To produce reconstituted rabbit embryos with fetal fibroblasts, the present study was evaluated the efficiencies of the different fusion and activation conditions as assessments of subsequent development and chromosome in the embryos. New Zealand White rabbits were used throughout the study. Fetal fibroblasts collected from 22-d of fetuses were cultured in DMEM + 10% FBS in 5% $CO_2$ in air. The culture was maintained for 10 passages. In every passage half of cell suspension were kept In frozen. From rabbits treated with FSH in 30% PVP solution and hCG, oocytes were surgically collected from oviducts at 14 h post-hCG injection and stripped off their cumulus cells by re-pipetting in a 300 IU hyaluronidase solution. Oocytes with an extruded first polar body and dense cytoplasm were enucleated by micromanipulation in Ham's F-10 medium+7.5 g/$m\ell$ cytochalasin B. Euncleation was confirmed under a fluorescence microscope after staining with 5 g/$m\ell$ bisbenzimide for 2 min. Each enucleated oocyte was injected with a fetal fibroblast into a perivitelline space. Reconstructed eggs were compared fusion rates either at 2.0 ㎸/cm or 1.6 ㎸/cm(60 sec, double pulses). After fusion, all eggs were activated with the combination of 5 M ionomycin (5 min) and 10 g/$m\ell$ cycloheximide (CHX, 3h), and cultured in CRlaa medium and transferred into TCM199+10% FBS on day 3. Although there was not significantly differ in fusion rate between treatments (60%, 2.0 ㎸/cm vs. 79.4%, 1.6 ㎸/cm), none of them in the eggs fused with 2.0 ㎸/cm developed to blastocyst. In comparison of development and chromosome status between different activation treatments (Group 1; 5 M ionomycin/10 g/$m\ell$ CHX, Group 2; 5 M ionomycin/5 g/$m\ell$ CHX + 2 mM DMAP after fusion with 1.6 ㎸/cm), there were not differ in cleavage and development rates (67.3% and 28.9% in Group 1; 67% and 33% in Group 2). All out of 8 embryos evaluated in Group 1 appeared a normal diploid chromosome sets and mean number of cells (Mean SEM) on day 4.5 of culture was 141.5 23.15 (n=8). It can be concluded that the use of cycloheximide has not happened in chromosome abnormalities, and fetal fibroblasts can be used for cloning in rabbit.
Our previous study showed that transgenic (TG) pigs harboring human EPO (hEPO) gene have been shown to have reproductive disorders, including low pregnancy rates, irregular estrus cycle and low little size. To investigate these reasons, we assessed estrus behavior (standing response) and plasma $17{\beta}$-estradiol ($E_2$) level, which partly reflect reproductive function, during the estrus cycles after synchronization and superovulation by hormone treatments. Then, we analysed blood composition and expression of hEPO gene in TG pigs. Pigs were injected with PG600. After 10 days, pigs were fed with Regumate porcine for 6 days. Blood samples were collected from jugular vein. Analysis of blood composition and $E_2$ level were measured by Hemavet 950 and $E_2$ ELISA kit, respectively. And, the expression of hEPO gene in reproductive organs was quantitated by real-time RT-PCR. The percentage of estrus behavior in TG was significantly decreased. Hematocrit (HCT), hemoglobin (Hb) concentration and red blood cell (RBC) number were significantly higher in TG than wild type (WT). On the other hand, high expression of hEPO gene in TG was observed in the mammary gland as well as in the uterus. Moreover, plasma $E_2$ level was significantly higher in TG than WT. These results suggest that nonspecific expression of hEPO gene in the other organs of TG may affect blood composition and plasma $E_2$ level, thereby causing reproductive disorders.
The objective of this study was to investigate the effects of abnormal ovarian cycles after superovulation treatment of Holstein Donor Cows. CIDRs were inserted into the vaginas of twenty two head of Holstein cows, regardless of estrous cycle. Superovulation was induced using folliclar stimulating hormone (FSH). For artificial insemination, donor cows were injected with $PGF_2{\alpha}$ and estrus was checked about 48 hours after the injection. Then they were treated with 4 straws of semen 3 times, with 12-hour intervals. Embryos were collected by a non-surgical method 7 days after the first artificial insemination. The cows were considered to have resumed ovarian cyclicity on the day of ovulation if followed by regular ovarian cycles. Seventy two point seven percentage of the cows(16/22) had normal resumption of ovarian cyclicity(resumption within 40 days after superovulation), and 27.3%(6/22) had delayed resumption(resumption did not occur until>40 days after superovulation). Delayed resumption Type II(first ovulation did not occur until ${\geq}40$ days after superovulation, i.e. delayed first ovulation 13.6%) were the most common types of delayed resumptions. The mean numbers of total ova from < 10 and $10{\leq}$ of corpora lutea(CL) was $7.8{\pm}1.8$ and $12.7{\pm}2.7$, respectively. The number of transferable embryos differed between < 10 and $10{\leq}$ CL was $5.4{\pm}1.3$ and $8.1{\pm}3.4$, respectively. Four point five percentage of the cows(1/22) did not resumption their ovarian cyclicity until 60 days after superovulation treatment. Diverse researches on the superovulation treatment method that is suitable for high-producing Holstein donor cows would contribute to preventing ovarian cyclicity disorder, as well as to the early multiplication of cows with superior genes by increasing the utilization value of donor cows.
This study was carried out not only to investigate the toxicities of 12 registered insecticides on different developmental stages, but also to determine the sublethal effects on longevity and reproduction of newly emerged adult female and development of the next generation in the rice skipper, Parnara guttata. Fenitrothion, fenthion, cartap hydrochloride, ethofenprox highly suppressed egg-hatch. All insecticides treated showed high larvicidal activity on the 1st to 2nd instar larva. The insecticides showed higher larvicidal activities on the 5th instar larva were fenitrothion, fenthion, ethofenprox, fipronil, methoxyfenozide, tebufenozide and Bt. var. kurstaki. The sublethal doses of fenthion, tebufenozide, cartap hydrochloride, methoxyfenozide, ethofenprox, imidacloprid and fipronil shortened the longevities of newly emerged adult female from the treated larva ($3{\sim}4$ instar). BPMC, imidacloprid, ethofenprox, fipronil and methoxyfenozide delayed the preoviposition periods of adult females and decreased the number of eggs laid when they were treated at the larval stages of the previous generation. Ethofenprox caused severe sublethal effects on P. guttata offspring, completely blocking the production. All insecticides except fenitrothion affected the egg viability, and all eggs from the adult females emerged from the survivors treated larvae with imidacloprid or fipronil fail to hatch. IGRs, methoxyfenozide and tebufenozide showed an adverse effect on the development of next generation larva.
This study identifies the symbolism of cross symptoms of different classes during the modernization process based on the imagery of and , both of which are Korean films made in the same era of the 1960s. The study referred to a number of Korean family movie films in the 1960s, in particular and , which are notably different in ways of expression and in social class of the subject matter, and identifies how the films express the mixed aspects of the cross modernization period. The film uses a grotesque expressionist method to voyeuristically look into a newly emerging middle class family, and the deconstructive aspect of a family unit whose excessive desire leads to its own ruin; thus, the film presents a new perspective into the various social issues that emerge in the near future. In contrast, engages realistic depictions to describe in detail the deep conflict of pain and hope within the hardships of life from the urban poor through a deep family conflict. Ultimately, the two films expose the various social issues through the familiar and the unfamiliar that appear from the mixed aspects of the cross modernization period, in other words, the conflicting images of the traditional and the modern.
Objective: Porcine respiratory disease is one of the most important health problems causing significant economic losses. To understand the genetic basis for susceptibility to swine enzootic pneumonia (EP) in pigs, we detected 102,809 single nucleotide polymorphisms in a total of 249 individuals based on genome-wide sequencing data. Methods: Genome comparison of susceptibility to swine EP in three pig breeds (Jinhua, Erhualian, and Meishan) with two western lines that are considered more resistant (Duroc and Landrace) using cross-population extended haplotype homozygosity and F-statistic (FST) statistical approaches identified 691 positively selected genes. Based on quantitative trait loci, gene ontology terms and literature search, we selected 14 candidate genes that have convincible biological functions associated with swine EP or human asthma. Results: Most of these genes were tested by several methods including transcription analysis and candidate genes association study. Among these genes: cytochrome P450 1A1 and catenin beta 1 (CTNNB1) are involved in fertility; transforming growth factor beta receptor 3 plays a role in meat quality traits; Wnt family member 2, CTNNB1 and transcription factor 7 take part in adipogenesis and fat deposition simultaneously; plasminogen activator, urokinase receptor (completely linked to AXL receptor tyrosine kinase, r2 = 1) plays an essential role in the successful ovulation of matured oocytes in pigs; colipase like 2 (strongly linked to SAM pointed domain containing ETS transcription factor, r2 = 0.848) is involved in male fertility. Conclusion: These adverse genes susceptible to swine EP may be selected while selecting for economic traits (especially reproduction traits) due to pleiotropic and hitchhiking effect of linked genes. Our study provided a completely new point of view to understand the genetic basis for susceptibility or resistance to swine EP in pigs thereby, provides insight for designing sustainable breed selection programs. Finally, the candidate genes are crucial due to their potential roles in respiratory diseases in a large number of species, including human.
Gonadal development, fertilization and egg development in the maternal body and reproductive cycle of ovoviviparous rockfish, Sebastes inermis, were investigated histologically. Gonadosomatic index(GSI) of male and female were increased from September and reached maximum values in December. In the male, GSI decreased from January, but in the female maintained high values till February and decreased from March. Hepatosomatic index(HSI) was related to GSI conversely. In both sex, HSI increased from February and reached maximum in August as the gonad were degenerating and resting, and began to decrease from September as gonad were glowing. This ovoviviparous rockfish copulates in December. Fertilization with sperms maintained between ovulated oocytes in the ovary occurs in January mainly. Egg development in the ovarian cavity and discharging of hatched preiarva occurs from January to February. The reproductive cycle includes the successive stages: Growing(September), Mature (October-November), Ripe and Fertilization(Decembr-Janua), Egg development and Discharging of hatched larva(January-February), Degeneration and Resting(February-August). According to the frequency distribution of egg diameter and histological observation, the ovoviviparous rockfish discharged the prelarva at a time in a spawning season. The sexual maturation is first attained at 2 ages. All females and males reaches first maturity at body length of 17.1cm and 15.1cm respectively. The mean number of the embryos increased with the increase of the total length of female.
Park, Kwan-Ha;Chung, Ee-Yung;Lee, Chang-Hoon;Kim, Sung-Han;Kim, Sung-Yeon;Seo, Won-Jae;Ryu, Dong-Ki
The Korean Journal of Malacology
/
v.27
no.3
/
pp.261-271
/
2011
The gametogenic cycle, the spawning season and the biological minimum sizes in female and male Protothaca (Notochione) jedoensis were investigated by quantitative statistical analysis. In females, monthly changes in the percents of the follicle areas to the ovarian tissue areas and the percents of the oocyte areas to the ovarian tissue areas increased in February and reached the maximum in April, and then gradually decreased from May to July, with the spawning peak between June and July. In males, monthly changes in the percents of the testicular tissue areas to total tissue areas and the percents of the spermatogenic stage areas to the testicular tissue areas increased in February and reached the maximum in April, and then showed a rapid decrease from May to July. From these data, it is apparent that the number of spawning seasons in female and male P. (N.) jedoensis occurred once a year, from May to July. Therefore, P. (N.) jedoensis in both sexes showed a unimodal gametogenic cycle during the year. Compared the gametogenic cycle by quantitative statistical analysis in 2007 with the previous qualitative results of this species, the results of the gametogenic cycle calculated by quantitative statistical analysis showed some differentiations in the spawning seasons evaluated by the gonad index by qualitative histological analysis. The intervals of the beginning of two spawning seasons showed one month between the results of quantitative and qualitative analyses. The biological minimum sizes (considering to 50% of group sexual maturity) in female and male clams by quantitative analysis of this species are 32.01 mm in shell length in females and 30.58 mm in males, respectively. According to the mean shell length fitted to von Bertalanffy's equation, 30.58 and 32.01 mm in shell length were considered to be two years old. Therefore, we assume that both sexes of this population begin reproduction from two years of age.
In patients with fully edentulous jaw, treatment of complete dentures should be carried out in many stages when following the conventional methods. Therefore there were disadvantages such as multiple visits to dental clinic is inevitable. In addition, errors caused by polymerization shrinkage, which happens during the fabrication of denture, and difficulties in reproduction of damaged or lost denture were considered as disadvantages. But nowadays, computer-aided design and computer-aided manufacturing (CAD/CAM) system is widely used in dentistry and it has begun to expand its spectrum in manufacturing complete dentures. Using CAD/CAM system to fabricate complete dentures can reduce the number of patient's visit and clinical chair time, since taking impression, recording jaw relation, and selection of artificial teeth are performed at the same time during the first visit, and delivering of dentures during the second visit is possible. In addition, because 3D-Printing technology is used, errors by polymerization shrinkage can be reduced. Among the companies that fabricate complete dentures using CAD/CAM system, DENTCA CAD/CAM denture (DENTCA Inc., Los Angeles, CA, USA) is the most commercialized company. In this case, we treated patients of complete dentures using conventional complete denture method and DENTCA CAD/CAM denture system in the same patient. We would like to report this case because we have achieved good results not only in functional aspects of pronunciation, chewing, and swallowing but also in aesthetic aspects.
To understand the effects of the cooling water system of the power plant on the succession of sessile zoobenthos, a series of experiments were carried out at the cooling water system of Samchonpo Power Plant from Hay 1985 to May 1986. The lowest species diversity of the zoobenthos was observed at the discharge canal. This is probably caused by both increased velocity and temperature of circulating water. It is also noticeable that species composition of the zoobenthos, to a certain degree, was affected due to the impingement of meroplankton during the entrainment process. While the number of species attached on the experimental substrata installed in the thermally altered area is smaller than that in the intake canal, the biomass is increased as the increased temperature eliminates some species and in turn stimulates reproduction and growth of the others. It is interesting that relatively high similarity exists between the summer communities in the intake canal and the spring communities in the thermally altered area, and the same relationship is also found between the fall communities in the intake canal and the winter communities in the thermally altered area. Ecological index curves of the zoobenthos of the intake canal follows the fluctuation pattern of the thermally altered area in one or two months of elapsed time, probably due to temperature increase by the inflow of heated water discharged from the power plant.
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