• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.9
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• pp.1257-1264
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• 2016

Extracts from Artemisia annua $Linn\acute{e}$ (AAE) have various functions (anti-malaria, anti-virus, and anti-oxidant). However, the mechanism of the effects of AAE is not well known. Thus, we determined the apoptotic effects of AAE in AGS human gastric carcinoma cells. In this study, we suggested that AAE may exert cancer cell apoptosis through the Akt/mammalian target of rapamycin (mTOR)/glycogen synthase kinase (GSK)-$3{\beta}$ signal pathway and mitochondria-mediated apoptotic proteins. Activation by Akt phosphorylation resulted in cell proliferation through phosphorylation of tuberous sclerosis complex 2 (TSC2), mTOR, and GSK-$3{\beta}$. Thus, de-phosphorylation of Akt inhibited cell proliferation and induced apoptosis through inhibition of Akt, mTOR, phosphorylation of GSK-$3{\beta}$ at serine9, and control of Bcl-2 family members. Inhibition of GSK-$3{\beta}$ attenuated loss of mitochondrial membrane potential and release of cytochrome C. Bax and pro-apoptotic proteins were activated by their translocation into mitochondria from the cytosol. Translocation of Bax induced outer membrane transmission and generated apoptosis through cytochrome C release and caspase activity. We also measured 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, lactate dehydrogenase assay, Hoechst 33342 staining, Annexin V-PI staining, 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethyl-imidacarbocyanine iodide staining, and Western blotting. Accordingly, our study showed that AAE treatment to AGS cells resulted in inhibition of Akt, TSC2, GSK-$3{\beta}$-phosphorylated, Bim, Bcl-2, and pro-caspase 3 as well as activation of Bax and Bak expression. These results indicate that AAE induced apoptosis via a mitochondrial event through regulation of the Akt/mTOR/GSK-$3{\beta}$ signaling pathways.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.15 no.3
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• pp.241-250
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• 1982

The structure of gonads, gametogenesis and reproductive cycle of the cockle, Fulvia mutice, were studied mainly by histological observation. The materials were monthly sampled in the southern area of Yeosu from October 1980 to September 1981. F. mutica was monoecious. The gonads were situated between the liver tissues and the outer fibronuscular layers compacted by the connective tissue fibers and muscle fibers beneath the outermost layer of simple cuboidal epithelium. The gonad was composed of a number of the ovarian sacs and the testicular tubules which form the tubular structure. Testicular tubules in the mature stage sometimes contained 'testis-ova' The undifferentiated mesenchymal tissues and the eosinophilic cells were abundantly distributed on the germinal epithelium in the early development stage. With the further development of the ovary and testis, these tissues and cells gradually disapprared. The undifferentiated mesenchymal tissues and the eosinophilic cells are related to the growing of the oocytes and spermatocytes . Early multiplicating oogonium was about $10{\mu}m$ in diameter. As the oocytes grow to $27-34\times50-58{\mu}m$ by increasing cytoplasm, the oocytes connected to the basement membrane by their egg-stalks. The ripe eggs were about $60{\mu}m$ in diameter and they were surrounded by gelatinous membrane. Most male germ cells in mature stage were transformed into the spermatozoa and they formed the sperm bundles. After spawning, undischarged ripe eggs and spermatozoa remained in the ovarian sac and the testicular tubule respectively for some time, then they finally degenerated. Especially the early spent ovarian sacs in May did not contract significantly and then they took part in the secondary maturation within two or three months during the summer season. The monthly changes of the fatness well agreed with the reproductive cycle. The reproductive cycle of F. mutica could be classified into six successive stages : multiplicative, growing, mature, spent, degenerative and recovery stage. It seems that the spawning season is closely rotated to the water temperature, and the spawning occurs from May to October at about $20^{\circ}C$ in water temperature. The peak spawning seasons appeared twice a year between June and July and in September. Acknowledgement The authors wish to express their gratitude to Dr. Kim, In Bae, Dr. Chun, Seh Kyu and Dr. Yoo, Sung Kyoo of National Fisheries University of Busan and Mr. Min, Byoung Seo of National fisheries Research and Development Agency for their critical reading of the manu script.

• Applied Microscopy
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• v.32 no.2
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• pp.91-105
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• 2002

Urea transport in the kidney is mediated by a family of transporter proteins that includes renal urea transporters (UT-A) and erythrocyte urea transporters (UT-B). The cDNA of five isoforms of rat UT-A, UTA1, UT-A2, UT-A3, UT-A4, and UT-A5 have been cloned. The purpose of this study was to examine the expression of UT-A (L194), which marked UT-A1, UT-A2 and UT-A4. Male Sprague-Dawley rats, weighing approximately 200 g, were divided into three group: control rats had free access to water, dehydrated rats were deprived of water for 3 d, and water loaded rats had free access to 3% sucrose water for 3 d before being killed. The kidneys were preserved by in vivo perfusion through the abdominal aorta with the 2% paraformaldehyde-lysine- periodate (PLP) or 8% paraformaldehyde solution for 10 min. The sections were processed for immunohistochemical studies using pre-embedding immunoperoxidase method and immunogold method. In the normal rat kidney, UT-A1 was expressed intensely in the cytoplasm of the inner medullary collecting duct (IMCD) cell and UT-A2 was expressed on the plasma membrane of the terminal portion of the shortloop descending thin limb (DTL) cells (type I epithelium) and of the long-loop DTL cells (type II epithelium) in the initial part of the inner medulla. Immunoreactivity for UT-A1 in the IMCD cells, was decreased in dehydrated animals whereas strongly increased in water loaded animals compared with control animals. In the short-loop DTL, immunoreactivity for UT-A2 was increased in intensity in both dehydrated and water loaded groups. However, in the long-loop DTL of the outer part of the inner medulla, immunoreactivity for UT-A2 was markedly increase in intensity in dehydrated group, but not in water loaded group. In conclusion, in the rat kidney, UT-A1 is located in the cytoplasm of IMCD cells, whereas UT-A2 is located in the plasma membrane of both the short-and long-loop DTL cells. Immunohistochemistry studies revealed that UT-A1 and UT-A2 may have a different role in urea transport and are regulated by different mechanisms.

• Journal of Korean Medical classics
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• v.12 no.2
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• pp.56-71
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• 1999

1. In the recohnition of cosmos true form, It is compared to the Boundless(無極) the Great Absolute(太極),Yin and Yang(兩儀) throungh the fertilazation process of spermatozoon and ovum. 2. It is explained that principle of unchange through the Form and Action(體 用) relation of the outer appearances and Number (象數) with matching the number of nine and ten to HaDoo(河圖) and RakSye(洛書). 3. Eigth divinations(八卦) being compared to the human body, Care presevation of pregnancy(養胎) is explained that head forms firstly(Gun-I 1乾一), secondly lung(Tae-E 兌二), heart(E-Sam 離三), liver(Jin-Sa 震四), gall bladder(Son-Oo 巽五), kidney(Gam-Yuk 坎六), intestines and stomach(Gan-Chill 艮七), lastly flesh forms(Gon-Pal 坤八). 4. It is explained that process of physiological change of $\ll$Nei Ching The Natural Truth in Ancient Times$\gg$(內經 上古天眞論) by matching boy at the age of 8 to Gan-divination(艮卦), and girl at the age of 7 to Tae-divination(兌卦). 5. The theory of six sons from Gun-Gon(乾坤六子論) is explained by relation of Apriority Eight-divination(先天八卦) obedience and disobedience-left and right. 6. It is explained that form of the human-body and the relationship of the Heart - the Kidney through the Gam(坎) Li(離) - divination 7. The effort of interpretating time and space of the Twelve Horary signs is explanined by season, direction, Five elements(五行), rise and decline, the Three Sum(三合), the Six Sum (六合), the six crash(六衡)'s relation. 8. the process of change from apriority(先天) to postery(後天) in the book of Changes(周易) is explanined by comparing to the phenomenum of nature and the human body. 9. The Energy Satus(氣位) are different from the direction of Eight-divination(八卦) and the properties of the good or bad of herb-drugs are differnt from the place of production. 10. The rightness of realizating the Overlapping-divinations(重卦) are compared to the phenomenum of nature through the Divination Virture(卦德). 11. The dependence-relations of The Twelve Meridians(十二經脈) are explained by-matching January with liver meridian, February with gall bladder meridian, march with heart pericardium meridiam, April with small intestine meridkan, August with lung spleen meridian, jury with stomach meridian, August with lung meridian, September with large intestine meridian, October with urinary bladder meridianm November with kidney meridian. December with triple energizer meridian throng The Twelve Byuk-divination. 12. The process of menstration cycle is explained by The Month symbolizing-divination(月候卦). 13. Through The Trade(交易) prove the reason of feverish sympotoms to use feverish Drug, mill sympotoms to use mill drug of prescription and Heart-Kidneys Consensus(心賢相交) and through The Change(變易), prove the chill and feverish consensus of forechill after feverish, fore feverish after chill and through. The Non-Change(不易) explain the reason of chill sympotoms to use feverish drug, feverish sympotoms to use chill drug of prscription. 14. Ho-divination(互卦) applicate Jxa Sa(佐使) herb drug match of Kun Sin Jwa Sa() theory. 15. According to the Hyo-position(爻位) match the ages, body form and drug by matching Ehight-divination(八卦) to the human body form and function in medicine and the book of Changes(周易) application emphasize the human body Ehight-divination(人身八卦). 16. Throgh the Order-divination(序卦) explain the rightness of Divination Image(卦象) arrangement and all things take shape by cosmo-energy conseusus(宇宙氣交). 17. Throgh the Mixing-divination(難卦) supply the vacancy of medicine and the book of Changes(周易) relationship in the foreword explian the human energy movements, sleep, vomitting, the energy arrival(逮氣), heart pericardium(心包), lung membrane(肺膜) etc.... Like the above sentence medicine and the book of Changes(周易) theory of scholar Tang on the viewpoint of easten-the way Western appliance(東道西器) researching abyss of medicine impart to descendants, so I think that the achievement of medicine and the book of Changes(周易) study is very excellant and I expect that the study Korean Oriental Medicine(韓醫學) theory by means of medicine the book of Changes(周易) reference, will be accelarated.

• Applied Microscopy
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• v.31 no.4
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• pp.355-365
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• 2001

This study was performed to investigate the distribution and differentiation on the immunoreacted cells of the ChAT (choline acetyltransferase) at the Meynert basal nucleus of the forebrains in the growth periods of rat, using the immunohistochemistric method. According to the cell shape and the ratio of long axis vs short axis of cell soma, the ChAT antibody reacted nerve cells in the Meynert basal nucleus of the rats were classified into six types. In the adult rats, the FD (frequency distributions) of round, oval and elongated cells were maximum. The FD of these types were shown to be progressively decreased during the postnatal development. In addition, the FD of elongated nerve cells in were observed in the adult rats respectively. This was thought to be the same phenomenon as those in the round and oval cells . The total mean volume of ChAT antibody reacted cell somata was lowest in the PND (postnatal days) 7 rats and was highest in the PND 21 rats. But, those were decreased to the adult. These results suggest that ChAT antibody reacted nerve cells grow up to PND 21 and then, differentiate into the various types by neurites outgrowing. On the electron micrography, the adult rat forebrain cells were obtained to be well developed ribosomes, polysomes , rough endoplasmic reticula and mitochondria. The immunreactivities were observed in ribosomes, polysomes, rough endoplasmic reticula and outer membrane of mitochondria. Golgi complexes were poorly developed and not showed jmmunreactivity. The ribosomes , polysomes and endoplasmic reticula are considered to be closely related to the inter cellular localization and biosynthesis of the ChAT but not Golgi complex. According to the results in the present study, it is considered that the ChAT-immunoreacted nerve cells in the Meynert basal nucleus of the rat forebrains are differentiated throughout the postnatal development with following processes of changes; 1) the cholinergic nerve cells develop postnatally 2) cell soma volumes gradually increase during the early postnatal days 3) and then, cells differentiate into the various types by projecting the neurites to the appropriate area after PND 21.

• The Korean Journal of Malacology
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• v.24 no.1
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• pp.41-50
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• 2008

The histochemical characteristics and ultrastructure of the mantle in the spiny top shell, Batillus cornutus were described using light and electron microscopy. The simple epidermal layer wrapped on the top and bottom of the centrally located connective tissue. And then the epidermal layer were divided into the outer epidermal layer near a shell and the inner epidermal layer closed to the visceral mass. The connective tissue layer was composed of the collagen fiber muscularfiber bundle and hemolymph sinus. Mucous cells in the apical mantle contained acid and neutral mucopolysaccaride, and acidic carboxylated mucopolysaccaride in the mid and marginal mantle. The mantle thickness, epidermal layer thickness and hemolymph sinus area displayed a trend of reduction from the marginal zone to the apical zone. From TEM observation, it was possible to distinguish epithelium, ciliated cell, absorptive cell and secretory cell in the epidermal layer. The epithelia were columnar and the nucleus was elliptical. The free surface were covered with microvilli. The lateral membranes of epithelium was con nected with neighboring cells by the zonular occludens, zonular adherens and membrane interdigitation. Ciliated cell on free surface had cilia and microvilli, and numerous mitochondria in the apical cytoplasm. In the epidermal layer, it observed 2 type cells having absorptive function. The absorptive cells were columnar in shape, and contained microvilli, pinocytotic vesicles, mitochondria and lysosomes of various electron density. Secretory cells can be divided into four types (A, B, C, D) depending on the cell shape and characteristics of secretory granules. These cells were unicellular glands and had similar characteristics to previously reported on the mantle of the gastropod and bivalves.

• Food Science of Animal Resources
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• v.23 no.2
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• pp.161-167
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• 2003

Identification of salmonellosis-infected commercial poultry flocks has become a pivotal component of efforts to reduce incidence of egg-associated transmission of S. enteritidis and S. typhimurium to humans. As a basic study for sanitary control of S. enteritidis and S. typhimurium, main food-borne pathogenic bacteria in eggs produced by domestic hens, commercial egg samples were tested for specific antibodies to whole cells of S. enteritidis and S. typhimurium and outer membrane protein(OMP) of S. typhimurium by ELISA to detect infection of S. enteritidis and S. typhimurium in various groups of hens. When the antibody titers of yolks from three commercial brand eggs were tested after diluting in the ratio from 1:100 to 1:1,600 with double dilution method, ELISA values of the specific antibodies could be shown as differences in dilution patterns by comparing with negative control egg. When the antibody titers of the yolks from two commercial brand eggs were tested after diluting in the ratio of 1:200 and 1:1,000, ELISA values of specific antibodies were different among same brand eggs. When the antibody titers of yolks from five eggs sampled randomly from twenty one commercial brand eggs were tested after diluting in the ratio of 1:1,000, ELISA value of the specific antibodies were shown generally high. ELISA values of 28.5, 30, and 28.5% of yolks from 21 brand eggs were shown low and similar to negative control egg in antibody titers to whole cells of S. enteritidis and S. typhimurium and OMP of S. typhimurium, respectively. The results demonstrated that ELISA test of egg yolk antibody could provide a highly sensitive indicator to detect contamination of S. typhimurium and S. enteritidis in poultry, and could be used effectively to reduce incidence of S. typhimurium and S. enteritidis infection in poultry.

• Journal of Oral Medicine and Pain
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• v.32 no.2
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• pp.137-150
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• 2007

Trees emit phytoncide into atmosphere to protect them from predation. Phytoncide from different trees has its own unique fragrance that is referred to as forest bath. Phytoncide, which is essential oil of trees, has microbicidal, insecticidal, acaricidal, and deodorizing effect. The present study was performed to examine the effect of phytoncide on Porphyromonas gingivalis, which is one of the most important causative agents of periodontitis and halitosis. P. gingivalis 2561 was incubated with or without phytoncide extracted from Hinoki (Chamaecyparis obtusa Sieb. et Zucc.; Japanese cypress) and then changes were observed in its cell viability, antibiotic sensitivity, morphology, and biochemical/molecular biological pattern. The results were as follows: 1. The phytoncide appeared to have a strong antibacterial effect on P. gingivalis. MIC of phytoncide for the bacterium was determined to be 0.008%. The antibacterial effect was attributed to bactericidal activity against P. gingivalis. It almost completely suppressed the bacterial cell viability (>99.9%) at the concentration of 0.01%, which is the MBC for the bacterium. 2. The phytoncide failed to enhance the bacterial susceptibility to ampicillin, cefotaxime, penicillin, and tetracycline but did increase the susceptibility to amoxicillin. 3. Numbers of electron dense granules, ghost cell, and vesicles increased with increasing concentration of the phytoncide, 4. RT-PCR analysis revealed that expression of superoxide dismutase was increased in the bacterium incubated with the phytoncide. 5. No distinct difference in protein profile between the bacterium incubated with or without the phytoncide was observed as determined by SDS-PAGE and immunoblot. Overall results suggest that the phytoncide is a strong antibacterial agent that has a bactericidal action against P. gingivalis. The phytoncide does not seem to affect much the profile of the major outer membrane proteins but interferes with antioxidant activity of the bacterium. Along with this, yet unknown mechanism may cause changes in cell morphology and eventually cell death.

• Journal of Life Science
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• v.26 no.8
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• pp.887-894
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• 2016

The extract from Artemisia annuain L.(AAE) is known as a medicinal herb that is effective against cancer. Apoptosis is the process of programmed cell death, and mitochondria are known to play a central role in cell death control. In this study, we evaluated the p53-independent apoptosis of extract of AAE through downregulation of Bcl-2 and the mitochondrial pathway in A549 (lung cancer cells). AAE may exert cancer cell apoptosis through regulating p-Akt, Cox-2, p53 and mitochondria-mediated apoptotic proteins. p-Akt/cox-2 is known to play an important role in cell proliferation and cell survival. The Bcl-2 pro-apoptotic proteins (such as Bax, Bak and Bim) mediate the permeabilization of the mitochondrial outer membrane. Treatment of AAE reduces p-Akt, p-Mdm2, cox-2 and anti-apoptotic proteins (such as Bcl-2), while tumor suppressor p53 and pro-apoptotic proteins. Activation of Bax/Bak releases cytochrome c from mitochondria to the cytosol to activate a caspase. Caspase-3 is the major effector caspase associated with apoptotic pathways. Caspase-3 generally exists in cytoplasm in the form of a pro-enzyme. In the initiation stage of apoptosis, caspase-3 is activated by proteolytic cleavage and activated caspase-3 cleaves poly (ADP-ribose) polymerase (PARP). We treated Pifithrin-α (p53 inhibitor) and Celecoxib (Cox-2 inhibitor) to learn the relationship between the signal transduction of proteins associated with apoptosis. These results suggest that AAE induces apoptosis through a p53-independent pathway in A549.

• Journal of Food Hygiene and Safety
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• v.39 no.4
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• pp.343-352
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• 2024

This study aimed to develop probiotics with anti-periodontitic effects to help treat inflammation in the tissues surrounding the teeth. We isolated Weisiella cibaria (W. cibareia) SPM402 and Lactobacillus paracasei (L. paracasei) SPM412 from homemade kimchi and used their cell-free supernatants. At a concentration of 10 mg/mL of L. paracasei SPM412 (LP412) inhibited the formation of Fusobacterium nucleatum (F. nucleatum) biofilm by 95.99±0.73%. In addition, 10 mg/mL of LP412 reduced the RQ value of fimA, an adhesin gene of Porphyromonas gingivalis (P. gingivalis) to 0.08±0.05, and the RQ value of radD, an adhesin gene of F. nucleatum, to 0.08±0.008. When the P. gingivalis outer membrane vehicle (Pg OMV) induced inflammation in YD-38 cells, the RQ value of TNF-α was increased to 36.68±1.85, but was reduced to 4.15±0.37 in the presence of 1 mg/mL of W. cibareia SPM402 (WC402). Similarly, in Pg OMV-induced inflammation in THP-1 cells, the RQ value of IL-1β increased to 2,330.65±204.61 but was reduced to 15.19±4.57 in the presence of 15 mg/mL of WC402. In F. nucleatum-induced inflammation in YD-38 cells, the RQ value of IL-8 increased to 15.10±1.11 and was decreased to 2.67±0.50 in the presence of 1 mg/mL of LP412. In conclusion, W. cibaria SPM402 and L. paracasei SPM412 showed potent anti-inflammatory effects against oral pathogenic bacteria and hold promise as functional probiotics with anti-periodontitic activity.