Cho, Jae Won;Lim, Chun Kyu;Shin, Mi Ra;Bang, Kyoung Hee;Koong, Mi Kyoung;Jun, Jin Hyun
Clinical and Experimental Reproductive Medicine
/
v.33
no.3
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pp.171-178
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2006
Objective: Human embryonic stem (ES) cells have a great potential in regenerative medicine and tissue engineering. The human ES cells could be differentiated into specific cell types by treatments of growth factors and alterations of gene expressions. However, the efficacy of guided differentiation and isolation of specific cells are still low. In this study, we characterized isolated cells from differentiated human ES cells by magnetic activated cell sorting (MACS) system using specific antibodies to cell surface markers. Methods: The undifferentiated hES cells (Miz-hESC4) were sub-cultured by mechanical isolation of colonies and embryoid bodies were spontaneously differentiated with DMEM containing 10% FBS for 2 weeks. The differentiated cells were isolated to positive and negative cells with MACS system using CD34, human epithelial antigen (HEA) and human fibroblast (HFB) antibodies, respectively. Observation of morphological changes and analysis of marker genes expression were performed during further culture of MACS isolated cells for 4 weeks. Results: Morphology of the CD34 positive cells was firstly round, and then it was changed to small polygonal shape after further culture. The HEA positive cells showed large polygonal, and the HFB positive spindle shape. In RT-PCR analysis of marker genes, the CD34 and HFB positive cells expressed endodermal and mesodermal genes, and HEA positive cells expressed ectodermal genes such as NESTIN and NF68KD. The marker genes expression pattern of CD34 positive cells changed during the extension of culture time. Conclusion: Our results showed the possibility of successful isolation of specific cells by MACS system from undirected differentiated human ES cells. Thus, MACS system and marker antibodies for specific cell types might be useful for guided differentiation and isolation of specific cells from human ES cells.
The morphological, anatomical and physiological traits were eximined for Populus alba ${\times}$ glandulosa which is an important planting species in Korea. The results obtained are as follows: 1. External characters in the leaf shape and chaff shape in the catkin were inherited as incomplete dominance but nectar gland was inherited as dominance. 2. Among the 15 selected clones, 9 clones were male, 2 clones female and 2 clones monoecious. 3. There were well-developed cork layers and bast fiber bundles in the bark. 4. Primordial leaves composed of 3 layers of cells and those undifferentiated into palisade and spongy parenchymas differed in its origin. 5. Leaf scare consisted of two kinds of tissues; one is connected to vascular bundle and the other not to vascular bundle. Tissues which had been connected to vascular bundle were isolated with only 2 or 3 layers of cork cells from the outside. 6. There was complicated arrangement in the vascular bundle of petioles. 7. Growth of the hybrid was sensitively influenced by external temperature, day-length and amount of light. In particular, it was apparent in height growth. 8. Flatness, loam soils and a $60{\times}60cm$ spacing might be best factors for the growth of P. alba ${\times}$ glandulosa. 9. The rooting of 15 clones was dependant upon external factors. 10. The growth of P. alba ${\times}$ glandulosa was best at around 80% of soil moisture content on the basis of plot water capacity. 11. Temperature difference between inside and outside stems below 100cm during the winter was the greatest at the south among seasons and among directions. 12. The sap movement was markedly influenced by air temperature, relative humidity in forest stand and moisture content in stem. 13. Total sugars in the cortex changed with season but did not differ in the dircetion of the stem. 14. Isoperoxidase variations in the leaf were different among 15 clones. Thus, it may be useful as a criterium for clonal identification. 15. The rate of soil moisture content decreased at a rapid slope was faster than that at a slow slope. Poor growth of P. alba ${\times}$ glandulosa at the slope was probably due to depletion of soil moisture.
This study aimed to find out suitable culture conditions for the differentiation of human amniotic membrane-derived stem cells(HAM) into hepatocyte-like cells. Almost homogenous population of fibroblast-like cells was successfully isolated from the amniotic membrane. In comparison to the non-coated plates and in the absence of insulin/transferrin/selenium(ITS), HAM cultured on the fibronectin-coated plates and in the presence of ITS showed the more intense immunocytochemical staining against the albumin. Addition of both fibroblast growth factor(FGF)-1 and -2 to the differentiation medium gave stronger staining compared to the treatment with FGF-1 or -2 alone. Periodic acid Schiff's base staining of glycogen and morphological turnover of fibroblast-like appearance of HAM into round shape matched the results of immunocytochemical studies. When the efficiency of two-step culture method was examined on the differentiation of HAM into hepatocyte-like cells, all of the results of immunocytochemical staining, periodic acid Schiff's staining and morphological change exhibited effective hepatic differentiation of HAM compared to the continuous culture method. Immunoblot analyses of HAM- conditioned media against the albumin showed that the culture of HAM in the presence of both ITS and fibronectin always gave a stronger staining intensity than those in the absence of them, and that the addition of ether mixture of FGF-4 and either FGF-2 or transforming growth $factor(TGF)-{\alpha}$ to the culture medium significantly enhanced the albumin secretion by HAM. Based on these observations, it is suggested that HAM could differentiate into hepatocyte-like cells under a culture condition consisting of fibronectin and ITS, and addition of FGF-4 with either one of FGF-2 or $TGF-{\alpha}$ could enhance the hepatic differentiation of HAM.
The importance of leaf area as related to transpiration and photosynthesis is generally recognized. In general, a compound leaf of soybean consist of one main leaflet and two side leaflets from each node of the stem. Takahashi and Fukuyama (1919) classified soybeans into three types, namely the long leaf type, round leaf type, and intermediate type, in which the last one had round leaves at the base and long leaves in the upper part of the stem. Nagai (1925) and Takahashi (1935). dealt with the genetics of the leaf form and association with other characters. The closely relationships, the correlation coefficients from 0.64 to 0.73, were shown between the leaf area and the soybean yield in the experiments by Nagai (1942). Nagata (1950) also tested the varietal differences of the variation of leaf length and its ratio to the leaf width on the nodes of stem, and finally divided varieties into five types. Three methods of measuring area of strawberry leaves were used by Darrow (1932). The first involved determining a factor to be used with length or length ${\times}$width measurements. The second method involved placing leaves on pieces of cardboard of known area cut to the shape of the leaves. Direct use of the planimeter on intact leaves was Darrow's third method. Miller (1938) enumerated several methods to determine the leaf surface area in plants, some of which were extremely laborious and required removing leaves from plants. They included tracing outlines of leaves on paper and measuring the enclosed area with a planimeter or cutting out the traced areas and comparing the weights obtained with the weight of a known paper. Another method involved placing the form of the leaf on sensitized paper with the area being determined by measuring or weighing as above. Miller further stated that the photoelectric cell can also be utilized to estitmate leaf area. Working with field beans, Davis (1940) found that 0.004517 (length ${\times}$ width) of the center leaflet was the most nearly accurate of four methods attempted. A simple procedure to measure leaf area in corn was devised 1 y Montgomery (1911) and used by Kiesselbach (1950). The formula was length ${\times}$ width ${\times}$ 0.75. Stickler et al. (1961) have successfully used length times width ${\times}$ 0.747 to estimate area of grain sorghum leaves. Bhan and Pande(1966) has also used length ${\times}$ width ${\times}$ 0.802 to determine leaf area of rice varieties. The main objectives of the present investigation were to develop an accurate, rapid method to determine leaf area in soybean varieties and to examine certain data associated with leaf area determinations.
This experiment was executed to investigate and compare qualitative and quantitative anatomical features in compression wood, opposite wood, and side wood formed in a bent stem, a straight branch, and an exposed horizontal root of Korean red pine(Pinus densiflora S. et Z.). The respective four discs containing compression wood taken at 20cm interval both in stem and branch as well as a disc containing well developed compression wood from horizontal root were analyzed. Percentage of compression wood and eccentricity showed decreasing tendency with the increasing distance in height direction of stem and length direction of branch. The qualitative anatomical features of compression wood appeared to differ from those of side and opposite wood in very gradual tracheid transition from earlywood to latewood, roundish tracheid shape on cross surface, tracheid distortion at tip on radial surface, existence of intercellular space, and helical cavity in tracheid wall. And the differences in these qualitative features among the compression wood, opposite wood, and side wood became less intensive with the decreasing trends in percentage of compression wood and eccentricity. The quantitative anatomical features in compression wood also appeared to be wider in that respective widths of fusiform and uniseriate ray than those of opposite and side wood, but the heights of fusiform and uniseriate ray in compression wood were smaller than in opposite and side wood. The number of horizontal resin canal(fusiform ray) and uniseriate ray, however, showed no differences among the compression wood, opposite wood, and side wood. And the number of vertical resin canal in unit area, $4{\pi}mm^2$ of compression wood was fewer than that in opposite wood, whereas numerous vertical resin canals contained in a growth ring. These rays of compression wood seemed to be characterized by smaller height and wider width than those of opposite and side wood.
It was revealed that the morphology and projection pattern of terminal arbors from single primary afferent are different among distinct fiber types, functional types and the different subdivision of trigeminal sensory nucleus complex(TSNC). But it was not identified the ultrastructural morphology and synaptic connections of terminal arbors from each primary afferent within TSNC. So we employed the intra-axonal horseradish peroxidase(HRP) injection technique to define the terminal arbors of primary afferent fiber from slowly adapting mechanoreceptors in the periodontal ligament of the cat, and examined 66 labeled terminal arbors within the rostrodorsomedial part(Vo.r) of the trigeminal nucleus oralis, electromicroscopically with 90nm serial sections. All the boutons labelled with HRP contained clear, spherical and uniform sized synaptic vesicles(diameter : $47.66{\pm}3.58nm$ ). Most of the labelled boutons were boutons en passant type and they were connected by unmyelinated axonal strand. In which neurofilament and microtubule was not developed but occasionally contained synaptic vesicle in contrast to the myelinated axon. The size of the labelled bouton was relatively small(long diameter : $1.46{\pm}0.24{\mu}m$, short diameter $0.85{\pm}0.26{\mu}m$, average diameter $1.15{\pm}0.24{\mu}m$) and the shape of which varied from dome to elongated shape, but scalloped glomerulus shape was not developed. Each primary ending in Vo.r made synapse with one or two neuronal propiles(average : $1.11{\pm}0.31$), of which, 89.4% of labelled boutons made synapse with only one neuronal pro pile, the remainder, 10.6% of labelled boutons, made synapse with two neuronal propile. So characteristically they made very simple synapse. Most of labelled boutons(80.03%) made asymmetrical synapse only with dendritic shaft or spine, and 6.1% of labelled boutons received symmetrical synapse from pleomorphic vesicle containing axonal ending(p-ending). So presynaptic inhibiton was relatively scarce. Synaptic triad, in which a p-ending is presynaptic both pre-and post-synaptic element of the axo-dendritic contact from the labelled primary ending was not observed.
Kim, Kyung Ah;Na, Kyung Soo;Seo, Seok Jin;Lee, Je Hee
The Journal of Korean Society for Radiation Therapy
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v.29
no.1
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pp.57-68
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2017
Purpose: The purpose of this study was to compare volumetric modulated arc therapy(VMAT) with fixed-field intensity modulated radiation therapy(IMRT) using non-coplanar beam when the shape of target is irregular and the location is adjacent to organ at risk(OAR). Materials and Methods: The subjects of this study were a total of 6 patients who had radiation therapy for whole scalp(2 patients), partial scalp(2 patients), and whole ventricle(2 patients) by True Beam STX(Varian Medical Systems, USA). VMAT plans consisted of coplanar or non-coplanar arcs which can minimize the volume of OAR included in beamlets. All fixed-field IMRT plans consisted of non-coplanar beams using more than 2 angles of Couch. Results: The VMAT and IMRT plans were compared with regard to the maximum dose of both lens, both optic nerves, optic chiasm, and brain stem and the mean dose of both eyeballs and hippocampus. VMAT plans showed higher dose than ncIMRT plans at more than 6 of all OARs in every patient, and the ratio was from 1.1 times to 8.2 times. In case of total scalp and partial scalp, the volume of brain which received more than 20 Gy in the VMAT plans was 2 times larger than the volume in the ncIMRT plans. In case of whole ventricle, there was no significant difference. Target coverage was satisfied in both plans($PTV_{100%}=95%$). The maximum dose in target volume and required monitor unit(MU) of ncIMRT were higher than them of VMAT plans. Conclusion: Even though ncIMRT is less efficient than VMAT with regard to required MU and treatment time, the dose to OARs is much lower than VMAT and PTV Coverage is similar with VMAT. If the shape of target is irregular and location is adjacent to OAR, comparison VMAT plan with ncIMRT plan deserves to be considered.
A maize line was selected in 1979 among 1000 Korean local maize lines collected in 1977. The selected maize line was characterized by having three to four tillers and eight to 10 ears on each individual plant. The line was assumed to have a great potential as a silage crop. The investigation was conducted as one of the serial studies on the Korean maize collected lines to provide basic information on the genetic variabilities of the multi-eared and tillered (MET) line and on other agronomic characters, prior to use the line as material for future breeding works for silage crop. The MET line and Suwon #19, single cross hybrid, as check variety were planted on May 1, 15 and 30, in three different levels of plant populations. The results obtained were summarized as follows: 1. The genetic variabilities of multi-ear and tillering habits were greater than environmental variabilities. 2. Total dry leaf weight of individual plant of MET line was also significantly higher than that of Suwon #19. 3. The mean number of tillers and ears bearing on the individual plant of MET line varied greatly with plant densities. The number of tillers and ears was on the average 2.9 and 7.0, respectively, when planted in 60cm. by 60cm. 4. The total dry matter and dried stem weight of the individual plant on MET line were comparable to those of Suwon #19. 5. The kernel weight from the individual plant of MET line was 5 to 40% less than that of Suwon #19, depending upon the plant densities. 6. The Kernel to stover ratio was higher for Suwon #19 than for the MET line. (41% to 35%). 7. The MET line had shown first tiller two weeks after planted on May 1. The second and third tillers appeared three to five days after the appearance of the first tiller. 8. The MET line was very specific in tillering habits. All the tillers were borne on the first few nodes of main stem below the soil surface. 9. The tillering habits of MET line were vigorous in the early part of the growing season, but less vigorous in the later part of the growing season. The number of efficient tillers bearing useable ears, was around two to three, when planted in 60cm. by 60cm. 10. The difference of plant height between main stem and first few tillers was around 10cm. 11. The ear size of MET line was around one-third of the major corn belt hybrids. The shape of ear of MET line was conical, with different diameter. 12. The kernel of the MET line was flinty with small soft starch patches on the endosperm part. 13. The 100 kernel weight was around 15gr., which is about one half of the major high yielding hybrids. 14. The ear height of MET line was comparatively higher than that of Suwon #19. 15. Significantly high and positive phenotypic correlation coefficients were obtained among major plant characters. 16. The growth rate of MET line was slower than that of Suwon #19. 17. MET line and Suwon #19 were both heavily infected with black streaked mosaic virus.
In this study, the major agronomic traits were investigated and RAPD technique was applied for the analysis of the genetic relations between the native ginsengs collected from Poonggi and Geumsan provinces in Korea. The main morphological traits were measured for a total of 54 collections of native ginseng from two areas based on UPOV standard. A total of 58 collections consisting of twenty-one native ginsengs collections from Poonggi area, twenty-nine collections from Geumsan area and four varieties of P. quinquefolium, P. japonicum, Chunpoong and Hwangsuk as controls were analyzed and clustered by RAPD. The results indicated that 01-9, 01-35 and 01-44 collections from Poonggi area were grouped into Geumsan area, while 332001, 332002 and 332003 collections from Geumsan area were grouped into Poonggi area. On comparison to the similarity of Poonggi collections (73-95%), the Geumsan collections showed 65-86% similarity in the population. Thus, the cluster should be applied according to the number of stem, number of leaves per stem and leaflet shape on the regionally native ginseng collections. The fourteen primers such as OPA02, OPA07, OPC08, OPD11, OPD20 and so on, will be used to select the native ginseng in the future studies.
Forage productivity of Job's tears (Coix lachryma-jobi L.) and adlay (C. lachryma-jobi var. mayuen (ROMAN.) STAPF) and a hybrid corn (Kwangog) and sorghum-sudangrass hybrid (Pioneer 988) was compared in the southern part of Korea where black streaked dwarf virus' (BSDV) is prevalent. Emergence of Job's tears and adlay was more delayed compared to a corn or sorghum-sudangrass hybrid under the drought field conditions. Early growth and regrowth of Job's tears and adlay were less than those of sorghum-sudangrass hybrid. Fresh weight, dry weight, and digestible dry matter of Job's tears and adlay cut once approximately 40 days after flowering were much higher than those of two cuts at every flowering stage. Dry weight of one-cut Job's tears and ad lay harvested 40 days after flowering was similar to the total dry weight of three-cut sorghum-sudangrass hybrid but higher than that of silage corn. Digestible dry matter production was greater in the order of sorghum-sudangrass, corn, one-cut and two-cut Job's tears and adlay. Corn was severely infected with BSDV, but digestible dry matter was relatively high due to its higher in vitro dry matter digestibility. Among the Job's tears and adlay varieties, a variety of Job's tears with globular fruit shape was best for forage production due to its taller plant height, thick stem, wider leaf blades, higher leaf blade/total dry weight ratio, and higher forage yield. However, hulled grain yield of ad lay varieties was higher than that of Job's tears due to a higher hulled/rough grain ratio although they had similar rough grain yields.
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