma were higher in cows treated with hCG on day 7(estrus=day 0) than in those untreated during 7∼43 days after insemination. The pregnancy rates were 65.5 and 54.6% for the hCG treated and untreated groups, respectively. In recipient group categorized with PUN concentration of <12 mg/이, the pregnancy rates were 68.8 and 46.7% for the hCG treated and untreated groups, respectively. The results suggest that hCG treatment at 7 days after insemination could be used to increase the pregnancy rate of embryo transfer, and transfer, and only the recipients with PUN concentration of <12 mg/dl were influenced by treatment with hCG.
Park, Yoon-Ki;Kim, Jong-Wook;Lee, Tae-Hyung;Park, Wan-Seok;Lee, Sung-Ho;Chung, Wun-Yong
Journal of Yeungnam Medical Science
/
v.2
no.1
/
pp.221-227
/
1985
Thirty-four patients with suspected ectopic pregnancy whose serum hCG levels had been measured by beta-hCG EIA before surgery were evaluated retrospectively. The results were as follows: 1. Final diagnosis of thirty-four patients with suspected ectopic pregnancy comprised twenty- eight tubal pregnancy, five ruptures of hemorrhagic corpus luteum and, one tubo-ovarian abscess. One of the five patients with rupture of hemorrhagic corpus luteum was accompanied by missed abortion. 2. Range of serum hCG levels in twenty-eight patients with tubal pregnancy was 59-21,980 mIU/ml and that of four patients with rupture of hemorrhagic corpus luteum and one patient with tubo-ovarian abscess was 0.6-6.6mIU/ml. Serum hCG level of a patient with rupture of hemorrhagic corpus luteum who was accompanied by missed abostion was 200 mIU/ml. 3. Serum hCG levels in twenty-two of twenty-eight patients with tubal pregnancy were lower than 3,000 mIU/ml. Low serum hCG level below 100mIU/ml and high serum hCG level above 6,500 mIU/ml were noticed in four and six patients with tubal pregnancy, respectively. 4. Mean serum hCG levels (${\pm}SD$) of twelve patients with tubal pregnancy who had intra-abdominal free blood of less than 500ml and sixteen patients with tubal pregnancy who had intra-abdominal free blood of more than 500ml were 4,131 (${\pm}7,801$) mIU/ml and 3,208 (${\pm}5,081$) mIU/ml, respectively. There was no statistical difference in the mean level of serum hCG between both group (P>0.05). 5. Mean serum hCG levels (${\pm}SD$) of thirteen patients with unruptured tubal pregnancy and fifteen patients with ruptured tubal pregnancy were 2,628 (${\pm}5,448$) mIU/ml and 4,449 (${\pm}6,938$) mIU/ml, respectively. Mean level of serum hCG was statistically higher in ruptured tubal pregnancy (P<0.01). 6. Positive rate of urine pregnancy test in the diagnosis of ectopic pregnancy was 64% (16/25) and mean range of serum hCG level of nine patients with false negative result were 353 mIU/ml and 59-933 mIU/ml.
Objective: This study was designed to evaluate the effects of endogenous LH surge, GnRH agonist (GnRH-a) or human chorionic gonadotropin (hCG) as ovulation trigger on pregnancy rate by intrauterine insemination (IUI). Method: Patients received daily 100 mg of clomiphene citrate (CC) for 5 days starting on the third day of the menstrual cycle followed by human menopausal gonadotropin (hMG) for ovulation induction. Follicles larger than >16 mm in diameter were present in the ovary, frequent LH tests in urine were introduced to detect an endogenous LH surge. Final follicular maturation and ovulation were induced by GnRH-a 0.1 mg (s.c.) or hCG $5,000{\sim}10,000$ IU (i.m.) administration except natural ovulation. Pregnancy was classified as clinical if a gestational sac or fetal cardiac activity was seen on ultrasound. Results: There were no differences in age, duration of infertility and follicle size, but more ampules of hMG were used in GnRH-a group compared to hCG 10,000 IU treated group (p<0.05). Lower level of estradiol ($E_2$) on the day of hCG or GnRH-a injection was observed in hCG 10,000 IU group than other treatment groups (p<0.01). The overall clinical pregnancy rate was 19.8% per cycle (32/162) and 22.2% per patient (32/144). Pregnancy rate was higher in natural-endogenous LH surge group (37.5%, 9/24) than GnRH-a (18.8%) or hCG treated group (20.9% & 13.9%), but this difference was not statistically significant. No patient developed ovarian hyperstimulation. Abortion rate was 22.2% (2/9) in hCG 5,000 IU group. Delivery or ongoing pregnancy rate was 37.5% (9/24), 18.8% (3/16), 16.3% (7/43) and 13.9% (11/79) in endogenous LH surge, GnRH-a, hCG 5,000 IU and hCG 10,000 IU treatment groups, respectively. Conclusion: These results support the concept that use of natural-endogenous LH surge in stimulated cycles may be more effective to obtain pregnancies by IUI than GnRH-a or hCG administration.
Background: An hCG regression curve has been used to predict the natural history and response to chemotherapy in gestational trophoblastic disease. We constructed hCG regression curves in high-risk gestational trophoblastic neoplasia (GTN) treated with EMA/CO and identified an optimal hCG level to detect EMA/CO resistance in GTN. Materials and Methods: Eighty-one women with GTN treated with EMA/CO were classified as primary high-risk GTN (n = 65) and single agent-resistance GTN (n = 16). The hCG levels prior to each course of chemotherapy were plotted in the 10th, 50th, and 90th percentiles to construct the hCG regression curves. Diagnostic performance was evaluated for an optimal cut-off value. Results: The median hCG levels were 264,482 mIU/mL mIU/mL and 495.5 mIU/mL mIU/mL for primary high-risk GTN and single agent-resistance GTN, respectively. The 50th percentile of the hCG level in primary high-risk GTN and single agent-resistance turned to normal before the 4th and the 2nd course of chemotherapy, respectively. The 90th percentile of the hCG level in primary high-risk GTN and single agent-resistance turned to normal before the 9th and the 2nd course of chemotherapy, respectively. The hCG level of ${\geq}118.6mIU/mL$ mIU/mL at the 5thcourse of EMA/CO predicted the EMA/CO resistance in primary high-risk GTN patients with a sensitivity of 85.7% and a specificity of 100%. Conclusion: EMA/CO resistance in primary high-risk GTN can be predicted by using an hCG regression curve in combination with the cut-off value of 118.6 mIU/mL at the 5thcourse of chemotherapy.
Recently, several researchers have attempted to combine human visual perception and computer graphics. Cole et. al. suggest the study results in which line drawing algorithm in computer graphics characterize the properties of line drawing made by artists. The paper also evaluates CG line drawing algorithms depending on whether people recognize effectively specific 3D shape from the image made by those CG line drawing algorithms. However, human recognizes the shape of objects more effectively in image made by BRDF shading model than line drawing algorithm. It means that the shading factor is important to recognize shape with human perception. In this paper, we analyze the correlation between shading made by human artists and that made by CG shading algorithms. The study is to characterize the mathematical properties of artists' shading and CG shading. This type of analysis can guide the future development of new CG shading algorithm in computer graphics for the purpose of shape perception.
Park, Jong-Ju;JarGal, Naidansuren;Yoon, Jong-Taek;Min, Kwan-Sik
Reproductive and Developmental Biology
/
v.34
no.1
/
pp.33-40
/
2010
Equine chorionic gonadotropin (eCG) is a heavily glycosylated glycoprotein composed of non-covalently linked $\alpha$- and $\beta$-subunits. To study the function and signal transduction of tethered recombinant-eCG (rec-eCG), a single chain eCG molecule was constructed, and the rec-eCG protein was prepared. In this study, we constructed 5 mutants (${\Delta}1$, ${\Delta}2$, ${\Delta}3$, ${\Delta}4$, and ${\Delta}5$) of rec-eCG using data about known glycoprotein hormones to analyze the role of specific follicle stimulating homone (FSH)-like activity. Three amino acids of certain specific sites were replaced with alanine. The expression vectors were transfected into CHO cells and subjected to G418 selection for 2~3 weeks. The media were collected and the quantity of secreted tethered rec-eCGs was quantified by ELISA. The LH- and FSH-like activities were assayed in terms of cAMP production by rat LH/CG and rat FSH receptors. Then, the metabolic clearance rate analyzed by the injection of rec-eCG (5 IU) into the tail vein was analyzed. The mutant eCGs (${\Delta}l$, ${\Delta}4$, and ${\Delta}5$) were transcripted, but not translated into proteins. Rec-eCG A2 was secreted in much lower amounts than the wild type. Only the rec-eCG ${\Delta}3$ ($\beta$-subunit: $Gln^{94}-Ile^{95}-Lys^{96}{\rightarrow}Ala^{94}-Ala^{95}-Ala^{96}$) was efficiently secreted. Although activity is low, its LH-like activity was similar to that of tethered $eCG{\beta\alpha}$. However, the FSH-like activity of rec-$eCG{\beta\alpha\Delta}3$ was completely flat. The result of the analysis of the metabolic clearance rate shoed the persistence of the mutant in the blood until 4 hours after the injection. After then, it almost disappeared at 8 hours. Taken together, these data suggest that 94~96 amino acid sequences in eCG $\beta$-subunit appear to be of utmost importance for signal transduction of the FSH receptor.
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