• The Korean Journal of Nuclear Medicine
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• v.37 no.6
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• pp.418-427
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• 2003

Objects: $^{99m}-lactosylated$ human serum albumin (LSA) is a newly synthesized radiopharmaceutical that binds to asialoglycoprotein receptors, which are specifically presented on the hepatocyte membrane. Hepatic uptake and blood clearance of LSA were evaluated in rat with acute hepatic injury induced by dimethylnitrosamine (DMN) and results were compared with corresponding findings of liver enzyme profile and these of histologic changes. Materials and Methods: DMN (27 mg/kg) was injected intraperitoneally in Sprague-Dawley rat to induce acute hepatic injury. At 3(DMN-3), 8(DMN-8), and 21 (DMN-21) days after injection of DMN, LSA injected intravenously, and dynamic images of the liver and heart were recorded for 30 minutes. Time-activity curves of the heart and liver were generated from regions of interest drawn over liver and heart area. Degree of hepatic uptake and blood clearance of LSA were evaluated with visual interpretation and semiquantitative analysis using parameters (receptor index : LHL3 and index of blood clearance : HH3), analysis of time-activity curve was also performed with curve fitting using Prism program. Results: Visual assessment of LSA images revealed decreased hepatic uptake in DMN treated rat, compared to control group. In semiquantitative analysis, LHL3 was significantly lower in DMN treated rat group than control rat group (DMN-3: 0.842, DMN-8: 0.898, DMN-21: 0.91, Control: 0.96, p<0.05), whereas HH3 was significantly higher than control rat group (DMN-3: 0.731,.DMN-8: 0.654, DMN-21: 0.604, Control: 0.473, p<0.05). AST and ALT were significantly higher in DMN-3 group than those of control group. Centrilobular necrosis and infiltration of inflammatory cells were most prominent in DMN-3 group, and were decreased over time. Conclusion: The degree of hepatic uptake of LSA was inversely correlated with liver transaminase and degree of histologic liver injury in rat with acute hepatic injury.

• The Korean Journal of Nuclear Medicine Technology
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• v.16 no.2
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• pp.162-166
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• 2012

Purpose : Vitamin $B_{12}$ and Folate are for anemia work-up which is well known for its sensitivity of light; the screening manual also specifies to be careful with light conditions. According to this, our laboratory minimized the exposure of light when inspecting the Vitamin $B_{12}$ and Folate, but the exposure cannot be wholly blocked due to other various factors such as when conducting specimen segregation. Thus, this inspection is to identify to what extent light can influence and whether the exclusion of light is mandatory during the Vitamin $B_{12}$/Folate test. Materials and Methods : We have conducted two experiments of identifying the extent of light's influence when conducting the Vitamin $B_{12}$/Folate test and also when specimens are under preservation. These experiments were progressed with various concentrations of patients' specimens which were requested to our hospital in March 2012. The first experiment is to verify the results on Vitamin $B_{12}$/Folate dependent on light exposure during the experiment. In the process, we have compared the results of light exposure/exclusion during the incubation process after the reagent division. The second experiment is about the impact of light exposure on the results on Vitamin $B_{12}$/Folate during the preservation. For 1, 2, 7 days the light on the specimen were wholly blocked and were preserved under $-15^{\circ}C$ temperature refrigeration. Then, we compared the results of light-excluded specimen and the exposed one. Results : When conducting first experiment, there were no noticeable changes in the Standard and specimen's cpm, but for Vitamin $B_{12}$, the average result of specimen exposed to light increased 7.8% compare to that of excluded one's. Furthermore, in the significant level 0.05, the significance probability or the p-value was 0.251 which means it has no impact. For Folate, the result being exposed to light decreased 5.4%, the significance probability was 0.033 which means it has little impact. For the second preservation, the result was dependent on the light exposure. The first day of preservation of Vitamin $B_{12}$, the clinical material exposed to light was 11.6%, second day clinical material exposed to light was 10.8%, seventh day clinical material exposed to light increased 3.8%, the significance probability of the $1^{st}$, $2^{nd}$, $7^{th}$ day is 0.372, 0.033, 0.144 respectively, and which indicates that the $1^{st}$ and $7^{th}$ day seems to have no impact. For Folate's case, the clinical material exposed to light has increased 1.4% but hardly had impact, $2^{nd}$ day clinical material being exposed to light was 6.1%, $7^{Th}$ day clinical material being exposed to light decreased 5.2%. The significance probability of Folate on the $1^{st}$, $2^{nd}$, $7^{th}$ day is 0.378, 0.037, 0.217 respectively, and the $1^{st}$ day and the $7^{th}$ day seems to have no impact. Conclusion : After scrutinizing the impact of light exposure/exclusion, Vitamin $B_{12}$ has no impact, while Folate seems to have no noticeable influence but light exclusion is recommended due to its significance probability of 0.033 when conducting experiment. During the preservation, the $2^{nd}$ day result depend on the light exclusion seems to have impact or influence. However, to consider the complication of the experimental process, the experiment including technical errors is predictable. Hence, it is likely to have no impact of light. Nevertheless, it is recommendable to exclude the light during the long preservation as per the significance probability (p-value) of $1^{st}$ and $7^{th}$ day has been diminished.

• Journal of Periodontal and Implant Science
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• v.37 no.sup2
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• pp.427-445
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• 2007

To improve osseointegration at the boneto-implant interface, several studies have been carried out to modify titanium surface. Variations in surface texture or microtopography may affect the cellular response to an implant. Osteoblast-like cells attach more readily to a rougher titanium surface, and synthesis of extracellular matrix and subsequent mineralization were found to be enhanced on rough or porous coated titanium. However, regarding the effect of roughened surface by physical and mechanical methods, most studies carried out on the reactions of cells to micrometric topography, little work has been performed on the reaction of cells to nanotopography. The purpose of this study was to examme the response of osteoblast-like cell cultured on blasted surfaces and alkali treated surfaces, and to evaluate the influence of surface texture or submicro-scaled surface topography on the cell attachment, cell proliferation and the gene expression of osteoblastic phenotype using ROS 17/2.8 cell lines. In scanning electron micrographs, the blasted, alkali treated and machined surfaces demonstrated microscopic differences in the surface topography. The specimens of alkali treatment had a submicro-scaled porous sur-face with pore size about 200 nm. The blasted surfaces showed irregularities in morphology with small(<10 ${\mu}m$) depression and indentation among flatter-appearing areas of various sizes. Based on profilometry, the blasted surfaces was significantly rougher than the machined and the alkali treated surfaces (p$TiO_2$) were observed on alkali treated surfaces, whereas not observed on machined and blasted surfaces. The attachment morphology of cells according to time was observed by the scanning electron microscope. After 1 hour incubation, the cells were in the process of adhesion and spreading on the prepared surfaces. After 3 hours, the cells on all prepared surfaces were further spreaded and flattened, however on the blasted and alkali treated surfaces, the cells exhibited slightly irregular shapes and some gaps or spaces were seen. After 24 hours incubation, most cells of the all groups had a flattened and polygonal shape, but the cells were more spreaded on the machined surfaces than the blasted and alkali treated surfaces. The MTT assay indicated the increase on machined, alkali treated and blasted surfaces according to time, and the alkali treated and blasted surfaces showed significantly increased in optical density comparing with machined surfaces at 1 day (p<0.01). Gene expression study showed that mRNA expression level of ${\alpha}\;1(I)$ collagen, alkaline phosphatase and osteopontin of the osteoblast-like cells showed a tendency to be higher on blasted and alkali treated surfaces than on the machined surfaces, although no siginificant difference in the mRNA expression level of ${\alpha}\;1(I)$ collagen, alkaline phosphatase and osteopontin was observed among all groups. In conclusion, we suggest that submicroscaled surfaces on osteoblast-like cell response do not over-ride the one of the surface with micro-scaled topography produced by blasting method, although the microscaled and submicro-scaled surfaces can accelerate osteogenic cell attachment and function compared with the machined surfaces.

• Journal of Animal Science and Technology
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• v.46 no.4
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• pp.645-658
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• 2004

To investigate the effects of conjugated linoleic acid added diet feeding on CLA accumulation and quality characteristics of manufactured press ham using CLA accwnulated pork loin meat. The CLA used to add in diet was chemically synthesized by alkaline isomerization method with com oil. Pigs were divided into 5 treatment groups(4 pigs/group) and subjected to one of five treatment diets(0, 1.25% CLA for 2weeks, 2.5% CLA for 2weeks, 1.25% CLA for 4weeks and 2.5% CLA for 4weeks, CLA diets; total fed diets) before slaughter. Pork loin were collected from the animals(110kg body weight) slaughtering at the commercial slaughter house. Manufacture press ham using CLA accumulated pork loin meat were vacuum packaged and then stored during 1, 7, 14, 21 and 28 days at 4$^{\circ}C$. Samples were analyzed for general compositions, physico-chemical properties(pH, color, shear force value), TBARS. pH value of CLA treatment(T4) was increased significantly than that of oontrol(P<0.05). pH of control and CLA treatments were increased significantly as the storage period passed(P< 0.05). Crude fat content of CLA treatment groups was significantly higher than the control pork (P<0.05). Meat color(CIE $L^*$, $a^*$$b^*$

• Tuberculosis and Respiratory Diseases
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• v.58 no.1
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• pp.31-42
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• 2005

Background : Peroxiredoxins (Prxs) are a relatively newly recognized, novel family of peroxidases that reduce $H_2O_2$ and alkylhydroperoxide into water and alcohol, respectively. There are 6 known isoforms of Prxs present in human cells. Normally, Prxs exist in a head-to-tail homodimeric state in a reduced form. However, in the presence of excess $H_2O_2$, it can be oxidized on its catalytically active cysteine site into inactive oxidized forms. This study surveyed the types of the Prx isoforms present in the pulmonary epithelial, macrophage, endothelial, and other cell lines and observed their response to oxidative stress. Methods : This study examined the effect of exogenous, excess $H_2O_2$ on the Prxs of established cell lines originating from the pulmonary epithelium, macrophages, and other cell lines, which are known to be exposed to high oxygen partial pressures or are believed to be subject to frequent oxidative stress, using non-reducing SDS polyacrylamide electrophoresis (PAGE) and 2 dimensional electrophoresis. Result : The addition of excess $H_2O_2$ to the culture media of the various cell-lines caused the immediate inactivation of Prxs, as evidenced by their inability to form dimers by a disulfide cross linkage. This was detected as a subsequent shift to its monomeric forms on the non-reducing SDS PAGE. These findings were further confirmed by 2 dimensional electrophoresis and immunoblot analysis by a shift toward a more acidic isoelectric point (pI). However, the subsequent reappearance of the dimeric Prxs with a comparable, corresponding decrease in the monomeric bands was noted on the non-reducing SDS PAGE as early as 30 minutes after the $H_2O_2$ treatment suggesting regeneration after oxidation. The regenerated dimers can again be converted to the inactivated form by a repeated $H_2O_2$ treatment, indicating that the protein is still catalytically active. The recovery of Prxs to the original dimeric state was not inhibited by a pre-treatment with cycloheximide, nor by a pretreatment with inhibitors of protein synthesis, which suggests that the reappearance of dimers occurs via a regeneration process rather than via the de novo synthesis of the active protein. Conclusion : The cells, in general, appeared to be equipped with an established system for regenerating inactivated Prxs, and this system may function as a molecular "on-off switch" in various oxidative signal transduction processes. The same mechanisms might applicable other proteins associated with signal transduction where the active catalytic site cysteines exist.

• The Korean Journal of Petroleum Geology
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• v.14 no.1
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• pp.1-11
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• 2008

As conventional oil and gas reservoirs become depleted, interests for oil sands has rapidly increased in the last decade. Oil sands are mixture of bitumen, water, and host sediments of sand and clay. Most oil sand is unconsolidated sand that is held together by bitumen. Bitumen has hydrocarbon in situ viscosity of >10,000 centipoises (cP) at reservoir condition and has API gravity between $8-14^{\circ}$. The largest oil sand deposits are in Alberta and Saskatchewan, Canada. The reverves are approximated at 1.7 trillion barrels of initial oil-in-place and 173 billion barrels of remaining established reserves. Alberta has a number of oil sands deposits which are grouped into three oil sand development areas - the Athabasca, Cold Lake, and Peace River, with the largest current bitumen production from Athabasca. Principal oil sands deposits consist of the McMurray Fm and Wabiskaw Mbr in Athabasca area, the Gething and Bluesky formations in Peace River area, and relatively thin multi-reservoir deposits of McMurray, Clearwater, and Grand Rapid formations in Cold Lake area. The reservoir sediments were deposited in the foreland basin (Western Canada Sedimentary Basin) formed by collision between the Pacific and North America plates and the subsequent thrusting movements in the Mesozoic. The deposits are underlain by basement rocks of Paleozoic carbonates with highly variable topography. The oil sands deposits were formed during the Early Cretaceous transgression which occurred along the Cretaceous Interior Seaway in North America. The oil-sands-hosting McMurray and Wabiskaw deposits in the Athabasca area consist of the lower fluvial and the upper estuarine-offshore sediments, reflecting the broad and overall transgression. The deposits are characterized by facies heterogeneity of channelized reservoir sands and non-reservoir muds. Main reservoir bodies of the McMurray Formation are fluvial and estuarine channel-point bar complexes which are interbedded with fine-grained deposits formed in floodplain, tidal flat, and estuarine bay. The Wabiskaw deposits (basal member of the Clearwater Formation) commonly comprise sheet-shaped offshore muds and sands, but occasionally show deep-incision into the McMurray deposits, forming channelized reservoir sand bodies of oil sands. In Canada, bitumen of oil sands deposits is produced by surface mining or in-situ thermal recovery processes. Bitumen sands recovered by surface mining are changed into synthetic crude oil through extraction and upgrading processes. On the other hand, bitumen produced by in-situ thermal recovery is transported to refinery only through bitumen blending process. The in-situ thermal recovery technology is represented by Steam-Assisted Gravity Drainage and Cyclic Steam Stimulation. These technologies are based on steam injection into bitumen sand reservoirs for increase in reservoir in-situ temperature and in bitumen mobility. In oil sands reservoirs, efficiency for steam propagation is controlled mainly by reservoir geology. Accordingly, understanding of geological factors and characteristics of oil sands reservoir deposits is prerequisite for well-designed development planning and effective bitumen production. As significant geological factors and characteristics in oil sands reservoir deposits, this study suggests (1) pay of bitumen sands and connectivity, (2) bitumen content and saturation, (3) geologic structure, (4) distribution of mud baffles and plugs, (5) thickness and lateral continuity of mud interbeds, (6) distribution of water-saturated sands, (7) distribution of gas-saturated sands, (8) direction of lateral accretion of point bar, (9) distribution of diagenetic layers and nodules, and (10) texture and fabric change within reservoir sand body.

• Korean Journal of Heritage: History & Science
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• v.37
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• pp.285-307
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• 2004

Synthesize and examine petrological characteristic and geochemical characteristic by weathering formation of rock and progress of weathering laying stress on stone cultural properties of Unjusa temple of Chonnam Hwasun county site in this research. Examine closely weathering element that influence mechanical, chemical, mineralogical and physical weathering of rocks that accomplish stone cultural properties and these do quantification, wish to utilize by a basic knowledge for conservation scientific research of stone cultural properties by these result. Enforced component analysis of rock and mineralogical survey about 18 samples (pyroclastic tuff; 7, ash tuff; 4, granite ; 4, granitic gneiss; 3) all to search petrological characteristic and geochemical characteristic by weathering of Unjusa temple precinct stone cultural properties and recorded deterioration degree about each stone cultural properties observing naked eye. Major rock that constitution Unjusa temple one great geological features has strike of N30-40W and dip of 10-20NE being pyroclastic tuff. This pyroclastic tuff is ranging very extensively laying center on Unjusa temple and stone cultural properties of precinct is modeled by this pyroclastic tuff. Stone cultural propertieses of present Unjusa temple precinct are accomplishing structural imbalance with serious crack, and because weathering of rock with serious biological pollution is gone fairly, rubble break away and weathering and deterioration phenomenon such as fall off of a particle of mineral are appearing extremely. Also, a piece of iron and cement mortar of stone cultural properties everywhere are forming precipitate of reddish brown and light gray being oxidized. About these stone cultural properties, most stone cultural propertieses show SD(severe damage) to MD(moderate damage) as result that record Deterioration degree. X-ray diffraction analysis result samples of each rock are consisted of mineral of quartz, orthoclase,plagioclase, calcite, magnetite etc. Quartz and feldspar alterated extremely in a microscopic analysis, and biotite that show crystalline form of anhedral shows state that become chloritization that is secondary weathering mineral being weathered. Also, see that show iron precipitate of reddish brown to crack zone of tuff everywhere preview rock that weathering is gone deep. Tuffs that accomplish stone cultural properties of study area is illustrated to field of Subalkaline and Peraluminous, $SiO_2$(wt.%) extent of samples pyroclastic tuff 70.08-73.69, ash tuff extent of 70.26-78.42 show. In calculate Chemical Index of Alteration(CIA) and Weathering Potential Index(WPI) about major elements extent of CIA pyroclastic tuff 55.05-60.75, ash tuff 52.10-58.70, granite 49.49-51.06 granitic gneiss shows value of 53.25-67.14 and these have high value gneiss and tuffs. WPI previews that is see as thing which is illustrated being approximated in 0 lines and 0 lines low samples of tuffs and gneiss is receiving esaily weathering process as appear in CIA. As clay mineral of smectite, zeolite that is secondary weathering produce of rock as result that pick powdering of rock and clothing material of stone cultural properties observed by scanning electron micrographs (SEM). And roots of lichen and spore of hyphae that is weathering element are observed together. This rock deep organism being coating to add mechanical weathering process of stone cultural properties do, and is assumed that change the clay mineral is gone fairly in stone cultural properties with these. As the weathering of rocks is under a serious condition, the damage by the natural environment such as rain, wind, trees and the ground is accelerated. As a counter-measure, the first necessary thing is to build the ground environment about protecting water invasion by making the drainage and checking the surrounding environment. The second thing are building hardening and extirpation process that strengthens the rock, dealing biologically by reducing lichens, and sticking crevice part restoration using synthetic resin. Moreover, it is assumed to be desirable to build the protection facility that can block wind, sunlight, and rain which are the cause of the weathering, and that goes well with the surrounding environment.

• Korean Journal of Poultry Science
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• v.20 no.2
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• pp.93-105
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• 1993

This study was carried out to determine the concentrations of previtamin D$_3$(PreD$_3$), lumisterol$_3$(L3), tachystero1$_3$(73), vitamin D$_3$(VD$_3$) and provitamin D$_3$(ProD$_3$) in leg skins of broiler chicks exposed to UVB lights (maximum intensity at 297 nm) with dose of 0.204 or 0.409 mJ/$\textrm{cm}^2$(30 or 60 min irradiation) . The broiler Hubbard line day old chicks(2 dose $\times$9 elapsed time $\times$4 replica＋10 control＝82) were fed VD-deficient diet for 31 days in a windowless subdued light room. The skin was collected at 0, 6, 12, 18, 30, 42, 66, 90 or 138 hr after UVB irradiation. The skin lipid was extracted by 9％ ethyl acetate/n-hexane, and the fraction of VD$_3$ and its analogues was purified by Sep-Pak silica cartridge. The straight phase HPLC was utilized to analyze ProD$_3$ and its products. The mole ％(absolute level expressed in ng/$\textrm{cm}^2$) of PreD$_3$ in leg skin (epidermis＋dermis) was 4.67％(44 ng/$\textrm{cm}^2$) or 3.97％(37 ng/$\textrm{cm}^2$) right after UVB irradiation by 0.204 or 0.409 mJ/$\textrm{cm}^2$(30 or 60 min) at 15 cm distance, respectively. It content in leg skin at 0 hr after exposure was 7.24％(12 ng/$\textrm{cm}^2$) or 0.92％(9 ng/$\textrm{cm}^2$), respectively. The increase in irradiation dose did not affect proportionally the If synthesis.73 concentration in leg skin was 0.58％(S ng/$\textrm{cm}^2$) or 0.57％(6 ng/$\textrm{cm}^2$), respectively 0 hr after irradiation. The VD$_3$ in leg skin of birds exposed to UVB light with dose of 0.204 or 0.409 mJ/$\textrm{cm}^2$ was 2.13％ (21 ng/$\textrm{cm}^2$) or 0.97％ (16ng/$\textrm{cm}^2$), respectively at 0 hr after exposure, 2.72％(26ng/$\textrm{cm}^2$) or 3.84％(37ng/$\textrm{cm}^2$), respectively at 6 hr, and 4.30％ ((33ng/$\textrm{cm}^2$) or 6.40％(76ng/$\textrm{cm}^2$), respectively at 12 hr. The peak concentration of VD$_3$ was presented at 18 or 30 hr when 0.204 or 0.409 mJ/$\textrm{cm}^2$) was treated, respectively. It was shown that 18~30 hr were necessary for the thermal conversion of PreD$_3$ into VD$_3$ in the leg skin of broiler chicks. The ProD$_3$ contents in leg skins of negative control, 0.204 mJ/$\textrm{cm}^2$ and 0.409 mJ/$\textrm{cm}^2$ treated birds were 966, 948 and 815 ng/$\textrm{cm}^2$, respectively at right before and after UVB exposure. It was estimated that 18 or 151 ng/$\textrm{cm}^2$ of ProD$_3$ was isomerized to PreD$_3$, L$_3$, T$_3$ and VD$_3$ when exposed to 0.204 or 0.409 mJ/$\textrm{cm}^2$, respective)y. Consequently it was shown that when double dose of UVB light was applied to irradiate the chick body, more but not double synthesis of VD$_3$ and its analogues was occured in leg skin of brolier chicks.

• Korean Journal of Soil Science and Fertilizer
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• v.10 no.3
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• pp.103-134
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• 1977

The physiological and biochemical role of potassium for upland crops according to recent research reports and the nutritional status of potassium in Korea were reviewed. Since physical and chemical characteristics of potassium ion are different from those of sodium, potassium can not completely be replaced by sodium and replacement must be limited to minimum possible functional area. Specific roles of potassium seem to keep fine structure of biological membranes such as thylacoid membrane of chloroplast in the most efficient form and to be allosteric effector and conformation controller of various enzymes principally in carbohydrate and protein metabolism. Potassium is essential to improve the efficiency of phoro- and oxidative- phosphorylation and involve deeply in all energy required metabolisms especially synthesis of organic matter and their translocation. Potassium has many important, physiological functions such as maintenance of osmotic pressure and optimum hydration of cell colloids, consequently uptake and translocation of water resulting in higher water use efficiency and of better subcellular environment for various physiological and biochemical activities. Potassium affects uptake and translocation of mineral nutrients and quality of products. potassium itself in products may become a quality criteria due to potassium essentiality for human beings. Potassium uptake is greatly decreased by low temperature and controlled by unknown feed back mechanism of potassium in plants. Thus the luxury absorption should be reconsidered. Total potassium content of upland soil in Korea is about 3% but the exchangeable one is about 0.3 me/100g soil. All upland crops require much potassium probably due to freezing and cold weather and also due to wet damage and drought caused by uneven rainfall pattern. In barley, potassium should be high at just before freezing and just after thawing and move into grain from heading for higher yield. Use efficiency of potassium was 27% for barley and 58% in old uplands, 46% in newly opened hilly lands for soybean. Soybean plant showed potassium deficiency symptom in various fields especially in newly opened hilly lands. Potassium criteria for normal growth appear 2% $K_2O$ and 1.0 K/(Ca+Mg) (content ratio) at flower bud initiation stage for soybean. Potassium requirement in plant was high in carrot, egg plant, chinese cabbage, red pepper, raddish and tomato. Potassium content in leaves was significantly correlated with yield in chinese cabbage. Sweet potato. greatly absorbed potassium subsequently affected potassium nutrition of the following crop. In the case of potassium deficiency, root showed the greatest difference in potassium content from that of normal indicating that deficiency damages root first. Potatoes and corn showed much higher potassium content in comparison with calcium and magnesium. Forage crops from ranges showed relatively high potassium content which was significantly and positively correlated with nitrogen, phosphorus and calcium content. Percentage of orchards (apple, pear, peach, grape, and orange) insufficient in potassium ranged from 16 to 25. The leaves and soils from the good apple and pear orchards showed higher potassium content than those from the poor ones. Critical ratio of $K_2O/(CaO+MgO)$ in mulberry leaves to escape from winter death of branch tip was 0.95. In the multiple croping system, exchangeable potassium in soils after one crop was affected by the previous crops and potassium uptake seemed to be related with soil organic matter providing soil moisture and aeration. Thus, the long term and quantitative investigation of various forms of potassium including total one are needed in relation to soil, weather and croping system. Potassium uptake and efficiency may be increased by topdressing, deep placement, slow-releasing or granular fertilizer application with the consideration of rainfall pattern. In all researches for nutritional explanation including potassium of crop yield reasonable and practicable nutritional indices will most easily be obtained through multifactor analysis.