• Clinical and Experimental Reproductive Medicine
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• 제19권1호
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• pp.71-79
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• 1992

In vitro fertilization and embryo transfer (IVF & ET) is widely used for the males with subnormal or abnormal semen quality, as this was recommended in view of the relatively small numbers of spermatozoa required for fertilization and subsequent pregnancies could be obtained. The aim of this study is to know how the various functional parameters of spermatozoa in semen analysis affect the outcome IVF. This study was carried out between 1988-1989, with male factor patients selected on the basis of the semen quality. The selection criteria was based upon the mean values of concentration,% motility and % normal morphology from at least two semen analysis. There is a significant decrease in the fertilization and embryo transfer rates in the study group compared with control group (35.9% vs. 68% and 48.6% vs. 85.5% respectively), however, there was no significant difference in the pregnancy or delivery rates (19.6% vs. 21.4% and 60.0% vs. 62.5% respectively) per embryo transfer cycles. Fertilization rate is variously affected by the type and degree of sperm defect. No pregnancy was occurred in triple defect group and asthenoteratospermia group. There is no significant increase in the abortion rate in the male factor group. Improvement have to be made with the fertilization rate, as the pregnancy rate per OPU cycle in male factor group is still lower than that of normal group (9.5% vs. 18.3%). In conclusion, IVF can be used as a treatment for male factor infertility and the preparation of the semen sample can be modified to improve sperm recovery and obtain fertilization from abnormal semen samples.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.75-75
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• 2003

Since the establishment of embryonic stem cell, pluripotency of the cells was known to allow differentiation of the cells into various cell types consisting whole body. Several protocols have been developed to induce expression of specific genes.. However, no precise protocol that will generate a single type of the cells from stem cells has been reported. In order to produce cells suitable for transplantion into brain of PD animal model, which arouse due to a progressive degeneration of dopaminergic neurons in midbrain, human embryonic stem cell (hESC, MB03) was transfected with cDNAs cording for tyrosine hydroxylase (TH). Successful transfection was confirmed by western immunoblotting. Newly transfected cell line (TH#2/MB03) was induced to differentiate by the two neurogenic factors retinoic acid (RA) and b-FGF. Exp. I) Upon differentiation using RA/ascorbic acid (AA), embryoid bodies (EB, for 4days) derived from hES cells were exposed to RA (10$^{-6}$ M)/AA (50 mM) for 4 days, and were allowed to differentiate in N2 medium for 7, 14, 21, or 28 days. Exp. II) When bFGF was used, neuronal precursor cells were selected for 8 days in N2 medium after EB formation. After selection, cells were expanded at the presence of bFGF (20 ng/ml) for another 6 days followed by a final differentiation in N2 medium for 7, 14, 21 or 28 days. By indirect immunocytochemical studies, proportion of cells expressing NF200 increased rapidly from 20% at 7 days to 70 % at 28 days in RA/AA-treated group, while those cells expressing NF160 decreased from 80% at 7 days to 10% at 28 days upon differentiation in N2 medium. However, in differentiation by RA/AA treatment system, there was a significant increase in proportion of neuron maturity (73%) at day 14 after N2 medium. TH#2/MB03 cells expressing TH are >90% when matured at the absence of either bDNF or TGF-$\alpha$. These results suggested that TH#2/MB03 cells could be differentiated in vitro into mature neurons by RA/AA.

• Journal of Plant Biotechnology
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• 제6권4호
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• pp.205-212
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• 2004

The objectives were to compare the salt tolerance levels in the parental rice cultivar, Dongjinbyeo, and induced mutagenesis derived its lines for plant height, MDA, ATPase, POD, and 2-dimensional protein electrophoresis pattern in NaCl-containing hydroponic nutrient solutions. Rice plants isolated from a population of rice (Oryza sativa L. cv. Dongjinbyeo) mutation lines, which were generated in combination with in vitro selection and gamma-ray, exhibited salt tolerance. Line No. 18 had the longest plant, whereas NaCl-sensitive line (No. 25) had the shortest plant. The parent, and the sensitive line showed severe damage from salt stress. Tolerant lines (No. 18, 50) had a lower malonaldehyde (MDA) content than the sensitive one (Dongjinbyeo, No. 25) during salt stress. Several proteins showed significant quantitative variation through 2DE; phosphoribulokinase, peroxidase, oxygen evolving enhancer 1 and the $H^+-ATPase$, which are known to be involved in salt tolerance. The effect of salt on peroxidase and $H^+-ATPase$ activity in the seedlings of two groups with contrasting genotypes of rice was studied. A greater activity was recorded in the tolerant lines as compared to the sensitive ones (P<0.05, Duncan's test). The results indicate that salt tolerant lines expressed more salt stress-inducible proteins associated with salt tolerance than the sensitive lines during salt stress.

• 농업과학연구
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• 제40권4호
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• pp.271-276
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• 2013

This experiment was carried out to determine optimal culture conditions for the production of tubers of Calla (Zantedeschia elliottiana 'Golden Affair' and 'Black Magic') in Korea highland. In vitro produced plantlets and tuberlets of Calla 'Golden Affair' and 'Black Magic' were planted plastic film greenhouse and grown for 100, 120, 140 days, with different night temperature treatments ($0{\sim}10^{\circ}C$ : no heating, 10, $15^{\circ}C$). In both cultivars, tuber size(tuber diameter, tuber height) and tuber weight increased with increasing cultivation period when the night temperature was maintained at $10^{\circ}C$. The largest tuber diameter in vitro produced plantlets was 5.8cm in 'Black Magic' and 3.2cm in 'Golden Affair', and daily tuber growth rate was 1.110g in 'Black Magic' and 0.092g in 'Golden Affair' under the culture conditions. Consequently we think that tuber harvest date was Oct. 30 and night temperature was $10^{\circ}C$ and no heating that was proper method of tuber production. However we had selection of $10^{\circ}C$ treatment for tuber production because it appeared freezing damage occasionally in highland late in October.

• Reproductive and Developmental Biology
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• 제36권1호
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• pp.71-77
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• 2012

The objective of this study was to examine the effect of thymidine treatment during $in$ $vitro$ maturation (IVM) of porcine follicular oocytes on blastocyst development. Porcine oocytes were treated with thymidine (10 mM, 20 mM and 30 mM) for 2 or 6 hr in the preiods of IVM I and/or II. The survival rates of the blastocysts in the 6 hr treatment groups of 10 mM and 20 mM during IVM I period were significantly higher than those of control group ($p$<0.05). However, the survival rate of the blastocysts in the 2 hr treatment group of 20 mM during IVM II period was significantly higher than control group ($p$<0.05). Furthermore, the survival rate of the blastocysts in the 6 hr treatment group of 30 mM during IVM II period was significantly lower than control group ($p$<0.05). Consistent with the previous result, blastocyst development of both IVM I and II treatment group was also showed as similar pattern. Total and apoptotic cell numbers of blastocysts derived from thymidine treated porcine oocytes were examined by using Tunel assay. The results showed that there was no significant differences in total cell number of blastocysts between thymidine treated and untreated groups. However, apoptosis-positive cells in the thymidine treated group (6 hr IVM I) were significantly lower than those of other groups ($p$<0.05). Taken together, these results indicate that high quality oocytes were selected by DNA synthesis mechanism according to high concentration thymidine treatment during porcine oocyte maturation. Therefore, we concluded that presumptive selected oocytes by thymidine treatment during maturation periods improved the further embryo development and embryonic quality of IVF embryos by decreasing the incidence of apoptosis in preimplantation porcine embryos.

• 한국동물위생학회지
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• 제37권1호
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• pp.1-9
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• 2014

Although it has been generally accepted that porcine reproductive and respiratory syndrome virus (PRRSV) induces weak and delayed protective immunity after infection, it is unclear that the same immunological features can be applicable to all PRRS viruses because huge genetic variation exists even among the same genotypes of PRRSV (Type 1 and 2). In the current study, two genetically distinct type 2 PRRSV strains (VR-2332 and JA142) which showed approximately 90% nucleotide homology based on ORF5 sequences were characterized by both in vitro and in vivo assessments to determine the immunological features of the viruses. For in vitro assessment, porcine alveolar macrophages (PAM) were infected with the viruses at $10^{-3}$ multiplicity of infection (MOI) and then supernatants and cells were collected separately at 36 hrs post infection to determine the relative expression levels of IL-$1{\alpha}$, IL-12, TNF-${\alpha}$ and INF-${\alpha}/{\beta}$ by quantitative RT-PCR. In addition, five PRRSV-free pigs were inoculated with either of JA142 or VR2332 for in vivo assessment. Serum samples were collected every week until 6 weeks post challenge. The serum samples were analyzed for the levels of viremia, PRRSV nucleocapsid-specific antibody and virus neutralizing antibody. Based on those assessments, the two viruses showed different patterns of cytokine expression in PAM and immune responses in pigs after infection. These results indicate that genetically distinct PRRSV strains have different immunological features, which might be criteria for virus classification and selection of candidate virus strains for vaccine development in the future.

• Journal of Microbiology and Biotechnology
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• 제17권8호
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• pp.1300-1307
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• 2007

This study assessed the possible role of different traits in selected plant growth-promoting rhizobacteria (PGPR) for improving wheat growth and yield under natural conditions. Rhizobacteria exhibiting 1-aminocyclopropane-1-carboxylate (ACC)-deaminase activity were isolated and screened for their growth-promoting activity in wheat under axenic conditions. Five isolates belonging to Pseudomonas and one Burkholderia caryophylli isolate that showed promising performances under axenic conditions were selected and characterized for in vitro ACC-deaminase activity, chitinase activity, auxin production, P solubilization, and root colonization. These isolates were then used as inocula for wheat cultivated under natural conditions in pot and/or field trials. Significant increases in root elongation, root weight, tillers per pot, 1,000-grain weight, and grain and straw yields were observed in response to inoculation with PGPR in the pot trials. Inoculation with these PGPR was also effective under field conditions and increased the wheat growth and yield significantly. However, the efficacy of the strains was inconsistent under the axenic, pot, and field conditions. Pseudomonas fluorescens ($ACC_{50}$), which exhibited a relatively high in vitro ACC-deaminase activity, chitinase activity, auxin production, and P solubilization and more intensive root colonization, was the most efficient isolate under the field conditions. Therefore, these results demonstrated that ACC-deaminase activity is an efficient parameter for the selection of promising PGPR under axenic conditions. However, additional traits of PGPR, including auxin production, chitinase activity, P solubilization, and root colonization, are also important for selecting PGPR as biofertilizers.

• Journal of Forest and Environmental Science
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• 제33권3호
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• pp.197-201
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• 2017

Populus. caspica L. is an Iranian indigenous poplar species which naturally distributed in the northern part of country. Unfortunately, overuse has removed many of the stems of better form, so that natural stands now usually appear small and crook. Therefore genetic variation for selection of new superior clone of this species is needed. Conventional hybridization system is currently used to induce genetic variation in poplar species but incompatibility barriers have been observed between them. In vitro ovule embryo culture was used to overcome incompatibility obstacle for interspecific hybridization between Populus caspica L. with Populus deltoids L.62/75. Female flowers of Populus caspica L. have artificially been pollinated with pollen grain of P. deltoides 62/75 in one direction using twig and pot crossing system. Ovaries at different ages (7, 14 and 21 days after pollination) were disinfected through 70% ethanol for 1 minute, 5% of sodium-hypochlorite solution for fifteen min followed by three time rising with sterile distil-water. Isolated ovaries were then transferred to MS hormone free medium containing 30 and 60 g/L sucrose for embryo development and germination. Collected data have been analyzed by two factorial experimental designs. The results indicated that there were significant differences between age of embryos for development and germination at ${\alpha}=0.01%$. Highest embryo germination (45%) was observed from 21 days old ovaries. No significant differences were observed between MS culture media containing 30 and 60 g/L for percentages of ovary-embryo germination and number of germinated embryo per ovary at ${\alpha}=0.05%$. Fourteen percentage of embryo germination obtained in MS medium supplemented with 60 g/L sucrose, while only 35% of isolated ovaries were able to germinate in MS containing 30 g/L sucrose. Induced plantlets in 4 cm height were transferred into pots containing soilless (1:1:1 peat, per lit and vermiculite) medium for acclimatization. After successful acclimatization, plants were delivered to nursery.

• 식물조직배양학회지
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• 제24권2호
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• pp.77-81
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• 1997

감자 실생개체들의 내서성계통을 조기 선발하기 위해 CIP575015 $\times$Katahdin CIP 575015 x B6603-6, 84 I 35-4 $\times$Katahdin, CIP 575015 $\times$Nooksack 그리고 CIP575015 $\times$Superior 등 5조합을 교배하여 획득된 총 750립의 진정종자를 기내에서 각각의 조합을 온도별로 처리하여 괴경 형성 반응을 조사한 결과 온도가 2$0^{\circ}C$에서 3$0^{\circ}C$로 증가할 경우 평균 괴경형성율이 2$0^{\circ}C$에 비해 평균 43% 정도가 감소하였으며 특히 CIP575015 $\times$Katahdin조합은 21%로 가장 낮았다. 반면에 CIP575015 $\times$B6603-6 조합은 3$0^{\circ}C$에서도 58%의 괴경형성율을 보여 고온에서도 괴경형성이 잘되는 내서성 조합으로 생각되었다. 기내 고온조건하에 선발된 계통을 고온기 포장에서 내서성을 검정하기 위해 기내소괴경에서 생산된 괴경을 저지대 난지인 강릉 포장에 재배하였다. 조합별 괴경특성 및 수량을 조사한 결과 2$0^{\circ}C$에서 선발된 계통 중 1계통, 3$0^{\circ}C$에서 선발된 계통중 3계통의 내서성계통을 선발하였다. 수량성에서는 89ML75-8 계통이 수미품종 대비 88%의 증가로 다수성이었으며, 89ML64-11 계통은 건물율이 18.8%로 수미 품종보다 11%가 증가되어 전분용으로 유망시 되었다.

• 미생물학회지
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• 제43권3호
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• pp.159-165
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• 2007

C형 간염바이러스(hepatitis C virus; HCV)증식을 효과적이며 특이적으로 제어할 수 있는 유전산물을 개발하기 위하여 HCV 중식조절이자인 NS5B RNA replicase 존재에 의해 allosteric하게 그 활성 이 조절될 수 있는 HCV internal ribosome entry site (IRES) 표적 hammerhead 리보자임을 개발하였다. 우선 HCV IRES 염기서열 중+382 nucleotide(nt) 부위가 리보자임에 의해 가장 잘 인식되었음을 관찰하였다. 이러한 allosteric 리보자임은 NS5B RNA replicase와 특이적으로 결합하는 RNA aptamer 부위, aptamer와 NS5B와의 결합에 의해 리보자임 활성을 유도할 수 있도록 구조적 변이를 전달할 수 있는 communication module부위 및 HCV IRES의 +382 nt를 인지하는 hammerhead 리보자임 등으로 구성되도록 설계하였다. 특히 in vitro selection기법을 활용하여 NS5B 의존적으로 리보자임 활성을 증가시킬 수 있는 communication module 염기서열을 밝혀내었다. 이러한 리보자임은 단백질이 없거나 대조 단백질인 bovine serum albumin이 존재할 때에는 절단반응을 유도하지 못하였으나 HCV NS5B 단백질이 존재할 매에만 효과적으로 NS5B 농도 의존적으로 절단 반응을 유도할 수 있음을 관찰하였다. 이러한 allosteric 리보자임은 HCV중식의 효과적인 증식 억제 선도물질 뿐만 아니라 HCV 치료선도물질의 스크리닝용 도구 및 HCV 조절 인자를 탐색할 수 있는 HCV 진단용 리간드로서도 활용될 수 있을 것이다.