Tooth movement by orthodontic force effects great tissue changes within the periodontium, especially by shifting the blood flow in the pressure side and resulting in a hypoxic state of low oxygen tension. The aim of this study is to elucidate the possible mechanism of apoptosis in response to hypoxia in MC3T3El osteoblasts, the main cells in bone remodeling during orthodontic tooth movement. MC3T3El osteoblasts under hypoxic conditions ($2\%$ orygen) resulted in apoptosis in a time-dependent manner as estimated by DNA fragmentation assay and nuclear morphology stained with fluorescent dye, Hoechst 33258. Pretreatment with Z-VAD-FMK, a pancaspase inhibitor, or Z-DEVD-CHO, a specific caspase-3 inhibitor, completely suppressed the DNA ladder in response to hypoxia. An increase in caspase-3-like protease (DEVDase) activity was observed during apoptosis, but no caspase-1 activity (YVADase) was detected. To confirm what caspases are involved in apoptosis, Western blot analysis was performed using anti-caspase-3 or -6 antibodies. The 10-kDa protein, corresponding to the active products of caspase-3, and the 10-kDa protein of the active protein of caspase-6 were generated in hypoxia-challenged cells in which the processing of the full length form of caspase-3 and -6 was evident. While a time course similar to this caspase-3 and -6 activation was evident, hypoxic stress caused the cleavage of lamin A, which was typical of caspase-6 activity. In addition, the stress elicited the release of cytochrome c into the cytosol during apoptosis. Furthermore, we observed that pre-treatment with SB203580, a selective p38 mitogen activated protein kinase inhibitor, attenuated the hypoxia-induced apoptosis. The addition of SB203S80 suppressed caspase-3 and -6-like protease activity by hypoxia up to $50\%$. In contrast, PD98059 had no effect on the hypoxia-induced apoptosis. To confirm the involvement of MAP kinase, JNK/SAPK, ERK, or p38 kinase assay was performed. Although p38 MAPK was activated in response to hypoxic treatment, the other MAPK -JNK/SAPK or ERK- was either only modestly activated or not at all. These results suggest that p38 MAPK is involved in hypoxia-induced apoptosis in MC3T3El osteoblasts.
Poly-P has been used to prevent decomposition of foods and has been shown to have inhibitory effect on the growth of gram positive bacteria. The purpose of this study was to evaluate the effect of poly-P on the growth of Porphyromonas endodontalis, a gram negative obligate anaerobic rod, endodontopathic bacterium. P. endodontalis ATCC 35406 was in BHI broth containing hemin and vitamin K with or without poly-P. Inhibitory effect of each poly-P which was added at the beginning(lag phase) or during(exponential phase) the culture, MIC(minimum inhibitory concentration) was determined by measuring the optical density of the bacterial cell at 540nm. Viable cell counts were measured to determined whether poly-P has a bactericidal effect. Leakage of intracellular nucleotides from P. endodontalis was determined at 260nm and morphological change of P. endodontalis was observed under the TEM(transmission electron microscope). Binding of 32P-labeled poly-P to P. endodontalis was examined. SDS-polyacrylamide gel electrophoresis and zymography were performed to observe the changes in protein and enzyme profiles of P. endodontalis, respectively. The results from this study were as follows : 1. The minimal inhibitory concentration(MIC) of poly-P to P. endodontalis appeared to be 0.04~0.05%. 2. Poly-P added to the P. endodontalis culture during the exponential phase of P. endodontalis was as much effective as poly-P added at the begining of the culture, suggesting that the antibacterial effect of poly-P is not much dependent on the initial inoculum size of P. endodontalis. 3. Poly-P are bactericidal to P. endodontalis, demonstrating the decrease of the viable cell counts. 4. Intracellular nucleotide release from the P. endodontalis, was not increased in the presence of poly-P and was not reversed by the addition of divalent cations like $Ca^{2+}$ and $Mg^{2-}$. 5. Under the TEM, it was observed that fine electro-dense materials were prominent in the poly-P grown P. endodontalis, appearing locally in the cell, and the materials were more abundant and more dispersed in the cell as the incubation time with poly-P increased. In addition, highly electron dense granules accumulated in many poly-P grown cells, most of which were atypical in their shape. 6. Binding of 32P-labeled poly-P to P. endodontalis appeared to be 32.8 and 45.5 and 53.4% at 30 minutes, 1 hours and 2 hours, respectively. 7. In the presence of poly-P. the synthesis of proteins with apparent molecular masses of 25, 27, 35, 45 was lost or drastically decreased whereas expression of a protein with an apparent molecular mass of 75 was elevated. 8. Proteolytic activity of P. endodontalis was decreased by poly-P. The overall results suggest that use of poly-P may affect the growth of P. endodontalis, and the anti-bacterial activity of poly-P seems largely bactericidal. Changes in shape, protein expression, and proteolytic activity of P. endodontalis by poly-P may be directly and indirectly attributed to the antibacterial effect of poly-P. Further studies will be needed to confirm the effect of poly-P.
Mycorrhiza refers to the association between a plant and a fungus colonizing the cortical tissue of the plant's roots during periods of active plant growth. The benefits afforded by plants from mycorrhizal symbioses can be characterized either agronomically, based on increased growth and yield, or ecologically, based on improved fitness (i.e., reproductive ability). In either case, the benefit accrues primarily because mycorrhizal fungi form a critical linkage between plant roots and the soil. The soilborne or extramatrical hyphae take up nutrients from the soil solution and transport them to the root. This mycorrhizae-mediated mechanism increases the effective absorptive surface area of the plant. There are seven major types of mycorrhizae along with mycoheterotrophy: endomycorrhizae (arbuscular mycorrhizae, AM), ectomycorrhizae (EM), ectendomycorrhizae, monotropoid, arbutoid, orchid, and ericoid. Endomycorrhizal fungi form arbuscules or highly branched structures within root cortical cells, giving rise to arbuscular mycorrhiza, which may produce extensive extramatrical hyphae and significantly increase phosphorus inflow rates in the plants they colonize. Ectomycorrhizal fungi may produce large quantities of hyphae on the root and in the soil; these hyphae play a role in absorption and translocation of inorganic nutrients and water, and also release nutrients from litter layers by producing enzymes involved in mineralization of organic matters. Over 4,000 fungal species, primarily belonging to Basidiomycotina and to a lesser extent Ascomycotina, are able to form ectomycorrhizae. Many of these fungi produce various mushrooms on the forest floor that are traded at a high price. In this paper, we discuss the benefits, nutrient cycles, and artificial cultivation of mycorrhizae in Korea.
MicroRNA-223-3p (miR-223-3p) is one of the potential microRNAs that have been shown to alleviate inflammatory responses in pre-clinical investigations and is highly encased in exosomes derived from bone mesenchymal stem cells (MSC-exosomes). MSC-exosomes are able to function as carriers to deliver microRNAs into cells. Autoimmune hepatitis is one of the challenging liver diseases with no effective treatment other than steroid hormones. Here, we examined whether MSC-exosomes can transfer miR-223-3p to treat autoimmune hepatitis in an experimental model. We found that MSC-exosomes were successfully incorporated with miR-223-3p and delivered miR-223-3p into macrophages. Moreover, there was no toxic effect of exosomes on the macrophages. Furthermore, treatments of either exosomes or exosomes with miR-223-3p successfully attenuated inflammatory responses in the liver of autoimmune hepatitis and inflammatory cytokine release in both the liver and macrophages. The mechanism may be related to the regulation of miR-223-3p level and STAT3 expression in the liver and macrophages. These results suggest that MSC-exosomes can be used to deliver miR-223-3p for the treatment of autoimmune hepatitis.
Tailings of Dukum mine in the vadose and saturated zone were investigated to reveal the mobility of metal elements and the condition of mineralogical solubility according to redox environments throughout the geochemical analysis, thermodynamic modelling, and mineralogical study for solid-samples and water samples(vadose zone; distilled water: tailings=5 : 1 reacted, saturated zone; pore-water extracted). In the vadose zone, sulfide oxidation has generated low-pH(2.72∼6.91) condition and high concentration levels of S $O_4$$^{2-}$(561∼1430mg/L) and other metals(Zn : 0.12∼l57 mg/L, Pb : 0.06∼0.83 mg/L, Cd : 0.06∼l.35 mg/L). Jarosite$(KFe_3(SO_4)_2(OH)_6)$ and gypsum$(CaSO_4{\cdot}2H_2O$) were identified on XRD patterns and thermodynamics modelling. In the saturated zone, concentration of metal ions decreased because pH values were neutral(7.25∼8.10). But Fe and Mn susceptible to redox potential increased by low-pe values(7.40∼3.40) as the depth increased. Rhodochrosite$(MnCO_3)$ identified by XRD and thermodynamics modelling suggested that $Mn^{4+}$ or $Mn^{3+}$ was reduced to $Mn^{2+}$. Along pH conditions, concentrations of dissolved metal ions has been most abundant in vadose zone throughout borehole samples. It was observed that pH had more effect on metal solubilities than redox potential. How-ever, the release of co-precipitated heavy metals following the dissolution of Fe-Mn oxyhydroxides could be the mechanism by which reduced condition affected heavy metal solubility considering the decrease of pe as depth increased in tile saturated zone.
Choi, Eun Kyung;Kim, Yun Seup;Park, Jae Seuk;Jee, Young Koo;Lee, Kye Young
Tuberculosis and Respiratory Diseases
/
v.58
no.1
/
pp.43-53
/
2005
Emphysema is characterized by air space enlargement and alveolar destruction. The mechanism responsible for the development of emphysema was thought to be protease/antiprotease imbalance and oxidative stress. A very recent study shows that alveolar cell apoptosis causes lung destruction and emphysematous changes. Thus, this study was performed to support the evidence for the role of apoptosis in the development of emphysema by characterizing cigarette smoke extract (CSE)-induced apoptosis in A549 (type II pneumocyte) lung epithelial cells. CSE induced apoptosis at low concentration (10% or less) and both apoptosis and necrosis at high concentration (20%). Apoptosis was demonstrated by DNA fragmentation using FACScan for subG1 fraction. Discrimination between apoptosis and necrosis was done by morphologic analysis using fluorescent microscopy with Hoecst 33342/propium iodide double staing and electron microscopy. Cytochrome c release was confirmed by using immunofluorescence with monoclonal anti-cytochrome c antibody. However, CSE-induced cell death did not show the activation of caspase 3 and was not blocked by caspase inhibitors. This suggests that CSE-induced apoptosis might be caspase-independent apoptosis. CSE-induced cell death was near completely blocked by N-acetylcystein and bcl-2 overexpression protected CSE-induced cell death. This results suggests that CSE might induce apoptosis through intracellular oxidative stress. CSE also activated p53 and functional knock-out of p53 using stable overexpression of HPV-E6 protein inhibited CSE-induced cell death. The characterization of CSE-induced cell death in lung epithelial cells could support the role of lung cell apoptosis in the pathogenesis of emphysema.
It was previously reported that red ginseng extract inhibited carcinogenesis by urethan, DMBA and aflatoxin $B_1E (Cancer Detection and Prevention, 6: 515-525, 1983). In an attempt to investigate the mechanism of the anticarcinogenic effect of ginseng, we assayed natural killer (N.K) activity in mice treated with urethan and benzo(a)pyrene. In our experiment newly born Swiss Webster mice, less than 24 hrs. old, were given a single subcutaneous injection of lmg of ure-than and 40ug of benzo(a)pyrene. The mice had been administered with ginseng since weaning, and sacrificed at various intervals. Major organs were examined both, with the naked eye and microscopically. N.K. activity of spleen cells was analyzed in a 12-hour $^{51}Cr^-release$ assay against YAC-1 cells. Administration of ginseng resulted in an increase of N.K. activity by $18\%$ at 4 weeks, $20\%$ (P < 0.05) at 6, $29\%$ (P < 0.05) at 12, and $13\%$ at 24 following a single injection of urethan. At the same time, significantly lower incidences of lung adenoma were noted at 6 weeks $(50\%)$ and 12 weeks $(27\%)$ following the administration of ginseng to urethan-injected mice. This result indicates that the enhancement of N.K. activity by ginseng makes a contribution to its anticarcinogenic effect. On the hand, N.K. activity was suppressed by benzo(a)pyrene during the time span of this experiment and it almost returned to the level of controls following the adminsitration of ginseng. However, the lung adenoma induced by benzo(a)pyrene began to occur at 48 weeks in which N.K. activity had naturally declined to a very low level in all experimental mice, and administration of ginseng did not decrease the incidence. In explanation of this result, we might propose that the recovery of the N.K. activity by ginseng had little effect on the incidence of lung adenoma because of the long latent period of carcinogenesis by benzo(a)pyrene. In conclusion, these results suggest that the anticarcinogenic effect of ginseng in urethan-treated mice may be related to the augmentation of N.K. activity.
Park, Gye-Young;Kim, Jae-Yeol;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo
Tuberculosis and Respiratory Diseases
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v.44
no.3
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pp.601-610
/
1997
Background : Monocytes/macrophages play a central role in determining the host response during Gram-negative infection through secretion of a variety of mediators after stimulation of LPS. Even though cytokine production has been shown to play an important role in host defense during sepsis, cytokine release may also lead to tissue injury. Thus, regulation of macrophage response to LPS is critical for host survival during Gram-negative sepsis. In animals exposed to nonlethal doses of endotoxin, a characteristic hyporesponsiveness to subsequent administration of endotoxin has been observed. This phenomenon was known as 'LPS tolerance'. However, little information is available regarding the underlying mechanism of LPS tolerance. Method : Peripheral blood monocyte(PBMC) was isolated from peripheral blood of normal volunteers by adhesion purification method. To evaluate the conditions to obtain LPS tolerance, preculture was carried out with LPS at 10ng/ml for 24 hours. For stimulation, culture plates were washed two times and were stimulated with LPS at $1{\mu}g/ml$ for 4, 6 and 26 hours. To assess the underlying mechanisms of LPS tolerance, autologous serum, PMA, anti-CD14 Ab, Indomethacin or $PGF_2$ were added to preculture solution respectively. Cytokine concentrations in culture supernatants were measured using ELISA for TNF-$\alpha$ and IL-8 and mRNA of TNF-$\alpha$ and IL-8 were determined by Northern blot analysis. Results : The exposure of PBMC to low dose of LPS suppressed the cytokine production and mRNA expression of TNF-$\alpha$, but not IL-8. Anti-CD14 Ab partially recovered production of TNF-$\alpha$ which was suppressed by preculture with low dose LPS. The preculture with PMA induces LPS tolerance, as preculture with low dose LPS. Conclusion : LPS tolerance to TNF-$\alpha$ is regulated pretranslationally and is influenced by protein kinase C pathway and CD14.
The Preservation Decree (1933) is the basic law relevant to the conservation of cultural property of colonial Chosun, and invoked clauses from the Old History Preservation Act (1897), the Historic Scenic Sites Natural Monument Preservation Act (1919), and the National Treasure Preservation Act (1929), which were all forms of Japanese Modern Cultural Heritage Law, and actually used the corresponding legal text of those laws. Thus, the fact that the Preservation Decree transplanted or imitated the Japanese Modern Cultural Heritage Law in the composition of the constitution can be proved to some extent. The main features and characteristics of the Preservation Decree are summarized below. First, in terms of preservation of cultural property, the Preservation Decree strengthened and expanded preservation beyond the existing conservation rules. In the conservation rules, the categories of cultural properties were limited to historic sites and relics, while the Preservation Decree classifies cultural properties into four categories: treasures, historic sites, scenic spots, and natural monuments. In addition, the Preservation Decree is considered to have advanced cultural property preservation law by establishing the standard for conserving cultural property, expanding the scope of cultural property, introducing explicit provisions on the restriction of ownership and the designation system for cultural property, and defining the basis for supporting the natural treasury. Second, the Preservation Decree admittedly had limitations as a colonial cultural property law. Article 1 of the Preservation Decree sets the standard of "Historic Enhancement or Example of Art" as a criteria for designating treasures. With the perspective of Japanese imperialism, this acted as a criterion for catering to cultural assets based on the governor's assimilation policy, revealing its limitations as a standard for preserving cultural assets. In addition, the Japanese imperialists asserted that the cultural property law served to reduce cultural property robbery, but the robbery and exporting of cultural assets by such means as grave robbery, trafficking, and exportation to Japan did not cease even after the Preservation Decree came into effect. This is because governors and officials who had to obey and protect the law become parties to looting and extraction of property, or the plunder and release of cultural property by the Japanese continued with their acknowledgement,. This indicates that cultural property legislation at that time did not function properly, as the governor allowed or condoned such exporting and plundering. In this way, the cultural property laws of the Japanese colonial period constituted discriminative colonial legislation which was selected and applied from the perspective of the Japanese government-general in the designation and preservation of cultural property, and the cultural property policy of Japan focused on the use of cultural assets as a means of realizing their assimilation policy. Therefore, this suggests that the cultural property legislation during the Japanese colonial period was used as a mechanism to solidify the cultural colonial rules of Chosun and to realize the assimilation policy of the Japanese government-general.
Kim, Eun Ji;Kim, Guen Tae;Kim, Bo Min;Lim, Eun Gyeong;Ha, Sung Ho;Kim, Sang-Yong;Kim, Young Min
Journal of Life Science
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v.26
no.7
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pp.764-771
/
2016
Extracts from Artemisia annua Linné (AAE) have been known to possess various functions, including anti-bacterial, anti-virus, and anti-oxidant effects. However, the mechanism of those effects of AAE is not well-known. The aim of this study was to analyze the inhibitory effects of AAE on cell proliferation of the human hepatoma cell line (Hep3B) and to examine its effects on apoptosis. Activation by phosphorylation of Akt is cell proliferation through the phosphorylation of TSC2, mTOR, and GSK-3β. We suggested that AAE may exert cancer cell apoptosis through Akt/mTOR/GSK-3β signal pathways and mitochondria-mediated apoptotic proteins. For this, we examined the effects of extracts of AAE on cell proliferation according to treatment concentration. Treatment with AAE not only reduced cell viability, but also resulted in the induced release of lactate dehydrogenase (LDH). These results were determined with a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and a lactate dehydrogenase (LDH) assay. Furthermore, we determined the effects of apoptosis through Hoechst 33342 staining, annexinⅤ-propidium iodide (PI) staining, 5,5′, 6,6′-tetrachloro-1,1′,3,3′-tetraethyl-imidacarbocyanine iodide (JC-1) staining, and Western blotting. Our study showed that the treatment of liver cancer cells with AAE resulted in the inhibition of Akt, TSC2, GSK-3β-phosphorylated, Bcl-2, and pro-caspase 3 and the activation of Bim, Bax, Bak, and cleaved PARP expressions. These results indicate that AAE induced apoptosis by means of a mitochondrial event through the regulate of Akt/mTOR/GSK-3β signaling pathways.
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