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.
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.
Vibrio parahaemolyticus (V. parahaemolyticus), which is commonly found in raw seafood, causes gastroenteritis in humans. Rapid and effective methods have been developed as culture methods require up to 5-7 days. In this study, real-time PCR was compared with the standard culture method for detecting V. parahaemolyticus in seafood and radish sprout samples. Five hundred grams of the samples were artificially contaminated with V. parahaemolyticus then divided into 20 samples. The samples were incubated in alkaline peptone water and then streaked onto thiosulfate-citrate-bile saltssucrose agar. Biochemical tests for suspicious colonies were performed using the API 20NE strip. In parallel, real-time PCR was performed targeting the toxR gene using the enrichment broth. The real-time PCR was sensitive in discriminating V. parahaemolyticus from other foodborne pathogens. The detection limit of the real-time PCR was $10^3\;CFU/mL$ in phosphate-buffered saline. Although the real-time PCR detected more positive samples (76 out of 180, 42%) than the culture method (66 out of 180, 37%), there was no significant statistical difference (p>0.05) between the two methods. In conclusion, real-time PCR assays could be an alternative to the standard culture method for detecting V. parahaemolyticus in seafood and radish sprouts, which has many advantages in terms of detection time, labor, and sensitivity.
This experiment was conducted to investigate the effects of concentrated pig slurry using membrane filter on growth of tomato in nutriculture. Pig slurry was filtered by ultra filtration and concentrated by reverse osmosis process. Filtration of pig slurry was necessary to prevent the hose clogging in nutriculture. The concentrated pig slurry (CS) and nutrient solution (NS) were mixed by six different mixing ratios of 0:100, 20:80, 40:60, 60:40%, 80: 20 and 100%:0% based on nitrogen content. The chemical nutrient solution was the solution of National Horticulture Research Station for the growth of tomato. The concentration of nutrient solution was adjusted a range of $1.6{\sim}2.0mS/cm$ in EC. The plant height of tomato treated with CS 20+NS 80% was similar with NS 100% control plot. Plant height was highest in the plot of CS 20+NS 80%. The treatment of 100% concentrated pig slurry was lowest in the gowth characteristics of tomato. Number of cluster was very lower in 100% concentrated pig slurry compared with plot of chemical nutrient solution. In the beginning of growth stage, SPAD reading value was reduced in plot treated with CS 100%, but CS 20+NS 80% plot was higher compared to 100% concentrated pig slurry. SPAD value of tomato leaves was decreased as the amount of CS was increased. The SPAD value also in treatment of concentrated pig slurry was lower in the middle growth stage compared to control plot. The dry weight of stem and leaf were 107.4, 104.2g in plot of NS 100% and CS 20%+NS 80%, respectively. The fruit number and weight were decreased at high application plots of concentrated pig slurry, The fruit setting of tomato showed lowest in the plot treated with 100% concentrated pig slurry, and the growth of tomato severely decreased after application of 100% CS treatment. In conclusion, the growth characteristics such as plant height and fruit weight of tomato were not significantly different between the plots treated with mixture of 20% CS +80%NS and 100% nutrient solution treatment. In conclusion, the mixture solution of 20% of concentrated pig slurry and 80% of nutrient solution could be used as a nutrition solution of tomato nuticulture.
Purpose: This study was undertaken to investigate the feasibility of using automated liquid-handling systems equipped with reusable fixed tips in Radioimmunoassays and to demonstrate that the use of an automated pipetting instrument can decrease in the typical daily workload. Materials and Methods: The precisions of the automated pipettor and the manual pipettor were determined gravimetrically (n=30). A total of 30 specimens for HBs Ag were repeatedly pipetted (three replicates) with the automated pipettor and then retested. PSA samples were simultaneously pipetted with either the automated pipettor or the manual pipettor and then analyzed (n=40). Sample carryover test assessed for CA19-9, AFP and HCG when the automated pipettor was used. Pipetting speed of the automated pipettor and the manual pipettor were compared by evaluation of each workload. Results: The precisions coefficients of variation (CV) were 2.1% for the automated pipettor and 1.6% for manual pipettor. The mean cpm and CV for each group of replicates were 41,203 cpm and 3.7% for HBs Ag positive specimens, and 99 cpm and 7.9% for HBs Ag negative specimens, respectively. PSA results showed no significant differences between automated pipettor and manual pipettor (p=0.15, r=0.999). Carryover for CA19-9, AFP and HCG analytes was <0.1 ppm or below the assay limit of detection. Pipetting speed was significantly improved by using the automated instrument. Conclusion: There was no evidence that the use of an automated pipettor adversely affected any of the performance characteristics of the assay. Indeed, routine use of the Tecan automated pipettor has resulted in a decrease in the typical daily workload.
Journal of the Korean Society of Food Science and Nutrition
/
v.40
no.10
/
pp.1353-1360
/
2011
This study was performed to investigate the antimicrobial effects against food-borne pathogens and antioxidant activity of Rhododendron brachycarpum ethanol-extract. The antimicrobial activity of the extract was determined using a paper disc-diffusion method, and the diameter of the clear zone was measured. The diameter of the clear zone in the presence of 10 mg of extract was maximal against Bacillus cereus among the three tested Gram-positive bacteria and against Escherichia coli O157:H7 among the five tested Gram-negative bacteria. Analysis of the minimum inhibitory concentration (MIC) showed that the extract exhibited a similar efficacy as that of sorbic acid, a well-known chemical preservative. The growth inhibitory effects of the extract at concentrations of 250, 500, 1,000, and 2,000 mg/L on food-borne pathogens were determined against Staphylococcus aureus, Listeria monocytogenes, Salmonella Typhimurium, and Escherichia coli O157:H7. Growth of the microorganisms was not affected by the extract at concentrations up to 250 mg/L, but it was significantly (p<0.05) inhibited by the extract at concentrations higher than 1,000 mg/L. The antioxidant effects of the extract were examined via measurement of DPPH radical scavenging activity, inhibition of reactive oxygen species (ROS) generation using fluorescent dichlorofluorescien (DCF) assay, and prevention of peroxyl radical- and hydroxyl radical-induced supercoiled DNA breakage. The $IC_{50}$ of the extract for DPPH radical scavenging activity was about half that of ${\alpha}$-tocopherol, which was used as a positive control. DCF fluorescence intensity decreased as the concentration of the extract increased, demonstrating that ROS generation was inhibited in a concentration-dependent manner. The ROS inhibitory effect of the extract was higher than that of ascorbic acid. The extract prevented supercoiled DNA strand breakage induced by peroxyl radical and hydroxyl radical. Thus, the results of the present study demonstrate that the extract exhibits antimicrobial effects against food-borne pathogens as well as potent antioxidant capacity, suggesting that R. brachycarpum could be used as a natural antibacterial agent and effective antioxidant in food.
In this study, predictive mathematical models were developed to predict the kinetics of Listeria monocytogenes growth in the mixed fresh-cut vegetables, which is the most popular ready-to-eat food in the world, as a function of temperature (4, 10, 20 and $30^{\circ}C$). At the specified storage temperatures, the primary growth curve fit well ($r^2$=0.916~0.981) with a Gompertz and Baranyi equation to determine the specific growth rate (SGR). The Polynomial model for natural logarithm transformation of the SGR as a function of temperature was obtained by nonlinear regression (Prism, version 4.0, GraphPad Software). As the storage temperature decreased from $30^{\circ}C$ to $4^{\circ}C$, the SGR decreased, respectively. Polynomial model was identified as appropriate secondary model for SGR on the basis of most statistical indices such as mean square error (MSE=0.002718 by Gompertz, 0.055186 by Baranyi), bias factor (Bf=1.050084 by Gompertz, 1.931472 by Baranyi) and accuracy factor (Af=1.160767 by Gompertz, 2.137181 by Baranyi). Results indicate L. monocytogenes growth was affected by temperature mainly, and equation was developed by Gompertz model (-0.1606+$0.0574^*Temp$+$0.0009^*Temp^*Temp$) was more effective than equation was developed by Baranyi model (0.3502-$0.0496^*Temp$+$0.0022^*Temp^*Temp$) for specific growth rate prediction of L.monocytogenes in the mixed fresh-cut vegetables.
Kim, Kyung-Hwan;Moon, In-Sung;Park, Jang-Sang;Koh, Yong-Bok;Ahn, Hyuk
Journal of Chest Surgery
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v.35
no.4
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pp.267-273
/
2002
Background: We performed a phase IV clinical trial to examine the usefulness of a continuous infusion of nicardipine hydrochloride to control hypertension in patients with acute aortic dissection. material and Method: Systolic/diastolic blood pressure, and heart rate were monitored before and after the intravenous administration of nicardipine in 31 patients with aortic diseases. The period of nicardipine administration in each patient was from 3 to 14 days. Efficacy was evaluated by determining the average amount of blood pressure reduction on the 3rd day of drug administration. The dosage of another antihypertensive agent was slowly tapered down, and ultimately replaced by the test drug. Result: 28 patients were diagnosed as acute aortic dissection, 2 patients as rupture of the aortic arch aneurysm, and 1 patient as traumatic aortic rupture. Mean age was 53.9 $\pm$ 14.9(29~89) years, and 21 patients(67.7%) were male. 14 patients(32.3%) had complications associated with underlying aortic disease: aortic insufficiency in 7, hemopericardium in 6, acute renal failure in 1, paraplegia in 1, lower extremity ischemia in 1, and hemothorax in 1. The time needed to reach the target blood pressure was within 15 minutes in 16, from 15 to 30 minutes in 10, from 30 to 45 minutes in 3 and from 45 to 60 minutes in 2, and their baseline average systolic, diastolic, and mean arterial blood pressures(mmHg) were 147$\pm$23, 82.3$\pm$ 18.6, and 104 $\pm$ 18, respectively. Average systolic, diastolic, and mean arterial blood pressures(mmHg) on the third day of nicardipine infusion were 119$\pm$ 12, 69$\pm$9, and 86$\pm$8, and they all showed statistically significant decrease(p<0.05). The average systolic, diastolic, and mean arterial blood pressure(mmHg) after the discontinuation of the nicardipine infusion were 119 $\pm$ 15, 71 $\pm$ 14, and 86$\pm$ 13, respectively. No significant difference was observed between the average pressures measured on the third day and those measured after the discontinuation of the nicardipine infusion, and no definite side effects were observed during the study period. Conclusion: Nicardipine hydrochloride was both effective and safe at controlling blood pressure in patients with acute aortic dissection.
Lung transplantation is the established treatment for the end stage lung disedse find preservation of the organ is a major obstacle In performing lung transplantation. For solving this problem, we evaluated the histopathologic changes for various preservation solutions. Male mongrel dogs of similar size and weight (15∼20 kg) were used. The dog lungs were flushed with 4fl normal saline(group 1 'n:5): Modified Euro-Collins solution(group 2 n:5) and University of Wisconsin solution (group 3 : n=6), 60m11kg through a catheter placed in the main pulmonary artery aft r flushing of PGE 1 (20ng1kg). The lungs were preserved for 60 hours and measured dry and wet weights. Histologic specimens were taken every 6 hours and %toed for light microscopic evaluation. The edema ratio of the lungs peaked in 12 hours although there was no difference between the groups. Histologically, alveolar septal changes developed in one case (20%) after 1 hour preservation with normal saline. In case of the University of Wisconsin solution, the alveolar septal distortions and swellings were seen in 1 cases (20%) after 6 hours preservation compared with 3 cases (60%) after 6 hours preservation with Modified Euro-Collins solution. Changes of the pneumocytes were observed after 24 hours preser- vation in group 1, after 48 hours preservation in group 2 and after 60 hours preservation in group 3. We conclude that University of Wisconsin solution might have a superior preservation effect compare to normal saline and Modified Euro-Collins solutions.
Separating the patient from hypothermic cardiopulmonary bypass(CPB) before achieving adequate rewarming often results in afterdrop, which can predispose to electrolyte disturbances, arrhythmia, hemodynamic alterations, and shivering-induced increase of oxygen consumption. In an attempt to find an adequate end point temperature of rewarming after hypothermic CPB, 50 pediatric cardiac surgical patients were r ndomly assigned for end point temperature of rewarming of 35.5$^{\circ}C$ (Group 1) or 37t (Group 2), rectal temperature. Thereafter the rectal temperature was measured half, one, four, eight, and 16 hour after arrival to the intensive care unit(ICU), with heart rate and blood pressure. Additionally the rectal temperature was compared with esophageal temperature during CPB, and axillary temperature luring stay in the ICU. Nonpulsatile perfusion with a roller pump was used in all patients and a membrane or bubble oxygenator was used for oxygenation. Both groups were comparable with respect to age, sex, body surface area, total bypass time, and rewarming time. There was no afterdrop in both groups, and there were no statistical differences in the rectal temperatures between two groups. There were also no statistical dilyerences with respect to the heart rate and blood pressure between two groups. At the end of rewarming the esophageal temperature was higher than the rectal temperature. The axil ary temperature measured in ICU was always lower than the rectal temperature. No shivering was noted in all patients. In conclusion, with restoration of rectal temperature above 35.5$^{\circ}C$ at the end of CPB in pediatric patients, we did not observe an afterdrop.
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