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Fluoxetine Simultaneously Induces Both Apoptosis and Autophagy in Human Gastric Adenocarcinoma Cells

  • Po, Wah Wah;Thein, Wynn;Khin, Phyu Phyu;Khing, Tin Myo;Han, Khin Wah Wah;Park, Chan Hee;Sohn, Uy Dong
    • Biomolecules & Therapeutics
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    • v.28 no.2
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    • pp.202-210
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    • 2020
  • Fluoxetine is used widely as an antidepressant for the treatment of cancer-related depression, but has been reported to also have anti-cancer activity. In this study, we investigated the cytotoxicity of fluoxetine to human gastric adenocarcinoma cells; as shown by the MTT assay, fluoxetine induced cell death. Subsequently, cells were treated with 10 or 20 µM fluoxetine for 24 h and analyzed. Apoptosis was confirmed by the increased number of early apoptotic cells, shown by Annexin V- propidium iodide staining. Nuclear condensation was visualized by DAPI staining. A significant increase in the expression of cleaved PARP was observed by western blotting. The pan-caspase inhibitor Z-VAD-FMK was used to detect the extent of caspase-dependent cell death. The induction of autophagy was determined by the formation of acidic vesicular organelles (AVOs), which was visualized by acridine orange staining, and the increased expression of autophagy markers, such as LC3B, Beclin 1, and p62/SQSTM 1, observed by western blotting. The expression of upstream proteins, such as p-Akt and p-mTOR, were decreased. Autophagic degradation was evaluated by using bafilomycin, an inhibitor of late-stage autophagy. Bafilomycin did not significantly enhance LC3B expression induced by fluoxetine, which suggested autophagic degradation was impaired. In addition, the co-administration of the autophagy inhibitor 3-methyladenine and fluoxetine significantly increased fluoxetine-induced apoptosis, with decreased p-Akt and markedly increased death receptor 4 and 5 expression. Our results suggested that fluoxetine simultaneously induced both protective autophagy and apoptosis and that the inhibition of autophagy enhanced fluoxetine-induced apoptosis through increased death receptor expression.

젖산균의 Plasmid DNA 분리방법 및 Electroporation에 의한 젖산균의 형질전환에 관한 연구

  • Kim, Seon-Gi
    • 한국유가공학회:학술대회논문집
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    • 1997.05a
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    • pp.41-61
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    • 1997
  • 젖산균의 유전자 연구를 촉진하기 위해 간단하고 신속한 plasmid DNA의 분리방법과 electro-poration을 이용하여 vector plasmid의 간단하고 신속한 전이방법을 얻기 위해 젖산균의 형질전환에 영향하는 요인에 대하여 연구하였으며 연구결과는 다음과 같다. 1. O'Sullivan과 Klaenhammer의 방법을 개선하여 젖산균 plasmid DNA의 분리에 좋은 결과를 얻을 수 있는 신속하고 쉬운 방법을 고안하였으며, genomic DNA 분리에 이용되는 guanidium thio-cyanate 처리방법을 plasmid의 분리에 적용할 수 있었다. 2. L. casei, L. acidophilus. L. delbruekii var. bulgaricus. L. brevis와 L. plantarum 균주에서 plasmid를 확인하였으며, 돼지 분에서 분리된 L. lactis ssp. lactis. L. fermentum과 L. plantarum에서도 plasmid를 분리 확인하였다. 3. Lactococci의 plasmid분리는 lactobacilli와는 달리 mutanolysin의 처리없이도 잘 되었으며, L. lactis ssp. lactis와 Ent. faecalis에서 plasmid를 확인하였다. 4. E. coli plasimd 분리에 이용되는 MPS membrane filter 방법으로 젖산균 plasmid pLZ12의 분리가 가능하였으나, 세포파편이 filter를 막아 사용에 어려움이 있는 것으로 확인되었다. 5. Plasmid 분리없이 electroporation을 이용한 세포 대 세포 전이법으로 간편하고 빠르게 E. coli DH5${\alpha}$에 E. coli Jm109의 plasmid pBX19, pBR322를 전이시켰다. 6. L. lactis ssp. lactis 균주에 lysozyme 처리시 30${\sim}$80%의 생존율을 보였으며, 대부분의 L. acidophilus 균주의 경우 약 70%의 생존율을 보였다. L. casei 102S의 경우는 45분간 처리 시에도 100%의 생존율을 보였다. 8. L. lactis ssp. lactis 균주에 pLZ12를 6.0kV에서 전이시킨 결과 12.5kV에서보다 형질전환 효율이 훨씬 높았으며 lysozyme 처리에 의해 형질전환 효율이 증가되었다. 9. L. acidophilus 균주에 pLZ12를 전이시 6.0kV에서는 전이가 모두 이루어졌으나, 12.5kV에서는 L. acidophilus WIESBY와 NCFM에서 전이가 이루어지지 않았으며, lysozyme 처리 후 pLZ12를 전이시켰을 때 12kV보다 6.0kV에서 형질전환 효율이 증가되었다. 10. Gene Pulser와 Progenitor II를 사용하여 pLZ12를 L. lactis ssp. lactis 균주에 전이하였을 때 Gene Pulser에 비해 Progenitor II의 형질전환 효율이 현저히 떨어졌다. L. acidophilus HY7008과 HY7001은 두 기기 모두 형질전환이 이루어졌으나, L. acidophilus WEISBY와 NCFM은 Progeni-tor II에서 전이가 일어나지 않았으며, Gene Pulser에서 전이균주를 얻어 두 electroporator간에 형질전환 효율의 차이를 보였다. 11. L. casei 102S에 pLZ12를 electroporation시 낮은 전압에서 형질전환 효율이 비교적 좋았으며, 배양 시기를 달리하여 전이시켰을 때 대수생장 말기의 세포가 형질전환 효율이 좋았다. 12. L. casei 102S세포를 각각 10% glycerol, EB, 2차 증류수 등에 녹여 electroporation을 실시하였을 때 각각 $3.8{\times}10^3$, $5.0{\times}10^2$,1.5${\times}10^2$cfu의 형질전환 효율을 보였으며, 1.0mM HEPES, TE buffer를 사용하였을 때에는 전이가 이루어지지 않았다. 13. Plasmid pLZ12의 농도를 달리하여 electroporation을 하였을 때 형질전환 효율이 농도에 비례하여 증가하였다. 14. L. casei 102S에 대수생장 말기의 세포를 채취하여 10% glycerol, 200 Ohms, 25 ${\mu}$FD, 10kV/cm로 plasmid pLZ12를 electroporation할 때 최대 형질전환 효율인 3.8${\times}$10$^{3}$cfu를 얻었으며, lysozyme 처리가 다른 젖산균과는 달리 형질전환 효율을 증가시키지 못하였다. 15. L. casei 102S 세포를 10% glycerol과 EB에 녹여 -20$^{\circ}C$에서 냉동시킨 다음 1일과 7일 후의 세포를 electroporation한 결과 냉동시 세포에 손상을 주는 것으로 인식되었다.

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Potential as a Geological Field Course of Mt. Geumdang located in Gwangju, Korea (광주광역시에 위치한 금당산의 지질학습장으로서 활용성)

  • Ahn, Kun Sang
    • Journal of the Korean earth science society
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    • v.34 no.3
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    • pp.235-248
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    • 2013
  • The purpose of this study is to investigate a feasibility of a small mountain as a field work site on geological features in Earth sciences classes at all levels. Mt. Geumdang with the height of 305 meters from the sea level is located in the metropolitan city of Gwangju, southern part of Korea. The study reviews the human and natural geography, geological features, geomorphic resources, landscapes, and conveniences of the mountain for a possibility of meaningful field work. The population within the distance of 5 km from the mountain stands at about 620,000 and 170,000 of them are students and teachers. Mt. Geumdang has a warm temperature climate with low rainfall throughout the year, so it seems suitable for a field survey. Road network and public transportation system around the area are well-developed and easily accessible. Mt. Geumdang shows various rock type and geological structures. The basement rock is Gwangju granite, which is plutonic body of the Jurassic period. Also, granophyre (micrographic granite) and various volcanic rocks distributed as bedded tuff, lapilli tuff, and rhyolite of the Cretaceous period. Many andesitic and felsic dykes were intruded into the rock by joint system. In Mt. Geumdang, many geomorphic resources are found such as U shaped mountain, joint, fault, lamination, gnamma, tor, cliff, groove, block stream and block field, regolith, and saprolite. It has a beautiful mountain scenery including the view of whole shape of Mt. Mudeung, panoramic view of the town, Pungam lake, World Cup stadium and sunrise and sunset. Furthermore, the area has ecologic study facilities related to geology, emergency medical and convenience facilities for field works. In conclusion, Mt. Geumdang is highly feasible for geological field studies at all levels.

Construction of a Lactococcal Shuttle/Expression Vector Containing a $\beta$-Galactosidase Gene as a Screening Marker (선별마커로써 $\beta$-Galactosidase 유전자를 포함한 Lactococcus용 셔틀/발현 벡터 제조)

  • Han Tae Un;Jeong Do-Won;Cho San Ho;Lee Jong-Hoon;Chung Dae Kyun;Lee Hyong Joo
    • Microbiology and Biotechnology Letters
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    • v.33 no.4
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    • pp.241-247
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    • 2005
  • A new lactococcal shuttle/expression vector for lactococci, pWgal13T, was constructed using a $\beta$-galactosi-dase gene (lacZ) from Lacfococcus lactis ssp. lactis ATCC 7962 as a screening marker. The pWgal 13T was introduced into Escherichia coli DH5a and L. lactis MG1363, and was easily detected by the formation of blue colonies on a medium containing X-gal without any false transformants. Also, the quantitatively lacZ activity of pWgal13T was measured in L. lactis ssp. cremoris MG1363, and was found to be four times higher than that of L. lactis ssp. lactis ATCC7962 grown on a medium containing glucose, which shows that the lacZ gene of pWgal13T can be used for the efficient screening of L. lactis on general media. The pWgal13T was equipped with a lactococcal replicon of pWV01 from L. lactis Wg2, the new promoter P13C from L. lactis ssp. cremoris LM0230, multiple cloning sites, and a terminator for the expression of a relevant gene. The vee-tor pWgal13T was used for the expression of the EGFP gene in E. coli and L. lactis. These results show that the lactococcal expression/shuttle vector constructed in the present study can be used for the production of foreign proteins in E. coli and L. lactis.

Analysis of Producing of Thermostable Alkaline Protease using Thermoactinomyces sp. E79 (Thermoactinomyces sp. E79를 이용한 내열성 Alkaline 단백질 분해효소 생산:환경인자의 영향)

  • 정상원;박성식;박용철;오태광
    • Microbiology and Biotechnology Letters
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    • v.28 no.3
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    • pp.167-171
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    • 2000
  • Analysis of Production of Thermostable Alkaline Protease using Thermoactinomyces sp. E79. Jung, Sang Won, Sung-Sik Park, Yong-Cheol Park" Tae Kwang Oh2, and Jin-Ho Seo*, Department of Food Science and Technology, Seoul National University, Suwon 441-744, Korea, 1lnterdisciplinary program [or Biochemical Engineering & Biotechnology, Seoul National Univer5it}~ Seoul 151 "7421 Koreal 2Microbial Enzyme RU, Korea Research Institute of Bioscience & Biotechnology, Po. Box 1151 Yusong, Taejon 305"6001 Korea - This research was undertaken to analyze fermentation properties of Thermoactinomyces sp. E79 for production of a thermostable alkaline protease, which is able to specifically hydrolyze defatted soybean meal (DSM) to amino acids. TIle optimum pH for cell growth and protease production was pH 6.7, Thermoactinomyces sp. E79 did not grow at pHlO Among carbon sources tested, soluble starch was the best for protease production, while glucose repressed protease production. Tryptone was found to be the best nitrogen source for cell growth and soytone was good tor protease production. Oxygen transfer rate played an important role in producing thermostable alkaline protease. Ma'<..imum values of 6.58 glL of dry cell weight and 43.0 UJmL of protease activity were obtained in a batch fermentation using a 2.5 L jar fermentor at 1.93 X 102 hr-l of volumetric oxygen transfer coeff'jcient (kLa). Addition of 200 mgIL humic acid to the growth medium resulted in 1.64 times higher protease activity and 1.77 times higher cell growth than the case without humic acid addition.

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Total Dietary Fiber and Mineral Absorption

  • Gordon, Dennis-T.
    • Journal of Nutrition and Health
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    • v.25 no.6
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    • pp.429-449
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    • 1992
  • The consumption of foods rich in TDF should not be associated with impaired mineral absorp-tion and long-term mineral status. In surveys of populations consuming high amounts of TDF e.g Third World populations and vegetarinas gross deficiencies in mineral nutrition have not been noted. If mineral status is low among these groups it is most likely caused by the inadequacy or imbalance of the diet and not by the TDF. The key word is interaction which should be inte-rpreted in dietary imbalances that produce nut-rient deficiencies. There are no strong data to support the concept that TDF inhibits mineral absorption through a binding chelation mechanism. Limited data sug-gest that positively charged groups on polymers such as chitosan and cholestyramine will decrease iron absorption in humans and animals. Because TDF does not contain positively charged groups future research should be directed at the possible role of protein consumed along with TDF and the combination of effects on mineral nutrition Phytic acid is acknowledged as a potent chela-tor of zinc. However its association with zinc and its propensity to lower Zn bioavaiability may enhance the absorption of other elements notably copper and iron. The importance of interactions among nutrients including TDF will gain addi-tional attention in the scientific community. Soluble and insoluble dietary fiber function di-fferently in the intestine. Insoluble fibers accele-rate movement through the intestine. Soluble die-tary fibers appear to regulated blood concentra-tions of glucose and cholesterol albeit by some unknown mechanism. In creased viscosity produ-ced by the SDF in the intestine may provide an explanation of how this class of polymers affects plasma glucose cholesterol and other nutrients. Employing a double-perfusion technique in the rat we demonstrated that viscosity produced by SDF will delay transfer of zinc into the circulatory system. This delayed absorption should not be interpreted as decreased utilization. A great deal of additional research is required to prove the importance of luminaly viscosity produced by SDF on slowing nutrient absorption or regulating bllod nutrient homeostasis. Increased intake of TDF in the total human diet appears desirable. A dietary intake of 35g/day should not be considered to have a negative effect on mineral absorption. It is important to educate people that an intake of more than 35g TDF/day may cause an imbalance in the diet that can adve-rsely affect mineral utilization. Acknowledgments. Appreciation is given to Dr. George V. Vahouny(deceased) who was intense a great competitor in and out of science and who gave the author inspiration Portions of this work were supported by the University of Missouri Ag-ricultural Station and by a grant from the Univer-rch Support Grant RR 07053 from the National Institutes of Health. Contribution of the Missouri Agriculatural Experiments Station Journal Series No. 10747.

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Identification of PM10 Chemical Characteristics and Sources and Estimation of their Contributions in a Seoul Metropolitan Subway Station (서울시 지하역사에서 PM10의 화학적 특성과 오염원의 확인 및 기여도 추정)

  • Park, Seul-Ba-Sen-Na;Lee, Tae-Jung;Ko, Hyun-Ki;Bae, Sung-Joon;Kim, Shin-Do;Park, Duckshin;Sohn, Jong-Ryeul;Kim, Dong-Sool
    • Journal of Korean Society for Atmospheric Environment
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    • v.29 no.1
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    • pp.74-85
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    • 2013
  • Since the underground transportation system is a closed environment, indoor air quality problems may seriously affect many passengers' health. The purpose of this study was to understand $PM_{10}$ characteristics in the underground air environment and further to quantitatively estimate $PM_{10}$ source contributions in a Seoul Metropolitan subway station. The $PM_{10}$ was intensively collected on various filters with $PM_{10}$ aerosol samplers to obtain sufficient samples for its chemical analysis. Sampling was carried out in the M station on the Line-4 from April 21 to 28, July 13 to 21, and October 11 to 19 in the year of 2010 and January 11 to 17 in the year of 2011. The aerosol filter samples were then analyzed for metals, water soluble ions, and carbon components. The 29 chemical species (OC1, OC2, OC3, OC4, CC, PC, EC, Ag, Al, Ba, Cd, Cr, Cu, Fe, Mn, Ni, Pb, Si, Ti, V, Zn, $Cl^-$, $NO_3{^-}$, $SO_4{^{2-}}$, $Na^+$, $NH_4{^+}$, $K^+$, $Mg^{2+}$, $Ca^{2+}$) were analyzed by using ICP-AES, IC, and TOR after proper pretreatments of each sample filter. Based on the chemical information, positive matrix factorization (PMF) model was applied to identify the $PM_{10}$ sources and then six sources such as biomass burning, outdoor, vehicle, soil and road dust, secondary aerosol, ferrous, and brakewear related source were classified. The contributions rate of their sources in tunnel are 4.0%, 5.8%, 1.6%, 17.9%, 13.8% and 56.9% in order.

Dynamic Characteristics on the CRDM of SMART Reactor (SMART 원자로 제어봉 구동 장치의 동특성해석)

  • Lee, Jang-Won;Cho, Sang-Soon;Kim, Dong-Ok;Park, Jin-Seok;Lee, Won-Jae
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.34 no.8
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    • pp.1105-1111
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    • 2010
  • The Korea Atomic Energy Research Institutes has been developing the SMART (System integrated Modular Advanced ReacTor), an environment-friendly nuclear reactor for the generation of electricity and to perform desalination. SMART reactors can be exposed to various external and internal loads caused by seismic and coolant flows. The CRDM(control rod drive mechanism), one of structures of the SMART, is a component which is adjusting inserting amount of a control rod, controlling output of reactor power and in an emergency situation, inserting a control rod to stop the reactor. The purpose of this research is performing the analysis of dynamic characteristic to ensure safety and integrity of structure of CRDM. This paper presents two FE-models, 3-D solid model and simplified Beam model of the CRDM in the coolant, and then compared the results of the dynamic characteristic about the two FE-models using a commercial Finite Element tool, ABAQUS CAE V6.8 and ANSYS V12. Beam 4 and beam 188 of simplified-model were also compared each other. And simplified model is updated for accuracy compare to 3-D solid.

Effects of PEP-1-FK506BP on cyst formation in polycystic kidney disease

  • Jo, Hyo Sang;Eum, Won Sik;Park, Eun Young;Ko, Je Young;Kim, Do Yeon;Kim, Dae Won;Shin, Min Jea;Son, Ora;Cho, Su Bin;Park, Jung Hwan;Lee, Chi Hern;Yeo, Eun Ji;Yeo, Hyeon Ji;Choi, Yeon Joo;Youn, Jong Kyu;Cho, Sung-Woo;Park, Jinseu;Park, Jong Hoon;Choi, Soo Young
    • BMB Reports
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    • v.50 no.9
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    • pp.460-465
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    • 2017
  • Polycystic kidney disease (PKD) is one of the most common inherited disorders, involving progressive cyst formation in the kidney that leads to renal failure. FK506 binding protein 12 (FK506BP) is an immunophilin protein that performs multiple functions, including regulation of cell signaling pathways and survival. In this study, we determined the roles of PEP-1-FK506BP on cell proliferation and cyst formation in PKD cells. Purified PEP-1-FK506BP transduced into PKD cells markedly inhibited cell proliferation. Also, PEP-1-FK506BP drastically inhibited the expression levels of p-Akt, p-p70S6K, p-mTOR, and p-ERK in PKD cells. In a 3D-culture system, PEP-1-FK506BP significantly reduced cyst formation. Furthermore, the combined effects of rapamycin and PEP-1-FK506BP on cyst formation were markedly higher than the effects of individual treatments. These results suggest that PEP-1-FK506BP delayed cyst formation and could be a new therapeutic strategy for renal cyst formation in PKD.

Fatigue-Decreased Proficiency(FDP) Boundary for Whole-Body Vibration Exposure in Passenger Car Driver (승용차 운전자의 전신진동노출에 대한 피로-감소숙달 경계)

  • Jeung ae Yeal;Lee Ki-Nam
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.6
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    • pp.1211-1216
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    • 2002
  • To evaluate whole-body vibration(WBV) exposure and fatigue-decreased proficiency(FDP) boundary in passenger car driver, several roads in Busan were divided into 3 types by the condition of road surface; Road 1 was partially damaged, Road 2 was normal without damage, and Road 3 was better than Road 2. The results were following: The highest passenger driver's exposures to whole-body vibration acceleration and fatigue-decreased proficiency boundary at 40km/h were 0.108m/s² and about 2099 minutes in Road 2 for xh axis, 0.134m/s² and about 1585 minutes in Road 2 for yh axis, and 0.183m/s² and about 1053 minutes in Road 2 for zh axis, respectively. The highest passenger driver's exposures to whole-body vibration acceleration and fatigue-decreased proficiency boundary at 80km/h were 0.219m/s² and about 830 minutes in Road 3 xh axis, 0.203m/s² and about 918 minutes in Road 3 for yh axis, and 0.622m/s² and about 195 minutes in Road 1 for zh axis, respectively. The highest vector sums of whole-body vibration exposure at 40km/h and 804km/h were 0.328m/s² in Road 2 and 0.730m/s² in Road 1, respectively. The highest crest factors at 40km/h were 4.25 in Road 1 for xh, 4.51 in Road 3 for yh, and 5.81 in Road 2 for zh, respectively. The highest crest factors at 80km/h were 5.57 in Road 1 for xh, 5.60 in Road 2 for yh, and 6.46 in Road 3 for zh, respectively. The highest transmissibilities of whole-body vibration from floor to seat at 40km/h and 80km/h were 0.89 in Road 3 and 0.82 in Road 3 for xh axis, 0.83 in Road 3 and 0.87 in Road 1 and 2 for yh, and 0.80 in Road 2 and 0.92 in Road 1 tor zh axis, respectively. The highest fatigue-decreased proficiency boundaries for whole-body vibration exposure of passenger car driver in floor and seat were 457 minutes in Road 3 and 583 minutes in Road 3 at 40km/h and 159 minutes in Road 2 and 251 minutes in Road 2 at 80km/h, respectively.