• Title/Summary/Keyword: B-MAC

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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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    • v.36 no.11
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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.

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

  • Lee, Eun-Jung;Kim, Tae-Hyung;Kim, Ha-Kun;Lee, Jung-Kee;Kwak, Hahn-Shik;Lee, Jong-Soo
    • Microbiology and Biotechnology Letters
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    • v.38 no.3
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    • pp.322-328
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    • 2010
  • Fermented skate is a traditional Korean food popular in Southwestern area of Korea. It has a characteristic flavor and alkaline pH. In this study we tried to determine the microbial flora in fermented skate using two different approaches. In culture-independent method, we amplified V2 region of 16S rRNA gene by PCR and cloned them into pUC18 plasmid to construct 16S rDNA fragment library. BLAST searches for the sequences obtained from this library revealed that uncultured bacterium clone 054E11.b was the most dominant flora in this fermented fish. In culture-dependent method, we diluted suspension of skate and spreaded on MRS, PCA, and MacConkey plates. We identified colonies grown on those plates by using PCR amplification of V2 region of 16S rRNA and DNA sequencing. BLAST searches of those DNA sequences resulted in totally different species with the observations from the 16S rDNA library analysis. Discrepancies of results obtained from both approaches suggest that the agar plates used in culture-dependent method may be different from the real condition of fermented skate. Therefore, results from culture-independent approach using 16S rDNA fragment library analysis may reflect real microbial flora in fermented skate.

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

  • Yu B. K;Kim H. J.;Oh K. Y.;Choe K. J.;Lee S. H.;Park H. J.;Kim B. K.
    • Journal of Animal Environmental Science
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    • v.11 no.1
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    • pp.11-16
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    • 2005
  • The round bale wrappers are generally used for rice straw after the harvesting of low land rice by combine harvester. In this situation, the bale wrappers should be well adapted under the travelling over raised borders and temporary ditches in soft soil of narrow rice fields. The study was conducted to improve the performance of bale wrapper through the new design for compact size, lowered gravity center and lowered power consumption. The prototype of round bale wrapper had been designed and assembled to tractor with three point hitch mounted. The machine type is one roll driving system with one roll for rotating and one roll for wrapping. The driving torque and work performance of the machines were measured and analysed. The torque requirement of the prototype and conventional type was 6kgf-m and 12kgf-m, respectively. The prototype shaved less friction resistance between bale driving roll and round bale. and the power requirement can also be reduced from 12kgf-m in the conventional to 6kgf-m in the prototype. The work efficiency of the new bale wrapper was $45\%$ higher than the conventional wrapper, and the working cost of the prototype can be reduced $17\%$ than that of the conventional.

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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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    • v.63 no.5
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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.

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

  • Kim Joo-Young;Lee Myung-Hag;Lee Kyu-Chan
    • Radiation Oncology Journal
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    • v.16 no.3
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    • pp.303-310
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    • 1998
  • Purpose : To evaluate the results of the treatment of locally advanced but resectable rectal cancers and to analyze prognostic factors. especially with the emphasis on the treatment time factor. Materials and Methods : There were 71 patients with rectal cancer who had been treated by curative surgical procedure and postoperative radiotherapy from August 1989 to December 1993. The minimum follow up period was 24 months and the median follow-up was 35 months Radiation therapy had been given by 6 MV linear accelerator by parallel opposing or four-box portals. Whole pelvis was treated up to 5040 cGy in most cases. Systemic chemotherapy had been given in 94$\%$ of the patients, mostly with 5-FU/ACNU regimen. Assessment for the overall and disease-free survival rates were done by life-table method and prognostic factors by Log-Rank tests. Results : Five-year overall survival, disease-free survival were 58.8$\%$ and 57$\%$, respectively. Two-year local control rate was 76.6$\%$. Stage according to Modified Astler-Coller (MAC) system, over 4 positive lymph nodes, over 6weeks interval between definitive surgery and adjuvant radiotherapy and over 7 days of interruption during radiotherapy period were statistically significant, or borderline significant prognostic factors. Conclusion : The treatment results of patients with rectal cancers are comparable to those of other large institutes. The treatment results for the patients with bowel wall penetration and/or positive regional lymph nodes were still discouraging for their high local recurrence rate for the patients with MAC 'c' stage diseases and high distant metastases rate even for the patients with node-negative diseases. Maybe more effective regimen of chemotherapy would be needed with proper route and schedule. To maximize postoperative adjuvant treatment. radiotherapy should be started at least within 6 weeks after surgery and preferably as soon as wound healing is completed. Interruption of treatment during radiotherapy course affects disease-free survival badly, especially if exceeds 7 days. So, the total treatment period trout definitive surgery to the completion of radiotherapy should be kept as minimal as possiable.

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

  • Kang, Dong-Kyoon;Kondo, Toru;Shin, Jong-Hee;Shin, Hye-Young;Sung, Jung-Hyun;Kang, Sang-Gu;Chang, Moo-Ung
    • Research in Plant Disease
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    • v.9 no.3
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    • pp.107-115
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    • 2003
  • Chinese yam (Dioscorea opposita cv. Jang-Ma) plants showing necrotic mosaic symptom were collected from their growing fields in Andong, Euisong, Gunwi and Daegu, Korea. Direct negative stainning method by electron microscope showed filamentous particles of about 660 nm in length. Immunosorbent electron microscopy (ISEM) revealed filamentous particles of 660nm decorated with antiserum of Chinese yam necrotic mosaic virus (ChYNMV). The virues purified partially were used to isolate viral RNA as template for RT-PCR to amplify about 1.2 kbp of 3'-terminal region (coat protein, 3'-UTR) with ChYNMV specific and oligo-dT primers. Amino acids sequences of amplified CP genes revealed that the viruses shared 97.9% similarity with ChYNMV (AB044386) wh ich was known as the member of Macluravirus. So the viruses from Chinese yam (D. opposita cv. Jang-Ma) plants were identified as ChYNMV. Comparing the CP amion acid sequences of ChYNMV strains with other macluraviruses such as Cardamon mosaic virus (CdMV), Narcissus latent virus (NLV) and Maclura mosaic virus (MacMV) revealed that N-terminal was the most varialbe region and conserved regions were present within the genus Macluravirus.

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

  • Pyo, Suhk-Neung;Baek, Soyoung;Kwak, Jang-Dong;Park, Dae-Sub;Joe, Sung-Jun;Lee, Hyun Ah
    • IMMUNE NETWORK
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    • v.3 no.1
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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.

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

  • Lee, Jae-Gu;Baek, Byeong-Geol;Lee, Sang-Bok
    • Parasites, Hosts and Diseases
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    • v.23 no.1
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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
    • Proceedings of the Korean Environmental Health Society Conference
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    • 2005.06a
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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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A STUDY OF THE MECHANISM OF IMPROVING ACID RESISTANCE OF BOVINE TOOTH ENAMEL AFTER PULSED Nd-YAG LASER IRRADIATION (펄스형 Nd-YAG 레이저 조사에 의한 법랑질 내산성 증가 기전에 관한 연구)

  • Lee, Young-Soon;Shon, Heung-Kyu
    • Journal of the korean academy of Pediatric Dentistry
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    • v.23 no.3
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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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