LBD-001, a recombinant human interferon $\gamma$ produced by genetically engineered yeast as a host system, was administered intraperitoneally to Sprague-Dawley male rats from premating to mating period at least for 60 days and to female rats from at least for 2 weeks before mating to early gestation period (from day 0 to 7 of gestation) at dose levels of $0.35\times10^6, 0.39\times10^6, and 1.38\times10^6$ I.U./kg/day. In the positive control group, ethynylestradiol ($EE_2$; 40 $\mu\textrm{g}$/kg/day) was subcutaneously administered only to female rats during the early gestation period. Effects of the test agents on reproductive performances of the male or female rats and embryonic development were as followings; (1) No significant changes by the treatment of LBD-001 were observed in general behaviors, body weight, food and water consumption, and necropsy of parent animals. However, significant decreases of body weight, food consumption, and water consumption were observed in ($EE_2$ -treated female rats. (2) Mating performances and fertility of parent animals were not significantly affected by the treatment of LBD-001. In ($EE_2$ -treated females, however, the fertility was completely inhibited. (3) No changes in resorption rate and external abnormality of F1 fetuses were observed by the treatment of LBD-001. The results show that LBD-001 at the dose of $1.38\times10^6$ I.U./kg/day or less does not affect general toxicity and reproductive function of parent animals and embryonic development of F1 fetuses.
Kim, Seul Ki;Han, E-Jung;Kim, Sun Mie;Lee, Jung Ryeol;Jee, Byung Chul;Suh, Chang Suk;Kim, Seok Hyun
Clinical and Experimental Reproductive Medicine
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제43권4호
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pp.233-239
/
2016
Objective: Uterine contraction induced by the embryo transfer (ET) process has an adverse effect on embryo implantation. The aim of this study was to determine the effect of oxytocin antagonist supplementation on the day of ET on in vitro fertilization outcomes via a meta-analysis. Methods: We performed a meta-analysis of randomized controlled trials (RCTs). Four online databases (Embase, Medline, PubMed, and Cochrane Library) were searched through May 2015 for RCTs that investigated oxytocin antagonist supplementation on the day of ET. Studies were selected according to predefined inclusion criteria and meta-analyzed using RevMan 5.3. Only RCTs were included in this study. The main outcome measures were the clinical pregnancy rate, the implantation rate, and the miscarriage rate. Results: A total of 123 studies were reviewed and assessed for eligibility. Three RCTs, which included 1,020 patients, met the selection criteria. The implantation rate was significantly better in patients who underwent oxytocin antagonist infusion (19.8%) than in the control group (11.3%) (n = 681; odds ratio [OR], 1.92; 95% confidence interval [CI], 1.25-2.96). No significant difference was found between the two groups in the clinical pregnancy rate (n = 1,020; OR, 1.57; 95% CI, 0.92-2.67) or the miscarriage rate (n = 456; OR, 0.76; 95% CI, 0.44-1.33). Conclusion: The results of this meta-analysis of the currently available literature suggest that the administration of an oxytocin antagonist on the day of ET improves the implantation rate but not the clinical pregnancy rate or miscarriage rate. Additional, large-scale, prospective, randomized studies are necessary to confirm these findings.
In order to study the implantation mechanism various methods for culture of endometrial cells in vitro have been attempted. However, a disadvantage is that primary cultures of stromal and epithelial cells do not have the ability to differentiate, and therefore cannot be reproduced in the same manner as in vivo endometrium. The object of this study is to establish a three dimensional culture of endometrial cells which are both morphologically and functionally identical to in vivo endometrium. Endometrial tissues obtained after hysterectomies were cut into thin slices and treated with collagenase and trypsin-EDTA. The stromal cells and the epithelial cells were separated by centrifugation and cultured for 24 hours in DMEM media containing 10% FCS, 100 nM progesterone, and 1 nM estradiol. The cultured stromal cells were mixed with collagen gel and solidified, after which it was covered with matrigel. Epithelial cells were inoculated on the top and then cultured for 3 days. The three dimensionally cultured endometrial cells were stained for integrin ${\alpha}1,\;{\alpha}4,\;{\beta}3$, and cyclooxygenase-l, -2 by immunohistochemistry, which all showed strong expression. The cultured epithelial cells showed the formation of microvilli, tight junctions and pinopodes by electron microscopy. Studies are currently under way utilizing this three dimensional culture model to ascertain the interaction between the embryo and human endometrial cells at the time of implantation, and it is thought that further studies into a new culture environment which would allow longer periods of culture will be necessary.
Since the blastocyst is broken and spreads out on a flat plastic culture dish (two dimensional culture) during in vitro development, it has been difficult to study the implantation process. It also has been difficult to analyse the interactions between endometrial epithelial and stromal cells because of the lack of a long-term in vitro model which can stimulate in vivo characteristics, as these cells eventually fail to proliferate or cease to express differentiated functions. Recently nontransformed cell lines, CUE-P and CUS-V2, derived from rat endometrial epithelium and stroma were reported. In this study, morphology of CUE-P and CUS-V2 was examined and oxytocin gene expression by CUE-P cells was demonstrated by RT-PCR. The CUE-P cells have a cuboidal morphology and CUS-V2 cells resemble fibroblast and exhibit a spindle-like morphology. In RT-PCR, same size of PCR products of oxytocin gene at hypothalamus, uterus and CUE-P cells were demonstrated. These results showed three dimensional culture system could be made by using the new cell lines.
Yang, Ning Jie;Seol, Dong-Won;Jo, Junghyun;Jang, Hyun Mee;Yoon, Sook-Young;Lee, Dong Ryul
Clinical and Experimental Reproductive Medicine
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제41권1호
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pp.1-8
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2014
Objective: Estrogen related receptor ${\beta}$ (Esrrb) is a member of the orphan nuclear receptors and may regulate the expression of pluripotencyrelated genes, such as Oct4 and Nanog. Therefore, in the present study, we have developed a method for delivering exogenous ESRRB recombinant protein into embryos by using cell-penetrating peptide (CPP) conjugation and have analyzed their effect on embryonic development. Methods: Mouse oocytes and embryos were obtained from superovulated mice. The expression of Oct4 mRNA and the cell number of inner cell mass (ICM) in the in vitro-derived and in vivo-derived blastocysts were first analyzed by real time-reverse transcription-polymerase chain reaction and differential staining. Then 8-cell embryos were cultured in KSOM media with or without $2{\mu}g/mL$ CPP-ESRRB protein for 24 to 48 hours, followed by checking their integration into embryos during in vitro culture by Western blot and immunocytochemistry. Results: Expression of Oct4 and the cell number of ICM were lower in the in vitro-derived blastocysts than in the in vivo-derived ones (p<0.05). In the blastocysts derived from the CPP-ESRRB-treated group, expression of Oct4 was greater than in the non-treated groups (p<0.05). Although no difference in embryonic development was observed between the treated and non-treated groups, the cell number of ICM was greater in the CPP-ESRRB-treated group. Conclusion: Treatment of CPP-ESRRB during cultivation could increase embryos' expression of Oct4 and the formation rate of the ICM in the blastocyst. Additionally, an exogenous delivery system of CPP-conjugated protein would be a useful tool for improving embryo culture systems.
To investigate the XIST gene expression and its effect in a Klinefelter's patient, we used Klinefelter's syndrome (XXY) patient with azoospermia and also used a normal male (XY) and a normal female (XX) as the control, We were performed cytogenetic analysis, Y chromosomal microdeletion assay (Yq), semi-quantitative RT-PCR, and the Northern blot for Klinefelter's syndrome (KS) patient, a female and a male control, We extracted total RNA from the KS patient, and from the normal cells of the female and male control subjects using the RNA prep kit (Qiagen), cDNA microarray contained 218 human X chromosome-specific genes was fabricated. Each total RNA was reverse transcribed to the first strand cDNA and was labeled with Cy-3 and Cy-5 fluorescein, The microarray was scanned by ScanArray 4000XL system. XIST transcripts were detected from the Klinefelters patient and the female by RT-PCR and Northern blot analysis, but not from the normal male, In the cDNA microarray experiment, we found 24 genes and 14 genes are highly expressed in KS more than the normal male and females, respectively. We concluded that highly expressed genes in KS may be a resulted of the abnormal X inactivation mechanism.
Endometrial cancer (EC) in young women tends to be early-stage and low-grade; therefore, such cases have good prognoses. Fertility-sparing treatment with progestin is a potential alternative to definitive treatment (i.e., total hysterectomy, bilateral salpingo-oophorectomy, pelvic washing, and/or lymphadenectomy) for selected patients. However, no evidence-based consensus or guidelines yet exist, and this topic is subject to much debate. Generally, the ideal candidates for fertility-sparing treatment have been suggested to be young women with grade 1 endometrioid adenocarcinoma confined to the endometrium. Magnetic resonance imaging should be performed to rule out myometrial invasion and extrauterine disease before initiating fertility-sparing treatment. Although various fertility-sparing treatment methods exist, including the levonorgestrel-intrauterine system, metformin, gonadotropin-releasing hormone agonists, photodynamic therapy, and hysteroscopic resection, the most common method is high-dose oral progestin (medroxyprogesterone acetate at 500-600 mg daily or megestrol acetate at 160 mg daily). During treatment, re-evaluation of the endometrium with dilation and curettage at 3 months is recommended. Although no consensus exists regarding the ideal duration of maintenance treatment after achieving regression, it is reasonable to consider maintaining the progestin therapy until pregnancy with individualization. According to the literature, the ovarian stimulation drugs used for fertility treatments appear safe. Hysterectomy should be performed after childbearing, and hysterectomy without oophorectomy can also be considered for young women. The available evidence suggests that fertility-sparing treatment is effective and does not appear to worsen the prognosis. If an eligible patient strongly desires fertility despite the risk of recurrence, the clinician should consider fertility-sparing treatment with close follow-up.
BACKGROUND: Selection an suitable light source is essential in the year-round production of horticultural crops in closed production systems such as plant factory with controlled environments. This study was investigated to confirm the effects of artificial light sources on growth of cherry tomato'CF Jelly'(Lycopersicon esculentum var.) under high-pressure sodium lamps (HPS), metal-halide lamps (MH), and LEDs. METHODS AND RESULTS: Light intensity of the light sources was controlled at 220±30 µmol/m2/s with 12 hrs of photoperiod for a day. Flower development was significantly faster in HPS and MH treatments compared to the LEDs. There was no significant difference between the leaf number and leaf shape under the HPS and MH treatments. Reproductive growth of cherry tomato was significantly promoted by the LEDs treatment of blue plus red lights. Fruit yield per plant also increased under the LEDs compared to the others. CONCLUSION(S): Growth, flowering, and fruit setting of the cherry tomato were accomplished by the artificial lights under plant factory conditions. The HPS treatment showed negative effect on fruit quality in terms of blossom-end rot incidence compared to the LEDs or MH treatment. Effect of the LEDs on promotion of fruit weight and yield was also proved. Additional research should be carried out for improving sugar metabolism or decreasing disease in the fruits under plant factory system using only artificial lights.
Adikari Arachchige Dilki Indrachapa Adikari;Malavi Arachchi Gamage Nayodya Dananjanee Malavi Arachchi Gamage;Wijesooriya Mudhiyanselage Nadeema Dissanayake;Jung Min Heo;Young-Joo Yi
농업과학연구
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제51권2호
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pp.109-121
/
2024
Mancozeb is a manganese and zinc-containing fungicide that belongs to the ethylene bisdithiocarbamate group and produces ethylene thiourea (ETU) after biotransformation or environmental degradation, which has toxicological hazard owing to its known antithyroid properties. Although mancozeb leads to negative changes in fertility capacity, the effects of ETU are less known. Therefore, this study examined the alteration of fertilization competence in boar spermatozoa exposed to ETU. The sperm motility, motion kinematics, viability, acrosome integrity, chromatin stability, and intracellular reactive oxygen species (ROS) production of sperm subjected to various ETU concentrations (10, 50, 100, and 200 µM) were evaluated after two different incubation times (30 min and 2 hrs). In addition, the relative mRNA expression of the sperm functional proteins was analyzed after exposure to ETU. A dose-dependent motility reduction was observed in sperm exposed to ETU during both incubation periods compared to the controls. The motion kinematics were reduced significantly in sperm incubated with ETU. Higher percentages of viable sperm were observed in the controls, while such viability was decreased significantly in sperm with 10 - 200 µM ETU. The acrosome integrity was particularly damaged on sperm incubated with 10 - 200 µM ETU for 30 min. Higher intracellular ROS levels were produced in sperm exposed to 200 µM ETU. In addition, lower relative levels of AKAP3, AKAP4, ODF2, and ZPBP2 expression were observed in sperm exposed to ETU compared to the controls. Mancozeb and ETU could adversely affect the reproductive functions of mammals. Hence, the effects of ETU on the reproductive system should be examined further.
Geray Reservoir is home to some fish species including Varicorhinus beso, Cyprinus carpio, Carasius carasius, Oreochromis niloticus, and Tilapia randelli. The V. beso is among the top fish-farming species for the reservoir. However, this data on total fish catch showed that the fish population is declining as a result of overexploitation of the stock. The purpose of this study was to set some management measures for the Geray reservoir using some biometric data on the most commonly caught fish-V. beso. A total of 170 fish samples (102 females and 68 males) ranging from 21 to 40.2 cm in total length (TL) and 94.6 to 618.4 g in total weight (TW) were gathered between October 2022 and May 2022 using a cast net with a mesh size of 6 cm. Nonetheless, it is thought that the sampling period is sufficient because the fish were spawning more during the month when the data was collected because of the high concentration of nutrients during that time. The overall male-to-female sex ratio in the population was 1:1.5, which differed considerably from the real ratio of 1:1 (x2 = 6.8; p < 0.01). V. beso in the Geray reservoir exhibited virtually an isometric growth (b = 3) considering the length-weight relationships of males (TW = 0.0122TL2.993) and females (TW = 0.0114TL3.0139). The V. beso had an absolute fecundity (AF) of 2,190 to 11,265 eggs. AF was significantly correlated with TL, TW, and gonad weight (p < 0.01). V. beso's reproductive season peaked between November and May, with the gonadosomatic index ranging from 0.02 to 20.12 for females and 0.03 to 5.67 for males. The average V. beso condition factors for both sexes were higher than 1, indicating that the fish in the reservoir had properly developed, which may be the result of favorable habitat conditions.
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