• Title/Summary/Keyword: Molecular Characteristics

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Biological Functions of N- and O-linked Oligosaccharides of Equine Chorionic Gonadotropin and Lutropin/Chorionic Gonadotropin Receptor

  • Min, K.S.
    • Korean Journal of Animal Reproduction
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    • v.24 no.4
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    • pp.357-364
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    • 2000
  • Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$ -subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$-subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t631 or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632~653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agoinst-occupied receptors ~2- and ~17- fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.

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Product Quality and Shelf-life Effect of Low-fat Functional Sausages Manufactured with Sodium Lactate and Chitosans During Storage at 15℃ (젖산나트륨과 키토산을 첨가한 저지방 기능성 소시지의 15℃에서 저장 중 품질 및 저장성 효과)

  • Chin, Koo-Bok;Kook, Sung-H.;Choi, Soon-H.
    • Journal of Animal Science and Technology
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    • v.47 no.4
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    • pp.655-666
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    • 2005
  • This study was performed to measure physicochemical and textural characteristics, and shelf-life effect of low-fat functional sausages(LFFSs) manufactured with sodium lactate(SL, 3.3%), lac pigment and various molecular weights(MWs) of chitosan (Low=1.5 kDa, Med=30-50 kDa and High=200 kDa) during storage at 15$^{\circ}C$ for 18 days. LFFSs had 73.7-76.0% moisture, lower than 3% fat and 14-15% protein, respectively. pH values were 6.05-6.44 and the control(150 ppm, $NaNO_2$) was the lowest among LFFSs (p<0.05). Increasing storage time decreased pH values, but no differences in pH values were observed up to 6 days of storage (p>0.05). LFFSs containing SL and low MW of chitosan improved water holding capacity (WHC) and different from those with SL and medium-MW chitosan. WHC was decreased with increased storage time and differences of WHC were observed from 18 days of storage. The addition of chitosan reduced both lightness and redness values, as compared to 150 ppm sodium nitrite(SN), and increased storage time decreased yellowness(p<0.05), especially at 12 days of storage. LFFSs with SL and medium-MW chitosan increased most textural properties compared to the control(p<0.05). The addition of SN of 150 ppm in LFFSs retarded microbial growth for E. coli 0157:H7, while those with SL tended to have an antimicrobial effect for Listeria monocytogenes(LM). The growth rate of LM was delayed by addition of various MW of chitosans in LFFSs, especially high MW chitosan, as compared to LFFSs containing SL alone. These results indicated that the functional, textural and antimicrobial effects of LFFSs were improved by addition SL and various MW of chitosan combinations. In addition, 0.05% lac pigment improved the cure color of LFFSs similar to those of 150 ppm SN.

Anaerobic Biodegradability of Leachates Generated at Landfill Age (매립년한에 따른 침출수의 혐기성 생분해 특성)

  • Shin, Hang-Sik;Lee, Chae-young;Kang, Ki-hoon
    • Journal of the Korea Organic Resources Recycling Association
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    • v.8 no.1
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    • pp.90-96
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    • 2000
  • The composition of leachates varies depending on the waste characteristics, landfill age and landfilling method. Generally, leachates contain high dissolved organic substance and ammonia nitrogen whereas phosphorus concentration was very low. Leachate A produced from young landfill is characterized by high BOD5/COD ratio (0.8) whereas leachate C produced from old landfill has lower BOD5/COD ratio (0.1). Maximum biochemical methane potential of leachate A, B (from medium landfill) and C were 271,106 and 4 ml CH4/g-COD, respectively. On the other hand, the maximum biodegradability of leachate A, B, and C were 75,30, and 1%, respectively. These results indicated that anaerobic treatment of leachate from young landfill was effective in removing organic pollutants. In case of leachate C, carbon might reside in the form of large molecular weight organic compounds such as lignins, humic acids and other polymerized compounds of soils, which are resistant to biodegradation. The lag-phase period increased with the increasing organic concentration in leachate. In case of leachate A of concentration greater than 25%, the lag-phase period increased sharply. This implied that the start-up period of anaerobic process using an unacclimated inoculum could be extended due to the higher concentration of leachate. This relatively long lag-phase is probably related to the fact that most of the inhibitory compounds have been diluted beyond their inhibitory concentrations of less than 50%. Furthermore, the ultimate methane yield and methane production rate decreased as leachate concentration increased. It was anticipated the potential inhibition was related with the steady-state inhibition as well as the initial shock load.

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Development of Transgenic Crops and Research Projects for Biotechnology Application (유전자 전환작물 개발 연구 현황과 과제)

  • 정태영
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.5
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    • pp.289-296
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    • 2001
  • The main objective of this topic is to establish strategies and to plan biotechnology researches which are related to the agricultural improvements especially focusing on the crop breeding in Korea. From 1960's to 1980's government policy had been emphasized to develope high yielding cultivars for the self sufficient supply of the staple food crops. As a result, considerable increase of rice production has been made with accumulating technology and man's powers. Recently genetically modified crops harboring useful characteristics have been developed using biotechnology and released in the developed countries. National research institutes and private companies have been developed biotechnology researches to establish competitive capabilities, however they have not been successfully used in commercialization. Therefore it is necessary to promote the practical. application by connecting molecular technology with conventional breeding. Proposed research projects are; (1) basic researches including plant genome studies, (2) developing new cultivars through gene transformation, (3) screening and producing antioxidants, secondary metabolite substances and edible vaccines. To set a government policy, both domestic and international research trends were reviewed and possibility of success based on the economic view point were discussed. The intellectual property and preservation of environment play a key role to decide the research priority. It is also necessary for us to make one step system for the distribution of research resources such as microorganisms, genes cloned, plant seeds and research informations for promoting research activities.

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Etiological Properties and Coat Protein Gen Analysis of Potato Virus Y Occuring in Potatoes of Korea (우리나라 감자에 발생하는 PVY의 병원학적 특성 및 외피단백질 유전자 분석)

  • ;Richard M. Bostock
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 1995.06b
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    • pp.77-96
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    • 1995
  • To obtain basic informations for the improvement of seed potato production in Korea, some etiological properties of potato virus Y(PVY) distributed in the major seed potato production area(Daekwanryeong) were characterized, and the nucleotide and amino acid sequences of the coat protein gene of the PVY strains isolated were analyzed. PVY strains in Daekwonryeong, an alpine area, were identified to be two strains, PVYo and PVYN by symptoms of indicator plants, and their distribution in potato fields was similar. Major symptom on potato varieties by PVY was grouped as either mosaic alone or mosaic accompanied with veinal necrosis in the lower leaves. The symptom occurrence of the two symptoms was similar with Irish Cobbler, but Superior showed a higher rate of mosaic symptom than the other. The PVY strain which was isolated from potato cv. Superior showing typical mosaic symptoms produced symptoms of PVY-O on the indicator plants of Chenopodium amaranticolor, Nicotiana tabacum cv. Xanthi nc and Physalis floridana, but no symptom o Capsicum annum cv. Ace. Moreover, results from the enzyme-linked immunosorbent assay with monoclonal and polyclonal antibodies showed that the isolated PVY reacts strongly with PYV-O antibodies but does not react specifically with PVY-T antibodies. The purified virus particles were flexious with a size of 730$\times$11nm. On the basis of the above characteristics, the strain was identified to be a PVY-O and named as of PVY-K strain. The flight of vector aphids was observed in late May, however, the first occurrence of infected plants was in mid June with the bait plants surrounded with PVY-infected potato plants and early July with the bait plants surrounded with PVY-free potato plants. PVY infection rates by counting symptoms on bait plants (White Burley) were 1.1% with the field surrounded with PVY-free potato plants and 13.7% the fields surrounded with PVY-infected potato plants, showing the effect of infection pressure. The propagated PVY-K strain on tobacco(N. sylvestris) was purified, and the RNA of the virus was extracted by the method of phenol extraction. The size of PVY-K RNA was measured to be 9, 500 nucleotides on agarose gel electrophoresis. The double-stranded cDNAs of PVY-K coat protein(CP) gene derived by the method of polymerase chain reaction were transformed into the competent cells of E. coli JM 109, and 2 clones(pYK6 and pYK17) among 11 clones were confirmed to contain the full-length cDNA. Purified plasmids from pYK17 were cut with Sph I and Xba I were deleted with exonuclease III and were used for sequencing analysis. The PVY-K CP gene was comprised of 801 nucleotides when counted from the clevage site of CAG(Gln)-GCA(Ala) to the stop codon of TGA and encoded 267 amino acids. The molecular weight of the encoded polypeptides was calculated to be 34, 630 daltons. The base composition of the CP gene was 33.3% of adenine, 25.2% of guanine, 20.1% of cytosine and 21.4% of uracil. The polypeptide encoded by PVY-K CP gene was comprised of 22 alanines, 20 threonines, 19 glutamic acids and 18 glycines in order. The homology of nucleotide sequence of PVY-K CP gene with those of PVY-O(Japan), PVY-T(Japan), PVY-TH(Japan), PVYN(the Netherlands), and PVYN(France) was represented as 97.3%, 88.9%, 89.3%, 89.6% and 98.5%, respectively. The amino acid sequence homology of the polypeptide encoded by PVY-K CP gene with those encoded by viruses was represented as 97.4%, 92.5%, 92.9%, 92.9%, and 98.5%, respectively.

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Incidental Benign Parotid Lesions on FDG-PET: Prevalence and Clinico-pathologic Findings (FDG-PET 검사시 우연히 발견한 양성 이하선 병변: 유병율과 임상 및 병리 소견)

  • Lim, Il-Han;Lee, Won-Woo;Chung, Jin-Haeng;Park, So-Yeon;Kim, Sang-Hee;Kim, Yu-Kyeong;Kim, Sang-Eun
    • Nuclear Medicine and Molecular Imaging
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    • v.41 no.5
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    • pp.359-363
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    • 2007
  • Purpose: Incidental parotid lesions on F-18 FDG-PET can mimic distant metastasis of underlying malignancy. The prevalence and the clinico-pathologic findings of PET positive parotid lesions have not been known. We investigated how often incidental parotid lesions are found on clinical FDG-PET studies and what the clinico-pathologic characteristics of those parotid lesions are in the present study. Materials and Methods: We retrospectively reviewed 3,344 cases of FDG-PET which had been obtained in our hospital from May 2003 to Dec 2006. The indications of FDG-PET were: evaluation of known/suspected cancer (n= 3,212) or screening of cancer in healthy subjects (n=132). Incidental parotid lesion on FDG-PET was defined as an un-expected FDG uptake in one of parotid glands which was not primary target lesion of current FDG-PET. FDG uptake was represented by maximum standardized uptake value (maxSUV). Final diagnosis was made by pathologic analysis or clinical follow-up assessment. Results: Fifteen (0.45% = 15/3,344) incidental parotid lesions were found and they were all benign lesions. The maxSUV ranged from 1.7 to 8.6 (mean${\pm}$s.d. = $3.7{\pm}1.9$). Final diagnoses of the incidental parotid lesions were; Warthin's tumor (n=2), pleomorphic adenoma (n=1), other un-specified benign lesion (n=1), and benign lesions under bases of imaging studies (n=3) and of clinical follow-up (n=8). Conclusion: All of incidentally found parotid lesions in clinical FDG-PET studies were confirmed as benign lesions with prevalence of 0.45%. Close follow up using PET or CT might be a reasonable approach for determining the nature of incidentally found parotid lesions.

Development of Rapid Analytical Method of Forbidden Medicines in Dietary Supplements Using LC-ESI-Tandem MS (LC-ESI-tandem MS를 이용한 기능성표방식품 중 부정유해물질 신속검사체계 개발)

  • Kim, Hee-Yun;Jang, Young-Mi;Joo, Hyun-Jin;Jung, Young-Hyun;Lee, Myoung-Sook;Park, Jong-Seok;Lee, Kwang-Ho;Lee, Hwa-Mi
    • Korean Journal of Food Science and Technology
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    • v.39 no.4
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    • pp.372-379
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    • 2007
  • A high-performance liquid chromatography-electrospray ionization (HPLC-ESI) tandem MS was developed for the rapid and simultaneous determination of forbidden medicines in dietary supplements. Thirteen medicinal components such as PDE-5 inhibitors and their analogues, and the newly identified dimethylsildenafil and xanthoanthrafil, were included in this study. After tentative standardization of molecular ions in both polarities using thirteen references on the mass spectrometer, with ESI-continuous infusion via the syringe pump method, the relative intensity of the ions present in the resulting spectra was quantitatively compared. From the results, the ion mode was selected depending on each reference's characteristics. A HPLC method coupled with the ESI mode was developed considering the matrix effect and interference depending on the type of sample. The validation test of the developed method was followed by carrying out precision, accuracy, recovery, sensitivity and linearity, etc. The method showed sufficiently high sensitivity, reproducibility, and specificity, and produced 4 times faster results when compared with the existing HPLC/UV method for the determination of forbidden compounds in dietary supplements.

Production of Poly-3-hydroxybutyrate from Xylose by Bacillus megaterium J-65 (Bacillus megaterium J-65에 의한 xylose로부터 poly-3-hydroxybutyrate 생산)

  • Jun, Hong-Ki;Jin, Young-Hi;Kim, Hae-Nam;Kim, Yun-Tae;Kim, Sam-Woong;Baik, Hyung-Suk
    • Journal of Life Science
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    • v.18 no.12
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    • pp.1625-1630
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    • 2008
  • A microorganism capable of producing high level of poly-3-hydoxybutyrate (PHB) from xylose was isolated from soil. The isolated strain J-65 was identified as Bacillus megaterium based on the morphological, biochemical and molecular biological characteristics. The optimum temperature and pH for the growth of B. megaterium J-65 were $37^{\circ}C$ and 8.0, respectively. The optimum medium composition for the cell growth was 2% xylose, 0.25% $(NH_4)_2SO_4$, 0.3% $Na_2HPO_4{\cdot}12H_2O$, and 0.1% $KH_2PO_4$. The optimum condition for PHB accumulation was same to the optimum condition for cell growth. Copolymer of ${\beta}$-hydroxybutyric and ${\beta}$-hydroxyvaleric acid was produced when propionic acid was added to shake flasks containing 20 g/l of xylose. Fermenter culture was carried out to produce the high concentration of PHB. In batch culture, cell mass was 9.82 g/l and PHB content was 35% of dry cell weight. PHB produced by B. megaterium J-65 was identified as homopolymer of 3-hydoxybutyric acid by GC and NMR.

Radiolabeling of NOTA and DOTA with Positron Emitting $^{68}$Ga and Investigation of In Vitro Properties (양전자 방출핵종 $^{68}$Ga을 이용한 NOTA와 DOTA의 표지 및 시험관내 특성 연구)

  • Jeong, Jae-Min;Kim, Young-Ju;Lee, Yun-Sang;Lee, Dong-Soo;Chung, June-Key;Lee, Myung-Chul
    • Nuclear Medicine and Molecular Imaging
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    • v.43 no.4
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    • pp.330-336
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    • 2009
  • Purpose: We established radiolabeling conditions of NOTA and DOTA with a generator-produced PET radionuclide $^{68}$Ga and studied in vitro characteristics such as stability, serum protein binding, octanol/water distribution, and interference with other metal ions. Materials and Methods: Various concentrations of NOTA 3HCl and DOTA 4HCl were labeled with 1 mL $^{68}$GaCl$_3$ (0.18$\sim$5.75 mCi in 0.1 M HCl in various pH. NOTA 3HCl (0.373 mM) was labeled with $^{68}$GaCl$_3$(0.183$\sim$0.232 mCi/0.1 M HCl 1.0 mL) in the presence of CuCl$_2$, FeCl$_2$, InCl$_3$, FeCl$_3$, GaCl$_3$, MgCl$_2$ or CaCl$_2$ (0$\sim$6.07 mM) at room temperature. The labeling efficiencies of $^{68}$Ga-NOTA and $^{68}$Ga-DOTA were checked by ITLC-SG using acetone or saline as mobile phase. Stabilities, protein bindings, and octanol distribution coefficients of the labeled compounds also were investigated. Results: $^{68}$Ga-NOTA and $^{68}$Ga-DOTA were labeled optimally at pH 6.5 and pH 3.5, respectively, and the chelates were stable for 4 hr either in the reaction mixture at room temperature or in the human serum at 37$^{\circ}C$. NOTA was labeled at room temperature while DOTA required heating for labeling. $^{68}$Ga-NOTA labeling efficiency was reduced by CuCl$_2$, FeCl$_2$, InCl$_2$, FeCl$_3$ or CaCl$_3$, however, was not influenced by MgCl$_2$ or CaCl$_2$. The protein binding was low (2.04$\sim$3.32%). Log P value of $^{68}$Ga-NOTA was -3.07 indicating high hydrophilicity. Conclusion: We found that NOTA is a better bifunctional chelating agent than DOTA for $^{68}$Ga labeling. Although, $^{68}$Ga-NOTA labeling is interfered by various metal ions, it shows high stability and low serum protein binding.

Usefulness of F-18 FDG PET/CT in Adrenal Incidentaloma: Differential Diagnosis of Adrenal Metastasis in Oncologic Patients (부신 우연종에서 F-18 FDG PET/CT의 유용성: 악성 종양 환자에서 부신 전이의 감별진단)

  • Lee, Hong-Je;Song, Bong-Il;Kang, Sung-Min;Jeong, Shin-Young;Seo, Ji-Hyoung;Lee, Sang-Woo;Yoo, Jeong-Soo;Ahn, Byeong-Cheol;Lee, Jae-Tae
    • Nuclear Medicine and Molecular Imaging
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    • v.43 no.5
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    • pp.421-428
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    • 2009
  • Purpose: We have evaluated characteristics of adrenal masses incidentally observed in nonenhanced F-18 FDG PET/CT of the oncologic patients and the diagnostic ability of F-18 FDG PET/CT to differentiate malignant from benign adrenal masses. Materials and Methods: Between Mar 2005 and Aug 2008, 75 oncologic patients (46 men, 29 women; mean age, $60.8{\pm}10.2$ years; range, 35-87 years) with 89 adrenal masses incidentally found in PET/CT were enrolled in this study. For quantitative analysis, size (cm), Hounsfield unit (HU), maximum standardized uptake value (SUVmax), SUVratio of all 89 adrenal masses were measured. SUVmax of the adrenal mass divided by SUVliver, which is SUVmax of the segment 8, was defined as SUVratio. The final diagnosis of adrenal masses was based on pathologic confirmation, radiologic evaluation (HU<0 : benign), and clinical decision. Results: Size, HU, SUVmax, and SUVratio were all significantly different between benign and malignant adrenal masses.(P < 0.05) And, SUVratio was the most accurate parameter. A cut-off value of 1.0 for SUVratio provided 90.9% sensitivity and 75.6% specificity. In small adrenal masses (1.5 cm or less), only SUVratio had statistically significant difference between benign and malignant adrenal masses. Similarly a cut-off value of 1.0 for SUVratio provided 80.0% sensitivity and 86.4% specificity. Conclusion: F-18 FDG PET/CT can offer more accurate information with quantitative analysis in differentiating malignant from benign adrenal masses incidentally observed in oncologic patients, compared to nonenhanced CT.