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Intra-laboratory Validation of an HPLC Post-column Oxidation Method for the Analysis of PSP Toxins in Oysters and Mussels (굴과 진주담치 중 마비성 패류독소 분석을 위한 HPLC post-column oxidation method의 시험소 내 유효성 검증)

  • Song, Ki Cheol;Lee, Ka-Jeong;Yu, Hong-Sik;Mok, Jong-Soo;Kim, Ji Hoe;Lim, Keun-Sik;Lee, Mi-Ae;Kim, Mee-Hye
    • Korean Journal of Food Science and Technology
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    • v.45 no.2
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    • pp.241-247
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    • 2013
  • AOAC Mouse Bioassay Analysis (MBA) has been the gold standard for the analysis of paralytic shellfish poisoning toxin (PSP toxin) for more than 50 years. However, this method has inaccurate limit of quantification and cannot be used to determine toxic profiles. An HPLC method (PCOX) was optimized for Korean shellfish to establish an alternative or supplementary method for PSP analysis and was intended to be used for the official monitoring and regulation of food. The recovery rate of the PCOX method was 83.5-112.1% and the limit of quantification for total toxin was about $8.6{\mu}g$/100 g. A long-term comparison study showed a good correlation of the PCOX results with the AOAC MBA results: the correlation factors were 0.9534 and 0.9109 for oyster and mussel matrices, respectively. The PCOX method may be used as an alternative or supplementary method for AOAC MBA to monitor the occurrence of PSP and to analyze PSP toxin profile in oysters and mussels.

Development of Multiplex PCR Assay for Identification of Eight Species from Meats in Korea (국내에서 유통되는 8종의 식육감별을 위한 multiplex PCR법 개발)

  • Heo, Eun-Jeong;Ko, Eun-Kyung;Yoon, Hyang-Jin;Kim, Yeon-Hwa;Kim, Young-Jo;Park, Hyun-Jung;Wee, Sung-Hwan;Moon, Jin-San
    • Journal of Food Hygiene and Safety
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    • v.31 no.1
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    • pp.28-35
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    • 2016
  • Species identification of animal tissues in meat products is an important issue to protect the consumer from illegal and/or undesirable adulteration; for economic, religious and health reasons. In this reason, accurate analytical methods are needed for the labeling of meat products with requiring simple and fast procedure. Recently, applications of PCR in food analysis have been increased because of their simplicity, specificity and sensitivity. Therefore, in this study, a multiplex PCR assay was developed for the simultaneous identification of eight species of cow, pig, chicken, duck, goat, sheep, horse and turkey from raw meats. The primers were designed in different regions of mitochondrial 16S RNA after alignment of the available sequences in the GenBank database. Two multiplex primer sets were designed as Set 1 (cow, pig, chicken, duck) and Set 2 (goat, sheep, horse, turkey), respectively. Total 274 samples from cow (n = 55), pig (n = 30), chicken (n=30), and duck (n = 30), goat (n = 40), sheep (n = 33), horse (n = 41), and turkey (n = 15) were tested. The primers generated specific fragments of 94, 192, 279, 477 bp (pig, chicken, cow, duck), 670, 271, 152, 469 bp (goat, sheep, horse, turkey) lengths for eight species, respectively. The animal species specificity was 100% in all eight samples in the multiplex PCR assay. The detection limit of the multiplex PCR assay showed from 100 fg to 1 pg of template DNA from extracted from raw meats. When applying multiplex PCR assays to sample from pork/beef and pork/chicken, beef/chicken tested raw mixed meats and heat-treated ($83^{\circ}C$ for 30min, $100^{\circ}C$ for 20min, and $121^{\circ}C$ for 10min) mixtures, detection limit was 0.1% level beef, pork and pork in beef and chicken in pork and 1.0% level pork in chicken. This study suggest that the developed multiplex PCR assay can be used for rapid and simultaneous species identification of cow, pig, chicken, duck, goat, sheep, horse and turkey from meats.

A Study on the Botanical Garden Visitors' Eco-consciousness and Motivation (식물원 이용객의 생태의식과 이용동기에 관한 연구)

  • Jeong, Jae-Man
    • Korean Journal of Environment and Ecology
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    • v.28 no.2
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    • pp.235-246
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    • 2014
  • The purpose of this study was to determine the correlation between botanical garden visitors' ecological consciousness and their needs, in order to provide some effective measures to manage them. For this purpose, 3 study points were set up: "botanical garden visitors' ecological consciousness and their needs", "differences of such consciousness depending on their demographic variables" and the "relationship between such consciousness and their needs". To this end, Botanical garden visitors were surveyed for an empirical analysis. The visitors' awareness about ecology was measured with Dunlap's 15-item NEP Inventory, while their needs were analyzed in reference to Maslow's 7-Step Human Desire Ladder. The survey was conducted at Botanical garden for 3 days. As a result, a total of 360 questionnaires were returned. The results of this study can be summarized as follows; First, the visitors' ecological consciousness and their needs were higher than normal level. In terms of their consciousness of ecology, their awareness of the ecological crisis potential and anti-humanism were the highest. In terms of their needs, the aesthetic need was the highest, followed by the cognitive need. On the other hand, the needs for self-achievement and self-esteem were the lowest; except them, the higher the needs were positioned at Maslow's ladder of desire, the more responsive the subjects became. As a result of analyzing the correlation between the subjects' consciousness of ecology and their needs, it was found that the correlation was negative in some sub-areas, while being positive in other sub-areas. After all, the ratio of the sub-areas having a positive correlation was 3 times higher than that of the sub-areas having a negative correlation. Even as for the correlation coefficient values, they were higher in the positive sub-areas, which suggests that the correlation between wetland visitors' ecological consciousness and their needs was positive, although at a lower level, in overall terms. As a result of comparatively analyzing visitors' needs by dividing them into 3 sub-groups depending on the levels of their ecological consciousness, it was found that the higher their consciousness of ecology was, the higher their needs were. Overall, botanical garden visitors' ecological awareness was higher than the normal level, and it was estimated that such awareness would continue to increase. Hence, it could be inferred that their needs, particularly their aesthetic and cognitive ones, would also continue to increase. Accordingly, it is important to manage the wetland landscape making use of its visual resources, while keep providing the visitors with the contents fulfilling their need for knowledge.

The Change of Peripheral Eosinophil Count after Bronchial Provocation with Inhaled Histamine in Bronchial Asthmatics (기관지 천식 환자에서 히스타민 기관지유발검사후 말초혈액 호산구수의 변화)

  • Kim, Chi-Hong;Kim, Young-Kyoon;Kwon, Soon-Seog;Kim, Kwan-Hyoung;Han, Ki-Don;Moon, Hwa-Sik;Song, Jeong-Sup;Park, Sung-Hak
    • Tuberculosis and Respiratory Diseases
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    • v.39 no.5
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    • pp.386-391
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    • 1992
  • Background: Recently, bronchial provocation of the airway of atopic asthmatic subjects with inhaled allergen has been shown to produce an initial peripheral blood eosinopenia followed by an eosinophilia occurring approximately 12 to 18 hrs after the challenge. However there are few studies about the change of peripheral eosinophil count (PEC) after bronchial provocation with nonspecific stimuli such as histamine or methacholine. Interestingly our preliminary study demonstrated a notable change of PEC during bronhial provocation with inhaled histamine in some asthmatic subjects. This study was designed to reevaluate our preliminary data and to further investigate the change of PEC during as well as after bronchial provocation with inhaled histamine in bronchial asthma tics. Methods: Sixteen asthmatic subjects participated in this study. Bronchial provocation with inhaled histamine was done between 9 AM and 12 MD. Blood samplings for PEC were done with 5 minutes intervals during the procedure, and repeated at 1 hour, 2 hours, 4 hours, 8 hours, 24 hours, and 48 hours after the procedure. Results: The results were as follows; 1) The patients were divided into two groups characterized by each pattern in the change of PEC during the procedure. A group (11 of sixten, group I) showed an increasing pattern of PEC and another group (5 of sixteen, group II) showed a decreasing pattern of PEC during the procedure. 2) Group I demonstrated a tendency to maintain continuously higher level of PEC than the baseline value until 48 hours after the procedure. 3) Group II demonstrated a tendency to maintain continuously lower level of PEC than the baseline value until 48 hours after the procedure. 4) There were no significant differences in their clinical parameters including baseline eosinophil count, baseline $FEV_1$, $PC_{20}$ of histamine, and serum IgE level between group I and group II. Conclusion: Our results suggest that the change of PEC produced by inhaled histamine in asthmatic subjects is much different from that produced by inhaled allergen, and that each patient may have their individual characteristics in the change of PEC in response to bronchial provocation with inhaled histamine. Alternatively these findings suggest that eosinophils may be partially involved in the early asthmatic reaction.

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Effect of Inoculation with Vesicular-Arbuscular Mycorrhizal (VAM) Fungi on the Early Growth of Strawberry Plantlets(Fragaria grandiflora Ehrn.) (딸기 묘(苗) 초기생육(初期生育)에 미치는 VA균근균(菌根菌)의 접종효과(接種效果))

  • Sohn, Bo-Kyoon;Huh, Sang-Man;Kim, Kil-Yong
    • Korean Journal of Soil Science and Fertilizer
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    • v.27 no.1
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    • pp.54-63
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    • 1994
  • Runner-derived(Expt.1) and tissue culture-derived strawbeery plantlets(Expt. 2) were grown in pots under greenhouse condition and inoculated with inocula of the vesicular-arbuscular mycorrhizal(VAM) fungi isolated from a field strawberry plants. Total biomass of mycorrhizal strawberry plants was significantly increased. There was a similar tendency in the number of cluster and flower at 20 weeks after inoculation, and VAM fungi inoculation positively influenced the leaf number, leaf length, leaf width and petiole length of strawberry plants in all investigated times. However, no difference was in the flowering time of strawberry plants. Leaf margin of non-inoculated strawberry plantlets turned into raddish brown(7.5R 4/8) from around 4 weeks after habituation. Inoculation of VAM fungi at the time of habituation was much more effective in stimulating plant growth. VA mycorrhizal dependency were 162.7 % in the runner-derived strawberry plants, Dependency with pre-and post-habituated incoulation in tissue culture-derived plants was respective 116.4% and 106.0%. The levels of mycorrhizal colonization were increased with plant growth and infection rates by endophytes at harvest time were 47.5% in Expt. 1, 56.4% in Expt. 2, respectively. Contents of phosphorus, potassium and calcium in mycorrhizal strawberry plants at harvest time were higher than non-mycorrhizal ones however, magnesium concentration was decreased. These experiments demonstrated that VAM fungi could be introduced into nursery stages of strawberry plantlets including the temporary planting period to improve growth and plant nutrients uptake by mycorrhizal plants.

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Limiting Nutrients of Cochlodinium polyklikoides Red Tide in Saryang Island Coast by Algal Growth Potential (AGP) Assay (조류성장잠재력 시험에 의한 사량도 연안 Cochlodinium polykrikoides 적조의 제한영양염)

  • KIM Hyung Chul;KIM Dong Myung;LEE Dae In;PARK Chung Kil;KIM Hak Gyoon
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.34 no.5
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    • pp.457-464
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    • 2001
  • Algal growth potential (AGP) assay using Cochlodinium polykrikoides was conducted in Saryang Island coast where C. polykrikoides red tide occurred annually from July to October 1998. The effects of macro- and micro-nutrients on the growth of C. polykrikoides were specifically evaluated by the algal assay method. Two different types of growth response of C. polykikoides for the addition of nutrients were clearly obseued. For both before and after C. polykrikoides occurrence, the growth of C. polykikoides was significantly stimulated by the addition of either nitrate or ammonium of $50{\mu]M$ with phosphate of $5{\mu}M$. The addition of a single nutrient had no clear effect on the growth of C. polyhikoides and the addition of trace metals, vitamins, and EDTA etc. did not stimulate the algal growth, also. This result indicates that both N and P potentially limited the growth of C. polyhikoides in this period. However, during a bloom of C. polyhikoides, the growth was unlikely to be stimulated by the addition of both macro- and micro-nutrients. At that time the nutrient concentration of Saryang Island coast was $24.33{\mu}M$ for ammonium, $1.61{\mu}M$ for phosphate, and $0.58{\mu}M$ for nitrate, respectively. The concentrations of nutrients increased, on average, 8.2-fold for ammonium and 4.8-fold for phosphate, decreased 3.3-fold for nitrate compared to both before and after the red tide. This result shows that the growth of C. polykikoides was not limited by the nutrients during the bloom in September. Therefore, our results suggest that the C. polykrikoides red tide may outbreak especially when the water is fertilized due to the increased N and P.

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Induction of Apoptosis and Expression of Apoptosis-related Gene Products in Response to Radiation in Murine Tumors (방사선에 대한 종양의 반응에서 아포프토시스의 유도와 이에 관련되는 유전자 발현)

  • Seong, Jin-Sil;Hunter, Nancv;Milas, Luka
    • Radiation Oncology Journal
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    • v.15 no.3
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    • pp.187-195
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    • 1997
  • Purpose : To analyze the involvement of apoptosis regulatory genes p53, $p21^{wart/cip1}$ bax and bcl-2 in induction of apoptosis by radiation in murine tumors. Materials and methods : The radiation-sensitive ovarian carcinoma OCa-1, and the radiation-resistant hepatocarcinorna HCa-I were used. Tumors, 8 mm in diameter, were irradiated with 25 Gy and at various times after irradiation, ranging from 1 to 48 h, were analyzed histologically for apoptosis and by western blot for alterations in the expression of these genes. The p53 status of the tumors were determined by the polymerase chain reaction-single strand conformation polymorphism assay. Results : Both tumors were positive for wild-type p53. Radiation inducesd apoptosis in OCa-1 but not in HCa-1. Apoptosis developed rapidly, peaked at 2 h after irradiation and returned to almost the background level at 48 h In OCa-1 radiation upregulated the expression of p53, $p21^{wart/cip1}$. and the bcl-2/bax ratio was decreased. In HCa-1 radiation increased the expression of both p53 and $p21^{wart/cip1}$, although the increase of the latter was small The bcl-2/bax ratio was greatly increased. In general the observed changes occurred within a few hours after irradiation, and either preceded or coincided with development of apoptosis Conclusions : The development of apoptosis required upregulation of both p53 and $p21^{wart/cip1}$ as well as a decrease in bcl-2/bax ratio. In contrast, an increase in bcl-2/bax ratio Prevented apoptosis in the presence of upregulated p53 and $p21^{wart/cip1}$ . These findings indentified the involvement of multiple oncogenes in apoptosis regulation in vivo and demonstrate the complexity that may be associated with the use of a single oncogene assessment for Predicting the outcome of cancer therapy with cytotoxic agents.

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Cellular Mechanism of Nicotine-mediated Intracellular Calcium Homeostasis in Primary Culture of Mouse Cerebellar Granule Cells (니코틴의 마우스 소뇌과립세포내 칼슘의 항상성 조절기전)

  • Kim, Won-Ki;Pae, Young-Sook
    • The Korean Journal of Pharmacology
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    • v.32 no.1
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    • pp.13-21
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    • 1996
  • Intracellular calcium concentration ($[Ca^{2+}]_i$) may play a crucial role in a variety of neuronal functions. Here we report that in primary culture of mouse cerebellar granule cells nicotinic acetylcholine receptors (nAChRs) are expressed in a specific developmental stage and involved in the regulation of intracellular calcium homeostasis. Nicotine-mediated calcium responses were measured using $^{45}Ca^{2+}$ or fluorometrically using the calcium-sensitive fluorescent dye fura-2. Maximal uptake of $^{45}Ca^{2+}$ evoked by nicotine in mouse cerebellar granule cells were revealed $8{\sim}12$ days in culture. In contrast, nicotine did not alter the basal $^{45}Ca^{2+}$ uptake in cultured glial cells. In cerebellar granule cells nicotine-evoked $^{45}Ca^{2+}$ uptake was largely blocked by the NMDA receptor antagonists. Glutamate pyruvate transaminase (GPT). which removes endogenous glutamate, also prevented nicotine effects, implying the indirect involvement of glutamate in nicotine-mediated calcium responses. Fluorometric studies using fura-2 showed two phases of nicotine-evoked $[Ca^{2+}]_i$ rises: the initial rising phase and the later plateau phase. Interestingly, the NMDA receptor antagonists and GPT appeared to inhibit only the later plateau phase of nicotine-evoked $[Ca^{2+}]_i$ rises. The present results imply that nicotine mediated $^{45}Ca^{2+}$ uptake and $[Ca^{2+}]_i$ rises are attributed to the calcium fluxes through both nAchRs and NMDA receptors in a time-dependent manner. Consequently, nAChRs may play an important role in neuronal development by being expressed in a specific developmental stage and regulating the intracellular calcium homeostasis.

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Studies on the CA Storage of Sweet Persimmon in Polyethylene Film Pack (Polyethylene film포장(包裝)에 의(依)한 단감의 CA저장(貯藏)에 관(關)한 연구(硏究))

  • Min, Byong-Yong;Oh, Sang-Lyong
    • Korean Journal of Food Science and Technology
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    • v.7 no.3
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    • pp.128-134
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    • 1975
  • For development of long-term storage method of sweet-persimmons using polyethylene film bags, basic experiment was conducted with 30 boxes of sweet-persimmons in 1973 and the same experiment was extended for industrial application with 2,500 boxes of the persimmons in the cold storage of Jinyoung Sweet-persimmon Association in 1974. Investigation was made on change of the quality by storage period. At the same time, persimmons put in the cold storage test were shipped to market at different time in order to monitor consumer response and commercial feasibility. The followings are conclusion obtained from the result of this experiment. 1. Storage of sweet-persimmon, Buyu, produced in Jinyoung, Kimhae was possible for 1 month at $2^{\circ}C{\pm}1^{\circ}C$ cold storage. This storage period was extended to 4 months until the end of February in case that the fruits were hermetically sealed in P.E. film bags of 0.08 to 0.1 mm thickness. 2. During the storage period of sweet-persimmons packed in the film bags, the loss of weight due to evaporation was effectively prevented with use of the film of bag thicker than 0.04 mm. 3. The storage ability of 3-5 persimmons per small bag was somewhat superior to that of many persimmons packed in the large box of 15kg capacity. 4. The thicker the film of bags, the more $CO_2$ gas was accumulated inside, however, from 1 month after beginning of the storage the rate of $CO_2$ accumulation became very low maintaining the stabilized level of 5-6% at the plot of 0.06-0.08mm thick bags. 5. While the persimmons were in storage, decreased was the content of total sugars, total acids, and vitamin C, of which the phenomenon was remarkable especially with the fruits of non-packed plot. 6. The sweet-persimmons in the film bags subjected to cold storage when shipped to market in their intact condition were more beneficial than when they were shipped out in unpacked condition. The intact fruits packed in the P.E. film bags were able to keep their commercial value for 10days in the outdoor situation. 7. The sweet-persimmons that were packed in the film bags and put in the cold storage had maintained promissing marketability and the economic feasibility was acknowledged when the experimental practice was applied to industrial scale.

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