• 제목/요약/키워드: B-MAC

검색결과 312건 처리시간 0.031초

Optimizing hormonal and amino acid combinations for enhanced cell proliferation and cell cycle progression in bovine mammary epithelial cells

  • Hyuk Cheol Kwon;Hyun Su Jung;Do Hyun Kim;Jong Hyeon Han;Seo Gu Han;Dong Hyun Keum;Seong Joon Hong;Sung Gu Han
    • Animal Bioscience
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    • 제36권11호
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    • pp.1757-1768
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    • 2023
  • Objective: The number of bovine mammary epithelial cells (BMECs) is closely associated with the quantity of milk production in dairy cows; however, the optimal levels and the combined effects of hormones and essential amino acids (EAAs) on cell proliferation are not completely understood. Thus, the purpose of this study was to determine the optimal combination of individual hormones and EAAs for cell proliferation and related signaling pathways in BMECs. Methods: Immortalized BMECs (MAC-T) were treated with six hormones (insulin, cortisol, progesterone, estrone, 17β-estradiol, and epidermal growth factor) and ten EAAs (arginine, histidine, leucine, isoleucine, threonine, tryptophan, lysine, methionine, phenylalanine, and valine) for 24 h. Results: Cells were cultured in a medium containing 10% fetal bovine serum (FBS) as FBS supplemented at a concentration of 10% to 50% showed a comparable increase in cell proliferation rate. The optimized combination of four hormones (insulin, cortisol, progesterone, and 17β-estradiol) and 20% of a mixture of ten EAAs led to the highest cell proliferation rate, which led to a significant increase in cell cycle progression at the S and G2/M phases, in the protein levels of proliferating cell nuclear antigen and cyclin B1, cell nucleus staining, and in cell numbers. Conclusion: The optimal combination of hormones and EAAs increased BMEC proliferation by enhancing cell cycle progression in the S and G/2M phases. Our findings indicate that optimizing hormone and amino acid levels has the potential to enhance milk production, both in cell culture settings by promoting increased cell numbers, and in dairy cows by regulating feed intake.

배양 의존적 및 배양 비의존적 방법에 의한 홍어회 서식 미생물의 다양성 분석 (Analysis of Bacterial Diversity in Fermented Skate Using Culture-dependent and Culture-independent Approaches)

  • 이은정;김태형;김하근;이정기;곽한식;이종수
    • 한국미생물·생명공학회지
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    • 제38권3호
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    • pp.322-328
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    • 2010
  • 발효홍어(홍어회)에 존재하는 박테리아 집단의 다양성을 확인하기 위해, 박테리아의 16S rDNA 절편들을 증폭하고 클로닝하여 라이브러리를 구축하였다. 삽입 서열의 염기서열을 결정한 후, BLAST 분석에 의해 미생물 동정을 하였다. 동일한 삽입서열 빈도를 계산하였을 때, 발효홍어(홍어회)에는 uncultured bacterium clone 054E11.b가 57.1% 나타남으로써 우점균으로 존재하고 있다고 추정하였다. 또한 발효홍어(홍어회) 현탁액을 한천배지에 도말하여 형성된 집락을 콜로니 PCR을 수행하였을 때, Pseudomonas sp. KC-EPS13 등 12 종의 박테리아가 동정되었다. 배양 의존적 방법과 배양 비의존적 방법으로 홍어회를 분석하였을 때, Psychrobacter sp. J466만이 두 가지 방법에서 모두 검출되었고 나머지 박테리아들의 균총은 상이하였다.

롤 구동 래핑암 회전식 원형베일래퍼의 구동 토크 분석 (Driving Torque Analysis of Role Driving & Wrapping Arm Rotation Type Round Bale Wrapper)

  • 유병기;김혁주;오권영;최광재;이성현;박환중;김병관
    • 한국축산시설환경학회지
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    • 제11권1호
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    • pp.11-16
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    • 2005
  • 1. 구획이 작고 논두렁이 많은 논의 볏짚 수집에 적합한 1축롤 구동 래핑암회전식 트랙터 3점히치 장착형 원형베일 래퍼를 개발하였다. 2. 2개의 구동롤을 구동하는 래핑암 회전식원형베일래퍼는 베일 적재시 베일에 변형이 발생하였으며 작업시 무게중심이 높아 기체 흔들림이 있었으나 시작기는 베일적재시 롤이 수평직선운동을 하여 베일의 변형이 적었으며 무게중심이 낮아 작업 안정성이 다소 높은 것으로 평가되었다. 3. 베일적재시에 두 개의 적재롤이 자유회전을 하여 적재시의 베일과 롤의 마찰저항을 구름저항으로 바꿈으로써 베일의 변형이 생기지 않았으며 베일을 회전시키기 위한 구동롤의 토크를 기존의 2축롤 구동식은 12 kgf-m 이었으나 새로 개발한 1축롤 구동식은 6 kgf-m로 낮았다. 4. 구동롤 토크가 낮아 1축롤 구동으로 베일래핑작업이 가능하였으며 기계구조를 단순화 킬 수 있었다. 5. 기존 회전테이블식에 비해 작업능률을 $45\%$ 향상, 작업비용은 $17\%$ 절감할 수 있었다.

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Enhanced supply of methionine regulates protein synthesis in bovine mammary epithelial cells under hyperthermia condition

  • Zhou, Jia;Yue, Shuangming;Xue, Benchu;Wang, Zhisheng;Wang, Lizhi;Peng, Quanhui;Xue, Bai
    • Journal of Animal Science and Technology
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    • 제63권5호
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    • pp.1126-1141
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    • 2021
  • Recent evidence has shown that methionine (Met) supplementation can improve milk protein synthesis under hyperthermia (which reduces milk production). To explore the mechanism by which milk protein synthesis is affected by Met supplementation under hyperthermia, mammary alveolar (MAC-T) cells were incubated at a hyperthermic temperature of 42℃ for 6 h in media with different concentrations of Met. While the control group (CON) contained a normal amino acid concentration profile (60 ㎍/mL of Met), the three treatment groups were supplemented with Met at concentrations of 10 ㎍/mL (MET70, 70 ㎍/mL of Met), 20 ㎍/mL (MET80, 80 ㎍/mL of Met), and 30 ㎍/mL (MET90,90 ㎍/mL of Met). Our results show that additional Met supplementation increases the mRNA and protein levels of BCL2 (B-cell lymphoma-2, an anti-apoptosis agent), and decreases the mRNA and protein levels of BAX (Bcl-2-associated X protein, a pro-apoptosis agent), especially at an additional supplementary concentration of 20 ㎍/mL (group Met80). Supplementation with higher concentrations of Met decreased the mRNA levels of Caspase-3 and Caspase-9, and increased protein levels of heat shock protein (HSP70). The total protein levels of the mechanistic target of rapamycin (mTOR) and the mTOR signalling pathway-related proteins, AKT, ribosomal protein S6 kinase B1 (RPS6KB1), and ribosomal protein S6 (RPS6), increased with increasing Met supplementation, and peaked at 80 ㎍/mL Met (group Met80). In addition, we also found that additional Met supplementation upregulated the gene expression of αS1-casein (CSN1S1), β-casein (CSN2), and the amino acid transporter genes SLC38A2, SLC38A3 which are known to be mTOR targets. Additional Met supplementation, however, had no effect on the gene expression of κ-casein (CSN3) and solute carrier family 34 member 2 (SLC34A2). Our results suggest that additional Met supplementation with 20 ㎍/mL may promote the synthesis of milk proteins in bovine mammary epithelial cells under hyperthermia by inhibiting apoptosis, activating the AKT-mTOR-RPS6KB1 signalling pathway, and regulating the entry of amino acids into these cells.

직장암의 수술 후 방사선치료의 성적 - 예후 인자와 전체 치료기간이 미치는 영향에 관한 고찰 - (Results of Postoperative Radiation Therapy of Rectal Cancers - with the Emphasis of the Overall Treatment Time -)

  • 김주영;이명학;이규찬
    • Radiation Oncology Journal
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    • 제16권3호
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    • pp.303-310
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    • 1998
  • 목적 : 국소적으로 진행된 직장암의 방사선치료에 있어 전통적인 치료방법으로 알려져 있는 근치적 수술 및 보조 방사선 및 항암치료의 성적을 알아보고 이에 영향을 미치는 예후 인자를 알아보기 위함이다. 대상 및 방법 : 1989년 7월 부터 1993년 12월 까지 항문상방 15 cm 이내에 존재하는 직장암으로 진단받고 근치적 수술을 받은 71명의 환자를 대상으로 후향적 분석을 시행하였다. 방사선치료는 6 MeV 선형가속기를 사용하여 주5회씩 5040 cGy 까지 조사되였고 수술후 21일에서 94일 사이에 시작되었고 5-FU와 ACNU chemotherapy가 4주 간격으로 시행되었다. 결과 : 전체환자의 5년 생존율과 5년 무병생존율은 각각 58.8$\%$및 57$\%$였다. 대상환자들의 2년 국소제어율은 76.6$\%$였다. 생존기간 및 무병생존기간의 중앙값은 각각 30개월 및 27개월이었다. 단변량 및 다변량분석시 무병생존율에 의미있게 예후인자로 작용했던 요인은 국소림프절로의 전이여부, 4개 이상의 임파선 전이, 6주이상의 수술과 방사선치료 시작 사이의 간격 및 7일 이상 지속되는 방사선치료 도중의 휴식기간 등이었다. 결론 : 국소적으로 진행된 직장암의 경우 근치적 수술 및 방사선치료 및 5-FU/ACNU를 기본으로 한 항암제를 시행하였을 때 B3 이상의 병기에서는 아직도 생존율이 저조하며 bowel wall penetration이 있거나 국소 림프절전이가 있을 때는 국소제어율도 환자의 절반 정도에서 밖에 얻을 수 없을 뿐만이 아니라 림프절전이가 없는 경우라도 원격전이율이 치료실패의 많은 부분을 차지함을 알 수 있었다. 이는 좀더 효과적인 항암치료제의 선택과 투여 방법이 방사선치료와 병행될 필요가 있음을 시사하며 수술 후 보조적인 방사선치료를 시행함에 있어서 방사선치료가 시작되는 기간이 지연될 때, 또 방사선치료가 7일 이상 중단될 때 치료결과에 영향을 줄 수 있음을 보여준다. 직장선암의 경우 수술과 방사선치료시작까지의 기간이 필요이상으로 연장되거나 치료중 휴식기간이 생겨 전체치료기간이 길어지지 않게 하여야 할 것이다.

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한국산 장마(Dioscorea oppasita cv. Jang-Ma)에서 분리한 Chinese yam necrotic mosaic virus (Chinese yam necrotic mosaic virus Isolated from Chinese Yam in Korea)

  • 강동균;곤도토루;신종희;신혜영;성정현;강상구;장무웅
    • 식물병연구
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    • 제9권3호
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    • pp.107-115
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    • 2003
  • 경상북도 안동, 의성, 군위, 대구의 장마 재배포장에서 모자이크와 괴저반점을 보이는 장마잎을 채집하였다. 이들 이병 잎 조직을 시료로 하여 D N 법 및 ISEM법에 의해 투과전자현미경으로 바이러스 입자를 관찰한 결과, 각각의 시료에서 ChYNMV 항혈청에 반응한 660nm의 사상형 입자가 확인되었다. 장마 잎에서 부분 정제한 바이러스에서 RNA를 추출하여 이것을 주형으로 ChYNMV 특이적 프라이머와 oligo-dT 프라이머를 이용하여 외피단백질 유전자와 3‘-말단 비전사부위를 포함하는 약 1.2kbp의 3’-말단을 증폭하였다. 외피단백질 아미노산서열은 Macluravirus로 알려진 ChYNMV (A B044386)와 97.9%의 상동성을 보여 장마에서 분리한 바이러스를 ChYNMV로 동정하였다. ChYNMV와 다른 Macluavirus의 외피 단백질 아미노산서열을 비교한 결과, M말단 영역에서 가장 많은 변이를 확인하였고 Macluravirus 속에는 잘 보존된 영역이 존재하였다.

종양 미세 환경 내 대식세포에서 혈관 신생 조절 인자로서의 TNF-α에 의한 IFN-γ의 분비 조절 (Production of IFN-γ by TNF-α in Macrophages from Tumor Micro Environment; Significance in Angiogenic Switch Control)

  • 표석능;백소영;곽장동;박대섭;조성준;이현아
    • IMMUNE NETWORK
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    • 제3권1호
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    • pp.53-60
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    • 2003
  • Background: The role of macrophages in tumor angiogenesis is known to be the production of angiogenic cytokines and growth factors including TNF-${\alpha}$. Recently, macrophage also can produce the INF-${\gamma}$ that is being studied to be involved in angiogenic inhibition. Thus, the importance of macrophages in tumor angiogenesis is might being an angiogenic switch. Thus, the hypothesis tested here is that TNF-${\alpha}$ can modulate the INF-${\gamma}$ production in the macrophages from tumor environment as a part of tumor angiogenic switch. Methods: Macrophages in tumor environment were obtained from the peritoneal cavity of C57BL/6 mice injected with B16F10 melanoma cell line for 6 or 11 days. $Mac1^+$-macrophages were purified using magnetic bead ($MACs^{TM}$; Milteny Biotech, Germany) and cultured with various concentrations of TNF-${\alpha}$ for various time points at $37^{\circ}C$. The supernatants were analyzed for IFN-${\gamma}$ or VEGF by ELISA kit (Endogen, Woburn, MA). Results: Residential macrophages from the peritoneal cavity did not respond to LPS or TNF-${\alpha}$ to produce INF-${\gamma}$. However, the cells from tumor environment produced IFN-${\gamma}$ as well as VEGF and upregulated by the addition of LPS or TNF-${\alpha}$. RT-PCR analysis revealed the external TNF-${\alpha}$-induced IFN-${\gamma}$ gene expression in the macrophages from tumor environment. Conclusion: The overall data suggest that the macrophages in tumor environment might have an important role not only in angiogenic signal but also in anti-angiogenic signal by producing related cytokines. And TNF-${\alpha}$ might be a key cytokine in tumor angiogenic switch.

간질증의 효소면역학적 진단 (Application of Micro-BLISA in Serodiagnosis of Fascioliasis in Cattle)

  • 이재구;백병걸;이상복
    • Parasites, Hosts and Diseases
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    • 제23권1호
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    • pp.95-101
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    • 1985
  • Fascioliasis in cattle is one of the most common and very serious trematode diseases in Korea. In the present study, the enzyme linked immunosorbent assay (ELISA) was applied in the diagnosis of fascioliasis using antigen of Fasciela hepatica, perokidase of conjugate anti-cattle Is G and orthophenylenediamine as a substrate by micro-method technique of Volley et at. (1976b) and MacLaren (1978) with a slight modification. Results obtained from the present study are as follows. 1. In assay for optimal dilution of stock antigen, the antigen (protein contents; 0. Bmgymz) was diluted from 1150 to 1/600 with carbonate buffer (pH 9.6), and then absorbance values were measured with 1/100 diluted sera. The regression equations between the OD values of ELISA and dilution of antigen were log Y: -0.181-0.00127X in infected sera, and log Y: -0.578-0. 000879X in normal sera. The significantly higher (p<0.05) OD value was observed in the former. 2. In assay for optimal dilution of sera, the sera were diluted from 1125 to 1/400 with in PBSJ Tween 20 (pH 7.4), and absorbance values were measured with 1/200 diluted antigen. The regression equation between the OD values of ELISA and dilution of sera were log Y: -0.1540-0.0007238X in infected sera and log Y: -0.4834-0.00116X in normal sera. The former was higher than the latter (p<0.05). 3. In the 27 cases of negative intradermal test, OD values of the ELISA are $0.447{\pm}0.144$, the 95% confidence interval (Mean+2 H SD) of the values was 0.735, and there was no case over the values. Therefore, the sensitivity of the antigen to diagnose fascioliasis was 100% in the negative case. The OD value 0.7 which is designed as a criterion (detection level of positive one) is useful for the performance of the ELISA in fascioliasis. 4. According to the OD value of criterion in the regression equations, the optimal dilutions of stock antigen and serum were 1/250 and 1/100, respectively. 5. In the 58 cases of fascioliasis from which the adult could be found in the bile ducts, the OD value was $0.846{\pm}0.224$. The 75% (44 cattle) among them had higher value with compared to the criterion, and the 60% (20 cattle) of the cases of proliferative cholangitis of 33 cattle which had been infected previousely with Fasciola sp. is higher than the criterion. 6. Prevalence of fascioliasis was 43.4% in the application of the ELISA to 272 cattle which were reared in Jeonbug district.

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Microbe Hunting: A Curious Case of Cryptococcus

  • Bartlett, Karen H.;Kidd, Sarah;Duncan, Colleen;Chow, Yat;Bach, Paxton;Mak, Sunny;MacDougall, Laura;Fyfe, Murray
    • 한국환경보건학회:학술대회논문집
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    • 한국환경보건학회 2005년도 국제학술대회
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    • pp.45-72
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    • 2005
  • C. neoformans-associated cryptococcosis is primarily a disease of immunocompromised persons, has a world-wide distribution, and is often spread by pigeons in the urban environment. In contrast, C. gattii causes infection in normal hosts, has only been described in tropical and semi-tropical areas of the world, and has a unique niche in river gum Eucalyptus trees. Cryptococcosis is acquired through inhalation of the yeast propagules from the environment. C. gattii has been identified as the cause of an emerging infectious disease centered on Vancouver Island, British Columbia, Canada. No cases of C. gattii-disease were diagnosed prior to 1999; the current incidence rate is 36 cases per million population. A search was initiated in 2001 to find the ecological niche of this basidiomycetous yeast. C. gaftii was found in the environment in treed areas of Vancouver Island. The highest percentage of colonized-tree clusters were found around central Vancouver Island, with decreasing rates of colonization to the north and south. Climate, soil and vegetation cover of this area, called the Coastal Douglas fir biogeoclimatic zone, is unique to British Columbia and Canada. The concentration of airborne C. gattii was highest in the dry summer months, and lowest during late fall, winter, and early spring, months which have heavy rainfall. The study of the emerging colonization of this organism and subsequent cases of environmentally acquired disease will be informative in planning public health management of new routes of exposure to exotic agents in areas impacted by changing climate and land use patterns. Cryptococcosis is an infection associated with an encapsulated, basidiomycetous yeast Cryptococcus neoformans. The route of entry for this organism is through the lungs, with possible systemic spread via the circulatory system to the brain and meninges. There are four cryptococcal serogroups associated with disease in humans and animals, distinguished by capsular polysaccharide antigens. Cryptococcus neoformans: variety grubii (serotype A), variety neoformans (serotype D), and variety gattii (serotypes B and C) (Franzot et at. 1999). C. neoformans variety gattii has recently been elevated to species status, C. gattii. C. neoformans val. grubii and var. neoformans have a world-wide distribution, and are particularly associated with soil and weathered bird droppings. In contrast, C. gattii (CG) is not associated with bird excrement, is primarily found in tropical and subtropical climates, and has a restricted environmental niche associated with specific tree species. (Ellis & Pfiffer 1990) Ellis and Pfeiffer theorize that, as a basidiomycete, CG requires an association with a tree in order to become pathogenic to mammals. In Australia, CG has been found to be associated with five species of Eucalypts, Eucalyptus camaldulensis, E. tereticornis, E. blakelyi, E. gomphocephala, and E. rudis. Eucalypts, although originally native to Australia, now have a world-wide distribution. CG has been found associated with imported eucalypts in India, California, Brazil, and Egypt. In addition, in Brazil and Columbia, where eucalypts have been naturalized, native trees have been shown to harbour CG (Callejas et al. 1998; Montenegro et al. 2000). In British Columbia, Canada, since the beginning of 1999, there have been 120 confirmed cases of cryptococcal mycoses associated with CG in humans, including 4 fatalities (data from British Columbia Centre for Disease Control), and over 200 cases in animal pets in BC (data from Central Laboratory for Veterinarians). What is remarkable about the BC outbreak of C. gattii-cryptococcosis is that all of the cases have been residents of, or visitors to, a narrow area along the eastern coast of Vancouver Island, BC, from the tip of the island in the south (Victoria) to Courtenay on the north-central island as illustrated in Figure 1. Of the first 38 human cases, 58% were male with a mean age of 59.7 years (range 20 - 82): 36 cases (95%) were Caucasian. Ten cases (26%) presented with meningitis, the remainder presented with respiratory symptoms. Cultures recovered from cases of cryptococcosis associated with the outbreak were typed as serogroup B, which is specific to CG (Bartlett et al. 2003). This was the first reported outbreak of CVG in Canada, or indeed, the world. Where infection with CG is endemic, for example, Australia, the incidence of cryptococcosis ranges from 1.8 - 4.7 per million between the southern and northern states (Sorrell 2001). However, the overall incidence of cryptococcosis in immunocompenent individuals has been estimated at 0.2 per million population per year (Kwon-Chung et al. 1984). The population of Vancouver Island is approximately 720,000,consequently, even if the organism were endemic, one would expect a maximum of 0.15 cases of cryptococcal disease annually.

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펄스형 Nd-YAG 레이저 조사에 의한 법랑질 내산성 증가 기전에 관한 연구 (A STUDY OF THE MECHANISM OF IMPROVING ACID RESISTANCE OF BOVINE TOOTH ENAMEL AFTER PULSED Nd-YAG LASER IRRADIATION)

  • 이영순;손흥규
    • 대한소아치과학회지
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    • 제23권3호
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    • pp.640-658
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    • 1996
  • The purpose of this study was to examine the mechanism of improving acid resistance of Nd-YAG laser irradiated tooth enamel and determine the most effective energy density for improving acid resistance. The bovine tooth enamel were lased with a pulsed Nd-YAG laser. The energy densities of exposed laser beam were varied from 10 to $70\;J/cm^2$. To investigate the degree of improving acid resistance by irradiation, all the samples were submerged to demineralize in 0.5 N $HClO_4$ solution for 1 minute. After 1 minute, 0.05 % $LaCl_3$ was added to the solution for interrupting the demineralization reaction. The amounts of dissolved calcium and phosphate in the solution were measured by using an atomic absorption spectrophotometer and the UV/VIS spectrophotometer, respectively. To examine the mechanism of improving acid resistance, X-ray diffraction analysis, infrared spectroscopy, and scanning electron microscopy were taken. The X-ray diffraction pattern of the samples were obtained in the $10^{\circ}{\sim}80^{\circ}2{\theta}$ range with $Cu-K{\alpha}$ radiation using M18HF(Mac Science Co.) with X-ray diffractometer operating at 40 KV and 300 mA. The infra-red spectra of the ground samples in 300 mg KBr pellets 10 mm diameter were obtained in the $4000cm^{-1}\;to\;400cm^{-1}$ range using JASCO 300E spectrophotometer. The scanning electron microscopy was carried out using JSM6400(JEOL Co.) with $500{\sim}2000$ times magnification. The results were as follow 1. The concentration of calcium dissolved from laser irradiated enamel with $50J/cm^2$ was significantly lesser than that of unlased control group (p<0.05) 2. From the result of the X-ray diffraction analysis, $\beta$-TCP, which increases acid solubility, was identified in lased enamel but the diffraction peaks of (002) and (004) became sharp with increasing energy density of laser irradiation. This means that the crystals in lased samples were grown through the c-axis and subsequently, the acid solubility of enamel decreased. 3. The a-axis parameter was slightly increased by laser irradiation, whereas the c-axis parameter was almost constant except for a little decrease at $50J/cm^2$. 4. In the infra-red spectra of lased enamels, phosphate bands ($600{\sim}500cm^{-1}$), B-carbonate bands (870, $1415{\sim}1455cm^{-1}$), and A-carbonate band ($1545cm^{-1}$) were observed. The amounts of phosphate bands and the B-carbonate bands were reduced, on the other hand, the amount of the A-carbonate band was increased by increase the energy density. 5. The SEM experiments reveal that the surface melting and recrystallization were appeared at $30J/cm^2$ and the cracks were observed at $70J/cm^2$. From above results, It may be suggested that the most effective energy density for improving acid resistance of tooth enamel with the irradiation of Nd-YAG laser was $50J/cm^2$. The mechanism of improving acid resistance were reduction of permeability due to surface melting and recrystallization of lased enamel and reduction of acid solubility of enamel due to decrease of carbonate content and growth of crystal.

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