• Title/Summary/Keyword: In-vivo Test.

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Effect of Chromium Dietary Supplementation on the Immune Response and Some Blood Biochemical Parameters of Transport-stressed Lambs

  • Al-Mufarrej, S.I.;Al-Haidary, I.A.;Al-Kraidees, M.S.;Hussein, M.F.;Metwally, H.M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.5
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    • pp.671-676
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    • 2008
  • Forty-eight Naemi lambs (avg. BW 31.7 kg) were transported by truck for a distance of 1,450 km from Al-Jouf to Riyadh, Saudi Arabia. On arrival day, the lambs were randomly allocated to four groups receiving diets supplemented with 0.0, 0.3, 0.6 and 0.9 ppm organic chromium (Cr). Each group consisted of four separately housed replicates of three lambs each. The animals were fed ad libitum on a grower diet for 84 days. Blood samples were obtained shortly before transportation, upon arrival and at weekly intervals thereafter from all lambs for analysis of plasma and serum. Plasma glucose and serum cortisol, total protein, albumin, urea-N and total cholesterol concentrations were determined. A cursory clinical examination of the lambs, along with rectal temperature, was undertaken at different intervals during the experiment. The lambs were inoculated each with 2 ml i.v. chicken red blood cells (CRBC) on days 0, 21, and 42. Serum total, IgG and IgM antibody titers were determined at weekly intervals post-immunization. An in vivo intradermal hypersensitivity test was carried out on 6 lambs from each group on days 10 and 70. Transportation of the lambs resulted in a significant (p<0.001) elevation of serum cortisol, total protein and albumin levels, as well as increased plasma glucose concentration, with corresponding decrease in total cholesterol, while blood urea-N remained largely unchanged. These constituents returned to normal levels during subsequent weeks, with no significant differences in their concentrations being observed between the Cr-supplemented groups and controls. Rise in rectal temperature after transportation was reduced to a greater extent (p<0.05) in Cr-supplemented versus control lambs. Total, IgG and IgM antibody titers against CRBC rose significantly (p<0.05) during immunizations in all groups, with significantly and linearly higher (p<0.05) total and IgG titers in Cr-supplemented versus control lambs. By contrast, no significant effect due to Cr supplementation was recorded in IgG titers, which increased equally in Cr-fed and control groups. Skin thickness in response to intradermal inoculation of phytohaemagglutinin (PHA) was also significantly (p<0.01) increased as a result of Cr supplementation. These results indicate that dietary Cr supplementation might be useful during stress especially for enhancing immune responses in transport-stressed lambs.

Initial Risk Assessment of Disodium Disulphite in OECD High Production Volume Chemical Program

  • Sanghwan Song;Park, Yoonho;Park, Hye-Youn;Kwon, Min-Jeoung;Koo, Hyun-Ju;Jeon, Seong-Hwan;Na, Jin-Gyun;Park, Kwangsik
    • Toxicological Research
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    • v.18 no.1
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    • pp.23-29
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    • 2002
  • Disodium disulphite, the HPV chemical, was assigned to Korea in order to implement OECD SIDS program in 1999. It was produced about 3,200 ton/year in 1998. This report evaluates the toxic potency of disodium disulphite based on the environmental and mammalian effects as well as human exposure. Oral $LD_{50}$ in rats is 1,540 mg/kg b.w. and effects was observed to the stomach, liver and the GI track that was filled with blood. For repeated dose toxicity, the predominant effect was the induction of stomach lesion due to local irritation. The no observed adverse effect lever for local (stomach irritation) was about 217 mg/kg bw/day. There is no evidence that disodium disulphite is genotoxic in vivo. No reproductive or developmental toxicty of disodium disulphite was observed for the period up to 2 yr and over three generation. In humans, urticaria and asthma with itching, edema, rhinitis, and nasal congestion were reported. Disodium disulphite is unlikely to induce respiratory sensitization but may enhance symptom of asthma in sensitive individuals. This chemical would be mainly transported to water compartment when released to environmental compartments since it is highly water soluble (470 g/l at 20). Low K oc (2.447) indicates disodium disulphite is so mobile in soil that it may not stay in the terrestrial compartment. The chemical has been tested in a limited number of aquatic species. hem acute toxicity test to fish, 96 hr-$LC_{50}$ was > 100 mg/1. For algae, 72 hr-$XC_{50}$ was 48.1 mg/1. For daphnid, the acute toxicity value of 48 hr-$EC_{50}$ was 88.76 mg/1, and chronic value of 21day-NOEC was > 10 mg/1. Therefore, PNEC of 0.1 mg/l for the aquatic organism was obtained from the chronic value of daphnid using the assessment factor of 100. Based on these data the disodium disulphite was recommended as low priority for further post-SIDS work in OECD.

Pathological Studies on the New Microsporidia K79 Isolated from the Silkworm, Bombyx mori L. II. Pathogenicity and Developmental Progress of the Microsporidia. (가잠에서 분리된 새로운 미포자충 K79의 병리학적 연구 II. 미포자충의 병원성 및 발육경과)

  • 윤재수;임종성
    • Journal of Sericultural and Entomological Science
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    • v.27 no.1
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    • pp.1-11
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    • 1985
  • In order to clarify the taxanomic position of a new microsporidia K79 which was isolated from the silkworm larvae, Bombyx mori L. in Korea in 1979, the following several experiments such as estimation of pathogenicity in different instar, histopathological examination under light and electronic microscope and examination of fine structure of the sporse were carried out and their result obtained are as follows. In the test of pathogenicity by oral inoculation, the new microsporidia K79 was lower than Nosema bombycis and the susceptibility of the new microsporidia to silkworm was getting lower as the silkworm larvae grew. The lesion of Silkworms' tissue which was infected with the new microsporidia K79 was found in the epithelial cells of trachea, fat body and silk gland cells. The developmental process of the new microsporidia K79 in vivo could be divided into the following five stages: sporoplasm, schizont, sporont, sporoblast, and spore. The process was just the same as the of N. bombycis, but its development was slower than that of N. bombycis. Several differences in the fine structure of the spore under electron microscope, which could be important keys for the classification of microsporidia, were obtained. Anchoring disk and polaroplast lamella of the new microsporidian spore were disclosed to be different from those of N. bombycis. An average number of polar filament coils of the new microporidian spore was 16 at an angle of 75$^{\circ}$. On the basis of various keys for the classification of microsporidia, the results obtained from various experiments proved that the newly isolated microsporidia should be classified into the Genus, "Nosema", nut is further classification for species should be conducted in the future. Therefore, it may be reasonable that the new microsporidia is temperally classified as Nosema sp. K79 considering the fact that it was discovered in Korea in 1979.a in 1979.

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Effects of Oral Intake of Kimchi-Derived Lactobacillus plantarum K8 Lysates on Skin Moisturizing

  • Kim, Hangeun;Kim, Hye Rim;Jeong, Bong jun;Lee, Seung Su;Kim, Tae-Rahk;Jeong, Ji Hye;Lee, Miyeong;Lee, Sinai;Lee, Jong Suk;Chung, Dae Kyun
    • Journal of Microbiology and Biotechnology
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    • v.25 no.1
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    • pp.74-80
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    • 2015
  • Skin is the soft outer covering of vertebrates that provides protection from pathogenic infection, physical damage, or UV irradiation, and controls body temperature and water content. In this study, we examined the effects of oral intake of kimchi-derived Lactobacillus plantarum K8 lysates on skin moisturizing. In an in vitro study, we observed that the hyaluronic acid content increased in HaCaT cells treated with L. plantarum K8 lysates. Oral administration of L. plantarum K8 lysates effectively attenuated the horny layer formation and decreased epidermal thickening in DNCB-treated SKH-1 hairless mice skin. The damage to barrier function was reduced after 8 weeks of oral administration of L. plantarum K8 lysates as compared with that in the atopic dermatitis mice. For the test with volunteers, we manufactured experimental candy containing 2.1% L. plantarum K8 lysates, while control candy did not contain bacterial lysate. A significant increase in hydration in the experimental candy-administered group as compared with the control candy-administered group was observed on the face after 4 and 8 weeks, and on the forearm after 4 weeks. Decreases in horny layer thickness and TEWL value were observed on the face and forearm of the experimental group. Together, the in vitro cell line and in vivo mouse studies revealed that L. plantarum K8 lysates have a moisturizing effect. A clinical research study with healthy volunteers also showed an improvement in barrier repair and function when volunteers took L. plantarum K8 lysates-containing candy. Thus, our results suggest that L. plantarum K8 lysates may help to improve skin barrier function.

Fruit and vegetable discards preserved with sodium metabisulfite as a high-moisture ingredient in total mixed ration for ruminants: effect on in vitro ruminal fermentation and in vivo metabolism

  • Ahmadi, Farhad;Lee, Won Hee;Oh, Young-Kyoon;Park, Keunkyu;Kwak, Wan Sup
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.3
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    • pp.446-455
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    • 2020
  • Objective: Our recent series of laboratory- and large-scale experiments confirmed that under aerobic and anaerobic conditions, sodium metabisulfite (SMB) was effective in preserving nutrients and antioxidant capacity of highly perishable fruit and vegetable discards (FVD). Hence, the purpose of this study was to examine how partial inclusion of SMB-treated FVD in total mixed ration (TMR) influences in vitro ruminal fermentation, whole-tract digestibility, nitrogen metabolism, blood metabolites, and voluntary feed intake of sheep. Methods: The FVD were mixed thoroughly with 6 g SMB/kg wet biomass and kept outdoors under aerobic conditions for 7 days. Four TMRs including four levels of SMB-treated FVD (as-fed basis) at 0%, 10%, 20%, and 30% (equaling to 0%, 1.9%, 3.8%, and 5.7% on dry matter basis, respectively), were prepared as replacement for corn grain. The ruminal fermentation metabolites were studied using an in vitro gas production test. Four mature male Corriedale sheep were assigned at random to the 4 diets for two separate sub-experiments; i) digestibility trial with four 21-d periods, and ii) voluntary feed intake trial with four 28-d periods. Results: Inclusion of SMB-treated FVD in the TMR tended to quadratically increase partitioning factor. No effect was seen on total-tract digestibility of organic matter, ether extract, crude protein, and acid detergent fiber, except for neutral detergent fiber digestibility that tended to linearly increase with increasing SMB-treated FVD in the TMR. The progressive increase of FVD preserved with SMB in the diet had no effect on nitrogen metabolism. Treatment had no effect on serum antioxidant capacity and blood metabolites assayed. Voluntary feed intake was not impaired by inclusion of SMB-treated FVD in the TMR. Conclusion: It appears that FVD preserved with SMB can be safely incorporated into TMR as replacement of corn grain without impairment of nutrient metabolism and feed intake.

Treatment of Dyrk1A-dependent Mental Retardation of Down Syndrome: Isolation of Human Dyrk1A-specific shRNA (다운증후군의 Dyrk1A 의존적 뇌기능저하의 치료: 인간 Dyrk1A 특이적 shRNA 발굴)

  • Jung, Min-Su;Kim, Yeun-Soo;Kim, Ju-Hyun;Kim, Joung-Hun;Chung, Sul-Hee;Song, Woo-Joo
    • Journal of Life Science
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    • v.19 no.3
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    • pp.317-321
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    • 2009
  • Down syndrome (DS) results from overexpressed genes on an extra copy of human chromosome 21. Among various phenotypes seen in DS patients, mental retardation, such as learning and memory deficits, is a major factor that prevents DS individuals from leading fully independent lives. The Dyrk1A gene that plays a critical role in neurodevelopment has been isolated from chromosome 21, and transgenic mice with over-expression of Dyrk1A show severe hippocampal dependent learning and memory defects. In the present study, as an initial step to test the treatment of Dyrk1A dependent mental retardation phenotypes in model animals, we isolated human Dyrk1A specific lentiviral short hairpin RNA (shRNA) that inhibits the exogenous human Dyrk1A expression, but not the endogenous mouse expression in transgenic mice with human Dyrk1A overexpression. This limited and specific repression of exogenous human Dyrk1A will prove to be valuable information, if Dyrk1A dependent learning and memory defects in DS patients could be treated or at least ameliorated in vivo.

In-vivo Studies on Effect of Lipo-PGE1 on Neoangiogenesis of Composite Graft in a Rabbit Model (가토모델에서 Lipo-PGE1이 복합조직이식편의 미세혈관신생에 미치는 영향)

  • Park, Ji-Ung;Eo, Su-Rak;Cho, Sang-Hun;Choi, Jong-Sun;Kim, Eo-Jin
    • Archives of Plastic Surgery
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    • v.37 no.6
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    • pp.721-725
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    • 2010
  • Purpose: The survival of composite graft is dependent on three steps, (1) plasmatic imbibitions, (2) inosculation, and (3) neovascularization. Among the many trials to increase the survival rate of composite graft, prostaglandin E1 (PGE1) has beneficial effects on the microcirculatory level with vasodilating, antithrombotic, anti-inflammatory and neoangiogenic properties. Lipo-PGE1 which is lipid microspheres containing PGE1 had developed to compensate the systemic and local side effects of PGE1. This study was proposed to determine whether Lipo-PGE1 administration enhanced the survival of composite graft through neovascularization quantitatively in a rabbit ear model. Methods: Fourteen New Zealand White Rabbits each weighing 3~4 kg were divided in two groups: (1) intravenous Lipo-PGE1 injection group and (2) control group. A $2{\times}1\;cm$ sized, full-thickness rectangular composite graft was harvested in each auricle. Then, the graft was reaaproximated in situ using a 5-0 nylon suture. For the experimental group, $3{\mu}g$/kg/day of Lipo-PGE1 ($5{\mu}g$/mL) was administered intravenously through the marginal vein of the ear for 14 days. The control group was received no pharmacologic treatment. On the 14th postoperative day, composite graft of the ear was harvested and immunochemistry staining used Monoclonal mouse anti-CD 31 antibody was performed. Neoangiogenesis was quantified by counting the vessels that showed luminal structures surrounded by the brown color-stained epithelium and counted from 10 random high-power fields (400x) by independent blinded observer. Statistical analysis (Wilcoxon Signed Ranks test for nonparametric data) was performed using SPSS v12.0, with values of p<0.05 considered significant. Results: The mean number of the microvessels was $15.48{\pm}8.65$ in the experimental group and $9.82{\pm}7.25$ in the control group (p=0.028). Conclusion: The use of Lipo-PGE1 facilitated the neoangiogenesis, resulted in the improvement of the survival rate of graft. On the basis of this results, we could support wider application of Lipo-PGE1 for more effective therapeutic angiogenesis and successful survival in various cases of composite graft in the human.

Comparison between the method of the measurement 25 Vitamin D3 (25 Vitamin D3 측정에 있어서 화학발광미세입자 측정법과 화학발광면역 측정법 간의 비교 및 고찰)

  • Kim, dae-won;Lee, jung-hee;Jung, an-na;Seo, so-yoen
    • The Korean Journal of Nuclear Medicine Technology
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    • v.19 no.2
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    • pp.112-114
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    • 2015
  • Purpose Vitamin D to Anti- Rickets both steroid compounds showing activity, By acting on bone tissue secretary and the key to maintain serum Ca homeostasis. The blood level of vitamin D is the largest in D3 that the concentration of the metabolite is reflected in the holding state of vitamin D in vivo. Sunlight to change the 7-dehydrocholesterol in the skin and through the skin to D3, In the liver in combination with the D2 and D3 D4 changes. The Radioimmunoassay(RIA) method is measuring the D 3, the sensitivity can be measured also difficult trace substance to measure the normal test because it is very sensitive, but recently, a check is possible, for the Total D3 in Chemimicroparticle immunoassay(CMIA) or Chemiluminescent immunoassay(CLIA) measuring using microparticle RIA and CMIA(Architect i2000SR) / use the CLIA(DXI-800) method to compare and evaluate the correlation between the tests in the same test items. Materials and Methods Commissioned from January 2014 to March 2015 patients were enrolled in a total of 273 people. 29 out of 273 people conducted by RIA were compared with CMIA, 244 patients were compared with CLIA. Using reagents and equipment were used RIA(Diasource), CMIA(Architect i2000SR, Abbott Diagnostics) / CLIA( Unicel DXi-800, Beckman coulter). Results Correlation of the RIA and CLIA was a R2 = 0.1844 (y = 0.7303x + 3.9005), and the correlation of RIA CMIA is R2 = 0.2762 (y = 0.8862x + 4.56) respectively. (According to statistics, during the same period RIA is Deficiency 4.31%, Insufficiency 90.53%, Sufficiency 5.16%, was Excess 0%, CLIA / CMIA is Deficiency 17.02%, Insufficiency 75.91%, Sufficiency 7.03%, indicating the distribution of 0.03 % Excess) Conclusion Serum vitamin D and parathyroid hormone that show an inverse relationship, the level above which are not parathyroid hormone and vitamin D reduced the increase. The density is different for each study, at most 20 is reported to be the maximum between 30 ng / ml. In Korea it requires a proposed standard of vitamin D deficiency, reference to the WHO lack the case more than 10ng/ml, 20ng/ml and defined by the lack of, if not more than, the IOM, but looking at 12ng/ml or less to the normal to lack, at least 20ng/ml, the reference do not match the deficit under 20ng/ml, 21-29ng/ml relative lack between, was also defined as a sufficient condition for more than 30ng/ml. Although not statistically is between RIA and CLIA two ways to vitamin D levels change according to season match, when seasonally seen in summer as commonly known (April to September), winter (October to March) relative to the increase measured than it was found. Finally, the study on the correlation between the two methods have been expected to result in a consistent and apply the same view high reference value on the graph is difficult. However, there may be differences between the test equipment and methods, and could be especially the case of RIA method using an organic solvent is difficult to compare different methods and correlated view similar trend in vitamin D deficiency and quarterly aspect ratio.

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Effects of Newly Synthesized Recombinant Human Amyloid-β Complexes and Poly-Amyloid-β Fibers on Cell Apoptosis and Cognitive Decline

  • Park, Soojin;Huh, Jae-Won;Eom, Taekil;Park, Naeun;Lee, Youngjeon;Kim, Ju-Sung;Kim, Sun-Uk;Shim, Insop;Lee, Sang-Rae;Kim, Ekyune
    • Journal of Microbiology and Biotechnology
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    • v.27 no.11
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    • pp.2044-2051
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    • 2017
  • The main pathological hallmark of Alzheimer's disease is the deposition of amyloid-beta ($A{\beta}$) peptides in the brain. $A{\beta}$ has been widely used to mimic several aspects of Alzheimer's disease. However, several characteristics of amyloid-induced Alzheimer's disease pathology are not well established, especially in mice. The present study aimed to develop a new Alzheimer's disease model by investigating how $A{\beta}$ can be effectively aggregated using prokaryotes and eukaryotes. To express the $A{\beta}42$ complex in HEK293 cells, we cloned the $A{\beta}42$ region in a tandem repeat and incorporated the resulting construct into a eukaryotic expression vector. Following transfection into HEK293 cells via lipofection, cell viability assay and western blotting analysis revealed that exogenous $A{\beta}42$ can induce cell death and apoptosis. In addition, recombinant His-tagged $A{\beta}42$ was successfully expressed in Escherichia coli BL21 (DE3) and not only readily formed $A{\beta}$ complexes, but also inhibited the proliferation of SH-SY5Y cells and E. coli. For in vivo testing, recombinant His-tagged $A{\beta}42$ solution ($3{\mu}g/{\mu}l$ in $1{\times}PBS$ containing $1mM\;Ni^{2+}$) was injected stereotaxically into the left and right lateral ventricles of the brains of C57BL/6J mice (n = 8). Control mice were injected with $1{\times}PBS$ containing $1mM\;Ni^{2+}$ following the same procedure. Ten days after the sample injection, the Morris water maze test confirmed that exogenous $A{\beta}$ caused an increase in memory loss. These findings demonstrated that $Ni^{2+}$ is capable of complexing the 50-kDa amyloid and that intracerebroventricular injection of $A{\beta}42$ can lead to cognitive impairment, thereby providing improved Alzheimer's disease models.

A QUANTITATIVE STUDY OF THE CHANCE OF CALCIUM, PHOSPHATE, FLUORIDE USING EPMA AFTER IN VITRO DEMINERALIZATION AND REMINERALIZATION OF HUMAN TOOTH ENAMEL (법랑질 표면의 탈회 및 재광화 후 EPMA (electron probe micro-analysis)를 이용한 칼슘, 인, 불소 변화의 정량적 분석)

  • Hong, Kyoung-Sik;Hur, Bock;Lee, Chan-Young;Keum, Ki-Yeon
    • Restorative Dentistry and Endodontics
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    • v.26 no.5
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    • pp.372-378
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    • 2001
  • The aim of this in vitro study was to analyze the composition of human tooth enamel in terms of three components, Ca, P, and F after demineralization and remineralization in acid buffer solution. A total of 8 human premolars without any defects and cracks were selected and buccal and lingual sides of the teeth were cleaned with an ultrasonic device and pumice without fluoride 5$\times$5mm windows were opened, and other areas were completely covered with 3-coats of nail varnish to prevent from being in contact with demineralized and remineralized solutions. After demineralization process, each tooth was sectioned into two slices, highly polished one of them with$\gamma$-alumina, and then analyzed the composition of the demineralized tooth with EPMA(electron probe micro-analyzer). The other slices were put into the remineralized solution for 10 days, polished, and analyzed in the same manner. These data were statistically analyzed with one sample t-test(p<0.05). The results were as follows. 1. Normal tooth enamel consists of 49.76% Ca, 39.80% P, and 0.28% F. 2. After demineralization, percentage of Ca and P ratio were decreased by about 5.57 and 5.07% respectively. Percentage of F ratio was also decreased by about 0.01%, which was not statistically significant. 3. After remineralization, percentage of Ca, P increased about by 4.47 and 4.35% respectively Percentage of F decreased by about 0.01%, which was not statistically significant. In conclusion, remineralized solution used in our study has the potential to induce the uptake the Ca and P into the pore sites of the demineralized enamel. But, in the oral cavity. there were rapid temperature change, organic matrix that inhibits the movement of the ions, and limitation of continuous contact with this remineralized solution. Therefore, further in vivo study is necessary.

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