• Title/Summary/Keyword: embryology

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Oocyte maturation under a biophoton generator improves preimplantation development of pig embryos derived by parthenogenesis and somatic cell nuclear transfer

  • Lee, DJoohyeong;Shin, Hyeji;Lee, Wonyou;Lee, Seung Tae;Lee, Geun-Shik;Hyun, Sang-Hwan;Lee, Eunsong
    • Korean Journal of Veterinary Research
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    • v.57 no.2
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    • pp.89-95
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    • 2017
  • This study was conducted to determine the effects of biophoton treatment during in vitro maturation (IVM) and/or in vitro culture (IVC) on oocyte maturation and embryonic development in pigs. An apparatus capable of generating homogeneous biophoton energy emissions was placed in an incubator. Initially, immature pig oocytes were matured in the biophoton-equipped incubator in medium 199 supplemented with cysteine, epidermal growth factor, insulin, and gonadotrophic hormones for 22 h, after which they were matured in hormone-free medium for an additional 22 hr. Next, IVM oocytes were induced for parthenogenesis (PA) or provided as cytoplasts for somatic cell nuclear transfer (SCNT). Treatment of oocytes with biophoton energy during IVM did not improve cumulus cell expansion, nuclear maturation, intraoocyte glutathione content, or mitochondrial distribution of oocytes. However, biophoton-treated oocytes showed higher (p < 0.05) blastocyst formation after PA than that in untreated oocytes (50.7% vs. 42.7%). In an additional experiment, SCNT embryos produced from biophoton-treated oocytes showed a greater (p < 0.05) number of cells in blastocysts (52.6 vs. 43.9) than that in untreated oocytes. Taken together, our results demonstrate that biophoton treatment during IVM improves developmental competence of PA- and SCNT-derived embryos.

Protective Effect of Melatonine Against Radiation Induced Nephrotoxicity in Rats

  • Kucuktulu, Eda;Yavuz, Aydin Ali;Cobanoglu, Umit;Yenilmez, Engin;Eminagaoglu, Selcuk;Karahan, Caner;Topbas, Murat;Kucuktulu, Uzer
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.8
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    • pp.4101-4105
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    • 2012
  • Purpose: The degree of radiation injury to kidneys which are located within the limits of radiotherapy area is determined by the volume and the dose of radiation to which the organ is exposed. When the tolerance dose of the kidney is exceeded after a latent period of 6 months acute nephritis develops and after 18 months chronic nephritis ensues. Melatonin is known to prevent the oxidative injury of toxins and radiotherapy with its free radical scavenging capacity. Methods and Materials: In this study 8 weeks old 24 Sprague -Dawley rats were allocated into 4 groups: Control group; Radiotherapy group (20 Gy bilaterally in 5 fractions); Melatonin group (10 mg/kg intraperitoneally), and Melatonin+radiotherapy group (20 Gy Radiotherapy in 5 fractions+ melatonin 10 mg/kg intraperitoneally). After a follow-up period of 6 months BUN was determined in all groups. After rats were euthanized the kidneys were removed for histopathological examination under both light and electron microscopes. Results: After 6 months follow-up, both at light and electron microscopy levels, the rats in radiotherapy+melatonin group were significantly protected against the radiation injury comparing to radiotherapy group (p<0.05). Conclusion: It was shown in this experimental model that melatonin has protective effects against radiation injury to kidneys.

Preventive Effects of a Major Component of Green Tea, Epigallocathechin-3-Gallate, on Hepatitis-B Virus DNA Replication

  • Karamese, Murat;Aydogdu, Sabiha;Karamese, Selina Aksak;Altoparlak, Ulku;Gundogdu, Cemal
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.10
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    • pp.4199-4202
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    • 2015
  • Background: Hepatitis B virus infection is one of the major world health problems. Epigallocatechin-3 gallate is the major component of the polyphenolic fraction of green tea and it has an anti-viral, anti-mutagenic, anti-tumorigenic, anti-angiogenic, anti-proliferative, and/or pro-apoptotic effects on mammalian cells. In this study, our aim was to investigate the inhibition of HBV replication by epigallocatechin-3 gallate in the Hep3B2.1-7 hepatocellular carcinoma cell line. Materials and Methods: HBV-replicating Hep3B2.1-7 cells were used to investigate the preventive effects of epigallocatechin-3 gallate on HBV DNA replication. The expression levels of HBsAg and HBeAg were determined using ELISA. Quantitative real-time-PCR was applied for the determination of the expression level of HBV DNA. Results: Cytotoxicity of epigallocathechin-3-gallate was not observed in the hepatic carcinoma cell line when the dose was lower than $100{\mu}M$. The ELISA method demonstrated that epigallocatechin-3 gallate have strong effects on HBsAg and HBeAg levels. Also it was detected by real-time PCR that epigallocatechin-3 gallate could prevent HBV DNA replication. Conclusions: The obtained data pointed out that although the exact mechanism of HBV DNA replication and related diseases remains unclear, epigallocatechin-3 gallate has a potential as an effective anti-HBV agent with low toxicity.

The Comparative Morphometric Study of the Posterior Cranial Fossa : What Is Effective Approaches to the Treatment of Chiari Malformation Type 1?

  • Hwang, Hyung Sik;Moon, Jae Gon;Kim, Chang Hyun;Oh, Sae-Moon;Song, Joon-Ho;Jeong, Je Hoon
    • Journal of Korean Neurosurgical Society
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    • v.54 no.5
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    • pp.405-410
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    • 2013
  • Objective : The objective of this study was to investigate changes in the posterior cranial fossa in patients with symptomatic Chiari malformation type I (CMI) compared to a control group. Methods : We retrospectively reviewed clinical and radiological data from 12 symptomatic patients with CMI and 24 healthy control subjects. The structures of the brain and skull base were investigated using magnetic resonance imaging. Results : The length of the clivus had significantly decreased in the CMI group than in the control group (p=0.000). The angle between the clivus and the McRae line (p<0.024), as the angle between the supraocciput and the McRae line (p<0.021), and the angle between the tentorium and a line connecting the internal occipital protuberance to the opisthion (p<0.009) were significantly larger in the CMI group than in the control group. The mean vertical length of the cerebellar hemisphere (p<0.003) and the mean length of the coronal and sagittal superoinferior aspects of the cerebellum (p<0.05) were longer in the CMI group than in the control group, while the mean length of the axial anteroposterior aspect of the cerebellum (p<0.001) was significantly shorter in the CMI group relative to control subjects. Conclusion : We elucidate the transformation of the posterior cranial fossa into the narrow funnel shape. The sufficient cephalocaudal extension of the craniectomy of the posterior cranial fossa has more decompression effect than other type extension of the craniectomy in CMI patients.

Neuroprotective Effects of Quetiapine on Neuronal Apoptosis Following Experimental Transient Focal Cerebral Ischemia in Rats

  • Yilmaz, Muhammet Bahadir;Tonge, Mehmet;Emmez, Hakan;Kaymaz, Figen;Kaymaz, Memduh
    • Journal of Korean Neurosurgical Society
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    • v.54 no.1
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    • pp.1-7
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    • 2013
  • Objective : This study was undertaken in the belief that the atypical antipsychotic drug quetiapine could prevent apoptosis in the penumbra region following ischemia, taking into account findings that show 5-hydroxytryptamine-2 receptor blockers can prevent apoptosis. Methods : We created 5 groups, each containing 6 animals. Nothing was done on the K-I group used for comparisons with the other groups to make sure adequate ischemia had been achieved. The K-II group was sacrificed on the 1st day after transient focal cerebral ischemia and the K-III group on the 3rd day. The D-I group was administered quetiapine following ischemia and sacrificed on the 1st day while the D-II group was administered quetiapine every day following the ischemia and sacrificed on the 3rd day. The samples were stained with the immunochemical TUNEL method and the number of apoptotic cells were counted. Results : There was a significant difference between the first and third day control groups (K-II/K-III : p=0.004) and this indicates that apoptotic cell death increases with time. This increase was not encountered in the drug groups (D-I/D-II : p=1.00). Statistical analysis of immunohistochemical data revealed that quetiapine decreased the apoptotic cell death that normally increased with time. Conclusion : Quetiapine is already in clinical use and is a safe drug, in contrast to many substances that are used to prevent ischemia and are not normally used clinically. Our results and the literature data indicate that quetiapine could help both as a neuronal protector and to resolve neuropsychiatric problems caused by the ischemia in cerebral ischemia cases.

Analysis of ENPP2 in the Uterine Endometrium of Pigs Carrying Somatic Cell Nuclear Transfer Cloned Embryos

  • Seo, Heewon;Choi, Yohan;Yu, Inkyu;Shim, Jangsoo;Lee, Chang-Kyu;Hyun, Sang-Hwan;Lee, Eunsong;Ka, Hakhyun
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.9
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    • pp.1255-1261
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    • 2013
  • Somatic cell nuclear transfer (SCNT) is a useful tool for animal cloning, but the efficiency of producing viable offspring by SCNT is very low. To improve this efficiency in the production of cloned pigs, it is critical to understand the interactions between uterine function and cloned embryos during implantation. Lysophosphatidic acid (LPA) is a lipid mediator that plays an important role in the establishment of pregnancy in pigs; however, LPA production in the uterine endometrium of pigs carrying SCNT-cloned conceptuses has not been determined. Therefore, we investigated expression of ENPP2, an LPA-generating enzyme, in the uterine endometrium of gilts with conceptuses derived from SCNT during the implantation period. Uterine endometrial tissue and uterine flushing were obtained from gilts carrying SCNT-derived conceptuses and from gilts carrying conceptuses resulting from natural mating on d 12 of pregnancy. Our results demonstrated no difference in the level of ENPP2 mRNA expression in the uterine endometrium between gilts carrying SCNT-derived conceptuses and gilts carrying naturally-conceived conceptuses, but secretion of ENPP2 protein into the uterine lumen did decrease significantly in pigs with SCNT-derived conceptuses. These results indicate that expression and secretion of ENPP2, which are critical for appropriate LPA production and successful pregnancy, are dysregulated in the uterine endometrium of pigs carrying SCNT-derived conceptuses.

Transgenic Alteration of Sow Milk

  • Wheeler, Matthew B.
    • Korean Journal of Animal Reproduction
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    • v.24 no.4
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    • pp.321-333
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    • 2000
  • High production of milk and its components are necessary to allow maximal growth of developing piglets. In this study, transgenic pigs were produced containing the $\alpha$ -lactalbumin gene, whose product is a potential limiting component in the production of milk. Two lines of transgenic pigs were produced to analyze the effects that overproduction of the milk protein $\alpha$ -lactalbumin may have on milk production and piglet growth. Transgenic pigs were produced through microinjection of the bovine $\alpha$ -lactalbumin gene. The gene construct contained 2.0 kb of 5'flanking region, the 2.0 kb coding region and 329 bp of 3'flanking region. Sows hemizygous for the trans gene produced as much as 0.9 g of bovine $\alpha$-lactalbumin per liter of pig milk. The production of the bovine protein caused approximately a 50% increase in the total $\alpha$ -lactalbumin concentration in pig milk throughout lactation. The concentration of bovine $\alpha$ -lactalbumin was highest on day 0 and 5 of lactation and decreased as lactation progressed. The ratio of bovine to porcine $\alpha$ -lactalbumin changed during the sow's lactation. This ratio was 4.3 to 1 on day 0 of lactation, but by day 20 of lactation the ratio was 0.43 to 1. This suggested that the bovine transgene and the endogenous porcine gene were under slightly different control mechanisms. The higher level of total $\alpha$-lactalbumin present on day 0 of lactation was correlated with higher lactose percentage on day 0 in transgenic sows (3.8%) as compared to controls (2.6%) (P<0.01). Although there was also a trend for higher lactose percentage in transgenic sows on day 5 and 10 of lactation, no significant differences were observed. These data suggest that $\alpha$ -lactalbumin is limiting early in lactation of swine. Furthermore, higher concentrations of $\alpha$ -lactalbumin early in lactation may boost milk output.

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Comparative Analysis of Stem Cell Research Policy Changes in UK, US, and South Korea: Application of Advocacy Coalition Framework Model (영국, 미국, 한국의 줄기세포연구에 관한 정책변동 비교 분석: Advocacy Coalition Framework 모형의 적용)

  • Bae, Green;Kang, Minah
    • Health Policy and Management
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    • v.23 no.4
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    • pp.314-325
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    • 2013
  • Background: Stem cell research competition is accelerating globally since President Obama signed an executive order, repealing Bush-era policy that limited use of federal tax dollars for embryonic stem cell research. Methods: In this paper, we conducted a comparative analysis of stem cell research policy changes in three countries, including the Human Fertilisation Embryology Act (HFEA) of UK, executive order 13,505 (removing barriers to responsible scientific research involving human stem cells) of USA, and Bioethics and Safety Act of South Korea. Debates on stem cell research are based on conflicts of fundamental beliefs that exist in the supporting and opposing coalitions. We compared regional characteristics of the advocacy coalitions in three countries and presented various factors that might be related to the policy changes. Results: The UK government, parliament, and the HFEA have sought expert consultations and public opinions to establish guidelines. UK has made social consensus through continued discussion for a long time. US President's veto power was one strongest factors influencing policy. South Korean policy was influenced by public opinion and policy brokers. Also, South Korea has not made social consensus. UK had a strong leadership and strong adjustment of coalitions but US and South Korea had not. Dr. Hwang's scandal has had one of the greatest impacts on policy decision in South Korea. Conclusion: The power of public opinion was critical in all three countries. In particular, the influence of public opinion was noticeable in South Korea. Also it turned out that in US and South Korea, the presence of a policy broker who could pursue his or her goals was the most powerful factor among the advocacy coalition factors.

Effects of High Dose Lysophosphatidic Acid Supplement during IVC on Preimplantation Development of Porcine Embryos

  • Jin, Minghui;Yu, Il-Jeoung;Jeon, Yubyeol
    • Journal of Embryo Transfer
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    • v.32 no.4
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    • pp.275-285
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    • 2017
  • Lysophosphatidic acid (LPA) is an important signaling molecule. Here, the effect and mechanism of LPA on the preimplantation development of porcine embryos during in vitro culture (IVC) was examined. Porcine embryos were cultured in porcine zygote medium (PZM-3) supplemented with $30{\mu}M$ LPA during different days. There was a significantly higher cleavage rate in Day 1-7 and significantly higher total cell number of blastocysts in Day 1-3 and Day 4-7. It was also found that messenger RNA (mRNA) expression level of PCNA, BCL2 and BAX in blastocysts obtained from D1-7 group were significantly higher and BCL2/BAX mRNA ratio in D1-3 group was significantly lower than control group but Day 4-7 and Day 1-7 groups were comparable with control group. Treatment with $20{\mu}M$ PLC inhibitor significantly decreased the embryo cleavage rate and blastocyst formation rate. Moreover, LPA as an activator of PLCs, enhanced the $30{\mu}M$ LPA + $20{\mu}M$ U73122 group embryo cleavage rate which similar with control group. In conclusion, the results suggest that treatment with LPA during IVC improves the porcine early embryo cleavage by activation of PLC signaling pathway and regulate the mRNA expression that contribute to total cell number of blastocysts during blastocyst formation.

Intracranial Pressure and Experimental Model of Diffuse Brain Injury in Rats

  • Blaha, Martin;Schwab, Juraj;Vajnerova, Olga;Bednar, Michal;Vajner, Ludek;Michal, Tichy
    • Journal of Korean Neurosurgical Society
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    • v.47 no.1
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    • pp.7-10
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    • 2010
  • Objective: In this study, we present a simple closed head injury model as a two-stage experiment. The height of the weight drop enables gradation of head trauma severity. Methods: The head injury device consists of three parts and there are three adjustable parameters-weight (100-600 g). height of fall (5-100 cm) and elasticity of the springs. Thirty male Wistar rats underwent monitoring of intracranial pressure with and without induction of the head injury. Results: The weight drop from 45 to 100 cm led to immediate seizure activity and early death of the experimental animals. Severe head injury was induced from 40 cm weight drop. There was 50% mortality and all surviving rats had behavioral deterioration. Intracranial pressure was 9.3${\pm}$3.76 mmHg. Moderate head injury was induced from 35 cm, mortality decreased to 20-40%, only half of the animals showed behavioral pathology and intracranial pressure was 7.6${\pm}$3.54 mmHg. Weight drop from 30 cm caused mild head injury without mortality and neurological deterioration. Intracranial pressure was slightly higher compared to sham group- 5.5${\pm}$0.74 mmHg and 2.9${\pm}$0.81 mmHg respectively. Conclusion: This model is an eligible tool to create graded brain injury with stepwise intracranial pressure elevation.