The cell cycle phase in which donor nuclei exist prior to nuclear transfer is an important factor governing developmental rates of reconstituted embryos. It was suggested that quiescent G0 and cycling G1 cells could support normal development of reconstituted embryos. In a quest of optimized donor nuclei treatment prior to nuclear transfer, this study was undertaken to examine the cell cycle characteristics of bovine fetal and adult somatic cells when cultured under a variety of culture treatments and the cell cycle change with the lapse of time after trypsinization. This was archived by measuring the DNA content of cells using flow cytometry, Cultured fetal fibroblast cells, adult skin and muscle cells, and cumulus cells were divided by 3 culture treatments; 1) grown to 60-70% confluency (cycling), 2) serum starved culture, 3) culture to confluency. Trypsinized cells were fixed by 70% ethanol and stained with propidium iodide. For one experiment, trypsinized cells were resuspended in DMEM+10% FBS and incubated for 1.5, 3 and 6 h with occasional shaking before ethanol fixation. Cell cycle phases were determined by flow cytometry enabling calculation of percentages of G0+G1, S and G2+M. The majority of cells were in G0+Gl stage regardless of origin of cells. Cultures that were serum starved or cultured to confluency contained significantly (P<0.05) higher percentages of cells in G0+G1 (89.5-95.4%). For every cell lines and culture treatments, percentages of cells in existing in G0+G1 increased with decreasing of the cell size from large to small. In the serum starved and confluency groups, about 98% of small cells were in G0+G1 Serum starved culture contained higher percentages of small-sized cells (38.5-66.9%) than cycling and confluent cultures regardless of cell lines (P<0.05). After trypsinization of fetal fibroblast and adult skin cells that were serum starved and cultured to confluency, the percentages of cells in G0+G1 significantly increased by incubation for 1.5(95.7-99.5%) and 3.0 h (95.9-98.6%). The results suggest that the efficient synchronization of bovine somatic cells in G0+G1 for nuclear transfer can be established by incubation for a limited time period after trypsinization of serum starved or confluent cells.
Frankliniella occidentalis and Thrips palmi are economically important pests of cucumber. The chemicals used to control them can be effective; however, they should not be used frequently on cucumbers that are consumed raw. Banker plants were selected to increase the control efficiency of Orius laevigatus, a natural enemy that can replace the chemicals. The hatching rate, emergence rate, nymph developmental period, survival rate, and reproductive capacity (fecundity and oviposition period) of O. laevigatus for four plants (moss rose, basil, broad bean, and cucumber) were investigated. The density of O. laevigatus on two selected banker plants was investigated in a cucumber greenhouse. The hatching rate of O. laevigatus eggs was highest at 92% in moss rose; however, there was no significant difference in survival rates between moss rose and basil. The fecundity and oviposition period of O. laevigatus were better in moss rose than in basil, with no significant difference between them. The flowering period of basil was longer than that of moss rose, from April to September. Therefore, basil has potential value as a banker plant for O. laevigatus and is expected to increase the biological control effect of O. laevigatus.
Purpose: Coronavirus disease 2019 (COVID-19) has posed a significant burden to infant and toddler's care globally, while the disease severity is generally mild in this age group. In this study, we aimed to assess epidemiological and clinical aspects of COVID-19 in infants and toddlers in Seoul, South Korea. Methods: We used Seoul Metropolitan Government's epidemiological investigation database to describe the epidemiological and clinical characteristics of COVID-19 in infants and toddlers, between March 2020 to December 2021. Results: A total of 5,025 infants and toddlers aged <5 years was diagnosed between the observed period. 2,720 (54.1%) had symptoms, and fever was the most common symptom in 1,941 (74.1%). Of the diagnosed cases, 96.4% did not have underlying diseases. In a district level, extended opening of childcare facility was associated with increased risk of COVID-19 in infants and toddlers. Conclusions: An efficient monitoring system, resembling routine clinical care, is crucial, considering the low rates of severe progression and fatality among infants and toddlers. Moreover, a well-grounded intervention based on scientific evidence, rather than unconditional closures, is necessary to establish a suitable childcare policy that ensures safety from infectious diseases while not overlooking the developmental aspects of social skills.
Tributyltin (TBT) is one of endocrine disrupters which are known as having similar function to sex steroid hormone inducing apoptosis in various tissues of rodents. Recently, it has been reported that TBT induces apoptosis in thymus causing the decreased thymic function, but little is known about the mechanism. To elucidate the mechanism, three-week-old SD female rats were orally administrated with TBT 1, 10, and 25 mg per body weight (kg) and sesame oil as a control for 7 days. On day 8, the thymi were obtained and weighed, and then the number of thymocytes was counted. We also performed H&E staining, TUNEL assay, and Annexin V flow cytometric analysis to examine the apoptosis rates and the structure in the thymus. Next, we investigated the adipogenesis and apoptosis-related mRNA expression levels in the thymi by real-time PCR. The thymic weight and the number of thymocytes were decreased by TBT in a dose-dependent manner. As a result of the H&E staining, the boundary between cortical and medullary area was blurred in the thymi of TBT treated rats compared to those of controls. In the results of TUNEL assay and Annexin V flow cytometric analysis, apoptosis rates in the thymus were increased after TBT treatment. The expression levels of thymic epithelial cell marker genes such as EVA, KGF, AIRE, and IL-7 were significantly decreased in the thymi of TBT treated rats, but $PPAR{\gamma}$, aP2, PEPCK, and CD36 were significantly increased. The expression of $TNF{\alpha}$ and TNFR1 as apoptosis-related genes also was significantly increased after TBT treatment. The present study demonstrates that TBT can increase the expression of adipogenesis and apoptosis-related genes leading to apoptosis in the thymus. These results suggest that the increased adipogenesis of thymus by TBT exposure might induce apoptosis in the thymus resulting in a loss in thymic immune function.
The objective of this study was to optimize the freezing/thawing method of in vitro produced Hanwoo blastocysts. Day 7 blastocysts after IVF were vitrified using EFS40 (40% ethylene glycol, 18% ficoll, 0.3 M sucrose and 10% FBS added m-DPBS) as a freezing solution and electron microscope (EM) grid (V-G) or straw (V-S) as an embryo container. In both method, freezing/thawing were treated by 2-step, treatment time was required in V-G method and V-S method, for 2 min / 3 min and 3.5 min / 10 min, respectively. Embryo survival was assessed as re-expanded and hatched rates at 24 h and 48 h after warming, respectively. The results obtained in these experiments were summarized as follows: when the effect of exposure in vitrification solution and chilling injury from freezing procedure on in vitro produced expanded blastocysts were examined, at 24 h after warming, embryo survival in exposure group (100.0%) was not different compared to that in control group (100.0%), although those results were significantly different with two vitrified groups (V-G: 87.8, V-S: 77.8%) (P<0.001). However, at 48 h after warming, hatched rates of V-G group (67.8%) were significantly higher than those of V-S group (53.3%) (P<0.05). In addition, this hatched rate in V-G group was not different with that in exposure group (73.3%). When the effects of embryo developmental stage (early, expanded and early hatching blastocysts) and embryo container (EM grid and straw) to the in vitro survival of vitrified-warmed day 7 Hanwoo blastocysts were simultaneously examined, fast developed embryos were indicated the better resistance to freezing than delayed developed one, irrespective of embryo containers (early; 57.1 & 24.4%, expanded; 84.7 & 60.6%, early hatching; 91.7 & 80.0%) (P<0.001). Especially, in expanded and early hatching blastocysts, embryo survival of V-G group (67.8, 95.0%) was significantly higher than those of V-S group (53.0, 65.0%) at 48 h post warming, respectively (P<0.05, P<0.001). Therefore, this study indicates that Hanwoo blastocysts can be cryopreserved more simple, efficient and successful by vitrification method using EM grid.
The present study was performed to improve the reproductive disturbance as well as the elimination of microbiological contamination for animals bred under conventional conditions followed by in vitro fertilization and embryo transfer techniques including embryo and sperm freezing, using a mouse strain(M. m. molossinus-tt@Kist) showing the abnormal behavior disorder derived from Korean wild mice (Mus musculus molossinus). Moreover, hematological and serum biochemical analyses were also carried out to obtain the basic data of this mouse strain The results are summarized as follows: 1. In comparison with hematological data, the numbers of RBC and platelet of this mouse strain were appeared as the higher value those that of the same aged inbred strains such as BALB/c, DBA/2, C57BL/6 and C3H /Hen. However, no differences were found in values of WBC, Hb and Ht. Moreover, total cholesterol of this strain showed a low value but triglyceride, total protein and albumin values were similar as in inbred strains. 2. The average numbers of superovulated oocytes treated with 2.5/2.5 IU and 5.0/5.0 IU of PMSG/hCG were 11.6 and 12.7, respectively. The fertilization rates of 2.5/2.5 IU PMSG /hCG treatment(87.9%) was higher than 5.0/5.0 IU treatment(52.0%) (p<0.05) and the developmental rate of 2 cell stage embryos were 외 so appeared as higher value 99.0% and 90.6%, respectively. 3. The rates of in vitro fertilization treated with frozen sperm(24.8%) was significantly lower than of that fresh sperm(87.9%), (p<0.05). 4. The five, six and ten heads of offspring were obtained from frozen-thawed 2 cell embryos by in vitro fertilized, 2 cell embryos from in vitro fertilized by frozen-thawed spermatozoa. and 2 cell embryos by in vitro fertilization, respectively. These offspring developed the expected disease about 2 weeks after birth, which was confirmed that the disease character of this mutant mouse strain was reliably reproduced. 5. MHV(Mouse hepatitis virus) and Staphylococcus aureus were successfully eliminated from conventional animals by in vitro fertilization-embryo transfer and the use of SPF recipient animals.
Kim, Eunseong;Choi, Bongki;Park, Youngjin;Cha, Ookhyun;Jung, Chungryel;Lee, Daeweon;Kim, Kwangho;Kim, Yonggyun
Korean journal of applied entomology
/
v.53
no.4
/
pp.355-365
/
2014
It has been unclear whether the diamondback moth, Plutella xylostella can overwinter in Korean field conditions. This study determined overwintering conditions of P. xylostella by conducting field exposure tests based on its cold tolerance and monitoring overwintering populations by direct examination of overwintering larval habitats and capturing adults with sex pheromone traps. In addition, the overwintering populations were analyzed using polymorphic genetic markers to trace their sources. When all immature stages of P. xylostella were exposed to $-5^{\circ}C$, which was the temperature much above their supercooling points, they significantly suffered with direct cold injuries, where larval stage was most tolerant to the cold injury. However, the exposure to $5^{\circ}C$ for a long period (4 weeks) did not give any significant cold injury to nonfeeding stages, while this treatment gave lethality to larval stage without diet. When all developmental stages of P. xylostella were exposed to open field conditions during winter, they exhibited significant decreases of survival rates. However, some protected and indoor conditions reduced the cold injuries and the diet provision significantly increased larval survival rates. Adult monitoring with sex pheromone during winter period indicated that the first captures were observed at similar periods at different locations (${\approx}260$ Km apart). The overwintering adults were captured until early April. Genetic variation of these overwintering populations was analyzed with polymorphic molecular markers, indicating significant genetic divergences among the overwintering populations. This study indicates that P. xylostella can overwinter in southern Korean fields or some protected greenhouses with host plants.
The aim of this study was to define the toxic effects of phenanthrene (PAHs) on the fertilization and normal embryogenesis rates in the two species of sea urchin (Hemicentrotus pulcherrimus and Mesocentrotus nudus). The sperm and fertilized eggs of both sea urchin species were exposed to serial dilutions of phenanthrene for 10 min and 48 hours, respectively. The fertilization rate and normal embryogenesis rate of H. pulcherrimus and M. nudus were decreased in a concentration-dependent manner. The EC50 for the fertilization rate of H. pulcherrimus and M. nudus was 17.48 mg L-1 and 16.21 mg L-1, and the EC50 for the normal embryogenesis rate was 2.99 mg L-1 and 0.36 mg L-1, respectively. Between the two species, H. pulcherrimus was more sensitive to phenanthrene exposure, and 48 h normal embryogenesis was the more sensitive endpoint. Therefore, the results of this study demonstrated that the exposure of both sea urchin species to phenanthrene caused alterations in egg fertilization and the early developmental stages.
To clarify the relationship between ambient water temperature and heading characteristics of rice (Oryza sativa L.), twelve of varieties rice were grown in a cold tolerance screening field where water temperature was controlled by continuous cold water irrigation system to test the cold tolerance of rice. When cold water was continuously irrigated with 5 cm of water depth, the water had stagnated for about three hours, and the water temperature increased gradually from inlet toward outlet in the experimental plot. The fluctuation of water temperature was well synchronized with that of ambient air temperature, and the water temperature near outlet became higher than the air temperature at the vegetative phase, while became lower at the reproductive phase of rice plant community. The leaf development rates on main culm increased by increased water temperature. The rice varities, Fukuhikari, Sangpungbyeo and YR3486-16-2 were more sensitive than the others in the response of leaf development to water temperature. However, Janack and Milyang 42 were comparatively less sensitive to water temperature in leaf development. Janack and Paro -white rices required longer days to develop one leaf on main culm at reproductive phase than at vegetative phase. Varietal difference in days required to develop one leaf on main culm of rice plant was more distinctive at the reproductive phase when water temperature was relatively lower than at the vegetative phase with relatively higher water temperature condition. No difference was found between the growth phases, vegetative and reproductive, in the response of average leaf developmental rates to water temperature under the similar air temperature condition. The estimated average days required to develop one leaf on main culm decreased by 1.3 day by 1$^{\circ}C$ increase in water temperature. Varietal differences in the total number of leaves on main culm depended upon the water temperature, in which the varieties such as Fukuhikari, Gwangmyeon-gbyeo, China 988, and YR3486-16-2 showed increased one leaf by increased water temperature, while Sobaekbyeo, Paro-white, Sangpungbyeo, Pungsanbyeo, Samgangbyeo, and Milyang 42 were kept at the same leaf number regardless of water temperature. However, the total leaf number on main culm and days to heading of Janack increased by increased water temperature. The other varieties showed the shortened days to heading by the increase in water temperature with noticeable varietal differences regardless of the variation in the total number of leaves on main culm.
The large amount of $CO_2$ emitted from mushrooms during incubation and developmental stages can be utilized in plant production systems as a $CO_2$ source. The objectives of this study were to measure the $CO_2$ emission and absorption rates of mushroom and lettuce, respectively, and to analyze the $CO_2$ concentrations at various ratios of mushroom and lettuce in a closed production system. The $CO_2$ emission rate of king oyster mushrooms (Pleurotus eryngii ( DC.) Qu$\acute{e}$l) and $CO_2$ absorption rate of lettuces (Lactuca sativa L. cv. Asia Heuk Romaine) were measured by using two closed acryl chambers ($1.0m{\times}0.8m{\times}0.5m$) in which indoor temperatures were maintained at $18^{\circ}C$ and $22^{\circ}C$, respectively. The lettuce was grown at a light intensity of PPF $340mol{\cdot}m^{-2}{\cdot}s^{-1}$ and with nutrient solution at EC $1.2dS{\cdot}m^{-1}$. The air was periodically circulated between the two chambers using a diaphragm pump. The $CO_2$ emission rate of the mushroom increased until the $15^{th}$ day after scratching (DAS) and then decreased. The rate also increased with increased indoor temperature. In particular, the $CO_2$ emission rate per fresh weight of fruit body increased by about 3.1 times after thinning compared to before thinning. In terms of $CO_2$ balance, the $CO_2$ emission rates from a bottle (950 mL) of the mushroom at 9, 12, and 14 DAS were equivalent to those of 3, 4.5, and 5.5 lettuce plants at 7, 10, and 12 DAT (days after transplanting), respectively. This work shows that balance in $CO_2$ concentration could be achieved using an appropriate ratio of the two crops in a closed production system.
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