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Amino Acid Biosynthesis and Gene Regulation in Seed (종자내 아미노산 합성 조절 유전자에 관한 연구)

  • ;;;;;Fumio Takaiwa
    • Proceedings of the Botanical Society of Korea Conference
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    • 1996.07a
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    • pp.61-74
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    • 1996
  • Human and monogastric animals can not synthesize 10 out of the 20 amino asids and therefor need to obtain these from their diet. The plant seed is a major source of dietary protein. It is particular important in their study to increase nutritional quality of the seed storage proteins. The low contents of lysine, asparagine and threonenein various cereal seeds and of cystein and methionine. In legume seeds is due to the low proportions of these amino acids in the major storage proteins, we have tried to apply the three strategies; (1) mutagenesis and selection of specific amino acid analogue resistance, (2) cloning and expression study of lysine biosynthesis related gene, (3) transfomation of lysine rich soybean glycinin gene. The 5-methyltryptophan (5MT) resistant cell lines, SAR1, SAR2 and SAR3 were selected from anther derived callus of rice (Oryza sativa L. "Sasanishiki"). Among these selected cell lines, two (SAR1 and SAR3) were able to grow stably at 200 mg/L of 5MT. Analysis of the freed amino acids in callus shows that 5MT resistant cells (SAR3) accumulated free tryptophan at least up to 50 times higher than those that of the higher than of SAS. These results indicated that the 5MT resistant cell lines are useful in studies of amino acid biosynthesis. Tr75, a rice (Oryza sativa L., var. Sasanishiki) mutant resistant to 5MT was segregated from the progenies of its initial mutant line, TR1. The 5MT resistant of TR75 was inherited in the M8 generations as a single dominant nuclear gene. The content of free amino acids in the TR75 homozygous seeds increased approximately 1.5 to 2.0 fold compared to wild-type seeds. Especially, the contents of tryptophan, phenylalanine and aspartic acid were 5.0, 5.3 and 2.7 times higher than those of wild-type seeds, respectively. The content of lysine is significantly low in rice. The lysine is synthesized by a complex pathway that is predominantly regulated by feedback inhibition of several enzymes including asparginase, aspatate kinase, dihydrodipicolinat synthase, etc. For understanding the regulation mechanism of lysine synthesis in rice, we try to clone the lysine biosynthetic metabolism related gene, DHPS and asparaginase, from rice. We have isolated a rice DHPS genomic clone which contains an ORF of 1044 nucleotides (347 amino acids, Mr. 38, 381 daltons), an intron of 587 nucleotides and 5'and 3'-flanking regions by screening of rice genomic DNA library. Deduced amino acid sequence of mature peptide domain of GDHPS clone is highly conserved in monocot and dicot plants whereas that of transit peptide domain is extremely different depending on plant specie. Southern blot analysis indicated that GDHPS is located two copy gene in rice genome. The transcripts of a rice GDHPS were expressed in leaves and roots but not detected in callus tissues. The transcription level of GDHPS is much higher in leaves indicating enormous chloroplast development than roots. Genomic DNA clones for asparaginase genes were screened from the rice genomic library by using plaque hybridization technique. Twelve different genomic clones were isolated from first and second screening, and 8 of 12 clones were analyzed by restriction patterns and identified by Southern Blotting, Restriction enzyme digestion patterns and Southern blot analysis of 8 clones show the different pattern for asparaginase gene. Genomic Southern blot analysis from rice were done. It is estimated that rice has at least 2-3 copy of asparaginase gene. One of 8 positive clones was subcloned into the pBluescript SK(+) vector, and was constructed the physical map. For transformation of lysine rich storage protein into tobacco, soybean glycinin genes are transformed into tobacco. To examine whether glycinin could be stably accumulated in endosperm tissue, the glycinin cDNA was transcriptionally fused to an endosperm-specific promotor of the rice storage protein glutelin gene and then introduced into tobacco genomic via Agrobacterium-mediated transformation. Consequently the glycinin gene was expressed in a seed-and developmentally-specific manner in transgenic tobacco seeds. Glycinin were targeted to vacuole-derived protein bodies in the endosperm tissue and highly accumulated in the matrix region of many transgenic plant (1-4% of total seed proteins). Synthesized glycinin was processed into mature form, and assembled into a hexamer in a similar manner as the glycinin in soybean seed. Modified glycinin, in which 4 contiguous methionine residues were inserted at the variable regions corresponding to the C - teminal regions of the acidic and basic polypeptides, were also found to be accumulated similarly as in the normal glycinin. There was no apparent difference in the expression level, processing and targeting to protein bodies, or accumulation level between normal and modified glycinin. glycinin.

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Physicochemical, Microbiological and Sensory Properties of Food Additive-Free Grilled Fork Products during Cold Storage (식품 첨가제 미함유 그릴 돈육햄의 냉장저장 중 물리화학, 미생물학 및 관능적 품질 특성)

  • Kim, Il-Suk;Jin, Sang-Keun;Park, Ki-Hoon;Jung, Gi-Jong;Kim, Dong-Hun;Yang, Mi-Ra;Hah, Kyung-Hee;Lee, M.
    • Food Science of Animal Resources
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    • v.26 no.3
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    • pp.269-275
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    • 2006
  • The objective of this study was to investigate the physicochemical, microbiological and sensory properties of food additive-free grilled pork products manufactured using loin (T1), tender loin (T2) and ham (T3). The samples were heated for 30 min at $60^{\circ}C$, and then 50 min for $150^{\circ}C$. After cooling, vacuum packaged grilled pork samples was stored at $4{\pm}1^{\circ}C$ for 40 days. The pH values of grilled pork samples ranged from 5.92 (T1) to 6.10 (T3) at the initial storage time, and from 6.28 (T1) to 6.60 (T3) after 40 days. The water holding capacities(%) was $85.99{\sim}93.24%$ for T1, $95.26{\sim}93.89%$ for T2 and $89.11{\sim}94.67%$ for T3, all of which were slightly higher than those of other pork products. The shear force values of T2 were significantly higher(p<0.05) than those of the other pork products throughout the storage period. The TBARS and VBN values of T2 were significantly higher(p<0.05) than those of T1 and T3. With regard to microorganisms, all grilled pork samples was in good condition, showing $1.93{\sim}3.48\;log_{10}$ CFU/g via total plate counts, and $1.74{\sim}3.48\;log_{10}$ CFU/g far lactic acid bacteria throughout the storage period. Regarding sensory evaluation, the scores of overall acceptability in all products were above 5.0 points through 40 days of storage.

Onion Favorably Affects Serum Markers of Ethanol-induced Fatty Liver in Rats (흰쥐에서 양파추출물의 알코올성 지방간 개선 작용)

  • Shin, Hye-Kyoung;Seo, Yun-Jung;Kim, Ju-Youn;Kim, Chang-Sun;Noh, Sang-K.
    • Food Science and Preservation
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    • v.14 no.6
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    • pp.662-668
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    • 2007
  • Studies have shown that onions exhibit a wide variety of health-promoting properties. The health benefits by the onion have been attributed to its ability to scavenge free radicals, to reduce blood lipids, to lower blood pressure, and to inhibit platelet aggregation. This study was performed to investigate whether onion extract supplementation would affect the blood markers of ethanol-induced fatty liver in rats. Initially, male Sprague-Dawley rats were housed singly in a room of controlled temperature and lighting and had free access to a nutritionally adequate AIN-93G and deionized water. The rats were trained for meal feeding to prevent a decline in food intake, as inevitably observed following an ethanol feeding. After the training period, rats were weight-matched and assigned to the following three groups: 1) a control group, fed the AIN-93G diet alone (control); 2) an ethanol group, fed the AIN-93G diet with ethanol at 4 g/day/kg body weight (ethanol); and 3) an onion group, fed the AIN-93G diet with ethanol plus supplemental freeze-dried onion powder at 500 mg/day/rat (ethanol + onion). All three group were meal-fed 7.0 g of their respective diets at 0900 h and 7.5 g at 1600 h for 28 days. At 0, 2, and 4 wk, blood was collected via the orbital sinus and organs were collected following overnight food deprivation. Both control and experimental groups continually gained weight throughout the study. No significant differences in the weights of the liver, kidneys, heart, spleen, and testis were observed. However, the serum level of triglycerides was significantly increased by ethanol but significantly decreased by onion extract. The activities of serum glutamate oxaloacetate transaminase (GOT), glutamate pyruvate transaminase (GPT) at 4 wk were significantly increased by ethanol feeding but were significantly decreased by onion supplementation. However, no differences among groups were observed in the serum levels of alkaline phosphatase, gamma-glutamyl transpeptidase, bilirubin, and protein. These results provide that onion extract favorably affect alcoholic fatty liver by decreasing the serum concentration of triglyceride and the activities of GOT and GPT.

Comparison of Taste Components of Giant Squid Architenthis dux via Processing Methods (대왕오징어의 가공방법에 따른 맛성분의 비교)

  • Park, Hee-Yeon;Jang, Joo-Ri;Nam, Gi-Ho;Lee, Doo-Seog;Yoon, Ho-Dong;Jang, Mi-Soon
    • Food Science and Preservation
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    • v.18 no.4
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    • pp.508-516
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    • 2011
  • The chemical components (e.g., proximate composition, nucleotide-related compounds, and amino-acid content) of raw, dried, and boiled giant squids were investigated. The moisture contents of raw, dried, and boiled giant squids were 75.65, 39.75, and 41.12%, respectively. The boiled giant squid had a higher moisture content than the dried giant squid. The crude protein contents of raw, dried, and boiled giant squids were 20.10, 56.25, and 49.58%, respectively, with that of the dried giant squid higher than that of the boiled giant squid. The crude lipid contents of the raw, dried, and boiled giant squids were 0.15, 0.57, and 1.35%, respectively, with the boiled giant squid having the highest crude lipid content. The crude ash content of raw, dried, and boiled giant squids were 1.68, 4.50, and 6.77%, respectively. IMP content was detected in the dried (1.21 mg/100 g) and boiled(0.25 mg/100 g) giant squids. In the sensory scores, the degree of bitterness, acidity, and aftertaste had lower values in the dried and boiled giant squids than in the raw giant squid. A total of 18 amino acids were detected in the samples, and most of the samples had high contents of glutamic acid, aspartic acid, proline, and arginine and low contents of histidine, tyrosine, and methionine. The free-amino-acid content was related to the taste component. The major free amino acid contained by the samples were hydroxyproline, alanine, arginine.

The Role of Pelvic Irradiation after Hysterectomy in Early Stage Cervical Carcinoma with Pelvic Nodal Metastasis (림프절 전이를 동반한 초기 자궁경부암에서 수술 후 방사선치료의 역할)

  • Suh, Hyun-Suk
    • Radiation Oncology Journal
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    • v.16 no.4
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    • pp.469-475
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    • 1998
  • Purpose : To determine the role of Pelvic irradiation in stage I or IIA cervical carcinoma with pelvic nodal metastasis after hysterectomy and bilateral pelvic lymphadenectomy. Materials and Methods : This is a retrospective study of 68 cervical carcinoma patients who were found to have pelvic nodal metastasis at hysterectomy and received pelvic irradiation from 1983 to 1996 at Inje University Seoul Paik Hospital. External beam radiation therapy to pelvis was delivered using 4 MeV Linac and intracavitary irradiation was given via vaginal ovoids or cylinders. Five-year survival and disease-free survival were estimated by Kaplan-Meier Method and prognostic factors related to survival were analysed by log-rank test and Cox proportional hazards regression model. Results : Median length of follow-up was 52months. Five-year overall survival and disease-free survival (DFS) were 81.8$\%$ and 81.7$\%$ respectively. Patients with endometrial invasion had a 57.1$\%$ 5-year DFS compared to 87.5$\%$ for those without endometrial invasion (p=0.0074). Multivariate analysis revealed endometrial invasion as an only statistically significant prognostic factor for 5-year DFS. Among total 15 (22$\%$) recurrences. pelvic recurrences occurred in 4 cases and distant metastases occurred in 13 cases. Conclusion : We have been able to confirm previous results demonstrating marked decrease in local recurrence after pelvic irradiation. In view of the high proportion of distant metastasis found in this study, a trial of aggressive adjuvant systemic therapy and irradiation in early stage cervical carcinoma patients with pelvic nodal metastasis, especially with endometrial invasion, appears to be warranted.

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Development of Vivorium, a new indoor horticultural ornamental plants via plant tissue culture techniques (식물조직배양 기술을 이용한 새로운 실내 원예 장식품인 비보리움(Vivorium)의 개발)

  • Hwang, Min Hee;Kim, Do Yeon;Cho, In Sun;Kim, Mi Hyung;Kwon, Hyun Sook;Kim, Jong Bo;Kim, Su Jung;Kim, Sun Hyung
    • Journal of Plant Biotechnology
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    • v.48 no.3
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    • pp.179-185
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    • 2021
  • Indoor gardening includes wall greening, terrariums, and flower arrangements. Among these types of indoor gardens, the terrarium is easy to access for the general public, but in Korea, because of the focus on esthetics, the original purpose of creating terrariums, which was to grow plants sustainably in an enclosed space, has been lost. In addition, miniaturization of plants is required to grow plants in an enclosed space. Since the available plant species suitable for a terrarium are limited, only plants such as succulents, cacti, and moss have been used. In this study, Bronze (X Graptosedum) was used, and these problems were solved using the following three methods: placement and growth of virus-free plants in the terrarium; extending the diversity of plants with minimal size that can be planted in terrariums; and reducing the price of in vitro plants with minimal size by achieving large-scale production. In particular, tissue-cultured succulents were developed into a Vivorium by replacing the tissue culture container and renewing the composition of the plant. This paper suggests a new indoor horticultural field, Vivorium, that can improve the current limitations of terrariums and make them more accessible to the general public. The introduction and popularization of new indoor gardening fields with the increase in single-person households and indoor activities in the Pandemic era can also improve psychological stability among people and in the society.

Screening of Natural Products for Anti-diabetic Activity and Analysis of Their Active Compounds (항당뇨 효능이 있는 천연물의 탐색 및 활성물질의 분석)

  • Hwa Sin Lee;Bo Bae Park;Sun Nyoung Yu;Min Ji Kim;Yun Jin Bae;Yi Rooney Lee;Ye Eun Lee;Si Yoon Kim;Yun Ho Shim;Soon Cheol Ahn
    • Journal of Life Science
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    • v.33 no.10
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    • pp.783-790
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    • 2023
  • Modern people have an increased incidence of metabolic diseases due to changed eating habits, and diabetes is considered the most significant metabolic disease. Given that existing diabetes treatments are accompanied by side effects, the aim of this study was to identify traditional natural products that have anti-diabetic activity. The potential anti-diabetic and antioxidant activities of natural products were examined using 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay, α-glucosidase assay, and protein tyrosine phosphatase 1B (PTP1B) inhibition assay. Methanol extracts of Ulmus davidiana var. japonica, Acer tegmentosum branches, Nelumbo nucifera seeds, and Carthamus tinctorius seeds were found to have high anti-diabetic activity and further fractionated with solvents using ethyl acetate and butanol. Consequently, the ethyl acetate fraction of C. tinctorius seeds (MG-11-E) with high α-glucosidase and PTP1B inhibitory activity was selected. MG-11-E was subjected to preparative thin layer chromatography, and fraction #6 showed high α-glucosidase and PTP1B inhibitory activity. Fraction #6 was analyzed and fractionated via high performance liquid chromatography with 50% methanol as the mobile phase, and anti-diabetic activity was observed in the sample that eluted after 4 min as a single peak. The α-glucosidase inhibitory activity exhibited by this sample seemed to be greater than the PTP1B inhibitory activity; thus, it was concluded that a greater anti-diabetic therapeutic effect may be achieved by combining this agent with natural products that inhibit PTP1B activity.

The Effect of the Gap between College Students' Perception of the Importance of Coffee Shops and Their Satisfaction after Patronizing Coffee Shops on Their Purchasing Behavior (대전원교학생대가배점중요성적감지화타문광고가배점지후적만의도지간적차거대타문구매행위적영향(大专院校学生对咖啡店重要性的感知和他们光顾咖啡店之后的满意度之间的差距对他们购买行为的影响))

  • Lee, Won-Ok
    • Journal of Global Scholars of Marketing Science
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    • v.19 no.4
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    • pp.1-10
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    • 2009
  • The purpose of this study was to categorize the gap between coffee shop 'importance' (as perceived by customers before patronizing the coffee shop) and 'satisfaction' (perception of customers after patronizing the coffee shop) as positive or negative and to analyze the effect of these gaps on purchasing behavior. To do this, I used the gap between importance and satisfaction regarding the choice of a coffee shop as the explanatory variable and performed an empirical analysis of the direction and size of the effect of the gap on purchasing behavior (overall satisfaction, willingness-to-revisit) by applying the Ordered Probit Model (OPM). A previous study that used IPA to evaluate the effects of gaps estimated the direction and size of a quadrant but failed to analyze the effect of gaps on customers. In this study, I evaluated the effects of positive and negative gaps on customer satisfaction and willingness-to-revisit. Using OPM, I quantified the effect of positive and negative gaps on overall customer satisfaction and willingness-to-revisit. Per-head expenditure, frequency of visits, and coffee-purchasing place had the most positive effects on overall customer satisfaction. Frequency of visits, followed by per-head expenditure and then coffee-purchasing place, had the most positive impact on willingness-to-visit. Thus per-head expenditure and frequency of visits had the greatest positive effects on overall satisfaction and willingness-to-revisit. This finding implies that the higher the actual satisfaction (gap) of customers who spend KRW5,000 or more once or more per week at coffee shops is, the higher their overall satisfaction and willingness-to-revisit are. Despite the fact that economical efficiency had a significant effect on overall satisfaction and willingness-to-revisit, college and university students still use coffee shops and are willing to spend KRW5,000 because they do not only purchase coffee as a product itself, but use the coffee shop for other activities, such as working, meeting friends, or relaxing. College and university students also access the Internet in coffee shops via personal laptops, watch movies, and study; thus, coffee shops should provide their customers with the appropriate facilities and services. The fact that a positive gap for coffee shop brand had a positive effect on willingness-to-revisit implies that the higher the level of customer satisfaction, the greater the willingness-to-revisit. A negative gap for this factor, on the other hand, implies that the lower the level of customer satisfaction, the lower the willingness-to-revisit. Thus, the brand factor has a comparatively greater effect on satisfaction than the other factors evaluated in this study. Given that the domestic coffee culture is becoming more upscale and college/university students are sensitive to this trend, students are attentive to brands. In most upscale coffee shops in Korea, the outer wall is built out of glass that can be opened, the interiors are exotic with an open kitchen. These upscale coffee shops function as landmarks and match the taste of college/university students. Coffee shops in Korea have become a cultural brand. To make customers feel that coffee shops are upscale, good quality establishments and measures to provide better services in terms of brand factor should be instituted. The intensified competition among coffee shop brands in Korea as a result of the booming industry indicates that provision of additional services is needed to differentiate competitors. These customers can also use a scanner free of charge. Another strategy that can be used to boost brands could be to provide and operate a seminar room for seminars and group study. If coffee shops adopt these types of strategies, college/university students would be more likely to consider the expenses they incur worthwhile and, subsequently, they would be more likely to be satisfied with the brands of these coffee shops, with an associated increase in their willingness-to-revisit. Gender and study year had the most negative effects on overall satisfaction and willingness-to-revisit. Female students were more likely to be satisfied and be willing to return than male students, and third and fourth-year students were more likely to be satisfied and willing-to-return than first or second-year students. Students who drink coffee, read books, and use laptops alone at coffee shops are easily noticeable. High-grade students tend to visit coffee shops alone in order to use their time efficiently for self-development and to find jobs. The economical efficiency factor had the greatest effect on overall satisfaction and willingness-to-revisit in terms of a positive gap. The higher the actual satisfaction (gap) of students with the price of the coffee, the greater their overall satisfaction and willingness-to-revisit. Economical efficiency with a negative gap had a negative effect on willingness-to-revisit, which implies that a less negative gap will result in a greater willingness-to-revisit. Amid worsening market conditions, coffee shops located around colleges/universities are using strategies, such as a point or membership card, strategic alliances with credit-card companies, development of a set menu or seasonal menu, and free coffee-shot services to increase their competitive edge. Product power also had a negative effect in terms of a negative gap, which indicates that a higher negative gap will result in a lower willingness-to-revisit. Because there are many more customers that enjoy coffee in this decade, as compared to previous decades, the new generation of customers, namely college/university students, want various menu items in addition to coffee, and coffee shops should, therefore, add side menu items, such as waffles, rice cakes, cakes, sandwiches, and salads. For example, Starbucks Korea is making efforts to enhance product power by selling rice cakes flavored in strawberry, wormwood, and pumpkin, and providing coffee or cream free of charge. In summary, coffee shops should focus on increasing their economical efficiency, brand, and product power to enhance the satisfaction of college/university students. Because shops adjacent to colleges or universities enjoy a locational advantage, providing differentiated services in terms of economical efficiency, brand, and product power, is likely to increase customer satisfaction and return visits. Coffee shop brands should, therefore, be innovative and embrace change to meet their customers' desires. Because this study only targeted college/university students in Seoul, comparative studies targeting diverse regions and age groups are required to generalize the findings and recommendations of this study.

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EFFECT OF OCTANOL, THE GAP JUNCTION BLOCKER, ON THE REGULATION OF FLUID SECRETION AND INTRACELLULAR CALCIUM CONCENTRATION IN SALIVARY ACINAR CELLS (흰쥐 악하선 세포에서 gap junction 봉쇄제인 octanol이 타액분비 및 세포내 $Ca^{2+}$ 농도 조절에 미치는 영향)

  • Lee, Ju-Seok;Seo, Jeong-Taeg;Lee, Syng-Il;Lee, Jong-Gap;Sohn, Heung-Kyu
    • Journal of the korean academy of Pediatric Dentistry
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    • v.26 no.2
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    • pp.399-415
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    • 1999
  • From bacteria to mammalian cells, one of the most important mediators of intracellular signal transduction mechanisms which regulate a variety of intracellular processes is free calcium. In salivary acinar cells, elevation of intracellular calcium concentration ($[Ca^{2+}]_i$) is essential for the salivary secretion induced by parasympathetic stimulation. However, in addition to $[Ca^{2+}]_i$, gap junctions which couple individual cells electrically and chemically have also been reported to regulate enzyme secretion in pancreatic acinar cells. Since the plasma membrane of salivary acinar cells has a high density of gap junctions, and these cells are electrically and chemically coupled with each other, gap junctions may modulate the secretory function of salivary glands. In this respect, I planned to investigate the role of gap junctions in the modulation of salivary secretion and $[Ca^{2+}]_i$, using mandibular salivary glands of rats. In order to measure the salivary flow rate, fluid was collected from the cannulated duct of the isolated perfused rat mandibular glands at 2 min intervals. $[Ca^{2+}]_i$, was measured from the cells loaded with fura-2 by spectrofluorometry. The results obtained were as follows: 1. CCh-induced salivary secretion was reversibly inhibited by 1 mM octanol, a gap junction blocker. 2. CCh-induced increase in $[Ca^{2+}]_i$, was also reversed by the application of 1 mM octanol. 3. Octanol did not block the initial increase in $[Ca^{2+}]_i$ caused by CCh, which suggested that the reduction of $[Ca^{2+}]_i$, caused by gap junction blockade was not resulted from the inhibition of $Ca^{2+}$ release from intracellular $Ca^{2+}$ stores. 4. Addition of octanol during stimulation with $1{\mu}M$ thapsigargin, a potent microsomal ATPase inhibitor, reduced $[Ca^{2+}]_i$, to the basal level. This suggested that inhibition of gap junction permeability closed plasma membrane $Ca^{2+}$ channels. 5. 2,5-di-tert-butyl-1,4-benzohydroquinone (TBQ) generated $[Ca^{2+}]_i$ oscillations resulting from periodic influx of $Ca^{2+}$ via plasma membrane. The TBQ-induced $[Ca^{2+}]_i$ oscillations were stopped by the application of 1mM octanol which implicated that gap junctions modulate the permeability of plasma membrane $Ca^{2+}$ channels. 6. Glycyrrhetinic acid, another well known gap junction blocker, also inhibited CCh-induced salivary secretion from rat mandibular glands. These results suggested that gap junctions play an important role in the modulation of fluid secretion from the rat mandibular glands and this was probably due to the inhibition of $Ca^{2+}$ influx through the plasma membrane $Ca^{2+}$ channels.

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The View of Life and Death in Jeon-gyeong (『전경』에 나타난 대순사상의 생사관)

  • Cheng, Chihming
    • Journal of the Daesoon Academy of Sciences
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    • v.27
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    • pp.79-132
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    • 2016
  • The view of life and death in Daesoonjinrihoe includes all the gods of Heaven and Earth, and the human heart is taken as the foundational key. Practitioners can realize their value according to how much they have cultivated themselves. This is regarded as the mythical use of a singularly focused mind (full dedication of one's heart). In other words, it focuses on the potentiality of humans who are able to enter a transcendental area of divinity through their self-cultivation. This view of life and death in Daesoonjinrihoe was established by the religious mission known as "Samgye Gongsa (the Reordering of Three Realms of Heaven, Earth, and Humanity)." Samgye Gongsa indicated a new opening of the Three Realms of Heaven, Earth, and Humanity. This new opening is a return to the original principle of Heavenly operation and also a new order for the universe. Heaven and Earth have their own underlying principle by which they operate. This act was directly initiated and manifested from Dao. Daesoonjinrihoe diagnoses that the underlying principle by which Heaven operates was damaged by human misconduct, and as a result, the human observance of that principle fell out of common usage. Therefore, Daesoonjinrihoe gives priority to the reestablishment of Dao as it existed originally and tries to bring about reconciliation between Heaven and Earth and Humanity. In short, it resolves the grievances accrued since time immemorial by correcting the order of Sindo (Divine Law). Furthermore, it shows that the Dao of Sangsaeng (mutual beneficence) was created by reordering the arrangement of Heaven and Earth so that human beings and divine beings could reach a state of perfection through self-realization. Humans not only communicate with Heaven and Earth, but also communicate with divine beings. Divine beings are transcendent living beings capable of communicating with humans through their heart-minds. In Daesoon thought, human beings are not swayed by the power of divine beings, but instead are able to control divine beings through the transcendent power of their heart-minds. Given this view, the aim of Daesoonjinrihoe lies in participating in the harmony of Heaven and Earth through the cultivation of the human heart. Also, it sees that the human heart-mind can be united with the universal Dao, and thus it is able to be united with the deities of Heaven and Earth. In order to actualize this, one does not rely on exterior rituals or magic but has to focus instead on cultivating the moral ethics of the heart-mind to reach perfection. In other words, one can reach a transcendent level in one's heart-mind through the cultivation of a singularly focused mind and be free from the contradiction of life and death and other such torments. Life and death is an inevitable process for humans. So they do not have to be happy for life and sad for death. They can rather be free from the fear of death by fulfilling the energetic zenith of the human heart-mind via training themselves to transcend their physical bodies. No aging and no death is not a pursuit of radical longevity or immortality for the physical body, but rather a pursuit of the essence of life and the realization of eternity on a spiritual level. Daesoonjinrihoe pursues the state of being unified with Dao by developing "Jeong·Gi·Sin (精·氣·神 the internal energies of essence, pneuma, and spirit)" and trying to reach the transcendent state of non-aging and radical longevity by spurring the practice of self-realization and the discovery one's own innate nature. Through the practice of human ethics, they can access the creative functions of Heaven and Earth and become one with Heavenly Dao thereby achieving harmony between temporal existence and eternity. In this way, humans transcend the life and death of their physical bodies. When "Doins (trainees of Dao)" reach the true state of unification with Dao through singularly focused cultivation, they not only realize self perfection as human beings, but also enable themselves the means to do away with all disasters and forms of suffering. They thereby attain ultimate happiness in their lives.