• Title/Summary/Keyword: Cell Observation

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Factors affecting pregnancy rates following transfer of bovine embryos derived from in vitro fertilization of oocytes matured in vitro (체외수정란 이식시 수태율에 미치는 요인에 관한 연구)

  • Kim, Sung-ki;Roh, Sang-ho;Lee, Eun-song;Lee, Byeong-chun;Hwang, Woo-suk
    • Korean Journal of Veterinary Research
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    • v.36 no.4
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    • pp.919-927
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    • 1996
  • In the last few years, methods for in vitro culture of early embryo stages from oocytes matured and fertilized in vitro using suitable cell culture systems have been established. But the factors affecting pregnancy rates following transfer of bovine embryos produced in vitro were not evaluated enough. So this study was performed to investigate the effects of quality and stage of embryos, parity and Corpus Luteum quality of recipients on pregnancy rates following non-surgical transfer of bovine embryos produced in vitro. Oocytes aspirated from small antral follicles of ovaries obtained at a local slaughter house were matured, fertilized with frozen-thawed semen and co-cultured for 6-7 days by utilizing co-culture system with bovine oviduct epithelial cell in vitro. After co-culture, embryos were transfered to recipients on day 7 (estrus=day 0). Recipients were monitored by ultrasonic scanning method or observation for estrus and rectal palpation after 50 days from transfer. The results of this study are follows. 1. Of the 70 recipients, 70%(49 of 70) had not showed estrus sign between day 0 and day 50, but 22.9%(16 of 70) was diagnosed not pregnant. Therefore the overall pregnancy rate of this study was 47.1%(33 of 70). 2. The pregnancy rate of recipients transfered with excellent(66.7%) and good(54.5%) embryos were higher than that of recipients transfered with fair embryos(15.8%) (p<0.05). 3. The pregnancy rate of recipients transfered with morula, compacted morula, blastocyst and expanded blastocysts were 46.2, 55.0, 62.5 and 50.0%, respectively. 4. The pregnancy rates of recipients transfered to heifer and cow were 54.5 and 55.2%, respectively. 5. The pregnancy rates of recipients with CL score I, II(66.7, 63.6%) were higher than those of recipients with CL score III (10%), (p<0.05). Success of transfer of embryos produced in vitro depends on many variables. The important factors identified in this study were the quality of embryos and the CL score of recipient animals after non-surgical transfer of embryos matured, fertilized and cultured in vitro.

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Identification and Characterization of an Endophytic Strain of Streptomyces from Rice Roots (Orysa sativa L.) (벼(Orysa sativa L.) 뿌리로부터 분리된 내생 Streptomyces 균주의 동정 및 특성)

  • Kim, Jae-Heon;Lee, Jun-Kwan
    • Korean Journal of Microbiology
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    • v.47 no.4
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    • pp.375-380
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    • 2011
  • We isolated an endophytic actionmycete from root tissues of rice plant collected from paddy field near Dankook University, Cheonan, Korea. Surface sterilized roots were laid on the selective agar plates and incubated. The powdery actinomycete colonies appeared on the root surface after four weeks incubation. We isolated a strain JK-5 among them and could determine its taxonomical position as Streptomyces diastaticus subsp. ardesiacus by using 16S ribosomal DNA sequencing. The chemotaxonomical and morphological studies confirmed the taxonomical position of the strain JK-5. The shape of aerial hyphae was flexible and they contained spore chains with more than 30 smooth spherical spores per chain. Cell walls contained LL-diaminopimelic acid. There was no characteristic sugar in whole-cell hydrolysates. The major fatty acids were anteiso-15:0, anteiso-17:0 and iso-16:0. The specific menaquinones, MK-9 ($H_6$), MK-9 ($H_8$), were detected. The GC content was 72%. Antifungal activities of the strain JK-5 were relatively strong against fungal plant pathogens. The endophytic growth of the strain JK-5 was confirmed by SEM observation of the root and stem of the infected rice plant.

Several Factors on Bulblets Regeneration from Callus Culture in Lilium longiflorum 'Celia' (백합 'Gelia' 캘러스로부터 자구 재분화에 미치는 제요인)

  • 박소영;김시동;신세균;이철희;백기엽
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.3
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    • pp.183-188
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    • 1997
  • Callus from scale segments of Lilium longiflorum 'Gelia' was effectively induced and maintained from unorganized tissue on the semi-solid medium by 0.42% Bacto agar with MS basal salts and vitamins of SH medium supplemented with 0.5 mg/L 2, 4-D, 1.0 mg/L NAA, 0.3 mg/L BA, and 3% sucrose. More than 5% of high sucrose level had inhibiting effect on regeneration capacity of formed callus and decreased callus growth. Various combinations of nitrogen did not effective to proliferate the ELC (Embryogenic-like callus), but friability of callus was increased in the medium containing only nitrate as nitrogen source. 5 mL conditioned medium into 30 mL fresh medium was good for cell growth. However friable cell aggregates during suspension culture had to form hard callus which hindered to establish suspention culture system. Addition of 2 g/L casein hydrolysate increased callus growth and friability of the hard callus. As a result of anatomical observation of callus, organogenesis such as shoots, roots and bulblets was independently induced from callus tissue. Somatic embryogenesis from callus tissue could be observed with low frequency.

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Expression analysis of RBMY1, CDY1, and VCY2 genes in Korean male infertility (한국 불임남성에 있어서 RBMY1, CDY1 그리고 VCY2 유전자의 발현 분석)

  • Huh, Jae-Won;Kim, Woo-Young;Kim, Dae-Soo;Ha, Hong-Seok;Park, Nam-Chul;Choi, Ook-Hwan;Nam, Ki-Man;Choi, Jin;Kim, Heui-Soo
    • Journal of Life Science
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    • v.17 no.5 s.85
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    • pp.613-618
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    • 2007
  • Azoospermia factor(AZFa, b, and c) regions have been focused on their involvement in the spermatogenic process by frequent observation of microdeletion in male infertility. Among the azoospermia factors, RBMY1, CDY1, and VCY2 genes are strongly associated with the male germinal cell differentiation and development in testis. Using RT-PCR approach, expression patterns of RBMY1, CDY1, and VCY2 genes are examined in testicular biopsy specimens from 42 Korean azoospermic patients. No expression of RBMY1, CDY1, and VCY2 genes appeared as 34%, 66%, and 27% of the male infertility, respectively. Patients who had no expression of RBMY1 and VCY2 genes also showed negative expression of the CDY1 gene in their testis tissues. All Sertoli cell-only syndrome patients showed no expression of the CDY1 gene. Taken together, the CDY1 gene expression seems to be necessary factor to complete spermatogenesis in Korean population.

Effects of the Duration of Highly Intensive Exercise on Lymphocyte Cell Death in Rats (고강도운동 지속시간이 rat의 림프구 세포사에 미치는 영향)

  • Kim, Hyeong-Soo;Hyun, Kyung-Yae
    • Journal of Life Science
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    • v.22 no.3
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    • pp.312-317
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    • 2012
  • The time-dependent effects of highly intensive exercise on the hematological properties of leukocytes, as well as $CD4^+$ and $CD8^+$ level changes as T-lymphocyte activation subsets and the cell death of lymphocytes in rats were studied in this research. Twenty, 60, and 120 min of highly intensive exercise was performed daily for 8 weeks. Total leukocyte counts in the blood of rats exercising for 20 min were elevated; they then decreased to less than the level of the control group up to 120 min. The patterns of lymphocyte level changes were directly influenced by exercise duration and the extents of alteration were similar to the total leukocytes counts. The levels of $CD4^+$ and $CD8^+$ in the blood of the exercising rats were not statistically different even when the exercise was continued for 120 min; thus, the exercise did not affect T-lymphocyte activation. Early- and late-stage lymphocyte apoptosis was not affected by the length of exercise, except that late-phase apoptosis was slightly increased at 120 min, suggesting that aging processes for lymphocyte apoptosis might be stimulated at that time. As the exercise time became longer, stimulated necrosis of lymphocytes was observed, so damage in lymphocytes and a potential loss of immunity might be presumed. The current observation suggests that long-term, highly intensive exercise might result in a loss of immunity that could be due to the damage of lymphocytes in terms of both their numbers and inflammation-related functions. The results suggest that under highly intensive exercise conditions, more than 20 min of exercise should not be suggested for health care purposes.

Effects of Bagging Periods on Pericarp Characteristics and Berry Cracking in 'Kyoho' Grape (Vitis sp.) (괘대시기가 '거봉' 포도의 과피 특성과 열과에 미치는 영향)

  • Son, In-Chang;Kim, Dae-Il
    • Horticultural Science & Technology
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    • v.28 no.3
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    • pp.381-386
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    • 2010
  • The berry growth and pericarp characteristics were characterized to confirm the effects of bagging periods on berry cracking during berry development in 'Kyoho' grape. The berry weight was the highest at 13.4 g in late period of bagging treated at 7 to 9 weeks after full bloom (WAFB) as compared with the lowest of 12.3 g in total period bagging. The berry cracking rate under critical turgor pressure in the non-bagging control was 53.3%, while those of bagging treatments were decreased in the order of 42.7%, 37.3%, 33.3%, and 18.7% in bagged during 3 to 9, 3 to 5, 5 to 7, and 7 to 9 WAFB, respectively. In the results of observation on histological characteristics of pericarp, berry lenticels of whole bagging treatments had smaller and normal shape compared with non-bagging control treatment. Especially on the pericarp of late period bagged during 7 to 9 WAFB, suberization around stomata and micro-cracking were not observed and structural strength of pericarp was increased with thicker sub-epidermal layer and cell wall. Therefore, the results indicate that bagging treatment for two weeks just before the veraison when the day length and daylight is relatively longer and stronger can effectively reduce berry cracking by strengthening structure of pericarp in 'Kyoho' grape.

Histopathological observation of the gill of the crucian carp, Carassius auratus by the leech, Limnotrachelobdella sinensis (거머리, Limnotrachelobdella sinensis의 기생으로 인한 붕어, Carassius auratus 아가미의 조직병리학적 관찰)

  • Park, Myoung-Ae;Kim, Seok-Ryel;Kim, Myoung-Sug;Kim, Jeong-Ho;Park, Jung-Jun
    • Journal of fish pathology
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    • v.23 no.3
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    • pp.399-407
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    • 2010
  • On the inner side of each operculum of the crucian carp, Carassius auratus (n=10), the leech, Limnotrachelobdella sinensis of 1-4 individuals were parasitic. The leeches had approximately 41.0 mm in total length and 11 mm in width. These body was composed with anterior sucker, neck, trunk and posterior sucker and average length was 2.3 mm, 7.2 mm, 23.3 mm and 8.7 mm respectively. To both sides of the trunk lateral vesicle of 11 pair existed. When observed by SEM, anterior sucker was hemisphere shape and the mouth where proboscis comes out existed with the its center. Proboscis was connected the esophagus directly. Under light microscopy, bloodsucking gill of C. auratus showed lamella fusion, hypertrophy the epithelial cell of the filament and lamella, increased mucocytes and congested capillaries. On the other hand, necrotic and hydropic degeneration epithelial cell of the lamella, and infiltration of the macrophages from some individuals were suggested the secondary infection with the bacteria or virus after bloodsucking activity of the leech.

Isolation and Antimicrobial Action of Growth Inhibitory Substance on Food-borne Microorganisms from Dryopteris crassirhizoma Nakai (관중(Dryopteris crassirhizoma Nakai)의 식중독 미생물 증식 억제 물질의 분리 및 항균작용)

  • Han, Ji-Sook;Lee, Ji-Young;Baek, Nam-In;Shin, Dong-Hwa
    • Korean Journal of Food Science and Technology
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    • v.33 no.5
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    • pp.611-618
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    • 2001
  • The ethanol extract of Dryopteris crassirhizoma Nakai showed strong growth inhibition against 5 strains of Listeria monocytogenes at the concentrations of $100{\sim}500$ ppm and the minimum inhibitory concentration of n-hexane fraction was under 50 ppm. The D8-2-5 fraction isolated from n-hexane fraction of Dryopteris crassirhizoma Nakai showed a strong bactericidal activity on 5 strains of L. monocytogenes at 20 ppm level in tryptic soy broth medium. At the level, the viable count was reduced $4{\sim}6$ log cycle compared to initial cell number. Observation by the measurement of adenosine triphosphate (ATP) contents and transmission electron microscope showed that disruptions of the cell wall and elution of intracellular ATP are assumed to be due to the bactericidal activity. In addition, the n-hexane fraction of Dryopteris crassirhizoma Nakai showed the strong growth inhibitions at 50 ppm on Vibrio parahaemolyticus and Bacillus cereus, and at 25 ppm on Staphylococcus aureus.

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Estimation of Daily Maximum/Minimum Temperature Distribution over the Korean Peninsula by Using Spatial Statistical Technique (공간통계기법을 이용한 전국 일 최고/최저기온 공간변이의 추정)

  • 신만용;윤일진;서애숙
    • Korean Journal of Remote Sensing
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    • v.15 no.1
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    • pp.9-20
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    • 1999
  • The use of climatic information is essential in the industial society. More specialized weather servies are required to perform better industrial acivities including agriculture. Especially, crop models require daily weather data of crop growing area or cropping zones, where routine weather observations are rare. Estimates of the spatial distribution of daily climates might complement the low density of standard weather observation stations. This study was conducted to estimate the spatial distribution of daily minimum and maximum temperatures in Korean Peninsula. A topoclimatological technique was first applied to produce reasonable estimates of monthly climatic normals based on 1km $\times$ 1km grid cell over study area. Harmonic analysis method was then adopted to convert the monthly climatic normals into daily climatic normals. The daily temperatures for each grid cell were derived from a spatial interpolation procedure based on inverse-distance weighting of the observed deviation from the climatic normals at the nearest 4 standard weather stations. Data collected from more than 300 automatic weather systems were then used to validate the final estimates on several dates in 1997. Final step to confirm accuracy of the estimated temperature fields was comparing the distribution pattern with the brightness temperature fields derived from NOAA/AVHRR. Results show that differences between the estimated and the observed temperatures at 20 randomly selected automatic weather systems(AWS) range from -3.$0^{\circ}C$ to + 2.5$^{\circ}C$ in daily maximum, and from -1.8$^{\circ}C$ to + 2.2$^{\circ}C$ in daily minimum temperature. The estimation errors, RMSE, calculated from the data collected at about 300 AWS range from $1.5^{\circ}C$ to 2.5$^{\circ}C$ for daily maximum/minimum temperatures.

Production of Transgenic Murine Embryos using Haploid Spermatids Transfected with EGFP Gene (EGFP 유전자가 도입된 반수체 정자세포에 의한 형질전환 설치류 난자의 생산)

  • Kang, K.Y.;Song, S.J.;Lee, H.T.;Chung, K.S.
    • Korean Journal of Animal Reproduction
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    • v.25 no.4
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    • pp.305-315
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    • 2001
  • In this study, the production of transgenic embryo was attempted by microinjection or round spermatid cultured with foreign DNA. At first, the expression of haploid spermatids specific gene, mTP1 in mouse and hPrm2 in hamster spermatids were investigated by RT-PCR method in testes of young mice and hamster testis. The specific gene expression first appeared at 18 days post partum (dpp) in mice spermatid and 20 dpp in hamster spermatid. Therefore, the round spermatids isolated from 17 dpp mice and 19 dpp hamster were used for the introduction of foreign EGFP gene into haploid round spermatids. For the introduction of EGFP gene haploid round spermatids suspended in medium including EGFP gene were treated with a different electric field strength at 0.11, 0.18 and 0.44 ㎸/cm. After electrical stimulation, viability of testicular sperm cells and 67.6%, 66.4% and 49.9%, in mice and 62.6%, 57.9% and 27% in hamster, respectively. These values were significantly lower than those of non-treated control groups 80.5% in mouse and 69.1% in hamster After 72 hrs culture, the highest expression rate of EGFP gene, 28.5% in mice and 32.1% in hamster were obtained from tile spermatogenic cells electroporated by the field strength or 0.18 ㎸/cm. Then, the ability of fertilization and embryonic development of haploid spermatids transfected with foreign EGFP gene were estimated by the microinjection of spermatids into hamster oocytes. The Irate pronuclear formation rate (77.5%) was lower than non-treated control (80%), and the cleavage rate of the treated group (58.8%) was lower than control (65%). To prove the foreign EGFP integration in hamster embryos, 2-cell stage hamster embryos were subjected to the observation under the fluorescence microscope, and the PCR analysis. As a result, about 44% of 2-cell embryos were showed the integration of EFGP gene into their genome. Therefore, These results suggest the possibility to produce transgenic hamsters by microinjection of haploid spermatid transfected with foreign DNA.

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