• The Korean Journal of Physiology and Pharmacology
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• v.11 no.5
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• pp.207-213
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• 2007

This study was designed to characterize ureteral smooth muscle motility and also to study the effect of forskolin(FSK) and isoproterenol(ISO) on smooth muscle contractility in murine ureter. High $K^+$(50 mM) produced tonic contraction by $0.17{\pm}0.06mN$(n=19). Neuropeptide and neurotransmitters such as serotonin($5{\mu}M$), histamine($20{\mu}M$), and carbarchol(CCh, $10{\sim}50{\mu}M$) did not produce significant contraction. However, CCh($50{\mu}M$) produced slow phasic contraction in the presence of 25 mM $K^+$. Cyclopiazonic acid(CPA, $10{\mu}M$), SR $Ca^{2+}$-ATPase blocker, produced tonic contraction(0.07 mN). Meanwhile, inhibition of mitochondria by protonophore carbnylcyanide m-chlorophenylhydrazone(CCCP) also produced weak tonic contraction(0.01 mN). The possible involvement of $K^+$ channels was also pursued. Tetraethyl ammonium chloride(TEA, 10 mM), glibenclamide($10{\mu}M$) and quinidine($20{\mu}M$) which are known to block $Ca^{2+}$-activated $K^+$ channels($K_{Ca}$ channel), ATP-sensitive $K^+$ channels($K_{ATP}$) and nonselective $K^+$ channel, respectively, did not elicit any significant effect. However, $Ba^{2+}$($1{\sim}2mM$), blocker of inward rectifier $K^+$ channels($K_{IR}$ channel), produced phasic contraction in a reversible manner, which was blocked by $1{\mu}M$ nicardipine, a blocker of dehydropyridine-sensitive voltage-dependent L-type $Ca^{2+}$ channels($VDCC_L$) in smooth muscle membrane. This $Ba^{2+}$-induced phasic contraction was significantly enhanced by $10{\mu}M$ cyclopiazonic acid(CPA) in the frequency and amplitude. Finally, regulation of $Ba^{2+}$-induced contraction was studied by FSK and ISO which are known as adenylyl cyclase activator and $\beta$-adrenergic receptor agonist, respectively. These drugs significantly suppressed the frequency and amplitude of $Ba^{2+}$-induced contraction(p<0.05). These results suggest that $Ba^{2+}$ produces phasic contraction in murine ureteral smooth muscle which can be regulated by FSK and $\beta$-adrenergic stimulation.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.2
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• pp.198-203
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• 1999

Gonadal development and reproductive cycle off Gomphina melanaegis collected in the coastal waters of Chumunjin, Korea were investigated monthly from April 1996 to April 1997. G. melanaegis was dioecious, The gonads were located between the digestive diverticula and muscle tissues of the foot, The ovary was composed of a number of ovarian sacs, and the testis was composed of several testicular tubules. The flesh weight rate was reached the maximum in August ($23.0\%$), and then decreased to $19.8\%$ in September. In March, the value was reached the minimum ($17.8\%$) and then increased, The size of mature oocyte was ranged $50\~60\mu$m in diameter and had a germinal vesicle with a nucleolus. Mature oocyte contained a large number of yolk granules and lipid granules in its cytoplasm. The spermatozoon was consisted of a conical nucleus with acrosome, a middle piece containing four mitochondria and proximal and distal centrioles, and a flagellum, Sex ratio (male/female) and minimum size for sexual maturation of G. melanaegis were 0.79 and about 25 mm in shell length, respectively. The reproductive cycle could be classified into five succesive stages: multiplicative (December to March), growing (April and May), mature(June), sprawning (July and August), and degenerative and resting (September to November) stages.

• Restorative Dentistry and Endodontics
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• v.13 no.2
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• pp.283-294
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• 1988

The purpose of this study was to investigate the characteristic features of the cells and tissues of the chronic periapical lesions using light microscope and electron microscope. Fifteen dental periapical lesions were obtained from the patients undergoing periapical surgery. Each specimen was divided into two parts along the tooth axis. One part was routinely processed for histopathologic examinations. 12 periapical lesions were diagnosed as granuloma and 3 periapical specimens as periapical cyst. The other part was fixed in 2.5% glutaraldehyde in 0.1M sodium cacodylate buffer at pH 7.4 and 1% osmic acid in same buffer. They were embedded in Epon 812. The semithin sections were used for the orientation of the lesions and the ultrathin sections were stained conventionally and examined with AEI Corynth 500 electron microscope. The results were as follows. 1. PMN and macrophages, which were dominant cell type, were scattered in small or large numbers throughout the central destructive area of granuloma. In the granulomatous area, plasma cells and lymphoytes were found in significant number and a lot of new capillary formation were revealed. Clefts caused by cholesterol were often seen in the connective tissue. Occasionally foam cells became collected in groups and epithelial proliferation were present. 2. In both granuloma and cyst, some plasma cells contained narrow cisternae of granular endoplasmic reticulum of which was tightly packed with electron dense materials, and other cells exhibited dilated profiles of granular endoplasmic reticulum. 3. In the area where plasma cells and lymphocytes were collected in groups, lymphocytes with well developed nucleolus and profuse cytoplasm were found and differentiating plasma cells were also present. 4. In the epithelial strands of the granulomatous area, epithelial cells contained enlarged endoplasmic reticulum, tonofilaments and ribosoms. Toward the intercellular space epithelial cells protruded a few microvilli. In the intercellular space, exudate-like electron dense materials, most of which was attached to the plasma membrane, appeared. 5. Some foam cells filled with numerous lipid droplets and others had lipid droplets and crystal-like structures. 6. Cyst epithelium consisted of bright cells and dark cells. The former had bright cytoplasm and small amounts of ribosoms, and the latter dark cytoplasm, many ribosoms, mitochondria and elongated microvilli. 7. Epithelial cells near the cyst lumen protruded a lot of long microvilli toward intercellular space and cyst lumen.

• Journal of the Korean Society of Food Science and Nutrition
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• v.20 no.4
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• pp.320-328
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• 1991

The present study was designed to evaluate the effects of dietary vitamin E and coenzyme $Q_{10}$ supplementation on adriamycin (ADR) -induced lipid petoxidation in rats. After feeding the experimental diets for e weeks. Ann treatment significantly decreased growth performance of rats. But this decrement was not modified by supplementation of vitamin E or coenzyme $Q_{10}$ . Lipid peroxide values of plasma and heart mitochondria were elevated by Ann treatment. But these values were significantly decreased according to vitamin E or coenzyme $Q_{10}$ supplementation. Adriamycin treatment elevated glutathione peroxidase (GSH-Px) activity of rats, but this increment was modified by vitamin E supplementation. There was a tendency of higher superoxide dismutase (SOD) activity in ADR-treated rats. However, vitamin E or coenzyme $Q_{10}$ administration reduced this enzyme activity. With ADR treatment, arachidonic acid (20 : 4) was greatly increased, but docosahexaenoic acid (22 : 6) was not detected. Arachidonic acid was decreased and docosahexaenoic acid increased by supplementation of higher level of vitamin E or coenzyme $Q_{10}$ . Present data showed that dietary vitamin E and coenzyme $Q_{10}$ influenced on ADR-induced lipid peroxidation in rats, and also the degree of antioxidative effect was greater in vitamin E-supplemented rats.

• Journal of the Korean Society of Food Science and Nutrition
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• v.14 no.2
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• pp.145-150
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• 1985

This study was conducted to examine the pharmacological effect of water soluble extract of Lichens (Parmelia, Physcia and Cladonia species) on liver-damaged rat by $CCl_4$ injection. Rat livers were damaged acutely and chronically by one-time injection of $CCl_4$ just prior to five days of experimental period and continuous injections in every three days for eight weeks of experimental period, respectively. During each period the experimental group was fed Lichens extract(5.5 mg of dry wt/ml) instead of water given to the control group. For both acute and chronic liver damage, the experimental group showed higher oxidative activity of hepatic mitochondria measured by state 3 respiration, P/O ratio, respiratory control and ATP synthesized, compard to the control group. Serum glucose was slightly higher in the experimental group but liver glycogen showed no significant difference between experimental and control groups. In experimental group, liver glucose-6-phosphatase activity was increased during first two days after acute liver damage, but not significantly different from control group during chronic damage. Liver lactate, malate plus fumarte and glutamate tended to be higher in the experimental group, especially for chronic liver damaged rat. It is concluded that Lichens extract stimulate cytoplasmic and mitochondrial oxidative activities and the possible mechanism of the latter is supposed to involve the preservation of membrane integrity by certain component(s) of water-soluble Lichens extract.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.5
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• pp.649-659
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• 2011

To examine the anti-cancer effects of Lepidium virginicum L., the anti-proliferative and pro-apoptotic effects of a water extract of L. virginicum leaves (WELVL) and of L. virginicum roots (WELVR) were investigated in HCT116 human colon carcinoma cells. The treatment of HCT116 cells with WELVL and WELVR resulted in the inhibition of growth and morphological changes in a concentration-dependent manner by inducing apoptosis. The growth inhibition and apoptosis induction by WELVR was stronger than that of WELVL thus, we determined that WELVR was the more optimal extract for this study. The increased apoptotic events in HCT116 cells caused by WELVR were associated with an up-regulation of Fas ligand, Bax, and Bad expression, a down-regulation of Bcl-2, Bcl-$_XL$, and Bid expression, and a decrease in the mitochondrial membrane potential (MMP, ${\Delta}{\psi}m$). WELVR treatment induced the proteolytic activation of caspase-3, -8, and -9, and the degradation of caspase-3 substrate proteins, such as poly (ADP-ribose) polymerase (PARP), ${\beta}$-catenin, and phospholipase C-${\gamma}1$ (PLC-${\gamma}1$). In addition, apoptotic cell death induced by WELVR was correlated with a down-regulation of inhibitors of the apoptosis protein (IAP) family, such as the X-linked inhibitor of apoptosis protein (XIAP), cIAP-1, and cIAP-2. These findings suggest that the WELVR-induced inhibition of cell proliferation is associated with the induction of apoptotic cell death. WELVR may be a potential chemotherapeutic agent for the control of HCT116 human colon carcinoma cells.

• Archives of Pharmacal Research
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• v.29 no.10
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• pp.859-865
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• 2006

With an approach to study the anti-tumor effects and mechanism of selenium compound, we investigated the anti-tumor activity and mechanism of $Na_5SeV_5O_{18}{\cdot}3H_2O$ (NaSeVO) in K562 cells. The results showed that $0.625{\sim}20\;mg/L$ NaSeVO could significantly inhibit the proliferation of K562 cells in vitro in a time- and concentration-dependent manner as determined by microculture tetrazolium (MTT) assay, the IC50 values were 14.41 (4.45-46.60) and 3.45 (2.29-5.22) mg/L after 48 hand 72 h treatment with NaSeVO respectively. In vivo experiments demonstrated that i.p. administration of 5, 10 mg/kg NaSeVO exhibited an significant inhibitory effect on the growth of transplantation tumor sarcoma 180 (S180) and hepatoma 22 (H22) in mice, with inhibition rate 26.8% and 58.4% on S180 and 31.3% and 47.4% on H22, respectively. Cell cycle studies indicated that the proportion of G0/G1 phase was increased at 2.5 mg/L while decreased at 10 mg/L after treatment for 24, 48 h. Whereas S phase was decreased at 2.5-5 mg/L and markedly increased at 10 mg/L after treatment for 48 h. After treatment for 24 h, 10 mg/L NaSeVO also markedly increased S and G2/M phases. Take together, the result clearly showed that NaSeVO markedly increased S and G2/M phases at 10 mg/L. The study of immunocytochemistry showed that the expression bcl-2 is significantly inhibited by 10 mg/L NaSeVO, and bax increased. Morphology observation also revealed typical apoptotic features. NaSeVO also significantly caused the accumulation of $Ca^{2+}$ and $Mg^{2+}$, reactive oxygen species (ROS) and the reduction of pH value and mitochondrial membrane potential in K562 cells as compared with control by confocal laser scanning microscope. These results suggest that NaSeVO has anti-tumor effects and its mechanism is attributed partially to apoptosis induced by the elevation of intracellular $Ca^{2+}$, $Mg^{2+}$ and ROS concentration, and a reduction of pH value and mitochondria membrane potential (MMP).

• The korean journal of orthodontics
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• v.24 no.2
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• pp.331-348
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• 1994

The purpose of this study was to investigate changes in the shape and ultrastructure of the articular disc of the rat mandibular joint with aging. Mechanical stress applied to the articular disc changes during neonatal, suckling, juvenile, adult and senile stages. Mandibular joints of 6 groups of rats(1-, 7-, 17-, 27-, 55-day and over-1-year groups) were removed en bloc and processed for light and electro microscopic study. The changes in the shape of articular disc were examined by light microscope in each group. Structural and ultrastructural changes in the articular disc were examined by light and electron microscope in each group. The results were as follows : In the 1-day and 7-day groups, the articular disc was long and slender in shape and the articular disc was not fitted with the shape of the mandibular fossa and condyle. However' after that time, the anterior and posterior portions of the articular disc were more bulged and the middle portion was shorter and biconcave. Thus the articular disc was well fitted with the shape of the mandibular fossa and condyle. The cell density decreased with aging. In the l -day and 7-day groups, the Golgi apparatus, rough endoplasmic reticulum and free ribosome, which are involved in the synthesis of intracellular and extracellular matrix, were developed. In the 17-day, 27-day and 55-day groups, not only the cell organelles involved in the synthesis of the intracellular and extracellular matrix but also the cell organelles involved in the remodeling of the extracellular matrix(i.e., finger-like cell process, lysosome and mitochondria)were well developed. With advancing age, intracytoplasmic microfilaments were more accumulated and condroid cells increased. In the over-1-year group, the majority of cells of the articular disc were chondroid cells. The majority of cytoplasmic compartment were filled with intracytoplasmic microfilaments and cell organelles were not developed. Therefore, metabolic activities of the cell was markedly reduced and cells contained structures enduring mechanical stress, and cells which were in the process of degeneration were observed occasionally.

• International Journal of Oral Biology
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• v.34 no.1
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• pp.7-14
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• 2009

Nitric oxide (NO) acts as an intracellular messenger at the physiological level but can be cytotoxic at high concentrations. The cells within periodontal tissues, such as gingival and periodontal fibroblasts, contain nitric oxide syntheses and produce high concentrations of NO when exposed to bacterial lipopolysaccharides and cytokines. However, the cellular mechanisms underlying NO-induced cytotoxicity in periodontal tissues are unclear at present. In our current study, we examined the NO-induced cytotoxic mechanisms in human gingival fibroblasts (HGF). Cell viability and the levels of reactive oxygen species (ROS) were determined using a MTT assay and a fluorescent spectrometer, respectively. The morphological changes in the cells were examined by Diff-Quick staining. Expression of the Bcl-2 family and Fas was determined by RT-PCR or western blotting. The activity of caspase-3, -8 and -9 was assessed using a spectrophotometer. Sodium nitroprusside (SNP), a NO donor, decreased the cell viability of the HGF cells in a dose- and time-dependent manner. SNP enhanced the production of ROS, which was ameliorated by NAC, a free radical scavenger. ODQ, a soluble guanylate cyclase inhibitor, did not block the SNP-induced decrease in cell viability. SNP also caused apoptotic morphological changes, including cell shrinkage, chromatin condensation, and DNA fragmentation. The expression of Bax, a member of the proapoptotic Bcl-2 family, was upregulated in the SNP-treated HGF cells, whereas the expression of Bcl-2, a member of the anti-apoptotic Bcl-2 family, was downregulated. SNP augmented the release of cytochrome c from the mitochondria into the cytosol and enhanced the activity of caspase-8, -9, and -3. SNP also upregulated Fas, a component of the death receptor assembly. These results suggest that NO induces apoptosis in human gingival fibroblast via ROS and the Bcl-2 family through both mitochondrial- and death receptor-mediated pathways. Our data also indicate that the cyclic GMP pathway is not involved in NO-induced apoptosis.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.1
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• pp.60-66
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• 2014

The aim of this study was to evaluate the antioxidant activity and protective effect of extracts from the stems and leaves of Chrysanthemum boreale (CBSL) on t-BHP induced oxidative stress in human liver cells (Chang cells). Antioxidant activities in the extracts were determined for various radical scavenging activities including ferric reducing antioxidant power, 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activity, and oxygen radical absorbance capacity (ORAC). CBSL showed a very good scavenging effect of DPPH radical ($IC_{50}$ $0.009{\pm}0.002$ mg/mL), alkyl radical ($IC_{50}$ $0.004{\pm}0.001$ mg/mL), and hydroxyl radical ($IC_{50}$ $6.742{\pm}0.152$ mg/mL). CBSL also showed a strong antioxidant effect in the ORAC assay. In the MTT assay on human liver cells (Chang cells), the extracts showed protective effects by increasing cell viability, decreasing ROS, and restoring mitochondria membrane potential upon t-BHP induced oxidative stress. Our findings suggest that CBSL extracts are a potential therapeutic with protective antioxidant effects upon oxidative stress.