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Cellular Localization and Translocation of Duplication and Alternative Splicing Variants of Olive Flounder Phospholipase C-δ1 (넙치 3가지 타입 인지질가수분해효소(PLC-δ1)의 세포 내 위치 및 이동)

  • Kim, Na Young;Kim, Moo-Sang;Jung, Sung Hee;Kim, Myoung Sug;Cho, Mi Young;Chung, oon Ki;Ahn, Sang Jung
    • Journal of Life Science
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    • v.27 no.11
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    • pp.1369-1375
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    • 2017
  • The purpose of this study was to investigate the cellular characterization of phospholipase C-${\delta}1$ in olive flounders (Paralichthys olivaceus). In general, phospholipase C signaling pathways are distributed in nuclei at plasma membranes and in cytoplasms, although the pathways' nuclear localization mechanisms are unclear. P. olivaceus duplicates type-A PoPLC-${\delta}1$ (PoPLC-${\delta}1A$), which has a high similarity to the human isoform PLC-${\delta}$; type-B PoPLC-${\delta}1$ (PoPLC-${\delta}1B$ [Sf]), which has a low similarity to the human isoform PLC-${\delta}$ and the alternative splice variant PoPLC-${\delta}1B$ (Lf), which has a nuclear localization signal (NLS) and a nuclear export signal (NES) for nuclear imports and exports, respectively. This study confirmed the effects of the cellular localization and translocation of GFP-tagged PoPLC-${\delta}1A$, PoPLC-${\delta}1B$ (Sf) and PoPLC-${\delta}1B$ (Lf). It administered treatments of $Ca^{2+}$ ionophore ionomycin and endoplasmic reticulum (ER)-$Ca^{2+}$ pump inhibitor thapsigargin to hirame natural-embryo (HINAE) cells. A laser-scanning confocal microscope was used. GFP-tagged PoPLC-${\delta}1A$ was distributed to the cellular organelles, rather than to the cytoplasms and cytomembranes, when PoPLC-${\delta}1B$ (Lf) and PoPLC-${\delta}1B$ (Sf) were localized at the plasma membranes. The treatments of ionomycin and thapsigargin showed the accumulation of PoPLC-${\delta}1A$ in the nuclei when PoPLC-${\delta}1B$ (Lf) nucleocytoplasmic shuttling and PoPLC-${\delta}1B$ (Sf) nucleocytoplasmic shuttling were not observed. The results were the first evidence that PoPLC-${\delta}1A$, which contains functional, intact NES sequences, has a main role in nucleocytoplasmic shuttling and translocation in fish.

Closed Integral Form Expansion for the Highly Efficient Analysis of Fiber Raman Amplifier (라만증폭기의 효율적인 성능분석을 위한 라만방정식의 적분형 전개와 수치해석 알고리즘)

  • Choi, Lark-Kwon;Park, Jae-Hyoung;Kim, Pil-Han;Park, Jong-Han;Park, Nam-Kyoo
    • Korean Journal of Optics and Photonics
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    • v.16 no.3
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    • pp.182-190
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    • 2005
  • The fiber Raman amplifier(FRA) is a distinctly advantageous technology. Due to its wider, flexible gain bandwidth, and intrinsically lower noise characteristics, FRA has become an indispensable technology of today. Various FRA modeling methods, with different levels of convergence speed and accuracy, have been proposed in order to gain valuable insights for the FRA dynamics and optimum design before real implementation. Still, all these approaches share the common platform of coupled ordinary differential equations(ODE) for the Raman equation set that must be solved along the long length of fiber propagation axis. The ODE platform has classically set the bar for achievable convergence speed, resulting exhaustive calculation efforts. In this work, we propose an alternative, highly efficient framework for FRA analysis. In treating the Raman gain as the perturbation factor in an adiabatic process, we achieved implementation of the algorithm by deriving a recursive relation for the integrals of power inside fiber with the effective length and by constructing a matrix formalism for the solution of the given FRA problem. Finally, by adiabatically turning on the Raman process in the fiber as increasing the order of iterations, the FRA solution can be obtained along the iteration axis for the whole length of fiber rather than along the fiber propagation axis, enabling faster convergence speed, at the equivalent accuracy achievable with the methods based on coupled ODEs. Performance comparison in all co-, counter-, bi-directionally pumped multi-channel FRA shows more than 102 times faster with the convergence speed of the Average power method at the same level of accuracy(relative deviation < 0.03dB).

Construction of Web-Based Database for Anisakis Research (고래회충 연구를 위한 웹기반 데이터베이스 구축)

  • Lee, Yong-Seok;Baek, Moon-Ki;Jo, Yong-Hun;Kang, Se-Won;Lee, Jae-Bong;Han, Yeon-Soo;Cha, Hee-Jae;Yu, Hak-Sun;Ock, Mee-Sun
    • Journal of Life Science
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    • v.20 no.3
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    • pp.411-415
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    • 2010
  • Anisakis simplex is one of the parasitic nematodes, and has a complex life cycle in crustaceans, fish, squid or whale. When people eat under-processed or raw fish, it causes anisakidosis and also plays a critical role in inducing serious allergic reactions in humans. However, no web-based database on A. simplex at the level of DNA or protein has been so far reported. In this context, we constructed a web-based database for Anisakis research. To build up the web-based database for Anisakis research, we proceeded with the following measures: First, sequences of order Ascaridida were downloaded and translated into the multifasta format which was stored as database for stand-alone BLAST. Second, all of the nucleotide and EST sequences were clustered and assembled. And EST sequences were translated into amino acid sequences for Nuclear Localization Signal prediction. In addition, we added the vector, E. coli, and repeat sequences into the database to confirm a potential contamination. The web-based database gave us several advantages. Only data that agrees with the nucleotide sequences directly related with the order Ascaridida can be found and retrieved when searching BLAST. It is also very convenient to confirm contamination when making the cDNA or genomic library from Anisakis. Furthermore, BLAST results on the Anisakis sequence information can be quickly accessed. Taken together, the Web-based database on A. simplex will be valuable in developing species specific PCR markers and in studying SNP in A. simplex-related researches in the future.

Development of Marker-free TaGlu-Ax1 Transgenic Rice Harboring a Wheat High-molecular-weight Glutenin Subunit (HMW-GS) Protein (벼에서 밀 고분자 글루테닌 단백질(TaGlu-Ax1) 발현을 통하여 쌀가루 가공적성 증진을 위한 마커프리(marker-free) 형질전환 벼의 개발)

  • Jeong, Namhee;Jeon, Seung-Ho;Kim, Dool-Yi;Lee, Choonseok;Ok, Hyun-Choong;Park, Ki-Do;Hong, Ha-Cheol;Lee, Seung-Sik;Moon, Jung-Kyung;Park, Soo-Kwon
    • Journal of Life Science
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    • v.26 no.10
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    • pp.1121-1129
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    • 2016
  • High-molecular-weight glutenin subunits (HMW-GSs) are extremely important determinants of the functional properties of wheat dough. Transgenic rice plants containing a wheat TaGlu-Ax1 gene encoding a HMG-GS were produced from the Korean wheat cultivar ‘Jokyeong’ and used to enhance the bread-making quality of rice dough using the Agrobacterium-mediated co-transformation method. Two expression cassettes with separate DNA fragments containing only TaGlu-Ax1 and hygromycin phosphotransferase II (HPTII) resistance genes were introduced separately into the Agrobacterium tumefaciens EHA105 strain for co-infection. Rice calli were infected with each EHA105 strain harboring TaGlu-Ax1 or HPTII at a 3:1 ratio of TaGlu-Ax1 and HPTII. Among 210 hygromycin-resistant T0 plants, 20 transgenic lines harboring both the TaGlu-Ax1 and HPTII genes in the rice genome were obtained. The integration of the TaGlu-Ax1 gene into the rice genome was reconfirmed by Southern blot analysis. The transcripts and proteins of the wheat TaGlu-Ax1 were stably expressed in rice T1 seeds. Finally, the marker-free plants harboring only the TaGlu-Ax1 gene were successfully screened in the T1 generation. There were no morphological differences between the wild-type and marker-free transgenic plants. The quality of only one HMW-GS (TaGlu-Ax1) was unsuitable for bread making using transgenic rice dough. Greater numbers and combinations of HMW and LMW-GSs and gliadins of wheat are required to further improve the processing qualities of rice dough. TaGlu-Ax1 marker-free transgenic plants could provide good materials to make transgenic rice with improved bread-making qualities.

A Study on Basic Plan for Upscaling Environmental Conservation Value Assessment Map(ECVAM) of National Land in South Korea (대축척 국토환경성평가지도 작성방안 연구)

  • Lee, Moung-Jin;Jeon, Seong-Woo;Lee, Chong-Soo;Kang, Byung-Jin;Song, Won-Kyong
    • Journal of Environmental Policy
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    • v.6 no.3
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    • pp.115-145
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    • 2007
  • This study was performed for developing upscaling Environmental Conservation Value Assessment Map(ECVAM) of National Land in South Korea and presenting the application method of ECVAM. This ECVAM adopted the least indicator method and uses a Geographic Information System(GIS). This map is made through evaluation of 67 items. As a result, the construction of ECVAM was defined as a process of identifying land use to scientifically assess the physical and environmental value of land and classify conservation value into several grades for the sustainable management of environmental resources. After applying ECVAM criteria of five degrees to the whole of study area, Grade I, showing the highest conservation value, accounted for 29.3% by land area of ECVAM. Grades II, III, IV and V likewise accounted for, respectively, 21.7%, 17.2%, 7.1% and the lowest conservation value of 24.7%. other result, ECVAM and land suitability assessment agreement rate is Grade I 33.05%, Grades II, III, IV and V likewise accounted for 12.92%, 15.05%, 36.93% and last value of 53.28% This study set up "the realization of the improvement ECVAM" as the vision of the advancing strategy. In order to accomplish the vision, this study established the purpose as follow; constructing strategic assessment value relation to ECVAM based on knowledge, arranging the foundation to upscaling assessment value And this study devised preparatory plans to achieve the vision and the purpose as next; construction on base theme map by 1:5,000 scalie, base on land register theme map and precision land cover map. Therefore, for applying the result of this study to the upscaling Environmental Conservation Value Assessment Map(ECVAM), it considers regularly the systematic categorization of preceding item, consideration issue of national environmental geographic information using the ECVAM.

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An Exploration on Public Perception of Social Welfare as a Discipline in Korea (사회복지학에 대한 한국인의 인식에 관한 연구)

  • Kang, Chul-Hee
    • Korean Journal of Social Welfare
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    • v.57 no.4
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    • pp.147-175
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    • 2005
  • Efforts to identify the public's perception of social welfare as an academic discipline have never been conducted in Korea since the establishment of social welfare department in 1947 at Ewha Womans University. Such efforts are very meaningful in identifying directions and tasks to strengthen Korean social welfare as well as in clarifying and promoting our understanding concerning status of the academic discipline. This study attempts to explore and describe the degree of the public's perception in Korea with analyzing data surveyed in 2004 by our interdisciplinary research team. This study develops and uses a questionnaire having a Likert scale format that is composed of 8 points and measures the public's perception in the following dimensions: (1) personal interests on academic discipline; (2) contribution of academic discipline; (3) prospect of academic discipline; (4) importance of academic discipline; (5) expertise of academic discipline; and (6) personal knowledge on academic discipline. To avoid social desirability and promote objectivity with comparative measurement, this study selects ten representative academic disciplines as follows: medicine; physics; biology; social welfare; economics; psychology; sociology; political science; library science; and communication & journalism. This study attempts to identify (1) the degree of the public's perception on ten academic disciplines; (2) the position of social welfare by comparing it with each academic discipline and by comparing mean of social welfare with overall mean of six social science disciplines in the six dimensions; (3) the differences in the public's perceptions of social welfare on six dimensions by the respondents' status factor(high school students, college and graduate students, and citizens) and gender factor by using MANCOVA, and (4) the differences in the public's perceptions of social welfare on six dimensions by major factor(social welfare, social science majors, and natural science majors) and gender factor of college and graduate school students by using MANCOVA. The results of data analysis are as follows: (1) while the 3,319 respondents gave relatively high rating on natural sciences in the dimensions of contribution and expertise, they did the same on social sciences in the dimensions of personal interests and personal knowledge; (2) in overall comparisons, while the 3,319 respondents gave relatively high rating on social welfare in the dimensions of contribution, prospect and importance, they gave the lowest rating on the expertise of social welfare; (3) in the comparisons with social science disciplines, while the 3,319 respondents gave relatively high rating on social welfare in the dimensions of contribution, prospect and importance, they gave the lowest rating on the expertise of social welfare; (4) when analyzing all the respondents, there were differences in the vector of personal interests, contribution, prospect, importance, expertise, and personal knowledge by status factor, gender factor, and interaction effect factor; and (5) when analyzing only the respondents in college and graduate schools, there were differences in the vector of personal interests, contribution, prospect, importance, expertise, and personal knowledge by only major factor and gender factor. The results provide empirical backgrounds for discussing current image, status and major characteristics of social welfare as a discipline in Korea. Indeed, this study provides new meaningful and thoughtful guide for further investigation on the topic. In addition, contributing to clarifying and broadening our understandings about the public's perception on social welfare in Korea, this study discusses the tasks for dealing with expertise issue that is the most vulnerable issue of Korean social welfare discipline and research directions to strengthen and promote social welfare discipline in Korea.

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Enhanced Growth Inhibition by Combined Gene Transfer of p53 and $p16^{INK4a}$ in Adenoviral Vectors to Lung Cancer Cell Lines (폐암세포주에 대한 p53 및 $p16^{INK4a}$의 복합종양억제유전자요법의 효과)

  • Choi, Seung -Ho;Park, Kyung-Ho;Seol, Ja-Young;Yoo, Chul-Gyu;Lee, Choon-Taek;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.1
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    • pp.67-75
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    • 2001
  • Background : Two tumor suppressor genes, p53 and p16, which have different roles in controlling the cell cycle and inducing apoptosis, are frequently inactivated during carcinogenesis including lung cancer. Single tumor suppressor gene therapies using either with p53 or p16 have been studied extensively. However, there is a paucity of reports regarding a combined gene therapy using these two genes. Methods : The combined effect of p53 and p16 gene transfer by the adenoviral vector on the growth of lung cancer cell lines and its interactive mechanism was investigated. Results : An isobologram showed that the co-transduction of p53 and p16 exhibited a synergistic growth in hibitory effect on NCI H358 and an additive effect on NCI H23. Cell cycle analysis demonstrated the induction of a synergistic G1/S arrest by a combined p53 and p16 transfer. This synergistic interaction was again confirmed in a soft agar confirmed in a soft agar clonogenic assay. Conclusion : These observations suggest the potential of a p53 and p16 combination gene therapy as another potent strategy in cancer gene therapy.

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Expression and Purification of Recombinant Human Interferon-gamma Produced by Escherichia coli (대장균이 생산한 재조합 인체 감마인터페론의 발현과 정제)

  • Park, Jung-Ryeol;Kim, Sung-Woo;Kim, Jae-Bum;Jung, Woo-Hyuk;Han, Myung-Wan;Jo, Young-Bae;Jung, Joon-Ki
    • KSBB Journal
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    • v.21 no.3
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    • pp.204-211
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    • 2006
  • For the production of the recombinant human interferon-gamma(rhIFN-${\gamma}$) in Escherichia coli, human glucagon and ferritin heavy chain were used as fusion partners. Even though rhIFN-${\gamma}$ is expressed as an inclusion body form in E. coli because of strong hydrophobicity of itself, over 50% of fused rhIFN-${\gamma}$ was expressed as soluble form in E. coli $Origami^{TM}$(DE3) harboring pT7FH(HE)-IFN-${\gamma}$ which encodes ferritin heavy chain-fused rhIFN-${\gamma}$. In the case of using glucagon-ferritin heavy chain hybrid mutant as a fusion partner, 6X His-tag was additionally introduced to N-terminus of GFHM(HE)-IFN-${\gamma}$ for enhancing purification yields of rhIFN-${\gamma}$. Fusion protein HGFHM(HE)-IFN-${\gamma}$ with two 6X His-tag was more effectively bound to Ni-NTA agarose bead than GFHM(HE)-IFN-${\gamma}$ with a 6X His-tag. rhIFN-${\gamma}$ was completely purified from enterokinase-treated HGFHM(HE)-IFN-${\gamma}$ by Ni-NTA affinity column. For high-level production of rhIFN-${\gamma}$, glucose was used as the sole carbon source with simple exponential feeding rate($2.4{\sim}7.2g/h$) in fed-batch process. The effective lactose concentration for the expression of the rhIFN-${\gamma}$ was $10{\sim}20mM$. Under the fed-batch culture conditions, rhIFN-${\gamma}$ production yield reached 11 g DCW/L for 6 hours after lactose induction.

Enhanced production of monacolin-K through supplement of monacolin-K precursors into production medium and cloning of SAM synthetase gene (metK) (Precursor제공 및 생합성 관련 유전자의 cloning을 통한 Monacolin-K 생산성 향상)

  • Lee, Mi-Jin;Jeong, Yong-Seob;Chun, Gie-Taek
    • KSBB Journal
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    • v.23 no.6
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    • pp.519-524
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    • 2008
  • Monacolin-K is a strong anti-hypercholesterolemic agent produced by Monascus sp. via polyketide pathway. High-yielding mutants of monacolin-K were developed through rational screening strategies adopted based on understanding of monacolin-K biosynthetic pathway. Through the experiments for investigating various amino acids as putative precursors for the monacolin-K biosynthesis, it was found that production level of monacolin-K was remarkably increased when optimum amount of cysteine was supplemented into the production medium. We suggested that these phenomena might be related to the special roles of SAM (S-adenosyl methionine), a putative methyl group donor in the biosynthetic pathway of monacolin-K, demonstrating close interrelationship between SAM-synthesizing primary metabolism and monacolin-K synthesizing secondary metabolism. Namely, increase in the intracellular amount of SAM derived from the putative precursor, cysteine which was extracellularly supplemented into the production medium might contribute to the significant enhancement in the monacolin-K biosynthetic capability of the highly mutated producers. On the basis of these assumptions derived from the above fermentation results, we decided to construct efficient expression vectors harboring SAM synthetase gene (metK) cloned from A. nidulans, with the hope that increased intracellular level of SAM could lead to further enhancement in the monacolin-K production through overcoming a rate-limiting step associated with monacolin-K biosynthesis. Hence, in order to overcome the plausible rate-limiting step associated with monacolin-K biosynthesis by increasing intracellular level of SAM, we transformed the producer mutants with an efficient expression vector harboring gpdA promoter of the producer microorganism, and metK gene. Notably, from the resulting various transformants, we were able to screen a very high-yielding transformant which showed approximately 3.3 fold higher monacolin-K productivity than the parallel nontransformed mutants in shake flask cultures performed under the identical fermentation conditions.

Korean Word Sense Disambiguation using Dictionary and Corpus (사전과 말뭉치를 이용한 한국어 단어 중의성 해소)

  • Jeong, Hanjo;Park, Byeonghwa
    • Journal of Intelligence and Information Systems
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    • v.21 no.1
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    • pp.1-13
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    • 2015
  • As opinion mining in big data applications has been highlighted, a lot of research on unstructured data has made. Lots of social media on the Internet generate unstructured or semi-structured data every second and they are often made by natural or human languages we use in daily life. Many words in human languages have multiple meanings or senses. In this result, it is very difficult for computers to extract useful information from these datasets. Traditional web search engines are usually based on keyword search, resulting in incorrect search results which are far from users' intentions. Even though a lot of progress in enhancing the performance of search engines has made over the last years in order to provide users with appropriate results, there is still so much to improve it. Word sense disambiguation can play a very important role in dealing with natural language processing and is considered as one of the most difficult problems in this area. Major approaches to word sense disambiguation can be classified as knowledge-base, supervised corpus-based, and unsupervised corpus-based approaches. This paper presents a method which automatically generates a corpus for word sense disambiguation by taking advantage of examples in existing dictionaries and avoids expensive sense tagging processes. It experiments the effectiveness of the method based on Naïve Bayes Model, which is one of supervised learning algorithms, by using Korean standard unabridged dictionary and Sejong Corpus. Korean standard unabridged dictionary has approximately 57,000 sentences. Sejong Corpus has about 790,000 sentences tagged with part-of-speech and senses all together. For the experiment of this study, Korean standard unabridged dictionary and Sejong Corpus were experimented as a combination and separate entities using cross validation. Only nouns, target subjects in word sense disambiguation, were selected. 93,522 word senses among 265,655 nouns and 56,914 sentences from related proverbs and examples were additionally combined in the corpus. Sejong Corpus was easily merged with Korean standard unabridged dictionary because Sejong Corpus was tagged based on sense indices defined by Korean standard unabridged dictionary. Sense vectors were formed after the merged corpus was created. Terms used in creating sense vectors were added in the named entity dictionary of Korean morphological analyzer. By using the extended named entity dictionary, term vectors were extracted from the input sentences and then term vectors for the sentences were created. Given the extracted term vector and the sense vector model made during the pre-processing stage, the sense-tagged terms were determined by the vector space model based word sense disambiguation. In addition, this study shows the effectiveness of merged corpus from examples in Korean standard unabridged dictionary and Sejong Corpus. The experiment shows the better results in precision and recall are found with the merged corpus. This study suggests it can practically enhance the performance of internet search engines and help us to understand more accurate meaning of a sentence in natural language processing pertinent to search engines, opinion mining, and text mining. Naïve Bayes classifier used in this study represents a supervised learning algorithm and uses Bayes theorem. Naïve Bayes classifier has an assumption that all senses are independent. Even though the assumption of Naïve Bayes classifier is not realistic and ignores the correlation between attributes, Naïve Bayes classifier is widely used because of its simplicity and in practice it is known to be very effective in many applications such as text classification and medical diagnosis. However, further research need to be carried out to consider all possible combinations and/or partial combinations of all senses in a sentence. Also, the effectiveness of word sense disambiguation may be improved if rhetorical structures or morphological dependencies between words are analyzed through syntactic analysis.