• Asian-Australasian Journal of Animal Sciences
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• 제26권4호
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• pp.488-500
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• 2013

The objective of this study was to compare the effectiveness of the protocols for superstimulation of follicular growth in Thai native heifers. Heifers (n = 20) were randomly divided into four groups of five heifers/group. Heifers were given a single dose by i.m. administration of 100 mg Follicle Stimulating Hormone dissolved in polyvinylpyrrolidone (FSHp) at 24 h. Ovum pick up (OPU) occurred at 72 h ($F_{24}O_{72}$ protocol; Group 1) or 96 h ($F_{24}O_{96}$ protocol; Group 2), and at 36 h and OPU at 72 h ($F_{36}O_{72}$ protocol; Group 3) or 96 h ($F_{36}O_{96}$ protocol; Group 4) after follicular ablation. The dynamics of ovarian follicular growth were monitored by twice-daily ultrasonographic examinations. Blood sample collections were performed every 12 h after initiation of treatment for assessment of FSH, E2 and P4 profiles. All heifers were subjected to eight repeated sequential sessions of OPU. The follicular deviation commenced $24{\pm}5.32$ h after follicular ablation in all groups. The circulatory FSH surged quickly from 24 to 36 h (>0.8 ng/ml) after follicular ablation and circulatory estrogen levels steadily increased from 36 h until OPU in all groups. At the end of the OPU sessions, the mean number of aspirated follicles/heifer/session in $F_{36}O_{72}$ protocol (Group 3) and $F_{36}O_{96}$ protocol (Group 4) were higher than in the two other groups (p<0.05). The number of cumulus-oocyte complexes (COCs), cleaved and day 8 blastocysts rates in the $F_{36}O_{72}$ protocol (Group 3) were higher than in the other groups (p<0.05). It can be concluded that a single dose i.m. administration of 100 mg FSHp at 36 h and OPU at 72 h after follicular ablation ($F_{36}O_{72}$ protocol; Group 3) was the most effective protocol for superstimulation of follicular growth for repeated OPU and subsequent in vitro embryo production in Thai native heifers.

• 한국수정란이식학회지
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• 제24권3호
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• pp.213-220
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• 2009

The objective of this study was to examine the effect of eCG and various concentrations (20, 40, and 80 ${\mu}g/ml$) of porcine FSH on nuclear maturation and intracellular glutathione (GSH) level of oocytes, and embryonic development after parthenogenetic activation (PA) and somatic cell nuclear transfer (SCNT) in pigs. Immature pig oocytes were matured in TCM-199 supplemented with porcine follicular fluid, cysteine, pyruvate, EGF, insulin, and hormones (10 IU/ml hCG and 10 IU/ml eCG or $20{\sim}80{\mu}g/ml$ FSH) for the first 22 h and then further cultured in hormone-tree medium for an additional 22 h. Nuclear maturation of oocytes ($85{\sim}89%$) was not influencem foreCG and various concentrations FSH. Embryonic development to the cleavage stage ($86{\sim}94%$) and mean number of cells in blastocyst ($33{\sim}37$ cells) after PA were not altered but blastocyst formation e-treignificaddlor(p<0.05) improvem forthe supplementation eith 80 ${\mu}g/ml$ FSHr(64%) compared to 47%, io8%, iand 47% in oocytes that were treated with eCG, 20,i and 40 ${\mu}g/ml$ FSH,i numectivelo. In SCNT, fusion ($78{\sim}83%$) of cell-cytoplast couplets and siosequent embryo cleavage ($82{\sim}88%$) were not influencem fordifferent gonadotropins but blastocyst formation tended to increase forthe supplementation eith 80 ${\mu}g/ml$ FSHr(25% vs. $11{\sim}18%$). Our nuults demonstrated that oocyte maturation and embryonic development after PA and SCNT e-frinfluencem fortype of gcem fortype of gits concentration. In this study, supplementation of maturation medium eith 80 ${\mu}g/ml$ FSHrimproved preimplantation development of PA and SCNT pig embryos, probably by increasing intracellular GSH concentration of matured oocytes.

• Journal of Genetic Medicine
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• 제2권1호
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• pp.11-15
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• 1998

This paper reports 3 cases with 46,XX sex reversed male. Three 46,XX hypogonadal subjects showed complete sex reversal and had normal phallus and azoospermia. We studied them under clinical, cytogenetic and molecular aspects to find out the origin of the sex reversal. Patients had markedly elevated serum follicle-stimulating hormone (FSH) and lutenizing hormone (LH) and decreased or normal range of serum testosterone. The testicular volumes were small (3-8ml). Testicular biopsy showed Leydig cell hyperplasia and atrophy of seminiferous tubules. We obtained the results of normal 46,XX, and the presence of Y chromosome mosaicism was ruled out through XY dual fluorescent in situ hybridization (FISH). By using polymerase chain reaction (PCR), we amplified short arm (SRY, PABY, ZFY and DYS14), centromere (DYZ3), and heterochromatin (DYZ1) region of the Y chromosome. PCR amplification of DNA from these patients showed the presence of the sex-determining region of the Y chromosome (SRY) but didn't show the centromere and heterochromatin region sequence. The SRY gene was detected in all the three patients. Amplification patterns of the other regions were different in these patients; one had four amplified loci (PABY+, SRY+, ZFY+, DYS14+), another had two loci (SRY+, ZFY+) and the other had two loci (PABY+, SRY+). We have found that each patient's translocation elements had different breakpoints at upstream and downstream of the SRY gene region. We conclude that the testicular development in 46,XX male patients were due to insertion or translocation of SRY gene into X chromosome or autosomes.

• 한국동물학회지
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• 제30권1호
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• pp.1-9
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• 1987

생쥐 난자-난구 복합체를 인공배양하면서 adenylate cyclase의 촉진제인 forskilin과 phosphodiesterase의 저해제인 3-isobutyl-1-methylxanthine(IBMX)을 배양액에 첨가하여 이들이 난구세포의 분산과 난자의 성숙(핵붕괴)에 미치는 효과를 관찰한 결과 다음과 같았다. 1. Forskilin은 0.001$\mu$M에서 부터 난구세포의 분산을 유도하기 시작하여 (36%) 0.1-10$\mu$M 구간에서 최대의 분산율을 나타내었고 (80-90%) 100$\mu$M에서는 그 효과가 줄어들었다(60%). 이때 난자의 핵 붕괴는 10$\mu$M까지 정상으로 일어나다 (75-80%) 100$\mu$M에서 부분적으로 억제되기 시작하였다(40%). 2. IMBX는 0.01$\mu$M에서 부터 난구세포의 분산을 유도하기 시작하여 (30%) 1-1,000$\mu$M의 전 구간에서 최대의 분산율(81-89%)을 나타내었다. 난자들은 10$\mu$M의 농도까지 정상적으로 핵 붕괴를 일으키었으나(90% 이상) 100$\mu$M 이상에서 급격히 억제되었다(14%). 3. 난구세포의 분산을 유도하는데 필요한 최소의 자극기간을 조사해 본 결과 HCG는 2분, FSH와 forskilin은 15-30분, IBMX는 2시간이었다. 위 결과로 부터 생쥐 난구세포에서 cAMP의 농도를 높이는데 adenylate cyclase 와 phosphodiesterase의 두 효소가 모두 중요한 기여를 하며 난구세포는 단 기간에 생긴 cAMP의 peak로서 분산이 유도될 수 있으나 난자의 성숙억제 과정에서는 지속적인 cAMP의 존재가 필요하다는 것을 알았다.

• Clinical and Experimental Reproductive Medicine
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• 제25권3호
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• pp.251-260
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• 1998

It is well known that the clinical test for responsibility of accurate fertilization capacity in male partners is very important to diagnose and treat the infertility. However, it has been reported that the traditional semen analysis cannot accurately predict fertilization and pregnancy potential. The present study was performed to evaluate the acrosomal reaction to ionophore challenge (ARIC) test as a prognostic indicator for fertilization of sperm and oocyte in an in vitro fertilization and embryo transfer (IVF-ET) program. From March 1996 to Februry 1997, 30 couples undergoing IVF program were allocated to this study group. All female partners in the study group were 35 years old or less and their serum level of basal follicle stimulating hormone (FSH) and estradiol $(E_2)$ were normal. All the male partners have normal parameters of semen analysis. The ARIC tests were performed on the day of ovum pick up and in vitro insemination in all the male partners. The controlled ovarian hyperstimulation (COH) using luteal long protocol of gonadotropin releasing hormone (GnRH) agonist was used in all couples for IVF-ET. The acrosomal reaction with $10{\mu}l$ of 10% DMSO was induced spontaneously in $10.1{\pm}9.8%$, and acrosomal reaction with calcium ionophore A 23187 was induced in $27.4{\pm}18.1%$, and the ARIC value was $17.4{\pm}16.2%$. There were no significant correlation between the ARIC value and the fertilization rate ($r^2$=0.044, p=0.268). There were also no significant correlation between the ARIC value and the percentage of the grade I, II embryos ($r^2$=0.046, p=0.261). On the basis of above results, it was suggested that ARIC test might not be a useful prognostic indicator for fertilization in IVF-ET in male partners with normal parameters of conventional semen analysis. We guessed that IVF-ET could be performed to the patients primarily without universal appilcation of ARIC test to all male partenrs, and if fertilization failure occurs, the micro assisted fertilization (MAF) such as intracytoplsmic sperm injection (ICSI) might be used as an alternative mode of treatment with acceptable success rate.

• Clinical and Experimental Reproductive Medicine
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• 제46권4호
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• pp.173-177
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• 2019

Objective: We investigated the clinical characteristics of men with testosterone replacement therapy (TRT)-induced hypogonadism and its effect on assisted reproductive technology (ART) in infertile couples. Methods: This study examined the records of 20 consecutive male patients diagnosed with azoospermia or severe oligozoospermia (< 5 × 10⁶/mL) who visited a single infertility center from January 2008 to July 2018. All patients were treated at a primary clinic for erectile dysfunction or androgen deficiency symptoms combined with low serum testosterone. All men received a phosphodiesterase 5 inhibitor and TRT with testosterone undecanoate (Nebido^®) or testosterone enanthate (Jenasteron^®). Patients older than 50 years or with a chronic medical disease such as diabetes were excluded. Results: The mean age of patients was 37 years and the mean duration of infertility was 16.3 ± 11.6 months. At the initial presentation, eight patients had azoospermia, nine had cryptozoospermia, and three had severe oligozoospermia. Serum follicle-stimulating hormone levels were below 1.0 mIU/mL in most patients. Three ongoing ART programs with female factor infertility were cancelled due to male spermatogenic dysfunction; two of these men had normal semen parameters in the previous cycle. After withholding TRT, serum hormone levels and sperm concentrations returned to normal range after a median duration of 8 months. Conclusion: TRT with high-dose testosterone can cause spermatogenic dysfunction due to suppression of the hypothalamic-pituitary-testicular axis, with adverse effects on infertility treatment programs. TRT is therefore contraindicated for infertile couples attempting to conceive, and the patient's desire for fertility must be considered before initiation of TRT in a hypogonadal man.

• Asian-Australasian Journal of Animal Sciences
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• 제30권11호
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• pp.1598-1604
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• 2017

Objective: The present study investigated the litter performance of multiparous sows fed 3% and 6% densified diets at farrowing to weaning during summer with mean maximum room temperature of $30.5^{\circ}C$. Methods: A total of 60 crossbred multiparous sows were allotted to one of three treatments based on body weight according to a completely randomized design. Three different nutrient levels based on NRC were applied as standard diet (ST; metabolizable energy, 3,300 kcal/kg), high nutrient level 1 (HE1; ST+3% higher energy and 16.59% protein) and high nutrient level 2 (HE2; ST+6% higher energy and 17.04% protein). Results: There was no variation in the body weight change. However, backfat thickness change tended to reduce in HE1 in comparison to ST treatment. Dietary treatments had no effects on feed intake, daily energy intake and weaning-to-estrus interval in lactating sows. Litter size, litter weight at weaning and average daily gain of piglets were significantly greater in sows in HE1 compared with ST, however, no difference was observed between HE2 and ST. Increasing the nutrient levels had no effects on the blood urea nitrogen, glucose, triglyceride, and creatinine at post-farrowing and weaning time. The concentration of follicle stimulating hormone, cortisol and insulin were not affected by dietary treatments either in post-farrowing or weaning time. The concentration of blood luteinizing hormone of sows in ST treatment was numerically less than sows in HE2 treatment at weaning. Milk and colostrum compositions such as protein, fat and lactose were not affected by the treatments. Conclusion: An energy level of 3,400 kcal/kg (14.23 MJ/kg) with 166 g/kg crude protein is suggested as the optimal level of dietary nutrients for heat stressed lactating sows with significant beneficial effects on litter size.

• Asian-Australasian Journal of Animal Sciences
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• 제30권10호
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• pp.1507-1514
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• 2017

Objective: The Bashang Long-tail chicken (BS), an indigenous Chinese breed, is considered cold tolerant. We selected BS, the Rhode Island Red (RIR), and their reciprocal crossbreds for the present study. The objectives were: i) to validate whether BS is cold tolerant and whether egg production and cold tolerance of crossbreds could be improved; and ii) to determine the physiological characteristics that underlie cold tolerance and favorable egg production performance in cold environments. Methods: A total of 916 chickens were reared in warm and natural cold environments (daily mean ambient temperature varied from $7.4^{\circ}C$ to $26.5^{\circ}C$ in the warm environment and from $-17.5^{\circ}C$ to $27.0^{\circ}C$ in the cold environment). To investigate their adaptability to the cold environment, the egg production performance and body weight were monitored and compared between breeds and environments. The cloacal temperature and serum biochemical parameters were monitored to reveal the physiological characteristics underlie cold tolerance and favorable egg production performance in the cold environment. Results: The warm environment experiment showed that RIR had the highest egg production performance, and that the reciprocal crossbreds had a higher egg production performance than BS. While in the cold environment RIR had the lowest egg production performance, and the reciprocal crossbreds had a higher egg production performance than BS. In the cold environment BS and reciprocal crossbreds had higher triiodothyronine, tetraiodothyronine levels than RIR. At 35 and 39 wk of age, when the ambient temperature was extremely low (varied from $-20^{\circ}C$ to $0^{\circ}C$), serum glucose, follicle-stimulating hormone, luteinizing hormone, estradiol of BS and crossbreds were higher than RIR. Conclusion: Bashang Long-tail chicken has a favorable cold tolerance ability. Crossbreeding with RIR and BS is an effective way to develop cold tolerant chickens with improved egg production performance.

• Toxicological Research
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• 제33권3호
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• pp.255-263
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• 2017

Chemotherapy is associated with male infertility. Cisplatin (cis-diamminedichloro-platinum (II) (CDDP) as a chemotherapy medication used to treat a number of cancers has been reported to most likely induce testicular toxicity. Administration of antioxidants, such as pentoxifylline (PTX) may reduce some Adverse Drug Reactions (ADRs) of CDDP. Therefore, this study investigated the potentially protective effects of PTX on CDDP-induced testicular toxicity in adult male rats. For this purpose, 42 male rats were randomly divided into 7 groups. The rats were orally pretreated with PTX at the 3 doses of 75, 150, and 300 mg/kg once a day for 14 successive days. On the $14^{th}$ day of the study, they were intraperitoneally (IP) administered with a single dose of CDDP (7 mg/kg). Finally, the sperm/testis parameters, serum levels of reproductive hormones, including testosterone, Luteinizing Hormone (LH), and Follicle Stimulating Hormone (FSH) as the pivotal endocrine factors controlling testicular functions, and histopathological changes of testis tissue were examined. Pretreatment with the two doses of 75 and 150 mg/kg PTX indicated significant increases in the sperm count and motility induced by CDDP administration. The right and significantly left testis weights were decreased following the treatment with 300 mg/kg of PTX plus CDDP. However, 75 mg/kg of PTX plus CDDP showed the best near-to-normal histopathological features. The results demonstrated that PTX alone enhanced some parameters, such as the sperm count, while reducing other parameters, including sperm fast motility and germ layer thickness. Furthermore, despite testosterone or LH levels, the mean serum FSH level was significantly augmented by the doses of 75 and 150 mg/kg. It was concluded that PTX administration cannot reduce CDDP-induced testicular toxicity even at high doses (e.g., 300 mg/kg), while it seemed to partially intensify CDDP toxicity effects at a dose of 75 mg/kg. Thus, further research is required in this regard.

• 한국수정란이식학회지
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• 제22권4호
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• pp.245-249
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• 2007

The aim of present experiment was to examine hatching rate as in vitro indicator of viability of porcine embryos before early stage embryo transfer such as zygotes or 2-cell stage embryos. Cumulus-oocyte complexes (COCs) collected from ovaries were matured in North Carolina State University 23 (NCSU-23) containing 10% porcine follicular fluid (pFF), 10 ng/ml epidermal growth factor (EGF), $10{\mu}g/ml$ follicle stimulating hormone (FSH), $35{\mu}g/ml$ luteinizing hormone (LH), and 1mg/ml cysteine. After 24 hours, the COCs were transferred to the same medium without hormones. After 65h of maturation, oocytes were exposed to phosphate buffered saline (PBS) with 7% ethanol (v/v) for 7 minutes, and then the oocytes were washed and cultured in tissue culture medium (TCM) 199 containing 5 ug/ml cytochalasin B for 5h at $38.5^{\circ}C$ in an atmosphere of 5% $CO_2$ and 95% air with high humidity. After cytochalasin B treatment, the presumptive parthenotes were cultured in porcine zygote medium (PZM)-5 and cleavage of the parthenotes was assessed at 72h of activation, Normally cleaved parthenotes were cultured for an additional 8 days to evaluate their ability to develop to blastocyst and hatching stages. The fetal bovine serum (FBS) were added at Day 4 or 5 with concentrations of 2.5, 5 or 10%. The blastocyst rates were ranged within $39.1{\sim}70%$ in each treatment. However hatching rate was dramatically decreased in non-addition group. In this experiment, embryo viability in female reproductive tract may be estimated before embryo transfer with in vitro culture adding FBS by hatching ability.