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Method Development for Determination of Multi-Mycotoxins in Chicken Liver and Kidney Tissues by LC-MS/MS (LC-MS/MS를 이용한 닭 간과 신장 중 곰팡이 독소 6종 동시분석법 개발)

  • Kim, Soohee;Kim, Kwang-Nam;Kim, Hyobi;Song, Jae-Young;Park, Sung-Won
    • Korean Journal of Poultry Science
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    • v.43 no.2
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    • pp.111-118
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    • 2016
  • Mycotoxins are secondary metabolites produced by molds, such as Aspergillus, Fusarium and Penicillium, that have adverse effects on animals and humans. Aflatoxin, ochratoxin, zearalenone, fumonisin and deoxynivalenol are the mycotoxins of greatest agro-economic importance and cause acute disease called mycotoxicoses. Mycotoxicosis in poultry birds results in decreased meat/egg production, immunosuppressant, and hepatotoxicosis. Some of toxins or their metabolites may be retained in animal or human tissues and induce health problems. This study was designed to develop a sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the simultaneous detection and quantification of mycotoxins, such as aflatoxin $B_1$, aflatoxin $M_1$, ochratoxin A, zearalenone, fumonisin B and deoxynivalenol, in chicken liver and kidney tissues. The mycotoxins were extracted and purified using modified QUECHERS methods, separated by LC and detected by an electrospray ionisation interface (ESI) and tandem MS. Good precision and linearity were observed for most of six mycotoxins. The recovery test for each mycotoxin in liver and kidney tissues mostly indicated good average recovery rates between 80.94% and 98.10% and the coefficient of variation mostly under 13.78%, except for aflatoxin $M_1$ and fumonisin $B_1$. The limit of detection (LOD) for six mycotoxins was $7.6{\sim}145.79{\mu}g/kg$ in liver tissues and $6.07{\sim}197.20{\mu}g/kg$ in kidney tissues. The quantification limits (LOQ) for 6 mycotoxins were in the range $23.04{\sim}441.78{\mu}g/kg$ in liver tissues and $18.40{\sim}597.59{\mu}g/kg$ in kidney tissues, respectively. The developed multi-mycotoxin method in this study permits simultaneous, simple, and rapid determination of several co-existing mycotoxins in chicken liver and kidney tissues.

Development and Evaluation of Validity of Dish Frequency Questionnaire (DFQ) and Short DFQ Using Na Index for Estimation of Habitual Sodium Intake (나트륨 섭취량 추정을 위한 음식섭취빈도조사지와 Na Index를 이용한 간이음식섭취빈도조사지의 개발 및 타당성 검증에 관한 연구)

  • Son, Sook-Mee;Huh, Gwui-Yeop;Lee, Hong-Sup
    • Korean Journal of Community Nutrition
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    • v.10 no.5
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    • pp.677-692
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    • 2005
  • The assessment of sodium intake is complex because of the variety and nature of dietary sodium. This study intended to develop a dish frequency questionnaire (DFQ) for estimating the habitual sodium intake and a short DFQ for screening subjects with high or low sodium intake. For DFQ112, one hundred and twelve dish items were selected based on the information of sodium content of the one serving size and consumption frequency. Frequency of consumption was determined through nine categories ranging from more than 3 times a day to almost never to indicate how often the specified amount of each food item was consumed during the past 6 months. One hundred seventy one adults (male: 78, female: 93) who visited hypertension or health examination clinic participated in the validation study. DFQ55 was developed from DFQ112 by omitting the food items not frequently consumed, selecting the dish items that showed higher sodium content per one portion size and higher consumption frequency. To develop a short DFQs for classifying subjects with low or high sodium intakes, the weighed score according to the sodium content of one protion size was given to each dish item of DFQ25 or DFQ14 and multiplied with the consumption frequency score. A sum index of all the dish items was formed and called sodium index (Na index). For validation study the DFQ112, 2-day diet record and one 24-hour urine collection were analyzed to estimate sodium intakes. The sodium intakes estimated with DFQ112 and 24-h urine analysis showed $65\%$ agreement to be classified into the same quartile and showed significant correlation (r=0.563 p<0.05). However, the actual amount of sodium intake estimated with DFQ112 (male: 6221.9mg, female: 6127.6mg) showed substantial difference with that of 24-h urine analysis (male: 4556.9mg, female: 5107.4mg). The sodium intake estimated with DFQ55 (male: 4848.5mg, female: 4884.3mg) showed small difference from that estimated with 24-h urine analysis, higher proportion to be classfied into the same quartile and higher correlation with the sodium intakes estimated with 24-h urine analysis and systolic blood pressure. It seems DFQ55 can be used as a tool for quantitative estimation of sodium intake. Na index25 or Na index14 showed $39\~50\%$ agreement to be classified into the same quartile, substantial correlations with the sodium intake estimated with DFQ55 and significant correlations with the sodium intake estimated with 24-h urine analysis. When point 119 for Na index25 was used as a criterion of low sodium intake, sensitivity, specificity and positive predictive value was $62.5\%,\;81.8\%\;and\;53.2\%$, respectively. When point 102 for Na index14 was used as a criterion of high sodium intake, sensitivity, specificity and positive predictive value were $73.8\%,\;84.0\%,\;62.0\%$, respectively. It seems the short DFQs using Na index 14 or Na index25 are simple, easy and proper instruments to classify the low or high sodium intake group.

A study on the search and selection processes of targets presented on the CRT display (컴퓨터 모니터에 제시된 표적의 탐색과 선택과정에 관한 연구)

  • 이재식;신현정;도경수
    • Korean Journal of Cognitive Science
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    • v.11 no.2
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    • pp.37-51
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    • 2000
  • The present study compared computer users target-selection response patterns when the targets were varied in terms of their relative location and distance from the current position of the cursor. In Experiment 1, where the mouse was used as an input device, the effects of different directions and distances of simple target(small rectangle) on target-selection response were investigated. The results of Experiment 1 can be summarized as follows: (1) Overshooting was more frequent than either undershooting or correct movement and (2) this tendency was more prominent when the targets were presented in the oblique direction or in farther location from the current cursor position. (3) Although the overshooting and undershooting were more frequent in the oblique direction, the degree of deviation was larger in horizontal and vertical direction. (4) Time spent in moving the mouse rather than that spent in planning, calibrating or clicking was found to be the most critical factor in determining total response time. In Experiment 2, effects of the font size and line-height of the target on target-selection response were compared with regard to two types of input devices(keyboard vs. mouse). The results are as follows: (1) Mouse generally yielded shorter target-selection time than keyboard. but this tendency was reversed when the targets were presented in horizontal and vertical directions. (2) In general, target-selection time was the longest in the condition of font size of 10 and line-height of 100%, and the shortest in the condition of font size of 12 and line-height of 150%. (3) When keyboard was used as the input device, target-selection time was shortest in the 150% line-height condition, whereas in the mouse condition, target-selection time tended to be increased as the line-height increased. which resulted in the significant interaction effect between input device and line-height. Finally, several issues relating to human-computer interaction were discussed based on the results of the present study.

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Surgical Treatment of Thoracoabdominal Aortic Aneurysm (흉복부 대동맥류의 외과적 치료)

  • Ahn, Hyuk;Kim, Jun-Seok
    • Journal of Chest Surgery
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    • v.29 no.2
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    • pp.177-184
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    • 1996
  • Between 1987 and 1994, 21 patients were treated surgically for aortic aneurysm involving the thoracoabdominal aorta. There were 11 males and 10 females, and their age ranged from 20 to 67 years old and mean age was 41.5 years. Many complained of back pain, chest pain or discomfort, and flank pain, but three patients were asymptomatic. 15 patients had chronic dissection (71.4%) and 6 had nondissecting fusiform or saccular aneurysm(28.5%), and of those 15 patients with chronic dissection, 6(28.5%) had atherosclerosis assniated with hypertension, 5 (23.8%) were Martian syndrome, and 2 (9.5%) were associated with pregnancy. The diameter of an aneurysm ranged from 6cm to 12cm, and their extent was classified as type I in 7(33.3%), type II in 8(38.1%), type III in 3(14.3%), and type IV in 3(14.3%) patients based on Crawford classification for TAA . Diseased aorta was replaced with artificial vascular graft in all but one patient. In whom the aortic tear site due to pseudoaneurysm was closed by primary suture. For the spinal cord protection during the operation, we used partial cardiopulmonary bypass (FV-FA or PA-FA bypass) in 12 patients (57.1%), Biopump (LA-FA bypass) in 4(19.0%), total circulartory arrest and CPB in 2 (9.5%), Gott's heparinized shunt in 1(4.7%), and simple aortic cross clamping in 2 (9.5%). The most common complication after the operation was hoarseness due to unilateral vocal cord palsy which onured in 5 patients (23.8%), and the next common complication was wound infection in 4 patients(19.0%), paraplegia in 2 patients (9.5%), chylothorax in 1 patient(4.7%). The hospital mortality rate was 9.5% (2deaths), and there was no late death. Our experience shows that the graft replacement of TAAA had reasonable rate of mortality, low rate of serious complication, and provided good post operative state of the pati nts, and since the thoracoabdominal aortic operation is not a high risky procedure anymore, we recommend a radical operation for the indicated patients.

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Investigation of the Signal Characteristics of a Small Gamma Camera System Using NaI(Tl)-Position Sensitive Photomultiplier Tube (NaI(Tl) 섬광결정과 위치민감형 광전자증배관을 이용한 소형 감마카메라의 신호 특성 고찰)

  • Choi, Yong;Kim, Jong-Ho;Kim, Joon-Young;Im, Ki-Chun;Kim, Sang-Eun;Choe, Yearn-Seong;Lee, Kyung-Han;Joo, Koan-Sik;Kim, Byung-Tae
    • The Korean Journal of Nuclear Medicine
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    • v.34 no.1
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    • pp.82-93
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    • 2000
  • Purpose: We characterized the signals obtained from the components of a small gamma camera using Nal(Tl)-position sensitive photomultiplier tube (PSPMT) and optimized the parameters employed in the modules of the system. Materials and Methods: The small gamma camera system consists of a Nal(Tl) crystal ($60{\times}60{\times}6mm^3$) coupled with a Hamamatsu R3941 PSPMT, a resister chain circuit, preamplifiers, nuclear instrument modules (NIMs), an analog to digital converter and a personal computer for control and display. The PSPMT was read out using a resistive charge division circuit which multiplexes the 34 cross wire anode channels into 4 signals (X+, X-, Y+, Y -). Those signals were individually amplified by four preamplifiers and then, shaped and amplified by amplifiers. The signals were discriminated and digitized via triggering signal and used to localize the position of an event by applying the Anger logic. The gamma camera control and image display was performed by a program implemented using a graphic software. Results: The characteristics of signal and the parameters employed in each module of the system were presented. The intrinsic sensitivity of the system was approximately $8{\times}10^3$ counts/sec/${\mu}Ci$. The intrinsic energy resolution of the system was 18% FWHM at 140 keV. The spatial resolution obtained using a line-slit mask and $^{99m}Tc$ point source were, respectively, 2.2 and 2.3 mm FWHM in X and Y directions. Breast phantom containing $2{\sim}7mm$ diameter spheres was successfully imaged with a parallel hole collimator. The image displayed accurate size and activity distribution over the imaging field of view Conclusion: We proposed a simple method for development of a small gamma camera and presented the characteristics of the signals from the system and the optimized parameters used in the modules of the small gamma camera.

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Development and Validation of Analytical Method for Determination of Biphenyl Analysis in Foods (식품 중 비페닐 분석법 개발 및 유효성 검증)

  • Kim, Jung-Bok;Kim, Myung-Chul;Song, Sung-Woan;Shin, Jae-Wook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.4
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    • pp.459-464
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    • 2017
  • Biphenyl is used as an intermediate in the production of crop protection products, a solvent in pharmaceutical production, and as a component in the preservation of citrus fruits in many countries. Biphenyl is not authorized for use and also does not have standards or specifications as a food additive in Korea. National and imported food products are likely to contain biphenyl. Therefore, control and management of these products is required. In this study, a simple analytical method was developed and validated using HPLC to determine biphenyl in food. These methods are validated by assessing certain performance parameters: linearity, accuracy, precision, recovery, limit of detection (LOD), and limit of quantitation (LOQ). The calibration curve was obtained from 1.0 to $100.0{\mu}g/mL$ with satisfactory relative standard deviations (RSD) of 0.999 in the representative sample (orange). In the measurement of quality control (QC) samples, accuracy was in the range of 95.8~104.0% within normal values. The inter-day and inter-day precision values were less than 2.4% RSD in the measurement of QC samples. Recoveries of biphenyl from spiked orange samples ranged from 92.7 to 99.4% with RSD between 0.7 and 1.7% at levels of 10, 50, and $100{\mu}g/mL$. The LOD and LOQ were determined to be 0.04 and $0.13{\mu}g/mL$, respectively. These results show that the developed method is appropriate for biphenyl identification and can be used to examine the safety of citrus fruits and surface treatments containing biphenyl residues.

The Association between Blood Pressure and Obstructive Sleep Apnea-Hypopnea Syndrome

  • Kim, Cheon-Sik
    • Korean Journal of Clinical Laboratory Science
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    • v.46 no.3
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    • pp.106-110
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    • 2014
  • Obstructive sleep apnea-hypopnea (OSAH) is known to be related to nocturnal blood pressure (BP) and hypertension. The aim of this study was to evaluate the prevalence of hypertension according to the apnea-hypopnea grading. A total of 2,210 adults with snoring and obstructive sleep apnea were referred to our sleep center from July 2009 to May 2013. Clinical blood pressure (BP) was measured before sleeping (bedtime BP) and immediately after waking up in the next morning (morning BP). Subjects were classified into four groups based on the apnea-hypopnea index (AHI) from PSG as follows: control group (n=470) simple snoring and with AHI<5; mild group (n=577) with $AHI{\geq}5$ and <15; moderate group (n=508) $AHI{\geq}15$ and <30; and severe group (n=655) with $AHI{\geq}30$. The differences and correlations between BP and PSG parameters according to the AHI groups were analyzed. Patient's were classified as nomentensive (blood pressure <120/90 mmHg, n=700), prehypertensive (blood pressure < $140-120{\leq}mmHg$, n=1297) hypertensive (blood pressure ${\geq}140/90mmHg$, n=214) according to the office blood pressure measurements. The comparison of sleep parameters showed that OSA groups had a significantly higher stage N1 (control group vs. moderate OSA, severe OSA; $66.4{\pm}30.7$ vs. $85.5{\pm}36.6$, $128.4{\pm}57.3$, p<0.001) and total arousal number (control vs. moderate OSA, severe OSA; $110.7{\pm}47.7$ vs. $150.8{\pm}56.6$, $236.6{\pm}95.8$, p<0.001) compared to control group. The comparison of sleep parameters showed that OSA groups had a significantly lower stage N2 (control group vs. moderate OSA, severe OSA; $172.6{\pm}47.2$ vs. $150.7{\pm}50.5$, $120.3{\pm}57.4$, p<0.001), stage N3 (control group vs. moderate OSA, severe OSA; $38.4{\pm}33.4$ vs. $27.4{\pm}26.0$, $56.1{\pm}27.5$, p<0.001), REM (control group vs. moderate OSA, severe OSA; $64.3{\pm}25.5$ vs. $56.1{\pm}27.5$, $47.3{\pm}25.9$, p<0.001) and mean SaO2% (control group vs. moderate OSA, severe OSA; $90.0{\pm}3.5$ vs. $82.5{\pm}5.5$, $70.0{\pm}8.8$, p<0.001) compared to control group. The Apnea-hypopnea index was significantly higher in OSA groups, increased systolic and diastolic blood pressure than in the nomentensive group (bed time systolic pressure vs. AHI; <120 vs. 120-139, 140-159, >159; $17.5{\pm}18.6$ vs. $24.9{\pm}21.0$, $31.0{\pm}25.7$, $42.3{\pm}31.7$, p<0.001), (bed time diastolic pressure vs. AHI; 60-79 vs. 80-89, 90-99, >99; $19.3{\pm}19.7$ vs. $22.4{\pm}20.3$, $29.8{\pm}23.3$, $38.8{\pm}28.5$, p<0.001). AHI was positively correlated with morning systolic pressure, diastolic pressure, bed time systolic pressure and diastolic pressure (r=0.314, 0.279, 0.233 and 0.200, respectively, p<0.001). We conclude that BMI, Age, neck circumference and AHI increase with the blood pressure.

Research Framework for International Franchising (국제프랜차이징 연구요소 및 연구방향)

  • Kim, Ju-Young;Lim, Young-Kyun;Shim, Jae-Duck
    • Journal of Global Scholars of Marketing Science
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    • v.18 no.4
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    • pp.61-118
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    • 2008
  • The purpose of this research is to construct research framework for international franchising based on existing literature and to identify research components in the framework. Franchise can be defined as management styles that allow franchisee use various management assets of franchisor in order to make or sell product or service. It can be divided into product distribution franchise that is designed to sell products and business format franchise that is designed for running it as business whatever its form is. International franchising can be defined as a way of internationalization of franchisor to foreign country by providing its business format or package to franchisee of host country. International franchising is growing fast for last four decades but academic research on this is quite limited. Especially in Korea, research about international franchising is carried out on by case study format with single case or empirical study format with survey based on domestic franchise theory. Therefore, this paper tries to review existing literature on international franchising research, providing research framework, and then stimulating new research on this field. International franchising research components include motives and environmental factors for decision of expanding to international franchising, entrance modes and development plan for international franchising, contracts and management strategy of international franchising, and various performance measures from different perspectives. First, motives of international franchising are fee collection from franchisee. Also it provides easier way to expanding to foreign country. The other motives including increase total sales volume, occupying better strategic position, getting quality resources, and improving efficiency. Environmental factors that facilitating international franchising encompasses economic condition, trend, and legal or political factors in host and/or home countries. In addition, control power and risk management capability of franchisor plays critical role in successful franchising contract. Final decision to enter foreign country via franchising is determined by numerous factors like history, size, growth, competitiveness, management system, bonding capability, industry characteristics of franchisor. After deciding to enter into foreign country, franchisor needs to set entrance modes of international franchising. Within contractual mode, there are master franchising and area developing franchising, licensing, direct franchising, and joint venture. Theories about entrance mode selection contain concepts of efficiency, knowledge-based approach, competence-based approach, agent theory, and governance cost. The next step after entrance decision is operation strategy. Operation strategy starts with selecting a target city and a target country for franchising. In order to finding, screening targets, franchisor needs to collect information about candidates. Critical information includes brand patent, commercial laws, regulations, market conditions, country risk, and industry analysis. After selecting a target city in target country, franchisor needs to select franchisee, in other word, partner. The first important criteria for selecting partners are financial credibility and capability, possession of real estate. And cultural similarity and knowledge about franchisor and/or home country are also recognized as critical criteria. The most important element in operating strategy is legal document between franchisor and franchisee with home and host countries. Terms and conditions in legal documents give objective information about characteristics of franchising agreement for academic research. Legal documents have definitions of terminology, territory and exclusivity, agreement of term, initial fee, continuing fees, clearing currency, and rights about sub-franchising. Also, legal documents could have terms about softer elements like training program and operation manual. And harder elements like law competent court and terms of expiration. Next element in operating strategy is about product and service. Especially for business format franchising, product/service deliverable, benefit communicators, system identifiers (architectural features), and format facilitators are listed for product/service strategic elements. Another important decision on product/service is standardization vs. customization. The rationale behind standardization is cost reduction, efficiency, consistency, image congruence, brand awareness, and competitiveness on price. Also standardization enables large scale R&D and innovative change in management style. Another element in operating strategy is control management. The simple way to control franchise contract is relying on legal terms, contractual control system. There are other control systems, administrative control system and ethical control system. Contractual control system is a coercive source of power, but franchisor usually doesn't want to use legal power since it doesn't help to build up positive relationship. Instead, self-regulation is widely used. Administrative control system uses control mechanism from ordinary work relationship. Its main component is supporting activities to franchisee and communication method. For example, franchisor provides advertising, training, manual, and delivery, then franchisee follows franchisor's direction. Another component is building franchisor's brand power. The last research element is performance factor of international franchising. Performance elements can be divided into franchisor's performance and franchisee's performance. The conceptual performance measures of franchisor are simple but not easy to obtain objectively. They are profit, sale, cost, experience, and brand power. The performance measures of franchisee are mostly about benefits of host country. They contain small business development, promotion of employment, introduction of new business model, and level up technology status. There are indirect benefits, like increase of tax, refinement of corporate citizenship, regional economic clustering, and improvement of international balance. In addition to those, host country gets socio-cultural change other than economic effects. It includes demographic change, social trend, customer value change, social communication, and social globalization. Sometimes it is called as westernization or McDonaldization of society. In addition, the paper reviews on theories that have been frequently applied to international franchising research, such as agent theory, resource-based view, transaction cost theory, organizational learning theory, and international expansion theories. Resource based theory is used in strategic decision based on resources, like decision about entrance and cooperation depending on resources of franchisee and franchisor. Transaction cost theory can be applied in determination of mutual trust or satisfaction of franchising players. Agent theory tries to explain strategic decision for reducing problem caused by utilizing agent, for example research on control system in franchising agreements. Organizational Learning theory is relatively new in franchising research. It assumes organization tries to maximize performance and learning of organization. In addition, Internalization theory advocates strategic decision of direct investment for removing inefficiency of market transaction and is applied in research on terms of contract. And oligopolistic competition theory is used to explain various entry modes for international expansion. Competency theory support strategic decision of utilizing key competitive advantage. Furthermore, research methodologies including qualitative and quantitative methodologies are suggested for more rigorous international franchising research. Quantitative research needs more real data other than survey data which is usually respondent's judgment. In order to verify theory more rigorously, research based on real data is essential. However, real quantitative data is quite hard to get. The qualitative research other than single case study is also highly recommended. Since international franchising has limited number of applications, scientific research based on grounded theory and ethnography study can be used. Scientific case study is differentiated with single case study on its data collection method and analysis method. The key concept is triangulation in measurement, logical coding and comparison. Finally, it provides overall research direction for international franchising after summarizing research trend in Korea. International franchising research in Korea has two different types, one is for studying Korean franchisor going overseas and the other is for Korean franchisee of foreign franchisor. Among research on Korean franchisor, two common patterns are observed. First of all, they usually deal with success story of one franchisor. The other common pattern is that they focus on same industry and country. Therefore, international franchise research needs to extend their focus to broader subjects with scientific research methodology as well as development of new theory.

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The SOFA Score to Evaluate Organ Failure and Prognosis in the Intensive Care Unit Patients (중환자실에 입원한 환자의 장기부전 및 예후 평가를 위한 SOFA 점수체계의 의의)

  • Kim, Su Ho;Lee, Myung Goo;Park, Sang Myeon;Park, Young Bum;Jang, Seung Hun;Kim, Cheol Hong;Jeon, Man Jo;Shin, Tae Rim;Eom, Kwang Seok;Hyun, In-Gyu;Jung, Ki-Suck;Lee, Seung-Joon
    • Tuberculosis and Respiratory Diseases
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    • v.57 no.4
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    • pp.329-335
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    • 2004
  • Background : The Sequential Organ Failure Assessment (SOFA) score can help to assess organ failure over time and is useful to evaluate morbidity. The aim of this study is to evaluate the performance of SOFA score as a descriptor of multiple organ failure in critically ill patients in a local unit hospital, and to compare with APACHE III scoring system. Methods : This study was carried out prospectively. A total of ninety one patients were included who admitted to the medical intensive care unit (ICU) in Chuncheon Sacred Heart Hospital from May 1 through June 30, 2000. We excluded patients with a length of stay in the ICU less than 2 days following scheduled procedure, admissions for ECG monitoring, other department and patients transferred to other hospital. The SOFA score and APACHE III score were calculated on admission and then consecutively every 24 hours until ICU discharge. Results : The ICU mortality rate was 20%. The non-survivors had a higher SOFA score within 24 hours after admission. The number of organ failure was associated with increased mortality. The evaluation of a subgroup of 74 patients who stayed in the ICU for at least 48 hours showed that survivors and non-survivors followed a different course. In this subgroup, the total SOFA score increased in 81% of the non-survivors but in only 21% of the survivors. Conversely, the total SOFA score decreased in 48% of the survivors compared with 6% of the non-survivors. The non-survivors also had a higher APACHE III score within 24 hours and there was a correlation between SOFA score and APACHE III score. Conclusion : The SOFA score is a simple, but effective method to assess organ failure and to predict mortality in critically ill patients. Regular and repeated scoring enables patient's condition and clinical course to be monitored and better understood. The SOFA score well correlates with APACHE III score.

Decrease of the Activation and Carbamylation of Rubisco by High CO2 in Kidney Bean (KidneyBean에서의 고 CO2 농도에 의한 Rubisco의 Activation과 Carbamylation의 감소)

  • 노광수;김재기
    • KSBB Journal
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    • v.11 no.3
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    • pp.295-302
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    • 1996
  • The measurements of rubisco parameters are important in photosynthetic studies. In this experiment, we used photometric assay method to detect these major parameters, such as activity, carbamylation and amount of rubisco. The main advantages of this method are very simple and as sensitive as conventional methods which usually produce radioactive waste. In this study, with kidney bean (Phaseolus vulgatis L.) leaves grown at normal $CO_2$ (350ppm) and high $CO_2$ (650 ppm), we investigated the effect of $CO_2$ concentration on activation and carbamylation of rubisco by measuring the rubisco activity, carbamylation rate and amount of rubisco using a dual beam (334nm and 405nm) spectrophotometer, and analyzed the polypeptide profiles of rubisco by SDS-PAGE. When $CO_2$ concentration was raised from 350ppm to 650ppm, all parameters of rubisco were decreased : $41.2{\mu}M/m^2/s and 52.2{\mu}M/m^2/s$ to $27.4{\mu}M/m^2/s and 46.1{\mu}M/m^2/s$ for initial and total rubisco activity, respectively ; from 79% to 58.9% for carbamylation rate ; from $1.94 {\mu}M/m^2$ to 1.58{\mu}M/m^2$ for amount of rubisco. These results suggests that the decrease in rubisco activity at high $CO_2$ was caused by carbamylation. The analysis of the preparation by SDS-PAGE showed two major polypeptides at 50 and 14.5 kD which were identified as the large and the small subunits of rubisco. There were no differences in the intensity compared high $CO_2$ to normal $CO_2$ in both 50 kD and 14.5 kD bands. We also found that these inhibitory effects of $CO_2$ were reversible. When high $CO_2$ was switched to normal $CO_2$, the parameters of rubisco changed were almost the same as normal rubisco parameters. These data provide an evidence that activity of rubisco was recovered by $CO_2$ concentration of 350 ppm.

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