• 대한미생물학회지
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• 제17권1호
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• pp.7-14
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• 1982

A model of induction of neoplasia by viruses has develpoed from experimental studies in animals and in cultured cells and oncogenic transformation of cells is the result of integration of viral genetic information into the cellular DNA. The evidence for these associations was derived primarily from seroepidemiologic investigation. However, data indicating that the relation between HSV-2 and cervical cancer fits the model derived from experimental animal studies are not yet sufficient to draw conclusion with regard to the etiologic role the virus in the development of the neoplasms. In other hand, the K562 tumor cell is highly susceptible target for natural killer cell lysis by the lymphocytes of human and murine periperal blood. The characteristics of this effector cell type has been investigated. A study on natural killer cell mediated cytotoxicity(NKMC) against $^{51}Cr$-K562 as target cell was studed in HSV-2 infected ICR mouse. We have studied for susceptibility of HSV-2 against mouse embryo fibroblast(MEF) cells and NKMC from HSV-2 infected mouse. The results obtained that the mouse embryo fibroblast cells culture, the number and size of the cells were markedly increased and formed a monolayers relatively rapid, and become complete monolayer sheet around 72 hrs. Duration of cytopathic effect on MEF cells was rapid by serial passing of HSV-2. The morphology of the HSV-2 infected cells appear to be mainly round, ovium, spindle form and some of them was forming large giant cells. The NKMC was decrease in mouse with HSV-2 and comparison between effector/target cells ratio as 25:1 and 50:1 respectively, the NKMC was found to be more significantly decreased than normal control we have concluded that the natural killer cell activity of the viral infected mouse was shown as a suppressed during the HSV-2 infection, day 7th and 14th.

• 원예과학기술지
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• 제32권4호
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• pp.550-557
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• 2014

Heloniopsis koreana and Heloniopsis tubiflora (Melanthiaceae) are endemic herbaceous species of the Korean Peninsula. The Melanthiaceae family has been described as having seeds with small, underdeveloped embryos at the time of dispersal, and morphological (MD) or morphophysiological dormancy (MPD). However, there are few reports on embryo growth, morphology, and seed germination in Heloniopsis species. The aims of this study were to investigate embryo growth and seed dormancy, and to determine the type of dormancy exhibited by these species. The effects of incubation temperatures, light conditions, and gibberellic acid ($GA_3$) on dormancy break and seed germination were tested. Freshly matured seeds of the two species had small embryos that occupied about 9-11% of the length of the endosperm, and which increased by more than 300% in length before radicle emergence, indicating that the embryos are underdeveloped at the time of dispersal. Embryos in the seeds grew under warm temperature regimes (between $25/15^{\circ}C$ and $30/20^{\circ}C$). $GA_3$ application (tested only in the light) overcame seed dormancy and promoted germination. Approximately 30% of the seeds of H. koreana and approximately 40% of the seeds of H. tubiflora germinated in suitable environmental conditions (light and temperature) within 4 weeks. Therefore, 30-40% of the seeds of the two species exhibited MD, and the rest of the seeds had non-deep simple MPD. Light was found to be one of the critical factors for germination because no seed of either of the two Heloniopsis species germinated under constant dark conditions, and thus, these species have the potential to form a persistent soil seed bank. Understanding these germination requirements will help in development of effective strategies to increase the establishment of seedlings in their native habitat.

• 한국수정란이식학회지
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• 제4권1호
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• pp.46-55
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• 1989

The effects of an antiprogesterone (RU 486) and an antiestrogen (tamoxifen) on ovulatory response and oocyte morphology were examined in pregnant mare serum gonadotropin (PMSG)-primed immatare female rats (28 days of age): a comparison has been made on two different regirnens primed with a "control" dose (4 IU) and a "superovulatory" dose (40 IU) of PMSG. Females for control control regimen received three consecutive injections of lmg RU486, lmg tamoxifen, or vehicle at 24, 36 and 48hr, and were killed at 72l'r after PMSG. Animals for superovalatory regimen received lmg RU486, 2.5mg tamoxifen, or vehicle fouowlag the injection schedule comparable to control regimen, and were killed at 60 and 72hr after PMSG. Compared to vehicle group, there was a significant reduction in ovulatory response as judged by the proportion of rats ovulating andi or by the mean number of oocytes per rat for each treatment of RU486 and tamoxifen in both regimens. The activity of tamoxifen in inhibiting the ovulatory response was greater in control, but less in superovulatory regimen than that of RU486 based on the dose employed for each antisteroid. In both regimens, RU 486 did not have any effect 6n the changes in the proportion of degenerate oocytes as well as ovarian weight, well tamoxifen treatment resulted in a marked promotion of oocyte degeneration as well as a great reduction in ovarian weight, compared to each parameter of vehicle group. RU486 treatment in each regimen did not alter the serum levels of any steroid hormones observed. Howerver, tamoxifen treatment was associated with significant increases in serum 17$\beta$-estradiol and decreases in progesterone in both regimens; also significant increases in androgens in superovulatory regimen. The results illustrate the relative inhibitory activity of RU486 and tamoxifen indicating major steroid hormone involved in PMSG-induced ovulation: 17$\beta$-estradiol for control and progesterone for superovulatory regimen. It also appears that tamoxifen-associated elevation of circulating 17$\beta$-estradiol andi or androgens could be in part, a contributing factor to the promotion of oocyte degeneration presumably by producing a hostile oviductal environment after ovulation.ent after ovulation.

• 한국가금학회지
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• 제12권2호
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• pp.135-143
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• 1985

관절염(또는 건초담), 발육부전증을 보이는 12주령 이하의 브로일러 종계와 브로일러에서 8주의 바이러스를 분리하여 avian reovirus로 동정하였다. 부리된 reovirus는 전자현미경에서 이중막을 갖는 구형으로 virion의 직경은 81nm였으며 한천겔침강반응에서 기지의 reovirus(S-1133 주) 및 항혈청(항 S-1133주 및 R-1주)과 반응하였다. 분리된 revirus는 혈구응집능력이 없었으며 chloroform, IUdR 및 열처리(56$^{\circ}C$, 30분)에 강한 저항성을 나타내었다. Reovirus의 감염성 측정시 계 태아섬유아세포 및 가세포배양과 Vero cell 배양에서 세포분주와 동시에 바이러스를 접종하거나 단층세포가 형성된 후접종하거나 간에 별다른 차이없이 감염 4-5일후에 end point에 도달되었다. 분리된 reovirus를 계태아간세포배양에 5-10대 계대배양한 후 $10^{5.0}$ TCI $D_{50}$의 바이러스를 10일령 발육계란의 장요막상에 접종하였을 때 평균치사시간이 국내분리주는 54-59시간인데 반하여 S-1133주는 73시간으로 약간 길었다.

• 한국수정란이식학회지
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• 제27권1호
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• pp.45-50
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• 2012

The objective of this study is to estimate the effect of adding TES to LEY and FGE freezing extender for the sperm viability, acrosomal morphology and DNA fragmentation from miniature pig sperm, we evaluated sperm characteristics in TFGE, TLE and LEY with various thawing condition ($37^{\circ}C$ for 20 sec, 45 sec and $75^{\circ}C$ for 5 sec, respectively), and in different concentration of glycerol at 1%, 1.5%, 3%. The sperm viability and normal acrosome intact(NAI) in TFGE (Viability : $60.3{\pm}2.4$, NAI : $58.6{\pm}2.2%$), TLE ($61.3{\pm}2.4$, $62.2{\pm}2.2%$) extender significantly(p<0.05) increased than that in LEY ($50.2{\pm}2.4$, $54.5{\pm}2.2%$) extender thawed at $75^{\circ}C$ for 5 sec. According to the results from glycerol concentration, the viability and NAI of miniature pig sperm in 1.5% glycerol TLE ($66.1{\pm}3.2$, $66.2{\pm}1.0%$) was highest among the experimental groups. In accordance with this, DNA fragmentation rates was the lowest in TLE ($43.3{\pm}0.5%$) while that in LEY ($63.5{\pm}2.3%$) is the highest. Therefore, these results suggest that TLE extender method for freezing- thawing of miniature pig sperm increased the viability after thawing.

• 한국수정란이식학회지
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• 제33권2호
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• pp.75-84
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• 2018

The cryopreservation has been extensively applied in many cells including spermatozoa (semen) during past several decades. Especially, the canine spermatozoa cryopreservation has contributed on generation of progeny of rare/genetically valuable dog breeds, genome resource banking and transportation of male germplasm at a distant place. However, severe and irreversible damages to the spermatozoa during cryopreservation procedures such as the thermal shock (cold shock), formation of intracellular ice crystals, osmotic shock, stress of cryoprotectants and generator of reactive oxygen species (ROS) have been addressed. According as a number of researches have been conducted to overcome these problems and to advance cryopreservation technique, several analytical methods have been employed to evaluate the quality of the fresh or cryopreserved canine spermatozoa in regards to the motility, morphology, integrity of membrane and DNA, mitochondrial activity, ROS generation, binding affinity to oocytes, in vitro fertilization potential and fertility potential by artificial insemination. Because the study designs with certain application of analytical methods are selective and varied depending on each experimental objective and laboratory condition, it is necessary to establish the normal reference data of the fresh or cryopreserved canine spermatozoa for each analytical method to monitor experimental procedure, to translate raw data and to discuss results. Here, we reviewed the recent articles to introduce various analytical methods for the canine spermatozoa as well as to establish the normal reference data for each analytical method in the fresh or cryopreserved canine spermatozoa, based on the results of the previous articles. We hope that this review contributes to the advancement of cryobiology in canine spermatozoa.

• 한국수정란이식학회지
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• 제18권3호
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• pp.179-186
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• 2003

Human embryonic stem (hES) cell lines have been derived from human blastocysts and are expected to have far-reaching applications in regenerative medicine. The objective of this study is to improve freezing method with less cryo-injuries and best survival rates in hES cells by comparing various vitrification conditions. For the vitrifications, ES cells are exposed to the 4 different cryoprotectants, ethylene glycol (EG), 1,2-propanediol (PROH), EG with dime-thylsulfoxide (DMSO) and EG with PROH. We compared to types of vehicles, such as open pulled straw (OPS) or electron microscopic cooper grids (EM grids). Thawed hES cells were dipped into sequentially holding media with 0.2 M sucrose for 1 min, 0.1 M sucrose for 5 min and holding media for 5 min twice and plated onto a fresh feeder layer. Survival rates of vitrified hES cells were assessed by counting of undifferentiated colonies. It shows high survival rates of hES cells frozen with EG and DMSO (60.8％), or EG and PROH(65.8％) on EM grids better than those of OPS, compared to those frozen with EG alone (2.4％) or PROH alone (0％) alone. The hES cells vitrified with EM grid showed relatively constant colony forming efficiency and survival rates, compared to those of unverified hES cells. The vitrified hES cells retained the normal morphology, alkaline phosphates activity, and the expression of SSEA-3 and 4. Through RT-PCR analysis showed Oct-4 gene expression was down-regulated and embryonic germ layer markers were up-regulated in the vitrified hES cells during spontaneous differentiation. These results show that vitrification method by using EM grid supplemented with EG and PROH in hES cells may be most efficient at present to minimize cyto-toxicity and cellular damage derived by ice crystal formation and furthermore may be employed for clinical application.

• 한국수정란이식학회지
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• 제18권3호
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• pp.187-193
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• 2003

본 연구는 한우에 GnRH ＋ PGF$_2$$\alpha$＋GnRH (Ov-synch)를 처리하여 배란동기화를 시켰으며, 2차 GnRH 투여후 배란시간, 수태율, 계절별 수태율, 산차별 수태율을 조사하였으며, 시험축은 2산 이상의 개체를 무작위로 선발하여 실험에 공시하였으며, 배란동기화 처리후 1회 인공수정을 실시하고 수태율을 환산하였다. 호르몬 처리방법으로는 GnRH ＋ PGF$_2$$\alpha$ ＋ Gn-RH(Ov-synch)법을 이용하였으며, 배란시간의 조사는 초음파를 이용하여 2차 GnRH 투여 후 24시간부터 31시간까지 2시간 간격으로 난소를 촬영하여 배란 여부를 조사하였다. 1. 배란동기화 처리후 24시간부터 31시간까지 2시간 간격으로 난소의 상태를 확인한 결과 28∼30시간 사이에 80%(20/25)로 가장 많이 배란된 것으로 나타났다. 2. 수태율에 있어서는 계통별 1회 수정 수태율은 고급육 계통이 48.1%(38/79)로 다소 높게 나타났다. 3. 산차에 따른 수태율은 1∼2, 3∼4산차에서 각각 44.3, 55%로 나타났다. 4. 계절별로는 여름보다는 봄과 가을에서 47.3%로 다소 높은 경향이였다.

• 한국수정란이식학회지
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• 제29권4호
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• pp.351-359
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• 2014

Phasianus colchicus is not only a beautiful bird but also a great value in science and under the threat of endanger. Hence, the aim of this study was to isolate the pheasant male germ cells (mGCs) and then induce them into elongated sperm-like cells in vitro. The mGCs were purified and enriched by a two-step plating method based on the different adherence velocities of mGCs and somatic cells. The percentage of the c-kit positive cells and c-kit negative cells examined by flow cytometry analysis (FCA) was 92.87% and 2.57%, respectively. Subsequently, the mGCs were induced for 48h in DMEM/F12 medium supplemented factors such as retinol acid, testosterone and bovine FSH, followed by 5 weeks in culture. We found that some elongated sperm-like cells appeared initially in vitro under inducement of stimulated factors. The elongated sperm-like cells showed in the expression of changed morphology and post-transcriptional marker such as spermatid associated (SPERT), spermatid perinuclear RNA binding protein (STRBP), round spermatid basic protein 1 (RSBN1) and SPER1L. Moreover, in DNA content identified assay, induced cells showed that the 1C DNA population markedly increased in differentiated group but it was not change in undifferentiated group. Successful in vitro differentiation of pheasant testicular germline cells into spermatids appears to offer extremely attractive potential for the conservation of endangered birds and treatment of male infertility.

• 한국수정란이식학회지
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• 제32권3호
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• pp.183-192
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• 2017

The mammary gland tumor (MGT) is the most common neoplasia in intact female dogs. Of these, 50% are malignant and metastasis to the other sites are often occurred. Therefore, it is very important for decision of treatment plan and prognosis to differentiate benign tumor from malignancies. Calcification of MGT is a very important imaging finding. The purpose of this study was to investigate the radiological and computed tomographic images of the MGT and the morphology and distribution of calcifications in the MGT using the Breast Imaging Reporting and Data System classification. A total of 42 dogs with MGT were included in this study. The dogs were divided into two groups into benign and malignant groups based upon histologic or cytologic results. The appearance of calcification in the tumor on radiographs and CT images was analyzed for the HU value of pre- and post-contrast injection, margin, surface, and shape of the tumor and the lymph node abnormalities. On radiographs, the positive predictive value of malignant and benign tumors was 72.72 and 85.71%, respectively. On CT examinations, the positive predictive value of malignant and benign tumors was the same value of 83.33%. The maximum diameter of the tumor and the presence of abnormal lymph nodes on CT images showed a strong correlation with malignancies. Therefore, it is thought that radiographs and CT provide useful information for evaluating MGT in dogs.