Journal of the korean academy of Pediatric Dentistry
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v.32
no.1
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pp.75-88
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2005
Ameloblastoma is the most commonly occurring odontogenic tumor in oral cavity. Although most are benign epithelial neoplasm, they are generally considered to be locally aggressive and destructive, exhibiting a high rate of recurrence. The biological behavior of this neoplasm is a slowly growing, locally invasive tumor without metastasis, therefore malignant neoplasm, changed its histological appearance to carcinoma or showed distant metastasis, is only defined clinically. In this study, we identified the differentially expressed genes(DEGs) in stages under benign or malignant ameloblastoma compared with normal patient using ordered differential display(ODD) reverse transcription(RT)-PCR and $GeneFishing^{TM}$ technology. ODD RT-PCR is rather effective when the investigation of samples containing very small amounts of total RNA must be accomplished. ODD RT-PCR used the means of amplification with anchored T-primer and adaptor specific primer. bearing definite two bases at their 3' ends and so this method could display differential 3'-expressed sequence taqs(ESTs) patterns without using full-length cDNAs. Compared with standard differential display, ODD RT-PCR is more simple and have enough sensitivity to search for molecular markers by comparing gene expression profiles, However, this method required much effort and skill to perform. $GeneFishing^{TM}$ modified from DD-PCR is an improved method for detecting differentially expressed genes in two or more related samples. This two step RT-PCR method uses a constant reverse primer(anchor ACP-T) to prime the RT reaction and arbitrary primer pairs(annealing control primers, ACPs) during PCR. Because of high annealing specificity of ACPs than ODD RT-PCR, the application of $GeneFishing^{TM}$ to DEG discovery generates reproducible, authentic, and long(100bp to 2kb) PCR products that are detectable on agarose gels. Consequently, various DEGs observed differential expression levels on agarose gels were isolated from normal, benign, and malignant tissues using these methods. The expression patterns of the some isolated DEGs through ODD RT-PCR and $GeneFishing^{TM}$ were confirmed by Northern blot analysis and RT-PCR. The results showed that these identified DEGs were implicated in ameloblastoma neoplasm processes. Therefore, the identified DEGs will be further studied in order to be applied in candidate selection for marker as an early diagnosis during ameloblastoma neoplasm processes.
Currently titanium is the material of choice for implants because of its biological acceptance. This high degree of biocompatibility is thought to result, in part, from the protective and stable oxide layer that presumably aids in the bonding of the extracellular matrix at the implant-tissue interface. Endosseous dental implants are interfaced with bone, connective tissue, and epithelium when implanted into the jaw bone. The soft tissue interface including connective tissue and epithelium is one of the most critical factors in the determination of implant maintenance and prognosis. For maintenance of failing or failed implants, it is essential to treat the implant fixture surface to remove bacterial endotoxins and make a surface tolerated by surrounding soft and hard tissues. In this study, the effect of mechanical treatment on titanium plasma sprayed implant on adhesiveness and proliferation of human gingival fibroblasts and changed surface characteristics were studied. titanium plasma sprayed discs manufactured by Friedrichsfeld company were treated with loaw speed stone bur, a rubber point and a jetpolisher. Its surface components were analyzed with Energy dispersive X-ray spectroscopy to evaluate whether the surface characteristics were altered or not. To observe the spreading pattern of the human gingival fibroblasts which attached to the all specimens author used the scanning electron microscope. The results were as follows : Pure titanium and plasma sprayed titanium, stone polished titanium showed titanium peak and small amout of aluminum, so there was no alteration on surface characteristics. Under the scanning electron microscopic examination in the initial attachment of human gingival fibroblast, there was a slight enhancement in pure titanium, stone polished titanium than plasma sprayed titanium. After 6 hours, the pure titanium and stone polished titanium showed human gingival fibroblasts were elongated and connected with numerous processes. Human gingival fibroblasts were more intimately attached on the pure titanium discs than on the other discs. The human gingival fibroblasts attached on the plasma sprayed titanium by thin and elongated processes. After 24 hours, the human gingival fibroblasts connected with each other via numerous processes and compeletly covered the pure titanium and stone polshed titanium discs. Human gingival fibroblasts had multiple point contacts with more long and thin lamellopodia and showed a little bare surface on plasma sprayed titanium discs.
Destruction of oral soft and hard tissues and resulting problems seriously affect the life quality of xerostomic patients. Although artificial saliva is the only regimen for xerostomic patients with totally abolished salivary glands, currently available artificial salivas give restricted satisfaction to patients. The purpose of this study was to contribute to the development of ideal artificial saliva through comparing viscosity and wettability between CMC solutions and human saliva. Commercially-available CMC is dissolved in simulated salivary buffer (SSB) and distilled deionized water (DDW). Various properties of human whole saliva, human glandular saliva, and a CMC-based saliva substitutes known as Salivart and Moi-Stir were compared with those of CMC solutions. Viscosity was measured with a cone-and-plate digital viscometer at six different shear rates, while wettability on acrylic resin and Co-Cr alloy was determined by the contact angle. The obtained results were as follows: 1. The viscosity of CMC solutions was proportional to CMC concentration, with 0.5% CMC solution displaying similar viscosity to stimulated whole saliva. Where as a decrease in contact angle was found with increasing CMC concentration. 2. The viscosity of human saliva was found to be inversely proportional to shear rate, a non-Newtonian (pseudoplastic) trait of biological fluids. The mean viscosity values at various shear rates increased as follows: stimulated parotid saliva, stimulated whole saliva, unstimulated whole saliva, stimulated submandibular-sublingual saliva. 3. Contact angles of human saliva on the tested solid phases were inversely correlated with viscosity, namely decreasing in the order stimulated parotid saliva, stimulated whole saliva, unstimulated whole saliva, stimulated submandibular-sublingual saliva. 4. Boiled CMC dissolved in SSB (CMC-SSB) had a lower viscosity than CMC-SSB (P < 0.01 at shear rate of $90s^{-1}$). 5. For human saliva, contact angles on acrylic resin were significantly lower than those on Co-Cr alloy (P < 0.01). 6. Comparing CMC solutions with human saliva, the contact angles between acrylic resin and human saliva solutions were significantly lower than those between acrylic resin and CMC solutions, including Salivart and Moi-Stir (P <0.01). The effectiveness of CMC solutions in terms of their rheological properties was objectively confirmed, indicating a vital role for CMC in the development of effective salivary substitutes.
The ultimate goal of periodontal therapy is the regeneration of periodontal tissue which has been lost due to destructive periodontal disease. To achieve periodontal regeneration, various kinds of methods have been investigated and developed, including guided tissue regeneration and bone graft. Bone graft can be catagorized into autografts, allografts, xenografts, bone substitutes. And materials of all types have different biological activity and the capacity for periodontal regeneration, but ideal graft material has not been developed that fits all the requirement of ideal bone graft material. Intensive research is underway to identity, purify, synthesize a variety biologic modulators that may enhance wound healing and regeneration of lost tissues in periodontal therapy. The present study evaluates the effects of ABM/P-15 on the periodontal regeneration in intrabony defects of human. We used thirty four 2-wall or 3-wall osseous defects in premolars and molars of chronic peridontitis patient that have more than 5mm pockets and more than 3mm in intrabony defect. 12 negative control group underwent flap procedure only, 11 positive control group received DFDBA graft with flap procedure, and 11 experimental group received ABM/P-15 graft with flap procedure. The changes of probing pocket depth, loss of attachment and bone probing depth following 6months after treatment revealed the following results: 1. The changes of probing pocket depth showed a statistically significant decrease between after scaling and 6months after treatment in negative control(2.0${\pm}$0.9mm), positive control(3.0${\pm}$0.9mm), and experimental group (3.4${\pm}$1.5mm) (P<0.01). Significantly more reduction was seen in experimental group compared to negative control group (P<0.05). 2. The changes of loss of attachment showed a statistically significant decrease between after scaling and 6months after treatment in positive control(2.0${\pm}$0.6mm), and experimental group (2.2${\pm}$l.0mm) except negative control group(0.1${\pm}$0.7mm) (P<0.01). Significantly more reduction was seen in both experimental and positive control group compared to negative control group(P<0.05). 3. The changes of bone probing depth showed a statistically significant decrease between after scaling and 6months after treatment in positive control(2.7${\pm}$l.0mm), and experimental group (3.4${\pm}$1.3mm) except negative control(0.l${\pm}$0.9mm) (9<0.01). Significantly more reduction was seen in both experimental and positive control group compared to negative control group (P<0.05). The results suggest that the use of ABM/P-15 in the treatment of periodontal intrabony defects can reduce loss of attachment and bone probing depth more than flap operation only. It suggests that ABM/P-15 may be an effective bone graft material for the regeneration of periodontal tissue in intrabony defects.
Kim Kye-Won;Ha Sun-Hwa;Cho Kang-Jin;Kim Eun-Ju;Lee Min-Kyung;Yu Jae-Ju;Kim Jong-Guk;Lee Shin-Woo
Journal of Plant Biotechnology
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v.32
no.3
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pp.167-173
/
2005
Three different cDNAS for cinnamate 4-hydroxylase (C4H) which are involved in the second step of the general phenylpropanoid pathway were isolated and designated as pc4h1 (1,755 bp), pc4h2 (1,655 bp), and pc4h3 (1,316 bp), respectively. The nucleotide sequence analysis revealed that both pc4h1 and pc4h2 clones encode polypeptides of 505 amino acids frame but pc4h3 clone was truncated at the 5'-end of coding region. The alignment of the deduced amino acid sequences showed that PC4H1 and PC4H2 are highly homologous (95.8% identical) with each other and contain three conserved domains which are typical in cytochrome P450 monooxygenase: proline-rich region, threonine-containing binding pocket for the oxygen molecule, and heme binding region. In addition, result of the phylogenic tree analysis revealed that both pepper C4Hs belong to Class 1. pc4h2 transcription was strongly induced in wounded fruit (400%) and root (200%) relative to its very low basal level but not in leaf or stem tissue. In case of pc4h1, the basal level of transcription was higher than pc4h2 but induction by wounding was lower in fruit and root while leaf and stem tissues did not respond to wounding. The basal level of pc4h3 transcripts was not, if any, detectable and response to wounding was not observed.
Park, Shin-Hyoung;Kim, Jung-In;Jeong, Yong-Kee;Choi, Yung-Hyun
Journal of Life Science
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v.21
no.6
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pp.796-804
/
2011
Microglia are central nervous system (CNS)-resident professional macrophages that function as the principal immune cells responding to pathological stimulations in the CNS. Activation of microglia, induced by various pathogens, protects neurons and maintains homeostasis in the CNS, but severe activation causes inflammatory responses secreting various neurotoxic molecules such as nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$) and pro-inflammatory cytokines. Allium fistulosum, a member of the onion family, is mainly cultivated for consumption, as well as medicinal use in Oriental medicine. It has been reported that A. fistulosum has various biological effects such as anti-oxidant, anti-platelet aggregation, anti-fungus and anti-cholesterol synthesis, however there has been no research about the anti-inflammatory effects of A. fistulosum extracts. In this study, it was undertaken to explore the functions of A. fistulosum as a suppressor of neuronal inflammation by using BV2 microglia cells. As a result, it was found that four kinds of extracts of A. fistulosum effectively reduced the expressions of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) at both mRNA and protein levels, and also attenuated pro-inflammatory cytokines such as tumor necrosis alpha (TNF-${\alpha}$), interleukin-$1{\beta}$ (IL-$1{\beta}$) and interleukin-6 (IL-6) at the mRNA level in BV2 stimulated by lipopolysaccharide (LPS). In addition, the extracts of A. fistulosum attenuated the release of NO markedly, as well as resulting in slight decreases of TNF-${\alpha}$ and IL-6 production, the effects of which were most significant when treated with ethyl alcohol extract from the whole A. fistulosum. In conclusion, the data indicated that the anti-inflammatory actions of A. fistulosum against BV2 microglia cells is through the down-regulation of iNOS, COX2 and pro-inflammatory cytokines such as TNF-${\alpha}$ and IL-6, and these effects are expected to help in the protection of nerve tissues by suppressions of neuronal inflammation in various neurodegenerative diseases.
Anatomical studies of sink tissues are required for better understanding the biological plant growth system and energy metabolism. Kinematics of root growth zones of two genotypes of tall fescue (Festuca arundinacea Schreb.) receiving 50 or 200 ppm N were determined. Longitudinal anatomy and cell dynamics of root growth zones were studied and calculated. The root growth zone is organized similarly to the leaf growth zone which has cell division, elongation, and maturation zones, but the root growth zone is only about 3.0 mm long compared to 25 to 30 mm for the leaf growth zone. The root cap extends about 0.4 to 0.5 mm from the apical initial, while the cell elongation zone for both cortical and metaxylem cells extends about 3.3 mm from the apical initial for both genotypes and N levels. Root cap cells elongate from an initial length of about 5$\mu{m}$ long to a final length of about 40$\mu{m}$ before being sloughed. Initial lengths of cortical and metaxylem cells were about 8.5 $\mu{m}$ and 13.0 $\mu{m}$, respectively. Elongation of cortex and metaxylem cell showed sigmoidal curves with final lengths of about 120 $\mu{m}$ for cortex cells and 650 $\mu{m}$ for metaxylem cells. Initial size and final size for both types were not affected by N level, but cell fluxes and cell elongation rates of cortical and metaxylem cells were about double in low N. Cell production rates were about 5 to 6 times higher in cortical cells than in metaxylem cells. Differences in N caused a larger change in cell production rate, duration of cell elongation, and relative cell elongation rate than did the genotypes. These data indicate that N application affects root growth longitudinally by changing cell production rate and elongation rate.
For the fundamental studies of radiation breeding in edible marine algae, the biological effects on conchospores of Porphyra species by gamma-irradiation were examined. Two varieties, Keun-cham-gim (Porphyra tenera Kjell. form tamatsuensis Miura) and Saga No.5, were chosen for this study, and their conchospores after r-irradiation($5.0{\sim}20.0$ KR) were cultured for 50 days. The results obtained were summarized as follows. 1. Gamma-irradiation in less than the dose of 20KR did not affect germination of conchospores, and almost all spores grew into two cells germ in 24 hours after irradiation, but withering germs were gradually increased in number according to higher exposure within 5 days old culture. 2. High irradiation caused the induction of giant cells, abnormal useless growth of hold-fast, lumpish thalli and callus-like lumpy tissues. 3. The liberation of neutral spores from young germs and carpospores from mature thalli were observed on the frond exposed at $10{\sim}20$ KR irradiation. All spores were normal in division and its size. 4. The best irradiation effect on growth of Keun-cham-gim was observed at 10 KR dose, whose growth-rates were $140\%$ in wet weight and $108\%$ in mean frond area, but only $48\%$ was recorded in wet weight at 20 KR exposure. Saga No.5 were in contrast with Keun-cham-gim, and their most growth-rate was $400\%$ in wet weight ($258\%$ in frond area) at 10 KR irradiation and the worst was $20\%$ at the dose of 20 KR. 5. The withering phenomenon to death by treatment of gamma-ray presented substantial difference between two varieties. Survival rate compared with control in Keun-cham-gim was $70.7\%$ at 20 KR, but that in Saga No.5 recorded $47.0\%$ at same dose. 6. Synthesizing the results of high and low r-irradiation, it was suggested tat high r-irradiation in more than 5.0 KR inhibited conspicuously the growth of germs derived from conchospores, and about half of them withered at 15.0 KR dose or more.
The aim of this study is to know how the rat submandibular gland changes under various emotional stress condition, using molecular biological methods. Restraint and chronic unpredictable mild stress (CUMS) experiment is conducted on fifty one 7-week old Sprague-Dawley rats (restraint stress experiment: 21, CUMS: 30). The rats were sacrificed, the submandibular glands were excised immediately at certain time, and examined by the use of immunohistochemistry and western blotting. In CUMS experiment, sucrose preference test, water intake change, weight change were implemented at 1 week interval for the experimental period The results are as follows: 1. The number of clusterin-secreting cells of restraint stress group compared to control group showed significantly decreasing tendency in all experimental groups except for the 1st hour group (p<0.001 in the 9th, 24th, 72nd, 120th, and 168th hour group). 2. The number of clusterin-secreting cells of CUMS group compared to control group showed significantly increasing tendency in the 2nd week group (p<0.01), and significantly decreasing tendency in the 4th and 5th week group (p<0.001). 3. Sucrose preference test in CUMS experiment showed significant difference between the 5th week experimental group and control group (p<0.01). 4. Weight change in CUMS experiment showed significant difference between the 5th week experimental group and control group (p<0.01), but water intake change didn't show significant difference compared to control group. 5. In western blot analysis, clusterin expression was decreased on a gradual basis in due time compared to the control group in the restraint stress group. As for CUMS group (chronic unpredictable mild stress group), it was increased till the 2nd week and decreased till the 5th week after that, which is similar to immunohistochemical analysis result and the decreasing tendency of sucrose preference and weigh changes. Through the test, it was proved that expression of clusterin in saliva glands decreases after receiving either acute or chronic stress, indicating relation with depression caused by chronic stress. Unlike other data, however, apoptotic tendency was hardly found in tissues. Diverse possibilities could be suggested on that: first, the stress was not enough to expedite apoptosis; second, apoptosis-related protein was already being secreted though not detected with microscope; third, clusterin, a major secretion molecule of saliva, decreased with saliva's malfunction due to stress. In the respect, it will be necessary to examine proteins expressed in case of cell death or other heat-shock proteins at the same time, in order to see whether any cellular change or death is caused by decreasing clusterin under high stress, and whether the original state is restored as time goes by under mild stress, through longer-term tests using even higher acute stress.
Journal of the Society of Cosmetic Scientists of Korea
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v.26
no.1
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pp.163-186
/
2000
We have previously screened 150 medicinal plants on the inhibition of elastase and found a significant inhibitory effects of the extracts of Areca catechu L. on the aging and inflammation against the skin tissues. To isolate and identify the compounds having biological activity, we was further purified by each of the solvent fractions, silica gel column chromatography, preparative TLC and reversed-Phase HPLC. Peak in HPLC, which coincided with the inhibitory activity against elastase, was identified as Phenolic substance using various colorimetric methods, UV, and IR. $IC_{50}$/ values of phenolic substance purified from Areca catechu were 26.9 $\mu\textrm{g}$/$m\ell$ for porcine pancreatic elastase (PPE) and 60.8 $\mu\textrm{g}$/$m\ell$ for human neutrophil elastase (HNE). This Phenolic substance showed more potent activity than those of reference compounds, oleanolic acid (76.5 $\mu\textrm{g}$/$m\ell$ for PPE, 219.2 $\mu\textrm{g}$/$m\ell$ for HNE) and ursolic acid (31.0 $\mu\textrm{g}$/$m\ell$ for PPE, 118.6 $\mu\textrm{g}$/$m\ell$ for HNE). According to the Lineweaver-Burk Plots, the inhibition against both PPE and HNE by this phenolic substance was competitive with substrate. Phenolic substance from Areca catechu exhibited high free radical scavenging effect ($SC_{50}$/ : 6 $\mu\textrm{g}$/$m\ell$) and inhibited effectively hyaluronidase activity ($IC_{50}$/: 210 $\mu\textrm{g}$/$m\ell$). These results suggest that the Phenolic substance Purified from Areca catechu showed anti-aging effect by protecting connective tissue proteins.
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