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Comparison of Property Changes of Black Jujube and Zizyphus jujube Extracts during Lactic Acid Fermentation (흑대추와 일반 건조대추의 추출 및 유산발효과정 중 특성 변화)

  • Auh, Mi Sun;Kim, Yi Seul;Ahn, Seung Joon;Ahn, Jun Bae;Kim, Kwang Yup
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.10
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    • pp.1346-1355
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    • 2012
  • This study was carried out to investigate the characteristics of black jujube and Zizyphus jujube extracts during lactic acid fermentation. Both extracts were fermented using Lactobacillus fermentum YL-3. As a result, viable cell number rapidly increased until 24 hours, after which it gradually decreased. Before lactic acid fermentation, the $IC_{50}$ of black jujube, which was 0.014 mg/mL, was lower than that of Zizyphus jujube. Further, black jujube showed stronger antioxidant activity (374.21 mg AA eq/g) than Zizyphus jujube. Contents of total polyphenolics in both extracts were 15.46 mg/g and 13.61 mg/g, respectively, whereas contents of total flavonoids were 374.21 ${\mu}g/g$ and 64.25 ${\mu}g/g$. After lactic acid fermentation, there was no significant increase in DPPH or ABTS free radical scavenging activity. Total polyphenolic content of Zizyphus jujube decreased to 12.39 mg/g upon fermentation, whereas flavonoid content significantly increased to 291.58 ${\mu}g/g$. Further, polyphenolic and flavonoid contents of black jujube increased from 15.46 mg/g to 17.46 mg/g and from 374.21 ${\mu}g/g$ to 1,135.29 ${\mu}g/g$, respectively. These results demonstrate that 9-Times Steamed and Dried increased functional components. Especially, lactic acid fermented black jujube showed remarkably high antioxidant activity. These results confirm the potential use of lactic acid fermented black jujube as a valuable resource for the development of functional foods.

The Late Quaternary Environmental Change in Youngyang Basin, South Eastern Part of Korea Penninsula (第四紀 後期 英陽盆地의 自然環境變化)

  • Yoon, Soon-Ock;Jo, Wha-Ryong
    • Journal of the Korean Geographical Society
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    • v.31 no.3
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    • pp.447-468
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    • 1996
  • The peat layer was deposited on the abandoned channel of incised meander of River Banbyuncheon with 7 meter thickness on Youngyang basin. The late Quaternary environmental change on the study area was discussed based on pollen anaalysis and radiocarbon-dating from this peat. The swamp which was caused to sediment the peat, was produced by which the fan debris from the adjacent slope damed the waterflow on the abandoned channel. The peat layer contains continuous vegetational history from 60,000y.B.P. to Recent. The peat deposit was divided into two layers by the organic thin sand horizon, which was sedimented at one time and made unconformity between the lower decomposed compact peat layers and the upper fresh fiberous peat layer. As the result of the pollen analysis, both peat layers from the two boring sites, Profile YY1 and Profile YY2 were divided into five Pollenzones(Pollenzone I, II, III, IV and V) and 12 Subzones which were mainly corresponded by the AP (Arboreal Pollen)-Dominance. The two profiles have some differences on the sedimentary facies and on the pollen composition as well. Therefore these were in common with the Pollenone III, however the Pollenzone I and II existed only on the Profile YY1 and the Pollenzone IV and V existed only on the Profile YY2. The lower layer containing the Pollenzone I, II and III revealed vegetational records of Pleistocene, which was characterized as tundra-like landscape and thin forested landscapes. It represented the NAP (Non-Arboreal Pollen)-period with a plenty of Artemisia sp., Sanguisorba sp., Umbelliferae, Gramineae and Cyperaceae. However a relatively high proportion of the boreal trees with Picea sp., Pinus sp. and Betula sp. as AP was observed in the lower layer. The upper layer contained the Pollenzone IVb and V and vegetational history in Holocene which was characterized by thick forested landscape with rich tree pollen. It represented AP-period with plenty of Pinus sp. and Quercus sp. as temperate trees. The temperature fluctuation supposed from the vegetational records is as follows; the Pollenzone I(Betula-Dominance, about 57,000y.B.P.) represents relatively cold period. The Pollenzone II(EMW-Domi-nance, 57,000-43,000y.B.P.)represents relatively warm period. This period is supposed to be Interstadial, the transi-tional stage from Alt- to Mittel Wurm. The Pollenzone III(Butula-, Pinus- and Picea-Dominace in turns, 43,000-15,000y.B.P.) reproesents cold period which had been built from Mittel-to Jung Wurm. Especially the Subzone IIId represents the coldest period throughout the Pollenzone III. It is corresponds to Wurm Glacial Maximu. It is supposed that the mean temperature in July of this period was coller about 10${^\circ}$C than present. The Pollenzone IV and V represent the vegetational history of Holocene. Tilia, Quercus and Pinus were dominant in turns during this period. Subzone IVb and Pollenzone I and II at east coastal plain of Korean penninsula reported by JO(1979).

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Fermentation Characteristics of Juice Pomace Feed by Horse Feces Microbes (말분변 미생물을 이용한 주스박 사료의 발효 특성)

  • Hwang, Won-Uk;Kim, Gyeom-Heon;Niu, Kai-Min;Lim, Joung-Ho;Woo, Jae-Hoon;Chae, Hyun-Seok;Park, Nam-Geon;Kim, Soo-Ki
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.37 no.4
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    • pp.290-300
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    • 2017
  • In vitro fermentation was conducted to figure out alternative fiber sources for horse feed. For the development of value-added products as a horse feed resource, the pomaces from apple, carrot, grape, and citrus were fermented under solid-state conditions in the presence of 60% soybean meal with 40% of each fruit pomace at 60% of moisture content. Lactobacillus plantarum SK3873, Lactobacillus plantarum SK3893, Weissella cibaria SK3880, and Bacillus subtilis SK3889 were isolated from the fermented fruit pomace by inoculation of horse feces. For the growth of Bacillus subtilis, Saccharomyces cerevisiae, and Lactobacillus plantarum, they were inoculated in 3-step order at 0, 12, and 24 h, respectively. The fruit pomace was fermented for 48 h at $35^{\circ}C$. The pH of the apple, carrot, grape, citrus and all mixed pomaces decreased from 5.45~6.25 to 4.40~4.77. Microbial growth was maintained at $10^8{\sim}10^9cfu/g$. After 12 and 24 h incubation, dry matter of carrot pomace were highest at 54.84 and 56.66%, respectively (P<0.05) and that of grape pomace was lower than others during fermentation (P<0.05). Dry matter was generally reduced by about 20%. NDF decreased gradually or maintained after 24 h, indicating the fiber degradation. Ash content tended to decrease during fermentation. After 48 hours fermentation, Bacillus, yeast and Lactobacillus showed an excellent growth by using juice by-products. These results suggest that fermented juice pomace has a potential as horse feedstuff with probiotics to maintain beneficial microflora in horse gut.

Relationship between Vaginal Cytology and Reproductive Hormone during the Estrous Cycle and Optimal Mating Time in Beagle Dogs (비글개에서 발정 주기 및 교배 적기 동안 질세포상과 번식 호르몬의 관계)

  • Lee K.C.;Kang H.G.;Cheun H.M.;Kim I.H.
    • Journal of Embryo Transfer
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    • v.21 no.2
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    • pp.109-119
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    • 2006
  • This study was conducted to examine relationship between vaginal cytology and reproductive hormones during the estrous cycle and to provide basic data to estimate for ovulation time and optimal mating time in 6 beagle dogs The duration of proestrus, estrus and diestrus were $8.5{\pm}1.4,\;10.0{\pm}1.4\;and\;54.0{\pm}2.8$ days at pregnant respectively, and $7.9{\pm}2.1,\;9.5{\pm}0.7\;and\;62.0{\pm}11.3$ days at non-pregnant respectively. The duration of interestrous intervals were $246.2{\pm}24.5$ days at pregnancy, and $175.3{\pm}34.5$ days at non-pregnancy. The duration of interestrous intervals at pregnancy was longer than that of non-pregnancy. A characteristic features of vaginal cytology during the estrous cycle were the high proportion of superficial cell, anuclear cell and erythrocyte in proestrus and estrus, parabasal cell, small intermediate cell and leukocyte in diestrus, and parabasal cell and small intermediate cell in anestrus, respectively. Cornification index (CI) in proestrus and estrus were significantly higher than that of CI in diestrus and anestrus. Plasma progesterone concentration was below 1.0 ng/ml at the first day of vulval bleeding at pregnancy and non-pregnancy, and then it was above 2.0 ng/ml at Day -2 in all bitches. When plasma progesterone concentration was first increased above 4.0 ng/ml, it was the second day after the first day of male acceptance. Plasma progesterone concentration showed above 40 ng/ml on Day $20{\sim}22$ in all bitches, and then it was gradually decreased until Day 35. Plasma progesterone concentration at pregnancy was higher than that of non-pregnancy from Day 35 to Day 63. Plasma estradiol-$17\;{\beta}$ concentration was above 9.0 pg/ml at the first day of vulval bleeding, and it showed 26.4 pg/ml on Day -2. When it was timed from the first day of male acceptance (Day 0), plasma estradiol-$17{\beta}$ concentration showed a peak on Day 0 and plasma progesterone concentration was first increased above 4.0 ng/ml on Day 2 which was the third day after plasma estradiol-$17{\beta}$ peak. CI was first increased above 80 and 90% on Day -1 and Day 1, respectively. CI was maintained above 80% from Day -1 to Day 8 (10 days) and above 90% from Day 1 to Day 6 (6 days), respectively. CI was maintained above 80% from Day 0 to Day 8 (9 days) and above 90% from Day 1 to Day 6 (6 days), respectively. Plasma progesterone concentration was first increased above 4.0 ng/ml on the second day after the day which CI was first increased above 90%. In conclusion, beagle bitches ovulated on the second day after the day which CI was first increased above 90% and on the day which plasma progesterone concentration was first increased 4.0 ng/ml, and it was estimated that the optimal mating time was the day which the second day after CI was first increased above 90% and plasma concentration was between $2{\sim}25ng/ml$. The measurement of plasma progesterone was used to determine of and accurate ovulation time and the optimal mating time, but vaginal cytology is low-priced and simple method to estimate estrous cycle, optimal mating time and ovulation time.

Analysis of Prognostic Factors Related to Survival Time for Patients with Small Cell Lung Cancer (소세포폐암 환자의 생존기간에 관련된 인자 분석)

  • Kim, Hee-Kyoo;Yook, Dong-Seung;Shin, Ho-Sik;Kim, Eun-Seok;Lim, Hyun-Jeung;Lim, Tae-Kwan;Ok, Chul-Ho;Cho, Hyun-Myung;Jung, Maan-Hong;Jang, Tae-Won
    • Tuberculosis and Respiratory Diseases
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    • v.54 no.1
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    • pp.57-70
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    • 2003
  • Background : Small cell lung cancer represents approximately 20% of all carcinomas of the lung, and is recognized as having a poor long term outcome compared to non-small cell lung cancer. Therefore, this study investigated the prognostic factors in small cell lung cancer patients in order to improved the survival rate by using the proper therapeutic methods. Material and method : The clinical data from 394 patients who diagnosed with small cell lung cancer and treated from 1993 to 2001 at the Kosin University Gospel Hospital, were analyzed. Result : There were 314 male patients (79.7%), and 80 female patients (20.3%). The number of those with limited disease was 177 (44.9%), and the number of those with extensive disease was 217 (55.1%). Overall, 366 out of 394 enrolled patients had died. The median survival time was 215 days (95% CI : 192-237days). The disease stage, Karnofsky performance state, 5% body weight loss for the recent 3 months, chemotherapy regimens, and the additive chest radiotherapy were identified as being statistically significant factors for the survival time. The median survival times of the supportive care group, one anticancer therapy, and two or more treatment groups were 17 days, 211 days, and 419 day, respectively (p<0.001). These data emphasize the importance of anticancer treatment to improve survival time for patients. The group of concurrent chemoradiotherapy (30 patients) showed significantly longer survival time than the group given sequential chemoradiotherapy (55 patients) (528 days versus 373 days, p=0.0237). The favorable prognostic factors of laboratory study were groups of leukocyte =8,000/mm3, ALP=200 U/L, LDH=450 IU/L, NSE=15 ng/mL, s-GOT=40 IU/L. In extensive disease, there was no difference according to the number of metastatic site. However, the median survival time of patients with ipsilateral pleural effusion had longer than patients having other metastatic sites. According to the survey periods, three groups were divided into 1993-1995, 1996-1998, and 1999-2001. The median survival time was significantly prolonged after 1999 in comparison to previous groups (177 days, 194 days, 289 days, p=0.001, 0.002, respectively). Conclusion: Disease stage and 5% body weight loss for recent 3 months at diagnostic state were significant prognostic factors. In addition, the performance status, serum ALP, LDH, NSE, CEA levels also appear to be prognostic factors. The survival time of those patients with small cell lung cancer has been prologned in recent years. It was suggested that the used of the EP (etoposied and cisplatin) chemotherapy method and concurrent chemoradiotherapy for patients with a limited stage contributed to the improved survival time.

THE EFFECTS OF THE PLATELET-DERIVED GROWTH FACTOR-BB ON THE PERIODONTAL TISSUE REGENERATION OF THE FURCATION INVOLVEMENT OF DOGS (혈소판유래성장인자-BB가 성견 치근이개부병변의 조직재생에 미치는 효과)

  • Cho, Moo-Hyun;Park, Kwang-Beom;Park, Joon-Bong
    • Journal of Periodontal and Implant Science
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    • v.23 no.3
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    • pp.535-563
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    • 1993
  • New techniques for regenerating the destructed periodontal tissue have been studied for many years. Current acceptable methods of promoting periodontal regeneration alre basis of removal of diseased soft tissue, root treatment, guided tissue regeneration, graft materials, biological mediators. Platelet-derived growth factor (PDGF) is one of polypeptide growth factor. PDGF have been reported as a biological mediator which regulate activities of wound healing progress including cell proliferation, migration, and metabolism. The purposes of this study is to evaluate the possibility of using the PDGF as a regeneration promoting agent for furcation involvement defect. Eight adult mongrel dogs were used in this experiment. The dogs were anesthetized with Pentobarbital Sodium (25-30 mg/kg of body weight, Tokyo chemical Co., Japan) and conventional periodontal prophylaxis were performed with ultrasonic scaler. With intrasulcular and crestal incision, mucoperiosteal flap was elevated. Following decortication with 1/2 high speed round bur, degree III furcation defect was made on mandibular second(P2) and fourth(P4) premolar. For the basic treatment of root surface, fully saturated citric acid was applied on the exposed root surface for 3 minutes. On the right P4 20ug of human recombinant PDGF-BB dissolved in acetic acid was applied with polypropylene autopipette. On the left P2 and right P2 PDGF-BB was applied after insertion of ${\beta}-Tricalcium$ phosphate(TCP) and collagen (Collatape) respectively. Left mandibular P4 was used as control. Systemic antibiotics (Penicillin-G benzathine and penicillin-G procaine, 1 ml per 10-25 1bs body weight) were administrated intramuscular for 2 weeks after surgery. Irrigation with 0.1% Chlorhexidine Gluconate around operated sites was performed during the whole experimental period except one day immediate after surgery. Soft diets were fed through the whole experiment period. After 2, 4, 8, 12 weeks, the animals were sacrificed by perfusion technique. Tissue block was excised including the tooth and prepared for light microscope with H-E staining. At 2 weeks after surgery, therer were rapid osteogenesis phenomenon on the defected area of the PDGF only treated group and early trabeculation pattern was made with new osteoid tissue produced by activated osteoblast. Bone formation was almost completed to the fornix of furcation by 8 weeks after surgery. New cementum fromation was observed from 2 weeks after surgery, and the thickness was increased until 8 weeks with typical Sharpey’s fibers reembedded into new bone and cementum. In both PDGF-BB with TCP group and PDGF-BB with Collagen group, regeneration process including new bone and new cementum formation and the group especially in the early weeks. It might be thought that the migration of actively proliferating cells was prohibited by the graft materials. In conclusion, platelet-derived growth factor can promote rapid osteogenesis during early stage of periodontal tissue regeneration.

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The Effect of Heat Shock Response on the Tumor Necrosis Factor-$\alpha$-induced Acute Lung Injury in Rats (Tumor Necrosis Factor-$\alpha$로 유도되는 백서의 급성 폐손상에 열충격반응이 미치는 효과)

  • Koh, Youn-Suck;Lim, Chae-Man;Kim, Mi-Jung;Cho, Won-Kyung;Jeoung, Byung-O;Song, Kyu-Young;Shim, Tae-Sun;Lee, Sang-Do;Kim, Woo-Sung;Kim, Dong-Soon;Kim, Won-Dong
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.6
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    • pp.1343-1352
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    • 1997
  • Background : Heat-treated cells are known to be protected from lysis by TNF, which is considered to play a central role in the pathogenesis of sepsis-induced acute lung injury. The objective of the study was to investigate the effect of heat shock response by heat-pretreatment on the acute lung injury of the rats induced by intratracheally administered TNF-$\alpha$, Methods : We intratracheally instilled either saline or TNF (R&D, 500ng) with and without heat pretreatment in Sprague-Dawley rats weighing 250~350 g. The heated rats were raised their rectal temperature to $41^{\circ}C$ and was maintained thereafter for 13 minutes at 18 h before intratracheal administration of saline or TNF. After 5 h of intratracheal treatment, lung leak, lung myeloperoxidase activity (MPO) and heat shock proteins were measured in rats. Lung leak index was defined as counts per minute of $I^{25}$ in the right lung divided by counts per minutes of $I^{25}$ in 1.0 ml of blood. All data are expressed as means ${\pm}$SE. Results : There is no difference in acute lung leak index ($0.099{\pm}0.024$ vs $0.123{\pm}0.005$) among the rats given saline intratracheally with and without heat pretreatment, but MPO activity showed a decreased tendency in heat-pretreated rats ($4.58{\pm}0.79\;U/g$) compared with heat-unpretreated rats ($7.32{\pm}0.97\;U/g$) (P=0.064). Rats administered TNF intratracheally with heat-pretreatment had decreased lung leak index ($0.137{\pm}0.012$) and lung MPO activity ($5.51{\pm}1.04\;U/g$) compared with those of heat-unpretreated and TNF-administered rats ($0.186{\pm}0.016$, $14.34{\pm}1.22\;U/g$) (P<0.05 in each). There were no significant difference of lung leak index and MPO activity between TNF-treated rats with heat-pretreatment and saline-treated rats with and without heat-pretreatment. Conclusion : The heat shock response attenuated neutrophil recruitment and acute lung leak induced by intratracheal instillation of TNF-in rats.

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Stock-Index Invest Model Using News Big Data Opinion Mining (뉴스와 주가 : 빅데이터 감성분석을 통한 지능형 투자의사결정모형)

  • Kim, Yoo-Sin;Kim, Nam-Gyu;Jeong, Seung-Ryul
    • Journal of Intelligence and Information Systems
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    • v.18 no.2
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    • pp.143-156
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    • 2012
  • People easily believe that news and stock index are closely related. They think that securing news before anyone else can help them forecast the stock prices and enjoy great profit, or perhaps capture the investment opportunity. However, it is no easy feat to determine to what extent the two are related, come up with the investment decision based on news, or find out such investment information is valid. If the significance of news and its impact on the stock market are analyzed, it will be possible to extract the information that can assist the investment decisions. The reality however is that the world is inundated with a massive wave of news in real time. And news is not patterned text. This study suggests the stock-index invest model based on "News Big Data" opinion mining that systematically collects, categorizes and analyzes the news and creates investment information. To verify the validity of the model, the relationship between the result of news opinion mining and stock-index was empirically analyzed by using statistics. Steps in the mining that converts news into information for investment decision making, are as follows. First, it is indexing information of news after getting a supply of news from news provider that collects news on real-time basis. Not only contents of news but also various information such as media, time, and news type and so on are collected and classified, and then are reworked as variable from which investment decision making can be inferred. Next step is to derive word that can judge polarity by separating text of news contents into morpheme, and to tag positive/negative polarity of each word by comparing this with sentimental dictionary. Third, positive/negative polarity of news is judged by using indexed classification information and scoring rule, and then final investment decision making information is derived according to daily scoring criteria. For this study, KOSPI index and its fluctuation range has been collected for 63 days that stock market was open during 3 months from July 2011 to September in Korea Exchange, and news data was collected by parsing 766 articles of economic news media M company on web page among article carried on stock information>news>main news of portal site Naver.com. In change of the price index of stocks during 3 months, it rose on 33 days and fell on 30 days, and news contents included 197 news articles before opening of stock market, 385 news articles during the session, 184 news articles after closing of market. Results of mining of collected news contents and of comparison with stock price showed that positive/negative opinion of news contents had significant relation with stock price, and change of the price index of stocks could be better explained in case of applying news opinion by deriving in positive/negative ratio instead of judging between simplified positive and negative opinion. And in order to check whether news had an effect on fluctuation of stock price, or at least went ahead of fluctuation of stock price, in the results that change of stock price was compared only with news happening before opening of stock market, it was verified to be statistically significant as well. In addition, because news contained various type and information such as social, economic, and overseas news, and corporate earnings, the present condition of type of industry, market outlook, the present condition of market and so on, it was expected that influence on stock market or significance of the relation would be different according to the type of news, and therefore each type of news was compared with fluctuation of stock price, and the results showed that market condition, outlook, and overseas news was the most useful to explain fluctuation of news. On the contrary, news about individual company was not statistically significant, but opinion mining value showed tendency opposite to stock price, and the reason can be thought to be the appearance of promotional and planned news for preventing stock price from falling. Finally, multiple regression analysis and logistic regression analysis was carried out in order to derive function of investment decision making on the basis of relation between positive/negative opinion of news and stock price, and the results showed that regression equation using variable of market conditions, outlook, and overseas news before opening of stock market was statistically significant, and classification accuracy of logistic regression accuracy results was shown to be 70.0% in rise of stock price, 78.8% in fall of stock price, and 74.6% on average. This study first analyzed relation between news and stock price through analyzing and quantifying sensitivity of atypical news contents by using opinion mining among big data analysis techniques, and furthermore, proposed and verified smart investment decision making model that could systematically carry out opinion mining and derive and support investment information. This shows that news can be used as variable to predict the price index of stocks for investment, and it is expected the model can be used as real investment support system if it is implemented as system and verified in the future.

The Monitoring on Plasticizers and Heavy Metals in Teabags (침출용 티백 포장재의 안전성에 관한 연구)

  • Eom, Mi-Ok;Kwak, In-Shin;Kang, Kil-Jin;Jeon, Dae-Hoon;Kim, Hyung-Il;Sung, Jun-Hyun;Choi, Hee-Jung;Lee, Young-Ja
    • Journal of Food Hygiene and Safety
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    • v.21 no.4
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    • pp.231-237
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    • 2006
  • Nowadays the teabag is worldwide used for various products including green tea, tea, coffee, etc. since it is convenient for use. In case of outer packaging printed, however, there is a possibility that the plasticizers which is used for improvement in adhesiveness of printing ink may shift to inner tea bag. In this study, in order to monitor residual levels of plasticizers in teabags, we have established the simultaneous analysis method of 9 phthalates and 7 adipates plasticizers using gas chromatography (GC). These compounds were also confirmed using gas chromatography-mass spectrometry (GC-MSD). The recoveries of plasticizers analyzed by GC ranged from 82.7% to 104.6% with coefficient of variation of $0.6\sim2.7%$ and the correlation coefficients of each plasticizer was $0.9991\sim0.9999$. Therefore this simultaneous analysis method was showed excellent reproducibility and linearity. And limit of detection (LOD) and limit of quantitation (LOQ) on individual plasticizer were $0.1\sim3.5\;ppm\;and\;0.3\sim11.5\;ppm$ respectively. When 143 commercial products of teabag were monitored, no plasticizers analysed were detected in filter of teabag products. The migration into $95^{\circ}C$ water as food was also examined and the 16 plasticizers are not detected. In addition we carried out analysis of heavy metals, lead (Pb), cadmium (Cd), arsenic (As) and aluminum (Al) in teabag filters using ICP/AES. $Trace\sim23{\mu}g$ Pb per teabag and $0.6\sim1718{\mu}g$ Al per teabag were detected in materials of samples and Cd and As are detected less than LOQ (0.05 ppm). The migration levels of Pb and Al from teabag filter to $95^{\circ}C$ water were upto $11.5{\mu}g\;and\;20.8{\mu}g$ per teabag, respectively and Cd and As were not detected in exudate water of all samples. Collectively, these results suggest that there is no safety concern from using teabag filter.

Multiplication of Infectious Flacherie and Densonucleosis Viruses in the Silkworm, Bombyx mori (가잠의 전염성 연화병 및 농핵병 바이러스 증식에 관한 연구)

  • 김근영;강석권
    • Journal of Sericultural and Entomological Science
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    • v.25 no.2
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    • pp.1-31
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    • 1984
  • Flacherie, as one of the most prevalent silkworm diseases, causes severe economic damage to sericultural industry and its pathogens have been proved to be flacherie virus (FV) and densonucleosis virus (DNV). Multiplications of the viruses in the larvae of the silkworm, Bombyx mori, were studied by the sucrose density gradient centrifugation and electron microscopy. The quantitative and qualitative changes of nucleic acids and proteins were investigated from the midgut and hemolymph in the silkworm larvae infected separately with FV and DNV. The histopathological changes of epithelial cells of infected midgut also were examined by an electron microscope. 1. Purified fractions of FV or DNV in a sucrose density gradient centrifugation yielded one homogenous and sharp peak without a shoulder, suggesting no heterogenous materials in the preparation. Electron microscopy also revealed that FV and DNV were spherical particles, 27nm and 21nm in diameter, respectively. 2. Silkworm larvae showed a decrease in body weight on the 6th day and in midgut weight on the 3rd day after inoculation with FV or DNV. 3. DNA content was higher in the midgut when infected with FV or DNV, but the hemolymph of the infected larvae showed no difference during first 6 days after inoculation, after which DNA concentration declined rapidly. 4. RNA synthesis of silkworm larvae infected separately with FV and DNV was stimulated in the midgut, but RNA content was reduced in the hemolymph at the early stage of virus multiplication. At the late stage of virus multiplication, however, it was extremely reduced in both midgut and hemolymph. 5. The concentration of protein in the midgut and hemolymph of silkworm larvae infected separately with FV and DNV showed no difference from that of the healthy larvae at the early stage of virus multiplication, but it was significantly reduced at the late stage of virus multiplication. 6. There was no difference in the electrophoretic patterns of RNAs extracted from the midgut of healthy or virus-infected larvae. 7. The electrophoresis of proteins extracted from the midgut infected with FV or DNV, when carried out on the 1st and 5th day after virus inoculation, showed no difference from that of the healthy larvae. But, there was an additional band with medium motility in the proteins on the 8th day after virus inoculation, while a band with low mobility shown in the proteins of healthy larvae disappeared in the infected larvae. However, a band with high mobility in the healthy larvae was separated into two fractions in the infected larvae. 8. The electrophoretic pattern of hemolymph proteins of the silkworm larvae infected separately with FV and DNV was similar to that of the healthy larvae, but the concentration of hemolymph proteins in the infected larvae was lower than that of the healthy larvae at the late stage. 9. Two types of inclusion bodies were shown by the double staining of pyronin-methyl green in the columnar cell of the midgut on the 8th day after FV inoculation. 10. Electron microscopy of the infected midgut revealed that the 'cytoplasmic wall' of the goblet cell thickened on the 5th day after FV inoculation and several types of the cytopathogenic structures, such as virus$.$specific vesicles, virus particles, linear structures, tubular structures, and high electron-dense matrices were observed in the cytoplasm of the goblet cell. The virus particles were also observed in the microvilli and the structures similar to spherical virus particles were observed around the virus-specific vesicles, suggesting the virus assembly in the cytoplasm. 11. Fluorescence micrograph of the infected midgut stained with acridine orange showed that the nucleus, the site of DNV multiplication in the columnar cell, enlarged on the 5th day after virus inoculation. 12. Electron microscopic examination of DNV infected midgut revealed that the nucleolus of the columnar cell was broken into granules and those granules dispersed into apical region of the nucleus on the 5th day after virus inoculation. On the 8th day after inoculation, it was also observed that the nucleus of the columnar cell was full with the high electron-dense virogenic stroma which were similar to virus particles. These facts suggest that the virogenic stroma were the sites of virus assembly in the process of DNV multiplication.

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