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Simultaneous Detection of Major Pathogens Causing Bovine Diarrhea by Multiplex Real-time PCR Panel (Multiplex real-time PCR을 이용한 송아지 설사병 원인 주요 병원체의 동시검출)

  • Kim, Won-Il;Cho, Yong-Il;Kang, Seog-Jin;Hur, Tai-Young;Jung, Young-Hun;Kim, Nam-Soo
    • Journal of Veterinary Clinics
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    • v.29 no.5
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    • pp.377-383
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    • 2012
  • Bovine diarrhea is a major economical burden to the bovine industry in Korea. Since multiple infectious agents can be involved in bovine diarrhea, differential diagnosis is essential for effective treatment. Therefore, a panel of two multiplex real-time PCR assays which can simultaneously detect six major bovine enteric pathogens [i.e., bovine viral diarrhea virus (BVDV), bovine coronavirus (BCoV), group A bovine rotavirus (BRV), Salmonella spp., Escherichia coli (E. coli) $K99^+$, and Cryptosporidium parvum] was developed and applied to test 97 fecal samples collected from cattle farms in Korea. In addition, microscopic examination was also preformed on the samples to detect Coccidium oocyst. The estimated sensitivity of the multiplex PCR was 0.1 $TCID_{50}$ for BVDV, BCoV and group A BRV, 5 and 0.5 CFU for E. coli $K99^+$ and Salmonella, respectively, and 50 oocysts for Cryptosporidium. The amplification efficiency of the multiplex PCR ranged between 0.97 and 0.99 for each pathogen. Among 97 samples, 36 samples were positive for at least one of the 6 major pathogens and 6 samples were simultaneously positive for 2 pathogens by the multiplex PCR assay. Coccidium oocysts were also detected in 48 samples, which were all collected from over 1 month old calves. In conclusion, the multiplex real-time PCR panel can be a useful tool for fast and accurate diagnosis of calf diarrhea associated with BVDV, BCoV, group A BRV, E. coli $K99^+$, Salmonella, and/or Cryptosporidium and Coccidium may be an important target which needs to be included in the multiplex PCR panel in the future.

Potentials of Synbiotics for Pediatric Nutrition and Baby Food Applications: A Review (소아 영양 및 유아식 응용을 위한 신바이오틱스의 잠재력: 총설)

  • Jung, Hoo Kil;Kim, Sun Jin;Seok, Min Jeong;Cha, Hyun Ah;Yoon, Seul Ki;Lee, Nah Hyun;Kang, Kyung Jin
    • Journal of Dairy Science and Biotechnology
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    • v.33 no.2
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    • pp.111-118
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    • 2015
  • Probiotic, prebiotic, and synbiotic substances as well as microorganisms were added to infant formula in an attempt to influence the intestinal microflora with an aim to stimulate the growth of lactic acid bacteria, especially bifidobacteria and lactobacilli. Over the last 10 years, new synbiotic infant formulas containing probiotics and prebiotics have been proposed in order to simulate the effect of breast-feeding on the intestinal microflora. Owing to their synergistic effect, the new synbiotics are expected to be more helpful than using probiotics and prebiotics individually. Maintenance of the viability of the probiotics during food processing and the passage through the gastrointestinal tract should be the most important consideration, since a sufficient number of bacteria ($10^8cfu/g$) should reach the intended location to have a positive effect on the host. Storage conditions and the processing technology used for the manufacture of products such as infant formula adversely affect the viability of the probiotics. When an appropriate and cost-effective microencapsulation methodology using the generally recognized as safe (GRAS) status and substances with high biological value are developed, the quality of infant formulas would improve. The effect of probiotics may be called a double-effect, where one is an immunomodulatory effect, induced by live probiotics that advantageously alter the gastrointestinal microflora, and the other comprises anti-inflammatory responses elicited by dead cells. At present, a new terminology is required to define the dead microorganisms or crude microbial fractions that positively affect health. The term "paraprobiotics" (or ghost probiotics) has been proposed to define dead microbial cells (not damaged or broken) or crude cell extracts (i.e., cell extracts with complex chemical composition) that are beneficial to humans and animals when a sufficient amount is orally or topically administered. The fecal microflora of bottle-fed infants is altered when the milk-based infant formula is supplemented with probiotics or prebiotics. Thus, by increasing the proportion of beneficial bacteria such as bifidobacteria and lactobacilli, prebiotics modify the fecal microbial composition and accordingly regulate the activity of the immune system. Therefore, considerable attention has been focused on the improvement of infant formula quality such that its beneficial effects are comparable to those of human milk, using prebiotics such as inulin and oligosaccharides and potential specific probiotics such as bifidobacteria, which selectively stimulate the proliferation of beneficial bacteria in the microflora and the indigenous intestinal metabolic activity of the microflora.

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The Effect of Seasoning on the Intestinal Absorption -Absorption by Passive Transport and the Effect of Red Pepper- (조미료가 창자 운동과 흡수기능에 미치는 영향 -소장의 피동적 흡수에 대한 고추의 영향-)

  • Shin, Dong-Hoon;Kim, Joong-Soo;Koh, Jae-Pyong;Ahn, Seung-Woon
    • The Korean Journal of Physiology
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    • v.7 no.1
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    • pp.23-31
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    • 1973
  • Numerous factors concern with the absorption of substances through the membrane of the gastrointestinal tract. To simplify the experimental condition, present work has been restricted to observe the disappearance rate of substance from the intestinal loop which was made in the jejunum, 70 cm apart from the pylorus of the adult rabbit. The purpose of the study is to clarify the absorption of urea through the jejunal wall is solely attributable to the concentration difference between the luminal fluid and plasma, and to observe the effect of adding red pepper upon the rate of absorption. The rabbits were anesthetized with nembutal, 35mg/kg I.V. Jejunal loop was made by ligating at 2 spots, 70 cm and 80cm apart from the pylorus. After rinsing with normal saline solution through the polyethylene tubing inserted from the end of the loop, 8 ml of test solution was placed through the same tubing. The test solution contained 200 mg% of urea and 150mg% of polyethylene glycol(M.W. 4,000) in normal saline solution. Right after placing the test solution the first specimen was taken through the tubing, and successive samplings were performed at 5, 10, 20, and 30 minutes. Logarithm of the difference of urea concentration between the luminal fluid and plasma was plotted against time elapsed after the onset of the experiment. If straight line is revealed, it would verify the nature of transport mechanism as diffusion, obeying the Fick's principle. The concentration of polyethylene glycol (PEG) was also measured in order to examine the change in the volume. PEG was used as the marker substance because it is not absorbable in the intestinal tract. Consequently the concentration of PEG relates inversely to the volume of the loop. Instantaneous concentration of urea in the loop times the volume will give the amount of urea remaining in the luminal fluid. The change in the amount of any substance is directly relate to the volume of the compartment and differs from the change in the concentration which is independent of the volume. After completion of the experiment without red pepper, it was added in the test solution and was centrifuged after thorough mixing. Supernatant of the mixture was placed in the loop and similar sampling were performed with the same time intervals that of previous run in order to observe the effects of the red pepper on the passive transport of the water soluble small substance, urea. The results obtained were as follows: 1. Logarithm of the concentration difference of urea between the luminal fluid and plasma was diminished exponentially as time elapsed. The decay constant in the experiment without red pepper was 0.0563/min. By adding red pepper in the test solution as much as the concentration rose to 4,000 mg% and 8,000 mg%, the decay constants were lowered to 0.0493/min and to 0.0506/min, respectively. The time interval by which the concentration difference dropped to one half of the initial value was prolonged. Without red pepper the half concentration time was 13.30 minutes, and by adding extract of red pepper, 15.31 minutes and 15.71 minutes were revealed. 2. The profile of the diminishing rate of tile amount of urea was quite different from that of the concentration because of the change in the volume of the loop during the observed period. 3. By adding the extract of red pepper, it slowed down the rate of absorption of urea in the intestinal loop, suggesting an increase in the diffusional barrier. 4. Larger dosage of red pepper brought an increase in the secretion of intestinal fluid with concomitant expansion of the luminal volume, and the retardation of the absorption of urea was noticed. This effect was largely dependent on the sensitivity of the individual animal to the red pepper, extract. The amount of urea remained after 10 minutes interval was 55.5% of the initial amount in the experiment without red pepper. On the other hand it was not consistent after administration of red pepper, showing 50.6% and 66.5% of the initial figures by adding 400 mg and 800 mg of red pepper in the test solution, respectively. It was postulated that symptom of diarrhea often encountered by taking a hot (red pepper) food might be attributable to the increase of secretion and the retardation of absorption in the intestinal tract.

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Study on Features of Software Cyborg in the Virtual Game -PS4 ocusing Game- (가상게임에 나타나는 소프트웨어 사이보그특징에 대한 고찰 -PS4 <언틸던> 게임을 중심으로-)

  • Kim, Dae-Woo
    • Cartoon and Animation Studies
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    • s.41
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    • pp.279-306
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    • 2015
  • This paper is a study of the changing nature of software for virtual Cyborg self and the virtual body that occur in the game from a philosophical point of view. Looking broadly, the cyborg concept refers to the combination of man and machine. Specifically, there is a hardware cyborg organism to combine human and restoration of machine In addition, there is software cyborg by electronic the human brain of converting a virtual body. Virtual games are cases software-Cyborg applied. In the game , There seems to have characteristics of virtual body and ego that different from general cyborg meaning. To analyze the features, I applied the concept of software-cyborg of Hans morabek and the multiple selves in cyberspace properties of Kim Sun-Hee. generally, software cyborg cloning the brain type tended to invalidate the body due to the nature of the virtual world. But If you look at third-person's view and the game character that made from real actors, it is pursuing the realism of photographic images and it stressed the need for a virtual body in order to maintain the psychological identity of the player. And, The game player crosses the eight characters to choose while completing the mission. This is a big role in the reality ego leads to the desired final ending with the selection and experience to be experienced as self-replication to multiple. These cyber multi-ego looks for an active and positive features compared to the multi-ego in the real world and highlights the advantages of the software cyborg. Game The characteristics of the final result varies depending on the selection of the player. The life and death of a friend is determined by the relationship between the characters friendship. In this case, the virtual self is empirically through trial and error, moral, and try to select the desired setting the standard for intuitive and self own choice. Also It can be fused to the knowledge of multiple selves as one step is formed by a high spiritual introspection. This process is a positive interpretation of the world and their own forms of mental reflection through self-overcoming human, Nietzsche is said that the process is Wibeomenswi.

A Study on Sea Water and Ocean Current in the Sea Adjacent to Korea Peninsula -III. Chemical Characteristics of Water Masses in the Polar Front Area of the Central Korean East Sea- (한반도 근해의 해류와 해수특성 -III. 한국 동해 중부 극전선역에 출현하는 수괴의 화학적 특성-)

  • YANG Han-Soeb;KIM Seong-Soo;KANG Chang-Geun;CHO Kyu-Dae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.24 no.3
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    • pp.185-192
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    • 1991
  • The vertical distribution and chemical characteristics of water masses were measured along two south-north transects in the polar front region of the central Korean East Sea. In February, a thermocline was present at depth between 50m and loom at the southern sites of a landward A-transect, and its depth was gradually deepened northward. At an outside B-transect, a thermocline was observed at significantly deep depth of 300m to 400m at two northern stations(Stn. 10 and 11), though the depth of the southward stations was nearly identical to that at the northward stations on a A-transect. In September, there were vertically more various water masses, i.e. the Tsushima Warm surface water(TWSW) or more than $20^{\circ}C$, the Tsushima Middle water(TMW) with a range of $12{\~}17^{\circ}C$, the North Korea Cold Water(NKCW) with $1{\~}7^{\circ}C$ temperature, the Japan Sea Proper Water(JSPW) of less than $1^{\circ}C$, and the mixed water. The North Korea Cold Water could be distinguishable from the other waters, especially from the mixed water of the Tsushima Middle Water and the Japan Sea Proper Water by the pattern of $T-O_2$ diagram. For instance, the North Korea Cold Water had higher oxygen by $1{\~}2ml/l$ than those in the mixed water, although both the two water masses ranged $1{\~}7^{\circ}C$ in water temperature. AOU value was the highest in the JSPW and the lowest in the TWSW. Also, AOU indicated a nearly linear and negative correlation with water temperature. However, AOU data for two masses, the NKCW and the TMW, in September departed remarkably from a regression line. Moreover, the ratio of $$\Delta P/\Delta AOU)$ in September was about $0.45{\mu}g-at/ml$ and higher than the value observed in the open sea. This high value could be elucidated by two factors; intrusion of the NKCW with high oxygen and molecular diffusion of dissolved oxygen from the surface into the lower layer. AOU would be a useful tracer for water masses in the polar front area of the Korean East Sea.

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A Study on the Actual Condition of the Adult-smoking in a Region (일 지역 성인의 흡연실태)

  • Chong Young-sook
    • Journal of Korean Public Health Nursing
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    • v.13 no.1
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    • pp.97-113
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    • 1999
  • To investigate the actual condition of the adult-smoking in Chinan County. I used self-reporting questionnaires among 923 residents living in nine districts selected at random among the sites of eleven eup-myons from December 28. 1998 to January 6. 1999. collected data and analyized using SPSS. The smoking rate of adults in Chinan County was $36.9\%$. There was a significant difference in smoking rate according to the age. gender, education and job among general characteristics. The smoking rate in group of above 40s was about $40\%$ and that of the male took $55.5\%$ which was higher than that of female and the smoking rate of the people having high-school education was the highest$(46.2\%)$. With regard to the career. the smoking rate of the farmers ranked first$(46.1\%)$. Therefore anti-smoking business for smokers should be focused on above 40 years old, males. people having high school education and farmers. As the result of the survey of smoking habits of 340 people who smoke currently, the average smoking begining age was 21.8 and $81.5\%$ among them was between 16-25 years old. $4.4\%$ was under 15. during around the elementary school. Most motives to smoke were as followed; curiosity or taste$(33.6\%)$, relief of stress $(31.2\%)$, peer presure$(26.5\%)$ And considering the amount of the cigarette which is smoked per day, the case which is less than a pack of cigarettes was highest as $75.5\%$ and the people who smoke over one pack of cigarettes took $24.5\%$. As for the kind of tobacco which is smoked, how to smoke and the desire for the smoking, most case was toxin was moderate$(47.8\%)$ or mild$(46.9\%)$. shallowly$(49.0\%)$ or deeply$(46.3\%)$ and under stress$(33.4\%)$. after meals$(27.8\%)$, during drink$(15.7\%)$ and so on. The highest point marked among the factors of smoking motives was 'the reduce of negative emotion' $(3.27\pm1.00)$. followed by 'uncomfortable habits' $(2.87\pm1.02)$, 'addiction' $(2.84\pm1.06)$. 'habit' $(2.74\pm1.12)$. 'pleasure' $(2.70\pm1.04)$. 'stimulus' $(2.59\pm.90)$, 'sensation-exercise satisfaction' $(2.42\pm.97)$. Smokers smoke to reduce the negative emotions when angry in most common case. depressed. anxious. uncomfortable. lone. ashamed or embarrased. and intend to solve the certain problem. etc. Other motives are uncomfortable habit. addiction. habit. pleasure and the pursuit of stimulus. The level of nicotine dependence of adults m Chinan County was 10.57 which amount to 'high' wholly. As the resulf of the level of nicotine dependence score. the people who are low in the level of nicotine was $33.5\%$. the people who are high was $48.2\%$. very high was $18.4\%$. The approach for anti-smoking for smokers should be conducted differently according to the level of the nicotine. For the people who are in low level of nicotin dependence the prohibition of the smoking should be guided through the approach to foster strong will. for those who are in 'high' by acquiring proper method for the prohibition of smoking. and for those who are 'very high' the anti-smoking should be induced by providing proper program because of the possibility of the suffer from abstinence syndrome. The difference of the level of nicotine with the general characteristics of the objects had not statistically significant difference. The difference of the level of nicotine dependence accompanied by smoking habit had statistically significant difference according to the amount of smoke, the kind of tobacos. smoke inhale habit. In other words, the group of heavy smokers had higher level of the nicotine dependence than that of the light smokers relatively and the group which smoke strong taboaco has higher level of nicotine than that of which smoke mild or moderate. And the group of smokers who smoke deeply has higher level of nicotine than that who smoke shallowly or nonswallow. Aa a result of the analysis of the correlation between smoking motive factors and the level of nicotine, there was the indication that people who smoke for the decrease of the negative emotion. habit, pleasure. stimulus. sensation-exercise satisfaction had high level of the nicotine dependence. As the result of the anti-smoking will of smokers. $65.0\%$ of them had prohibition of smoking will. $29.3\%$ had no will to quit smoke. The most important reason for anti-smoking was health. $67.9\%$ had experience to try to quit smoke and the biggest reason to fail to quit smoking was the lack of the will power to keep anti-smoking. $52.8\%$ of them were advised to stop smoking from their spouses or children. only $2.8\%$ were by medical. The people who have the opinion to need anti-smoking education were $69.6\%$. Therefore when the business for the hygine of the mouth for adult is set. it should be centered on the people who have intention of prohibition of the smoking and help to quit smoking by way of other affirmative counter-program not smoking under stress.

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A study on the present condition of family planning and the status of health and nutrition in Wido islanders (낙도주민(落島住民)의 가족계획(家族計劃) 현황(現況)과 건강(健康) 및 영양실태(營養實態)에 관(關)한 연구(硏究) (I))

  • Lee, Geum-Yeong;Kong, Yeong-Ja
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.8 no.1
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    • pp.51-62
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    • 1979
  • Due to the relatively high number of members and low income in each family, Wido inhabitants take very low nutrition. The result indicates that nutrition at the present time is generally lower than that of 1973 with the exception of fat. Consequently, the degree of body development of primary and junior high school students has not generally reached the standard in Korea, that was collected from Gyeongido, Korea and from Japan, of the corresponding grade students as well as the Hb content is significantly insufficient. As the body development of 12-year-old children grown at home and nursery, and attending the sports school are compared with that of the Korean standard and Wido students, the body development of Wido students is similar to the Korean standard, and slighly better than that of the sports school students by three years. Health is important as said that health power is nation's power. In order to maintain good health, the normal level of body development and Hb content is necessary for which the recommended daily nutrition for each person should be taken. In addition, the present status and importance of body development and nutrition should be fully educated the primary and junior high school students as well as directly to the inhabitants.

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Studies on the Control of the Pine Bark Beetle (Myelophilus pinipedera LINNE) (I) -Analysis of the carotenoids in important pine species in Korea- (소나무좀 방제(防除)에 관한 연구(硏究)(제1보(第1報)) -한국산(韓國産) 중요(重要)소나무류(類) 잎에 함유(含有)되어 있는 Carotenoid 분석(分析)-)

  • Kim, Young Ho
    • Journal of Korean Society of Forest Science
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    • v.27 no.1
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    • pp.15-21
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    • 1975
  • According to Yoshihisa Kuroki's report that the total amount of carotenoid was more in the susceptible to pine bark beetle, than in the resistant ones, carotenoids were extracted from needle leaves of one year old seedlings of Pinus thunbergii, Pinus desiflora, Pinus rigida, Pinus koraiensis, Pinus rigitaeda and Pinus taeda which are all important pines in Korea, to find their resistant ratio to the insect. The carotenoids were analyzed and compared using the spectra of them by spectrophotometer. The results were as follows: 1. The visible absorption spectra of carotenoids in those pine trees were proved to be very similar. 2. The total amount of carotenoids in needles differed with the tree species and the contents were arranged in decreasing order P. koraiensis>P. rigida>P. thunbergii>P. rigitaeda>P. taeda, it seemed that there was corelation between the cartenoid content and the extent of harm caused by the pine bark beetles except P. koraiensis. 3. But carotenoids were contained in Robinia pseudoacacia, Castanea crenata, Chamaecyparis obtusa and Cedrus deodra leaves too. 4. The total amounts of carotenoids in these pines of 9 species were arranged in decreasing order Robinia pseudoacacia>Pinus koraiensis>Pinus rigida>Pinus thunbergii>Castanea crenata>Pinus rigitaeda>Pinus taeda>Chamaecyparis obtusa>Cedrus deodara. Therefore, it was proved that there was no correlation between carotenoid cotent and extent of resistance to the insect. 5. In the thin-layer chromatography of these carotenoids, 13 kinds of components in P. densiftora, P. koraiensis and P. rigida and 12 kinds of spots in other pines, were detected respectively, under ultra-violet fluorescent lamp $3,600{\AA}$ and $2,537{\AA}$. 6. The eighth spots from the bottom in P. densiflora, P. koraiensis and P. rigida were not found in other pines and other 4 species (Robinia pseudoacacia, Castanea crenata, Chamacyparis obtusa, Cedrus deodra). Especially the spot in P. densiflora fluoresced strong cobalt blue-fluorescence under ultra-violet fluorescent lamp $2,537{\AA}$.

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Effect of Natural Plant Mixtures on Behavioral Profiles and Antioxidants Status in SD Rats (자생식물 혼합 추출물이 SD 흰쥐에서의 행동양상 및 항산화 체계에 미치는 영향)

  • Seo, Bo-Young;Kim, Min-Jung;Kim, Hyun-Su;Park, Hae-Ryong;Lee, Seung-Cheol;Park, Eun-Ju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.9
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    • pp.1208-1214
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    • 2011
  • Caffeine, a psychoactive stimulant, has been implicated in the modulation of learning and memory functions due to its action as a non-selective adenosine receptors antagonist. On the contrary, some side effects of caffeine have been reported, such as an increased energy loss and metabolic rate, decrease DNA synthesis in the spleen, and increased oxidative damage to exerted on LDL particles. Therefore, the aim of this study was to develop a safe stimulant from natural plants mixture (Aralia elata, Acori graminei Rhizoma, Chrysanthemum, Dandleion, Guarana, Shepherd's purse) that can be used as a substitute for caffeine. Thirty SD rats were divided into three groups; control group, caffeine group (15.0 mg/kg, i.p.), and natural plants mixture group (NP, 1 mL/kg, p.o.). The effect of NP extract on stimulant activity was evaluated with open-field test (OFT) and plus maze test for measurement of behavioral profiles. Plasma lipid profiles, lipid peroxidation in LDL (conjugated dienes), total antioxidant capacity (TRAP) and DNA damage in white blood, liver, and brain cells were measured. In the OFT, immobility time was increased significantly by acute (once) and chronic (3 weeks) supplementation of NP and showed a similar effect to caffeine treatment. Three weeks of caffeine treatment caused plasma lipid peroxidation and DNA damage in liver cells, whereas there were no changes in the NP group. NP group showed a higher plasma HDL cholesterol concentration compared to the caffeine group. The results indicate that the natural plants mixture had a stimulant effect without inducing oxidative stress.

The Analysis of Management and the Method of Cultivation of Lentinus edodes I. for Full-Development of Mycelium in Bed Logs (표고재배(栽培)의 관리분석(管理分析)과 종균활착(種菌活着)을 위한 골목관리(管理)에 관(關)한 연구(硏究))

  • Joo, Myoung Chil
    • Journal of Korean Society of Forest Science
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    • v.85 no.4
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    • pp.596-604
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    • 1996
  • This study was carried out to offer the successive method of cultivation and increase the productivity of mushroom yield with good quality through the elevation of rate of spawn development for Lentinus edodes. Studied about the analysis of current management of actural cultivation, a base of these, researched and presented for the upward method of productivity through an experiment of the high rate of spawn development and cultivation, putting first cultural environment. The results obtained were as follows ; 1. As the result of the analysis of current management in actural cultivation, many cultivators had a tendency to neglect managements of cultivation. These were reason for the deficiency of labour, funds and the lack of knowledge of cultivation, etc. 2. Water contents in bed logs according to the date of inoculation was shown as the decreasing order of 28.63%(3/12), 25.20%(3/25) and 23.19%(4/10). For the purpose of the maintenance of the water contents, the full-development of mycelium in bed logs and the dispersion of labour, the date of inoculation should be started in the early March. 3. The difference of the rate of spawn development among species was not shown, 100%(Mori 465). 98.98%(Mori 3046) on the spawn in high temperature and 98.97%(Mori 290) on the spawn in low temperature. The relative rate of spawn development was 97.70%(Mori 465), 82.45%(Mori 3046) on the spawn in high temperature and 88.87%(Mori 290) on the spawn in low temperature, it showed the difference. The spawn should be selected carefully in the future, as the spawn of cultivater's preference showed the difference for the development of mycelium. 4. The rate of spawn development following the date of inoculation was 100.0%(3/12), 98.98%(3/25) and 96.79%(4/10) on the spawn in high temperature and 99.09%(3/12), 98.97%(3/25) and 97.89% (4/10) in low temperature, it showed little difference. And the relative rate of spawn development was 97.70%(3/12), 82.45%(3/25) and 81.42%(4/10) on the spawn in high temperature and 93.27%(3/12), 89.67%(3/25) and 88.87%(4/10) that in low temperature, As the result of the relative rate, the time of inoculation of spawn should begin in the early March. 5. The height of stock logs on temporary placing should be less than 60cm at most on the surface, because of the low rate of water contents.

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