A specific analytical method able to identify and quantify traces of six glucocorticoids residues in eggs were developed. The extraction and clean-up parameters for simultaneous analysis were evaluated and HPLC and spectrometric conditions were also established. For determination of glucocorticoids, 5 g of egg was transferred into a test tube, adjusted pH 5.2 with acetate buffer and was $\beta$-glucuronidase/arylsulfatase from Helix pomatia added. The mixture was centrifuged and supernatant was extracted twice with 20 mL n-hexane. The extraction was performed with HLB cartridge using methanol, followed by clean-up with silica cartridge using methanol/ethyl acetate (4/6, v/v). The analytes were determined by HPLC/ESI-MS/MS operating in the negative ion mode. Validation studies with fortified egg samples for established method were performed. The result of method validation gave good efficiency, linearity, accuracy and precision. The correlation coefficients ($r^2$) of the calibration curves appeared to be higher than 0.99 in egg, indicating excellent linearity. LOD was ranged 0.09 to $0.17{\mu}g/kg$, and recoveries for most compounds were in the range of 55.7-69.8%. This method can be used to determine ${\mu}g/kg$ levels of glucocorticoids in eggs.
Background: It is well known that preoperative administration of combined antiplatelet agents can have an impact on the postoperative bleeding, the requirement for transfusion and the need for reexploration during on-pump coronary artery bypass surgery. Yet its effects have not been well evaluated in the case of off-pump coronary artery bypass surgery. Material and Method: We performed a retrospective study of nineteen patients who underwent OPCAB from March 2003 to December 2004. All the patients had taken antiplatelet agents until 12 hours before operation. The patients were divided into bo groups as an aspirin group and a combined (aspirin+clopidogrel) group. The perioperative platelet count, the hemoglobin level, the hematocrit, the prothrombin time and the aPTT were compared between both groups. The amount of postoperative bleeding, the transfusion requirement and the need for re-exploration to control bleeding were also compared between both groups. Result: There was no difference of operation time and the intraoperative ACT between the aspirin group and the combined group. The amount of blood loss through the chest tube for 24 hours was not different between the aspirin group $(697{\pm}271mL)$ and the combined group $(944{\pm}432mL)$. The number of patients who received blood transfusion was also not different between both groups. There was no patient who required reexploration for bleeding control in both groups. The perioperative hemoglobin level and hematocrit were also not different between both groups, but the postoperative hemoglobin level and hematocrit were decreased significantly in the group. Conclusion: The Preoperative combined antiplatelet (aspirin+clopidogrel) therapy group was not different from the aspirin group for the amount of postoperative bleeding, the amount of blood transfusion and the need for reexploration during off-pump coronary artery bypass grafting. This subject needs further evaluation because of small population in our study.
BACKGROUND: In order to select a fungicide that can effectively control anthracnose disease in Japanese plum fruit, mycelial growth inhibition effect and spore germination inhibition effect of six fungicides were tested in vitro against six isolates of Colletotrichum acutatum and five isolates of C. gloeosporioides that were isolated from diseased Japanese plum fruit. METHODS AND RESULTS: Inhibitory effects of fungicides on mycelial growth were investigated after inoculating each isolate on potato dextrose agar amended with four discriminatory concentrations of each fungicide for 7 days at $25^{\circ}C$. For spore germination inhibitory effect, each isolate of the Colletotrichum spp. was cultured in potato dextrose agar for 7-14 days at $25^{\circ}C$. After adjusting the concentration of spores of each isolate to $1{\times}10^6mL^{-1}$ by diluting with 0.025% PDB, the spore suspension was mixed with each fungicide (1:4, v/v), and $60{\mu}L$ aliquots were dispensed to sterile hole slide glass. Hole slide glasses were placed in a humidified box and incubated for 15 hours at $25^{\circ}C$. Then, spore germination was observed under an optical microscope. At recommended concentration of fungicide prochloraz manganese showed the highest mycelial growth inhibitory effect and dithianon showed the lowest mycelial growth inhibition. The $EC_{50}$ values for the inhibition of spore germination by dithianon and pyraclostrobin were $0.069-0.126{\mu}g/mL$ and $0.37-1.59{\mu}g/mL$, respectively. Although benomyl, prochloraz manganese, azoxystrobin, and tebuconazole did not inhibit the spore germination, they appeared to restrain mycelial growth by abnormal growth of germ tube and mycelium after germination. CONCLUSION: Dithianon seemed to have preventive effect. Prochloraz manganese, azoxystrobin, and tebuconazole were likely to have control effect. Pyraclostrobin is considered to have both preventive and control effect against anthracnose disease of Japanese plum fruit.
Background : Airway inflammation and hyperresponsiveness are recognized as major characteristics of bronchial asthma. Airway inflammation has usually been assessed by invasive methods, e.g. BAL or bronchial biopsy, but recent studies proposed induced sputum as another reliable and non-invasive tool to investigate airway inflammation in asthmatic patients. Thus, the relationship between airway inflammation assessed by induced sputum and airway hyperresponsiveness was investigated in asthmatic patient. Method : Airway responsiveness was determined by the concentration that caused a 20% decrease in $FEV_1$($PC_{20}$) after inhaling incremental concentrations of methacholine. The numbers of inflammatory cells and the concentration of eosinophilic cationic protein(ECP) were assessed in induced sputum obtained by inhalation of hypertonic saline(3%). Result: We analyzed sputum induced in 15 stable asthmatic patients. The differential cell count(%) of macrophages, neutrophils, eosinophils and lymphocytes in induced sputum were $39.1{\pm}27.0%$, $29.6{\pm}21.0%$, $28.8{\pm}18.8%$, $1.3{\pm}3.1%$ respectively. The mean value of baseline FEV1(predicted) and ECP were $76.3{\pm}30.3%$ and $1,101{\pm}833{\mu}g/L$ respectively. The geometric mean value of $PC_{20}$ was 0.56 mg/mL. The relationships between the sputum eosinophil and ECP in induced sputum, and between sputum eosinophil and degree of airway responsiveness($PC_{20}$) were found to be significantly correlated (r=0.81, p<0.05 and r=-0.78, p<0.05, respectively). Sputum neutrophils and $PC_{20}$ were not correlated to each other (r=0.11, p=0.69) and a significant negative correlation was found between ECP and baseline $FEV_1$(predicted)(r=-0.62, p<0.05). Conclusion : The results of this study suggest that an induced sputum via a inhalation of hypertonic saline is useful to determine a patient's status of airway inflammation, and airway inflammation is one of the major causal factors in the development of bronchial hyperresponsiveness in asthmatic patients.
Background: Extubation is recommended to be performed at minimum pressure support (PSmin) during the pressure support ventilation (PSV). In field, physicians sometimes perform additional 1 hr T-piece trial to the patient at PSmin to reduce re-intubation risk. Although it provides confirmation of patient's breathing reserve, weaning could be delayed due to increased airway resistance by endotracheal tube. Methods: To investigate the effect of additional 1 hr T-piece trial on weaning outcome, a prospective study was done in consecutive 44 patients who had received mechanical ventilation more than 3 days. Respiratory mechanics, hemodymic, and gas exchange measurements were done and the level of PSmin was calculated using the equation (PSmin=peak inspiratory flow rate $\times$ total ventilatory system resistance) at the 15cm $H_2O$ of pressure support. At PSmin, the patients were randomized into intervention (additional 1 hr T-piece trial) and control (extubation at PSmin). The measurements were repeated at PSmm, during weaning process (in cases of intervention), and after extubation. The weaning success was defined as spontaneous breathing more than 48hr after extubation. In intervention group, failure to continue weaning process was also considered as weaning failure. Results: Thirty-six patients with 42 times weaning trial were satisfied to the protocol. Mean PSmin level was 7.6 (${\pm}1.9$)cm $H_2O$. There were no differences in total ventilation times (TVT), APACHE III score, nutritional indices, and respiratory mechanics at PSmin between 2 groups. The weaning success rate and re-intubation rate were not different between intervention group (55% and 18% in each) and control group (70% and 20% in each) at first weaning trial. Work of breathing, pressure time product, and tidal volume were aggravated during 1 hr T-piece trial compared to those of PSmin in intervention group ($10.4{\pm}1.25$ and $1.66{\pm}1.08$ J/L in work of breathing) ($191{\pm}232$ and $287{\pm}217$cm $H_2O$ s/m in pressure time product) ($0.33{\pm}0.09$ and $0.29{\pm}0.09$ L in tidal volume) (P<0.05 in each). As in whole, TVT, and tidal volume at PSmin were significantly different between the patients with weaning success ($246{\pm}195$ hr, $0.43{\pm}0.11$ L) and the those with weaning failure ($407{\pm}248$ hr, $0.35{\pm}0.10$L) (P<0.05 in each). Conclusion : There were no advantage to weaning outcome by addition of 1 hr T-piece trial compared to prompt extubation to the patient at PS min.
A duplex RT-PCR (dRT-PCR) assay was developed for the simultaneous detection and discrimination of non-virulent and virulent Newcastle disease virus (NDV) in a single PCR tube. Primers targeting the large polymerase protein (L) gene and the fusion protein (F) gene of NDV were designed to detect all NDVs (by common type PCR primers) and virulent NDVs (by pathotype PCR primers), respectively and evaluated experimentally with reference NDV strains and other poultry viral pathogens. PCR products of the expected size of 386 bp were amplified from all NDV samples whereas PCR products of the expected size of 229 bp were amplified from virulent NDV samples alone. Cross reaction was not observed with other avian viral pathogens. The detection limit of NDV by the dRT-PCR was estimated to be $10^3$ 50% egg infectious dose/0.1 mL. In the dRT-PCR using field isolates of NDV, the pathotype PCR primers detected specifically all of virulent field isolates of NDV from Malaysia, Pakistan and China whereas common type PCR primers detected 94.4% (51/54) of field isolates of NDV from China. Three Chinese NDV isolates with false negative result were non-virulent viruses. Our results indicate that the dRT-PCR might provide a rapid and simple tool for rapid simultaneous detection and discrimination of non-virulent and virulent NDVs. Therefore the developed dRT-PCR assay provides a powerful novel means for the rapid diagnosis of Newcastle disease.
The main purpose of this research is to prepare and provide basic materials for the propagational strategy of eelgrass by investigating on the morphological adaptation of Korean Zostera marina to ocean currents. An eelgrass plant mainly consists of rhizome, leaf sheath, leaves and roots. The rhizome is the horizontal stem of the plant that serves as the backbone from which the leaves and roots emerge. The leaf sheath is the bundle at the base of the leaves that holds the leaves together, protecting the meristem, the primary growth point of the shoot. Leaves originate from a meristem which is protected by a sheath at the actively growing end of the rhizome. As the shoot grows, the rhizome elongates, moving across or within the sediment, forming roots as it progresses. The aggregated leaves from the leaf sheath are found to have two cell layers on one side and multiple layers of airy tissues called aerenchyma on the other. The aerenchyma tissues are developed in multi-layered cell structures surrounding the veins which are formed in the leaf sheath. Generative shoots are made of rhizomes, which are circular or ovoidal, stem, and spathe and spadix. The transverse section of rhizome and the stem and central floral axis is found to be circular, ovoid and in the shape of convex respectively, and the vascular bundle, which is a part of transport system, has one large tube in the center and two small tubes on both sides. The layers of collenchyma cells numbered from 12 to 15 in the stem, and from 7 to 12 in the rhizome. The seed coat is composed of sclereids, small bundles of sclerenchyma tissues, which prevent the influx of sea water from the outside and help endure the environmental stress. In conclusion, alternative multi-layer structure in circular, convex type aggregated leaf base are interpreted to morphological adaption as doing tolerable elastic structure through movement of seawater. The generative shoots develop long slim stem and branches in circular or ovoidal shapes to minimize the adverse impacts of sea current, which can be interpreted as the plant's morphological adaptation to its environment.
In this study, the atmospheric vertical structure (AVS) associated with summertime (June, July, and August) heavy rainfall in Seoul was classified into three patterns (Loaded Gun: L, Inverted V: IV, and Thin Tube: TT) using rawinsonde soundings launched at Osan from 2009 to 2018. The characteristics of classified AVS and precipitation property were analyzed. Occurrence frequencies in each type were 34.7% (TT-type), 20.4% (IV-type), 20.4% (LG-type), and 24.5% (Other-type), respectively. The mean value of Convective Available Potential Energy (1131.1 J kg-1) for LG-types and Storm Relative Helicity (357.6 ㎡s-2) for TT-types was about 2 times higher than that of other types, which seems to be the difference in the mechanism of convection at the low level atmosphere. The composited synoptic fields in all cases showed a pattern that warm and humid southwesterly wind flows into the Korean Peninsula. In the cases of TT-type, the low pressure center (at 850 hPa) was followed by the trough in upper-level (at 500 hPa) as the typical pattern of a low pressure deepening. The TT-type was strongly influenced by the low level jet (at 850 hPa), showing a pattern of connecting the upper- and low-level jets. The result of analysis indicated that precipitation was intensified in the first half of all types. IV-type precipitation induced by thermal instability tended to last for a short term period with strong precipitation intensity, while TT-type by mechanical instability showed weak precipitation over a long term period.
Purpose: Adipose tissue is located beneath the skin, around internal organs, and in the bone marrow in humans. Its main role is to store energy in the form of fat, although it also cushions and insulates the body. Adipose tissue also has the ability to dynamically expand and shrink throughout the life of an adult. Recently, it has been shown that adipose tissue contains a population of adult multipotent mesenchymal stem cells and endothelial progenitor cells that, in cell culture conditions, have extensive proliferative capacity and are able to differentiate into several lineages, including, osteogenic, chondrogenic, endothelial cells, and myogenic lineages. Materials and Methods: This study focused on endothelial cell culture from the adipose tissue. Adipose tissues were harvested from buccal fat pad during bilateral sagittal split ramus osteotomy for surgical correction of mandibular prognathism. The tissues were treated with 0.075% type I collagenase. The samples were neutralized with DMEM/and centrifuged for 10 min at 2,400 rpm. The pellet was treated with 3 volume of RBC lysis buffer and filtered through a 100 ${\mu}m$ nylon cell strainer. The filtered cells were centrifuged for 10 min at 2,400 rpm. The cells were further cultured in the endothelial cell culture medium (EGM-2, Cambrex, Walkersville, Md., USA) supplemented with 10% fetal bovine serum, human EGF, human VEGF, human insulin-like growth factor-1, human FGF-$\beta$, heparin, ascorbic acid and hydrocortisone at a density of $1{\times}10^5$ cells/well in a 24-well plate. Low positivity of endothelial cell markers, such as CD31 and CD146, was observed during early passage of cells. Results: Increase of CD146 positivity was observed in passage 5 to 7 adipose tissue-derived cells. However, CD44, representative mesenchymal stem cell marker, was also strongly expressed. CD146 sorted adipose tissue-derived cells was cultured using immuno-magnetic beads. Magnetic labeling with 100 ${\mu}l$ microbeads per 108 cells was performed for 30 minutes at $4^{\circ}C$ a using CD146 direct cell isolation kit. Magnetic separation was carried out and a separator under a biological hood. Aliquous of CD146+ sorted cells were evaluated for purity by flow cytometry. Sorted cells were 96.04% positivity for CD146. And then tube formation was examined. These CD146 sorted adipose tissue-derived cells formed tube-like structures on Matrigel. Conclusion: These results suggest that adipose tissue-derived cells are endothelial cells. With the fabrication of the vascularized scaffold construct, novel approaches could be developed to enhance the engineered scaffold by the addition of adipose tissue-derived endothelial cells and periosteal-derived osteoblastic cells to promote bone growth.
With the improvement of living standard and socioenvironmental change, esophageal stricture due to acetic acid, Hcl, insecticides and lye has led to decrease remarkably. Esophageal stricture has decreased because of immediate treatment and adequate management. However there are still sporadic incidents of esophageal stricture due to inadequate treatment and uncooperation of patients. The authors recently have experienced seven severe cases of esophageal stricture. All six patients were treated with 18-51 French Sippy esophageal dilating bougie. One patient was transfered to thoracic surgery department due to complete esophageal obstruction. Case 1. A 23 year old housewife who ingested Hcl for suicide. After Witzel's operation, she visited to dilate esophageal first constriction stircture due to swallowing difficulty postoperation 2 months later. We were treated successfully. Case 2. A case of esophageal stricuture in the second and third physiologic constriction part. The patient was 51 years old man who ingested lye accidently, and was dilated by bougination. Case 3. The patient was 43 year-old man who ingested acetic acid as a mistake and was treated inadequately at hopsital. Inspite of treatment, esophageal stricture developed at the third physiologic part. We are trying to dilate the esophagues now. Case 4. The patient was 55 year-old woman who had ingested Hcl for the purpose of suicidal attempt. 2 months later gastroduodenal anastomoisis due to pyloric region stenosis, the first physiologic constriction stricture was dilated successfully. Case 5. The patient was 41 year-old woman who ingested Hcl for suicide 4 months ago. There was indwelled orogastric tube for 1 month but esophageal stricture developed at the first and 3rd constriction part. She was treated by using a bougination. Case 6. An athlete 21 year old man, ingested acetic acid 2 spoonful per 3 days by purpose in order to soften the bone for last 14 months. There was complete esophageal obsturction in esophagogram and transferred to the thoracic surgery department. Case 7. A 3 year-old girl was ingested lye at a accident. She had a bougination for 16 months under the general anesthesia for dilation due to whole irregular esophageal stricture. She developed lower esophageal perforation, but healed eventually.
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