• Title/Summary/Keyword: Molecular identification

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Isolation of Isoflavones and Soyasaponins from the Germ of Soybean (콩 배아로 부터 Isoflavone과 Soyasaponin의 동시 분리)

  • Kim, Sun-Lim;Lee, Jae-Eun;Kim, Yul-Ho;Jung, Gun-Ho;Kim, Dea-Wook;Lee, Choon-Ki;Kim, Mi-Jung;Kim, Jung-Tae;Lee, Yu-Young;Hwang, Tae-Young;Lee, Kwang-Sik;Kim, Wook-Han;Kwon, Young-Up;Kim, Hong-Sig;Chung, Ill-Min
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.58 no.2
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    • pp.149-160
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    • 2013
  • The objective of present study was to simultaneously isolate of isoflavone and soyasaponin compounds from the germ of soybean seeds. Soy germ flours were defatted with hexane for 48h at room temperature, and methanolic extracts were prepared using reflux apparatus at $90^{\circ}C$ for 6h, two times. After extraction, extracts were separated with preparative RP-$C_{18}$ packing column ($125{\AA}$, $55-105{\mu}m$, $40{\times}150mm$), and collected 52 fractions were identified with TLC plate (Kieselgel 60 F-254) and HPLC, respectively. Among the identified isoflavone and soyasaponin fractions, isoflavone fractions were re-separated using a recycling HPLC with gel permeation column (Jaigel-W252, $20{\times}500mm$). Final fractions were air-dried, and the purified compounds of two isoflavones (ISF-1-1, ISF-1-2) and four soyasaponins (SAP-1, SAP-2, SAP-3, SAP-4) were obtained. Two isoflavone compounds (ISF-1-1, ISF-1-2) were acid-hydrolyzed for the identification of their aglycones, and confirmed by comparing with 12 types of isoflavone isomers. While the four kinds of soyasaponins were identified by using a micro Q-TOF mass spectrometer in the ESI positive mode with capillary voltage of 4.5kV, and dry temperature of $200^{\circ}C$. Base on the obtained results, it was conclude that ISF-1-1 is the mixture isomers of daidzin (43.4%), glycitin (47.0%), and genistin (9.6%), but ISF-1-2 is the single compound of genistin (99.8% <). On the other hand, soyasaponin SAP-1 is the mixture compounds of soyasaponin A-group (Aa, Ab, Ac, Ae, Af); SAP-2 is soyasaponin B-group (Ba, Bb, Bc) and E-group (Bd, Be); SAP-3 is soyasaponin B-group (Ba, Bb, Bc), E-group (Bd, Be), and DDMP-group (${\beta}g$); SAP-4 is soyasaponin B-group (Ba, Bb, Bc), E-group (Bd, Be), and DDMP-group (${\beta}g$, ${\beta}a$), respectively.

Characterization of a Korean Domestic Cyanobacterium Limnothrix sp. KNUA012 for Biofuel Feedstock (토착 남세균 림노트릭스 속 KNUA012 균주의 바이오연료 원료로서의 특성 연구)

  • Hong, Ji Won;Jo, Seung-Woo;Kim, Oh Hong;Jeong, Mi Rang;Kim, Hyeon;Park, Kyung Mok;Lee, Kyoung In;Yoon, Ho-Sung
    • Journal of Life Science
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    • v.26 no.4
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    • pp.460-467
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    • 2016
  • A filamentous cyanobacterium, Limnothrix sp. KNUA012, was axenically isolated from a freshwater bloom sample in Lake Hapcheon, Hapcheon-gun, Gyeongsangnam-do, Korea. Its morphological and molecular characteristics led to identification of the isolate as a member of the genus Limnothrix. Maximal growth was attained when the culture was incubated at 25℃. Analysis of its lipid composition revealed that strain KNUA012 could autotrophically synthesize alkanes, such as pentadecane (C15H32) and heptadecane (C17H36), which can be directly used as fuel without requiring a transesterification step. Two genes involved in alkane biosynthesis-an acyl-acyl carrier protein reductase and an aldehyde decarbonylase-were present in this cyanobacterium. Some common algal biodiesel constituents-myristoleic acid (C14:1), palmitic acid (C16:0), and palmitoleic acid (C16:1)-were produced by strain KNUA012 as its major fatty acids. A proximate analysis showed that the volatile matter content was 86.0% and an ultimate analysis indicated that the higher heating value was 19.8 MJ kg−1. The isolate also autotrophically produced 21.4 mg g−1 phycocyanin-a high-value antioxidant compound. Therefore, Limnothrix sp. KNUA012 appears to show promise for application in cost-effective production of microalga-based biofuels and biomass feedstock over crop plants.

Development of Position Encoding Circuit for a Multi-Anode Position Sensitive Photomultiplier Tube (다중양극 위치민감형 광전자증배관을 위한 위치검출회로 개발)

  • Kwon, Sun-Il;Hong, Seong-Jong;Ito, Mikiko;Yoon, Hyun-Suk;Lee, Geon-Song;Sim, Kwang-Souk;Rhee, June-Tak;Lee, Dong-Soo;Lee, Jae-Sung
    • Nuclear Medicine and Molecular Imaging
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    • v.42 no.6
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    • pp.469-477
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    • 2008
  • Purpose: The goal of this paper is to present the design and performance of a position encoding circuit for $16{\times}16$ array of position sensitive multi-anode photomultiplier tube for small animal PET scanners. This circuit which reduces the number of readout channels from 256 to 4 channels is based on a charge division method utilizing a resistor array. Materials and Methods: The position encoding circuit was simulated with PSpice before fabrication. The position encoding circuit reads out the signals from H9500 flat panel PMTs (Hamamatsu Photonics K.K., Japan) on which $1.5{\times}1.5{\times}7.0\;mm^3$ $L_{0.9}GSO$ ($Lu_{1.8}Gd_{0.2}SiO_{5}:Ce$) crystals were mounted. For coincidence detection, two different PET modules were used. One PET module consisted of a $29{\times}29\;L_{0.9}GSO$ crystal layer, and the other PET module two $28{\times}28$ and $29{\times}29\;L_{0.9}GSO$ crystal layers which have relative offsets by half a crystal pitch in x- and y-directions. The crystal mapping algorithm was also developed to identify crystals. Results: Each crystal was clearly visible in flood images. The crystal identification capability was enhanced further by changing the values of resistors near the edge of the resistor array. Energy resolutions of individual crystal were about 11.6%(SD 1.6). The flood images were segmented well with the proposed crystal mapping algorithm. Conclusion: The position encoding circuit resulted in a clear separation of crystals and sufficient energy resolutions with H9500 flat-panel PMT and $L_{0.9}GSO$ crystals. This circuit is good enough for use in small animal PET scanners.

Role of FDG-PET in the Diagnosis of Recurrence and Assessment of Therapeutic Response in Cervical Cancer and Ovarian Cancer Patients: Comparison of Diagnostic Report between PET, Abdominal a and Tumor Marker (자궁경부암 및 난소암 환자 재발진단과 치료반응평가에 있어서 FDG-PET의 역할: 양전자방출단층촬영, 복부전산화단층촬영 및 종양표지자 판독의 비교 분석)

  • Han, You-Mie;Choe, Jae-Gol;Kang, Bung-Chul
    • Nuclear Medicine and Molecular Imaging
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    • v.42 no.3
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    • pp.201-208
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    • 2008
  • Purpose: We aimed to assess the role of positron emission tomography using fluorodeoxyglucose (FDG-PET) in the diagnosis of recurrence or the assessment of therapeutic response in cervical and ovarian cancer patients through making a comparison between FDG-PET, abdominal computed tomography (CT) and serum tumor marker. Materials and methods: We included 103 cases (67 patients) performed FDG-PET and abdominal CT. There were 42 cervical cancers and 61 ovarian cancers. We retrospectively reviewed the interpretations of PET and CT images as well as the level of tumor marker. We calculated their sensitivity, specificity, positive predictive value and negative predictive value for these three modalities. And then we analyzed the differences between these three modalities. Results: Tumor recurrences were diagnosed in 37 cases (11 cervical cancers and 26 ovarian cancers). For PET, CT and tumor marker, in cervical cancer group, sensitivity was 100% (11/11), 54.5% (6/11) and 81.1% (9/11), respectively. And specificity was 93.6% (29/31), 93.6% (29/31) and 100% (31/31). In ovarian cancer group, sensitivity was 96.2% (25/26), 84.6% (22/26) and 80.8% (21/26), and specificity was 94.3% (33/35), 94.3% (33/35), 94.3% (33/35), PET was highly sensitive to detect the intraperitoneal and extraperitoneal metastasis with the help of the CT images to localize the lesions. However, CT had limitations in differentiation of the recurrent tumor from benign fibrotic tissue, identification of viable tumors at the interface of tissues, and detecting extraperitoneal lesions. Conclusion: FDG-PET can be an essential modality to detect the recurrent or residual tumors in gynecologic cancer patients because of its great field of the application and high sensitivity.

Genetic Differences and Variations in Freshwater Crab(Eriocheir sinensis) and Swimming Crab(Portunus trituberculatus) (참게(Eriocheir sinensis)와 꽃게(Portunus trituberculatus)의 유전적 차이와 변이)

  • Yoon, Jong-Man
    • Development and Reproduction
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    • v.10 no.1
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    • pp.19-32
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    • 2006
  • Genomic DNA isolated from two species of Korean freshwater crab(Eriocheir sinensis) and swimming crab(Portunus trituberculatus) was amplified several times by PCR reactions. The seven arbitrarily selected primers OPA-05, OPA-13, OPA-16, OPB-06, OPB-15, OPB-17 and OPD-10 were used to generate the identical, polymorphic, and specific fragments. 505 fragments were identified in the freshwater crab species, and 513 in the swimming crab from Buan: 81 specific fragments(16.0%) in the freshwater crab species and 100(19.5%) in the swimming crab. 165 identical fragments, with an average of 23.6 per primer, were observed in the freshwater crab species. 66 fragments, with an average of 9.4 per primer, were identified in the swimming crab species. The numbers of polymorphic fragments in the freshwater crab and swimming crab were 50 and 14, respectively. The oligonucleotides decamer primer OPB-17 generated identical DNA fragments, approximately 300 bp, in both the freshwater crab and swimming crab species. Compared separately, the average genetic difference was higher in the swimming crab than in the freshwater crab species. The average genetic difference was $0.726{\pm}0.004$ between the freshwater crab and swimming crab species. The dendrogram obtained by the seven primers indicates four genetic clusters: cluster 1(FRESHWATER 01), cluster 2(FRESHWATER 02, 03, 04, 05 and 06), cluster 3(FRESHWATER 07, 08, 09, 10 and 11), and cluster 4(SWIMMING 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 and 22). The shortest genetic distance displaying significant molecular difference was between individuals SWIMMING no. 18 and SWIMMING no. 17 from swimming crab(0.096). Ultimately, individual no. 02 of the freshwater crab was most distantly related to freshwater crab no. 03(genetic distance = 0.770). As stated above, the potential of RAPD-PCR to identify diagnostic markers for the identification of two crab species has been demonstrated.

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The Relationship between Intracellular Protein Kinase C Concentration and Invasiveness in U-87 Malignant Glioma Cells (교모세포종 세포주 U-87에서 세포내 PKC 농도와 종양침습성과의 상관 관계)

  • Ji, Cheol;Cho, Kyung-Keun;Lee, Kyung Jin;Park, Sung Chan;Cho, Jung Ki;Kang, Joon Ki;Choi, Chang Rak
    • Journal of Korean Neurosurgical Society
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    • v.30 no.3
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    • pp.263-271
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    • 2001
  • Objective : Glioblastomas, the most common type of primary brain tumors, are highly invasive and cause massive tissue destruction at both the tumor invading edges and in areas that are not in direct contact with glioma cells. As a result, patients with high-grade gliomas are faced with a poor prognosis. Such grim statistics emphasize the need to better understand the mechanisms that underlie glioma invasion, as these may lead to the identification of novel targets in the therapy of high grade gliomas. Protein kinase C(PKC) is a family of serine/threonine kinases and an important signal transduction enzyme that conveys signals generated by ligand-receptor interaction at the cell surface to the nucleus. PKC appears to be critical in regulating many aspects of glioma biology. The purpose of this study was to assess accurately the role of PKC in the invasion regulation of human gliomas based on hypothesis that protein kinase C(PKC) is functional in the process of glial tumor cell invasion. Method : To test this hypothesis, U-87 malignant glioma cell line intracellular PKC levels were up and down regulated and their invasiveness was tested. Intracellular PKC level was characterized using PKC activity assays. Invasion assays including barrier migration and spheroid confrontation were used to study the relationship between PKC concentration and invasiveness. Result : The cell line which were treated by PKC inhibitor tamoxifen and hypericin exhibited decreased PKC activity and decreased invasive abilities dose dependently both in matrigel invasion assay and tumor spheroid fetal rat brain aggregates(FRBA) confrontation assay. However, the cell line that was treated by PKC activator 12-O-tetradecanylphorbol-13acetate(TPA) did not exhibit increases in either PKC activity or invasive ability. Conclusion : These studies suggest that PKC may be a useful molecular target for the chemotherapy of glioblastoma and other malignancies and that a therapeutic approach based on the ability of PKC inhibitors may be helpful in preventing invasion.

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Antigen analysis of Toxoplasma gondii Iysate and excretory-secretory materials by enzyme-linked immunoelectrotransfer blot (EITB) (효소면역 전기영동이적법에 의한 톡소포자충 용해물 및 분비 항원의 분석)

  • 안명희;손혁진
    • Parasites, Hosts and Diseases
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    • v.32 no.4
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    • pp.249-258
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    • 1994
  • Recently, the importance of toxoplasmosis is raised as a complication in immunosuppressed or AIDS patients. Our study focused on the identification of a variety of Toxoplasma antigens by immunoblotting. Rabbits and BALB/c mice were immunized with Toxoplosmo Iysate (RH strain) , frozen tachyzoites (RH strain) or cysts (Beverly and Fukaya strain) . Blood were collected from ear vein, heart or orbital plexus for detecting the serum antibody levels. For excretory-secretory (E.S) antigens, T gondii (RH) tachyzoite were cultured in CHL (Chinese hamster lung) cells with MEM containing of 5% FCS. After 72hrs, culture supernatant was collected. BALB/c mice were inoculated with RH tachyzoite intraperitoneally and peritoneal fluids were extracted three days later. E.S antigens were detected in culture supernatant and infected mouse peritoneal fluid by EITB. Serum IgG levels in rabbit were 1 :512 of 10 days after primary immunization, 1 : 2,048 of 10 days after secondary immunization, 1: 1,024 of 20 days after secondary immunization by IFAT, respectively. Serum IgG levels of immunized mice were 1:128 after 7 weeks. Tachyzoite antigens of the RH strain were detected 25 protein bands ranging 10 kDa-220 kDa of molecular weights with Coomassie blue stain. Toxoplcsma major antigens corresponding to n of 24 kDa, 27 kDa,30 kDa, 35 kDa, 38 kDa were recognized by IgG and IgM antibodies. Excretory-secretory antigens present in culture supernatant with M. W. of 20, 30 kDa and in infected mouse peritoneal fluid with M.W. of 33 (P30), 45 kDa. When RH tachyzoite antigen was probed with different mice sera immunized with 2 strains of T gondii, the IgG antibody bud of Fukaya and Beverly strain (8 week-serum) is identical to those of RH strain. It is considered that the 30 kDa polypeptide detected in excretory- secretory materials and Iysate was important major antigen of T gondii (RH).

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THE EFFECT OF ALTERED FUNCTIONAL FORCE ON THE EXPRESSION OF SPECIFIC MRNAS IN THE DEVELOPING MOUSE MANDIBLE (하악골의 발육중인 생쥐에서 기능력의 변화가 특이-유전자 발현에 미치는 영향)

  • Kim, Hyung-Tae;Park, Joo-Cheol;Lee, Chang-Seop;Park, Heon-Dong
    • Journal of the korean academy of Pediatric Dentistry
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    • v.30 no.2
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    • pp.308-319
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    • 2003
  • Mechanical forces are known to have an effect on bone formation, maintenance and remodeling, and there is evidence that the development of the mandibular condyle in the rat or mouse is influenced by altered functional force. However, studies are lacking in molecular-biologic mechanism such as the identification of differentiation factor induced from functional force. Here a mouse model was used to investigate the functional stress-responsive gene or factors which is related to the altered force by comparing the expression genes of functional state and hypo-functional state of the mouse mandible. ICR mice were provisioned with either a soft, mushy diet (soft-diet group) or hard rat pellets (hard-diet group) beginning at weaning for the alteration of functional force and subsequently sacrificed at 89 days of age. Incisor of mice in group 1 were trimmed twice a week to reduce occlusal forces. After killing the animals, mandibular bone including condyle were collected for RNA extraction, subtractive hybridization, northern blot analysis and mRNA in-situ hybridization. The results as follows; 1. A total of 39 clones were sequenced, and 11 individual sequence types were subsequently identified by subtractive hybridization, as 28 clones were represented twice in the analyzed sets. 2. Consequently four candidate clones, FS-s (functional stress-specific)2, -5, -18, and -22 were identified and characterized by homolgy search and northern analysis. Four of these clones, FS-s2, -5, -18, and -22, were shown to be expressed differentially in the hard-diet group. 3. Histologic sections showed that osteoblastic activity along the bone trabeculae and active bone remodeling were significantly lower in soft than in hard diet animals. A soft diet seems to enable a longer period of endochondral ossification in the mandibular condyle. 4. Although the mRNAs of FS-s2, -5, -18, and -22 were expressed rarely by cells of the soft-diet group, highest expression was detected in the cells of the hard-diet group. Together with the above results, it is suggested that FS-s2, -5, -18, and -22 could act as an important factors controlling the tissue changes in response to functional stress. The exact functional significance of these findings remains to be established.

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Isolation and Structure Identification of Photosensitizer from Perilla frutescens Leaves Which Induces Apoptosis in U937 (들깻잎(Perilla frutescens)으로부터 U937 세포에 apoptosis를 유도하는 광과민성 물질의 분리 및 구조동정)

  • Ha, Jun Young;Kim, Mi Kyeong;Lee, Jun Young;Choi, Eun Bi;Hong, Chang Oh;Lee, Byong Won;Bae, Chang Hwan;Kim, Keun Ki
    • Journal of Life Science
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    • v.25 no.1
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    • pp.53-61
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    • 2015
  • In this study, we tried to separate the photosensitizer that induces apoptosis of leukemia cells (U937) from perilla leaves. Perilla leaves (Perilla frutescens Britt var. japonica Hara) are a popular vegetable in Korea, being rich in vitamins (A and E), GABA, and minerals. Dried perilla leaves were extracted with methanol to separate the photosensitizer by various chromatographic techniques. The structure of the isolated compound (PL9443) was identified by 1D-NMR, 2D-NMR, and FAB-mass spectroscopy. Absorbance of the UV-Vis spectrum was highest at 410 nm and was confirmed by the 330, 410, and 668 nm. PL9443 compound was determined to be pheophorbide, an ethyl ester having a molecular weight of 620. It was identified as a derivative compound of pheophorbide structure when magnesium comes away from a porphyrin ring. Observation of morphological changes in U937 cells following cell death induced by treated PL9443 compound revealed representative phenomena of apoptosis only in light irradiation conditions (apoptotic body, vesicle formation). Results from examining the cytotoxicity of PL9443 substance against U937 cells showed that inhibition rates of the cell growth were 99.9% with the concentration of 0.32 nM PL9443. Also, the caspase-3/7 activity was 99% against U937 cells with the concentration of 0.08 nM of PL9443 substance. The result of the electrophoresis was that a DNA ladder was formed by the PL9443. The PL9443 compound is a promising lead compound as a photosensitizer for photodynamic therapy of cancer.

Effectiveness of Radionuclide Cisternography to Detect the Leakage Site of CSF in Spontaneous Intracranial Hypotension; Preliminary Report (자발성 두개강내 저뇌압증 환자의 뇌척수액 누출부위 진단에 방사성동위원소 뇌조조영술의 유효성: 예비결과 보고)

  • Kim, Seong-Min;Kim, Jae-Moon
    • Nuclear Medicine and Molecular Imaging
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    • v.40 no.3
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    • pp.148-154
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    • 2006
  • Purpose: Although radionuclide cisternography (RNC) is an useful study to detect cerebrospinal fluid (CSF) leakage in the patient with spontaneous intracranial hypotension (SIH), it sometimes fails to demonstrate the site of CSF leakage. The aim of the study is to improve the detection of leakage site of CSF and to reduce time for the study in RNC using modified protocol (m-RNC). Materials & methods : The study consists of 8 studies of 7 patients ($38{\pm}8$ years, M:F=2:5) with SIH, who underwent m-RNC following administration of 185-222 MBq of $^{99m}Tc$-DTPA into the lumbar subarachnoid space. Sequential images were obtained the whole spine with the head including urinary bladder at 10 minute, 30 minute, 1 hour, 2 hour, 4 hour and 6 hour. Radioactivity of extradural space and urinary bladder was evaluated. Results: Leakage site of CSF was identified in all 8 cases by m-RNC. Leakage site was cervicothoracic junction (CTJ, n=3), CTJ with C1-2 (n=2), CTJ with thoracic spine, thoracolumbar spine and lumbar spine (each n=1). All cases presented leakage sites within 1 hour and multiple sites, where CTJ was included in 6 cases. Only one case presented additional site in 6 hour image. Early radioactivity within the urinary bladder was noted in 6 cases, but that was fellowing after identification of the leakage site. Conclusion: Radionuclide cisternography is sensitive to detect the leakage site of CSF and is expected to improve the detection of CSF leakage site and reduce time for the study using modified protocol.