• Title/Summary/Keyword: Positive stain

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The Diagnostic Usefulness of Endoscopic Findings and Detection Rates of CLO and HpKit Test for Helicobacter pylori Infection in Children (소아의 Helicobacter pylori 감염에 있어서 위내시경 소견의 진단적 의의와 CLO 및 HpKit 검사의 검출률)

  • Lee, Seung Yeon;Yoo, Ji Hyung;Chung, Ki Sup
    • Clinical and Experimental Pediatrics
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    • v.45 no.4
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    • pp.482-488
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    • 2002
  • Purpose : We performed this study to evaluate the diagnostic usefulness of endoscopic finding of nodular gastritis, CLO and HpKit test for H. pylori infection in children. Methods : Gastroduodenal endoscopy and mucosal biopsy were performed on 212 children who visited our hospital between Jul. 1999 and May 2000 due to abdominal pain. We performed CLO and HpKit test for H. pylori with the time interval of 15, 30 minutes, 1, 2, 3, 24, 48, 72, 96, 120 and 144 hours. Histological examination of H. pylori was made by H-E or Alcian yellow stain with biopsy specimens. Sensitivity, specificity, positive predictive and negative predictive value of nodular gastritis, CLO and HpKit test were calculated from the analysis of above data. Results : Sensitivity and specificity of 3 hour-CLO test was 68.4% and 100% respectively. Sensitivity and specificity of 3 hour-HpKit test was 65.8% and 100% respectively. No significant difference in sensitivity and specificity was found between in 3 hour-CLO and HpKit test(P>0.05). Sensitivity of CLO test increased as time lapsed, but corresponding specificity did not decrease as time lapsed(sensitivity and specificity at 144 hours : 89.5% and 94.8% respectively). However, sensitivity of HpKit test increased as time lapsed, but specificity markedly decreased. Sensitivity and specificity of the nodular gastritis was 78.9% and 93.7% respectively. Conclusion : Both CLO and HpKit test have relatively low sensitivity and specificity for the detection of H. pylori in 3 hours of testing in children. The endoscopic finding of nodular gastritis is another good standard in the diagnosis of H. pylori infection in children.

Induction of Autophagy by Low Dose of Cisplatin in H460 Lung Cancer Cells (폐암세포주에서 저용량 시스플라틴에 의해 유도된 자가포식)

  • Shin, Jeong-Hyun;Jang, Hye-Yeon;Chung, Jin-Soo;Cho, Kyung-Hwa;Hwang, Ki-Eun;Kim, So-Young;Kim, Hui-Jung;Lee, Sam-Youn;Lee, Mi-Kung;Park, Soon-Ah;Moon, Sun-Rock;Lee, Kang-Kyu;Jo, Hyang-Jeong;Yang, Sei-Hoon
    • Tuberculosis and Respiratory Diseases
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    • v.69 no.1
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    • pp.16-23
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    • 2010
  • Background: Most lung cancer patients receive systemic chemotherapy at an advanced stage disease. Cisplatin-based chemotherapy is the main regimen for treating advanced lung cancer. Recently, autophagy has become an important mechanism of cellular adaptation under starvation or cell oxidative stress. The purpose of this study was to determine whether or not autophagy can occurred in cisplatin-treated lung cancer cells. Methods: H460 cells were incubated with RPMI 1640 and treated in $5{\mu}M$ or $20{\mu}M$ cisplatin concentrations at specific time intervals. Cells surviving cisplatin treatment were measured and compared using an MTT cell viability assay to cells that underwent apoptosis with autophagy by nuclear staining, apoptotic or autophagic related proteins, and autophagic vacuoles. The development of acidic vascular organelles was using acridine orange staining and fluorescent expression of GFP-LC3 protein in its transfected cells was observed to evaluate autophagy. Results: Lung cancer cells treated with $5{\mu}M$ cisplatin-treated were less sensitive to cell death than $20{\mu}M$ cisplatin-treated cells in a time-dependent manner. Nuclear fragmentation at $5{\mu}M$ was not detected, even though it was discovered at $20{\mu}M$. Poly (ADP-ribose) polymerase cleavages were not detected in $5{\mu}M$ within 24 hours. Massive vacuolization in the cytoplasm of $5{\mu}M$ treated cells were observed. Acridine orange stain-positive cells was increased according in time-dependence manner. The autophagosome-incorporated LC3 II protein expression was increased in $5{\mu}M$ treated cells, but was not detected in $20{\mu}M$ treated cells. The expression of GFP-LC3 were increased in $5{\mu}M$ treated cells in a time-dependent manner. Conclusion: The induction of autophagy occurred in $5{\mu}M$ dose of cisplatin-treated lung cancer cells.

Clincal Manifestations of Patients Dying of Severe Community Acquired Pneumonia (중증 원외폐렴으로 사망한 환자의 임상적 분석)

  • Choi, Won-Il;Sohn, Jeong-Ho;Kwun, Oh-Yong;Heo, Jeong-Sook;Whang, Jae-Seok;Han, Seong-Beom;Jeon, Young-June
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.5
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    • pp.537-545
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    • 1994
  • Background: In 1987, the British Thoracic Society (BTS) subjected an extensive list of patient variables to statistical analysis in a prospective study of prognosis in 453 adults with community-acquired pneumonia and, subsequently published guidelines for management of severe community acquired pneumonia. It was hoped that those at risk of dying from community acquired pneumonia could be easily identified and treated appropriately, thereby reducing mortality. To date, severe community acquired pneumonia has not been well studied in Korea. Therefore, we studied retrospectively 10 patients dying of severe community acquired pneumonia in Dongsan Hospital to see clinical manifestations of dying of severe community-acquired pneumonia. Methods: Between July 1987 and july 1993, 498 patients were admitted to Keimyung University Dongsan Hospital with community acquired pneumonia, and 77 of them received intensive care. Of the 77 patients, 10 patients died. We reviewed medical records of these patients. Results: 1) The mean age of the patients was 56.2 years(range, 25 to 75 years). There were 7 men and 3 women. Seven patients(70%) were older than 60years of age. 2) The clinical features on admission were as follows: tachypnca, hypoxemia, mental change, cyanosis, leukopenia, leukocytosis, azotemia, hypotension, hypoalbuminemia in order of frequency. Three patients had one abnormal physical finding, 3 patients had 2, 2 patients had 3, and 2 patients had none of these abnormal physical findings. All patients had at least one of the abnormal laboratory findings. 3) A potential bacterial pathogen was isolated in sputum culture from 2 patients. One was E.coli, the other Enterobacter species. Sputum stain were positive in eight cases (G(+)cocci in six, G(+) cocci and G(-)bacilli in two). 4) Features of respiratory failure were the main reasons for ICU transfer, but two patients were transferred only following a cardiac or respiratory arrest in the general ward. 5) The mean of 2.7 different antibiotics were given to the patients. The aminoglycoside and first generation cephalosporin were the most frequently prescribed antibiotics, followed by the third generation cephalosporin and vancomycin. The most frequently prescribed antibiotics combination was a 1st generation cephalosporin plus an aminoglycoside. 6) Seven patients death(70%) occured after admission within the first five days, and a mean duration of hospitaliztion was 11.2 days. Conclusion: As the results show most death occured within the first five days after admission and aged patients; consequently, an aggressive intensive treatment should be provided early to the patients with severe community acquired pneumonia, and we should pay more attention to the aged patients.

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The Effect of 2-Week Dual Therapy with Proton Pump Inhibitor and Amoxicillin in the Eradication of Helicobacter pylori Infection for Treatment of Children with Recurrent Abdominal Pain (만성 반복성 복통 환아에서 Helicobacter pylori 감염에 대한 Proton Pump Inhibitor 및 Amoxicillin 2주 치료 효과)

  • Lee, Young-Mock;Lee, Chang-Han;Chung, Ki-Sup
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.3 no.1
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    • pp.23-29
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    • 2000
  • Purpose: It has recently been recognized that Helicobacter pylori (H. pylori) is an important factor in the pathogenesis of recurrent abdominal pain (RAP) in children. But, the best treatment for H. pylori infection is still unsettled. This study was performed to evaluate the efficacy of 2 weeks dual therapy with proton pump inhibitor (PPI) and amoxicillin for children with H. pylori infection associated with RAP. Method: Our study included 24 children with RAP who were H. pylori positive assessed by CLO test and histologic examination (silver stain). We used the regimen consisted of PPI (omeprazole, 0.7 mg/kg/day) and amoxicillin (50 mg/kg/day) for 2 weeks to eradicate H. pylori. Eradication of H. pylori was determined 4 weeks after the termination of treatment using the CLO test and histologic examination. Results: The endoscopic diagnoses of patients were nodular gastritis in 11 cases, superficial gastritis in 7 cases, peptic ulcer in 4 cases and normal finding in 2 cases. H. pylori was eradicated in 12 cases by omeprazole and amoxicillin dual therapy for 2 weeks and the eradication rate was 50%. In 4 of 12 children in whom H. pylori had not been eradicated with that regimen, we successfully eradicated H. pylori with other regimens of which 2 or 3 drugs among omeprazole, amoxicillin, clarithromycin, colloidal bismuth subcitrate ($Denol^{(R)}$) and metronidazole were used. Conclusion: The dual therapy with PPI and amoxicillin for 2 weeks had no clear advantage over other regimens for the eradication of H. pylori infection in children. We concluded that the combi-nation of PPI and amoxicillin for 2 weeks is not so good for H. pylori eradication as other commonly used regimens.

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Effects of Matrix Metalloproteinase Inhibitor on Ventilator-Induced Lung Injury in Rats (기계환기로 인한 백서의 급성 폐손상에서 Matrix Metalloproteinase Inhibitor의 효과)

  • Kim, Je-Hyeong;Park, Soo-Yeon;Hur, Gyu-Young;Lee, Seung-Heon;Lee, Sang-Yeub;Park, Sang-Myeon;Suh, In-Bum;Shin, Chol;Shim, Jae-Jeong;In, Kwang-Ho;Kang, Kyung-Ho;Yoo, Se-Hwa
    • Tuberculosis and Respiratory Diseases
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    • v.53 no.6
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    • pp.619-634
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    • 2002
  • Background : Many inflammatory mediators and collagenases are involved in the pathogenesis of acute lung injury (ALI) and acute respiratory distress syndrome (ARDS). The increase of matrix metalloproteinase-9 (MMP-9, gelatinase-B) produced mainly by inflammatory cells was reported in many ALI models and connective tissue cells. In this study, the expression of MMP-9 in ventilator-induced lung injury (VILI) model and the effects of matrix metalloproteinase inhibitor (MMPI) on VILI were investigated. Methods : Eighteen Sprague-Dawley rats were divided into three groups: low tidal Volume (LVT, 7mL/Kg tidal volume, 3 $cmH_2O$ PEEP, 40/min), high tidal volume (HVT, 30mL/Kg tidal volume, no PEEP, 40/min) and high tidal volume with MMPI (HVT+MMPI) groups. Mechanical ventilation was performed in room air for 2 hours. The 20 mg/Kg of CMT-3 (chemically modified tetracycline-3, 6-demethyl 6-deoxy 4-dedimethylamino tetracycline) was gavaged as MMPI from three days before mechanical ventilation. The degree of lung injury was measured with wet-to-dry weight ratio and acute lung injury score. Expression of MMP-9 was studied by immunohistochemical stain with a mouse monoclonal anti-rat MMP-9 $IgG_1$. Results : In the LVT, HVT and HVT+MMPI groups, the wet-to-dry weight ratio was $4.70{\pm}0.14$, $6.82{\pm}1.28$ and $4.92{\pm}0.98$, respectively. In the HVT group, the ratio was significantly higher than other groups (p<0.05). Acute lung injury score measured by five-point scale was $3.25{\pm}1.17$, $12.83{\pm}1.17$ and $4.67{\pm}0.52$, respectively. The HVT group was significantly damaged by VILI and MMPI protects injuries by mechanical ventilation (p<0.05). Expression of MMP-9 measured by four-point scale was $3.33{\pm}2.07$, $12.17{\pm}2.79$ and $3.60{\pm}1.95$, respectively, which were significantly higher in the HVT group (p<0.05). Conclusion : VILI increases significantly the expression of MMP-9 and MMPI prevents lung injury induced by mechanical ventilation through the inhibition of MMP-9.

Immunoelectron Microscopic Study on the Paneth Cell of Rabbit after the Common Bile Duct Ligation (총담관결찰후 집토끼 Paneth세포의 변화에 대한 면역전자현미경적 연구)

  • Park, Kyung-Ho;Cho, Hwee-Dong;Yang, Nam-Gil;Ahn, E-Tay;Ko, Jeong-Sik;Kim, Jin-Gook
    • Applied Microscopy
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    • v.24 no.2
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    • pp.78-92
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    • 1994
  • Lysozyme has been reported to be present in the secretory granules of the Paneth cell, and lysozyme immunoreactivity has been detected by immunogold method in Paneth cells of the intestine of human, mouse and rat. The present study was aimed at clarifying the intracellular distribution and changes of the lysozyme immunoreactivity in rabbit Paneth cell after common bile duct ligation of rabbit, using the electron microscope immunogold technique. Healthy adult rabbits weighing about 2kg body weight were divided into normal and bile duct ligated groups. Common bile duct ligation was performed aseptically under ether anesthesia. Experimental animals were sacrificed on the 1st, the 3rd, the 5th, the 7th and the 14th day after the operation. Mucosal specimens from the intestinal gland of ileum were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde, followed by 1% osmium tetroxide, embedded in araldite mixture, cut with LKB-V ultratome. Ultrathin sections were placed on parlodion coated nickel grids (200mesh). The section-bearing grids were floated upside down on the added substance in a moist chamber at room temperature except for the primary antibody step, which was at $4^{\circ}C$. Sections were etched with a saturated solution of sodium m-periodate for 60min. After etching, sections were pretreated with 0.02M tris buffered saline (TBS), pH 8.4, with 1% bovine serum albumin (BSA, Sigma) for 60min, then treated polyclonal rabbit anti-human lysozyme (Dakipatts) diluted 1 : 50 in TBS with 0.1% BSA for 20hr. Subsequently, grids were incubated 60min in biotinylated goat anti rabbit IgG (Amersham) diluted 1 : 100 in TBS with 0.1% BSA. After this, sections were incubated 60min on streptavidin gold G10 (Amersham) diluted 1 : 50 in TBS with 0.1% BSA. After each step, the grids were briefly rinsed with TBS with 0.1% BSA. After the strepavidin gold step, the sections were jet washed with distilled water. Counterstain of the sections performed by uranyl acetate and lead citrate, and observed with JEM 100 CX II electron microscope. Observed results were as follow; 1. Secretory granules of mouse Paneth cells have a lysozyme immunoreactivity and also eosinophil leucocyte of rabbit applied for the positive-control stain, are well labeld with gold particles. 2. Normal rabbit Paneth cells have a lysozyme immunoreactivity restricted on the secretory granules. 3. Amount lysosomes containing myelin figures in the Paneth cells were significantly increased from 5th day after the common bile duct ligation. 4. Immunoreactivity of Paneth cell secretory granules were more activated on the 3rd day after the common bile duct ligation as compared with those of the normal animal. But the lysozyme immunoreactivity were decreased from the 5th day after the common bile duct ligation. 5. Considering the above finding, lysozyme contained Paneth cell are affected following of common bile duct ligation, whereas lysosomes containing myelin-figure do not exhibit any immunoreactive relationship with those of secretory granules.

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Expression of Hypoxia-inducible Factor-1 $\alpha$ in Esophageal Squamous Cell Carcinoma: Relationship to Prognosis and Tumor Biomarkers (식도 편평세포암에시 Hypoxia-inducible Factor-1 $\alpha$의 발현: 예후와 종양표지자와의 상관성)

  • 양일종;김종인;이해영;천봉권;조성래
    • Journal of Chest Surgery
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    • v.37 no.8
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    • pp.691-701
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    • 2004
  • Background: Tissue hypoxia is a characteristic of many human malignant neoplasms, and hypoxia inducible factor-1 (HIF-1) plays a pivotal role in essential adaptive response to hypoxia, and activates a signal pathway for the expression of the hypoxia-regulated genes, resulting in increased oxygen delivery or facilitating metabolic adaptation to hypoxia. Increased level of HIF-1 a has been reported in many human malignancies, but in esophageal squamous cell carcinoma, the influence of HIF-1 a on tumor biology, including neovascularization, is not still defined. Material and Method: The influence of HIF-1 a expression on angiogenic factors, correlation between the tumor proliferation and HIF-1 a expression, interaction of HIF-1 a expression and p53, and correlation between HIF-1 a expression and clinicopathological prognostic parameters were investigated, using immunohistochemical stains for HIF-1 a, VEGF, CD34, p53, and Ki-67 on 77 cases of resected esophageal squamous cell carcinoma. Result: HIF-1 a expression in cancer cells was found in 33 of 77 esophageal squamous cell carcinoma cases. The 33 cases (42.9%) showed positive stain for HIF-1 a. High HIF-1 a expression was significantly associated with several pathological parameters, such as histologic grade (p=0.032), pathological TMN stage (p=0.002), the depth of tumor invasion (p=0.022), regional lymph node metastasis (p=0.002), distant metastasis (p=0.049), and lymphatic invasion (p=0.004). High HIF-1 a expression had significant VEGF immunoreactivity (p=0.008) and Ki-67 labeling index (p<0.001), but was not correlated with microvascular density within tumors (p=0.088). The high HIF-1 a expression was correlated with aberrant p53 accumulation with a marginal significance (p=0.056). The overall 5-year survival rate was 34.9%. The survival rate of patients with a high HIF-1 a expression was worse than that of patients with low-expression tumors (log-rank test, p=0.0001). High HIF-1 a expression was independent unfavorable factors although statistical significance is marginal in multivariate analysis. Conclusion: It is suggested that (1) high HIF-1 a expression in esophageal squamous cell carcinoma is associated with tumor hypoxia, or with genetic alteration in early carcinogenesis and progressive stages, (2) high HIF-1 a expression may be associated with intratumoral neovascularization through HIF-VEGF pathway, and (3) high HIF-1 a expression is associated with poor prognosis in patients with esophageal squamous cell carcinoma and may playa role as biomarker for regional lymph node metastasis.

An Analysis for Effects of Stain Family Drugs on Osteogenic Differentiation using Human Periosteum-derived Mesenchymal Stem Cells (스타틴(statin) 약물이 성체줄기세포의 골분화에 미치는 영향)

  • Moon, Dong Kyu;Yun, Jeong-Won;Kim, Bo Gyu;Lee, A Ram;Moon, Sun Young;Byun, June-Ho;Hwang, Sun-Chul;Woo, Dong Kyun
    • Journal of Life Science
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    • v.29 no.12
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    • pp.1337-1344
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    • 2019
  • Osteoporosis is characterized by a reduction in bone mass and typically manifests as an increase in fractures. Because this disease is common in elderly populations and lifespans are rapidly increasing, the incidence of osteoporosis has also grown. Most drugs currently used for osteoporosis treatment target osteoclasts in the bone tissue to prevent absorption. However, these medications also cause certain side effects and, furthermore, cannot increase bone mass. Thus, in order to control osteoporosis, regenerative medicine that utilizes adult stem cells and osteoblasts has been extensively studied. Statins, also known as 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors, are cholesterol-lowering drugs that have been widely prescribed for cardiovascular diseases. Interestingly, recent studies have reported the beneficial effects of various statins on bone formation via the activation of osteoblasts. Thus, the current study investigated the effects of seven statin-family drugs on osteoblast activity during osteogenic differentiation using adult stem cells from human periosteal tissue. Specifically, statin effects on alkaline phosphatase activity, an early marker of bone cell differentiation, and on calcium deposit, a late marker of bone cell differentiation, were assessed. The results demonstrate that some statins (for example, pitavastatin and pravastatin) have a weak but positive effect on bone formation, and the findings therefore suggest that statin treatments can be a novel modulator for osteogenic differentiation and regenerative medicine using periosteal stem cells.

The pathogenicity of Streptococcus parauberis isolated from cultured olive flounder Paralichthys olivaceus (양식 넙치, Paralichthys olivaceus에서 분리한 연쇄구균증 원인균, Streptococcus parauberis의 병원성)

  • Choi, Hee-Jung;Cho, Mi-Young;Lee, Jae-Il;Kwon, Mun-Gyeong;Choi, Dong-Lim;Kim, Jin-Woo;Han, Myoung-Chul;Lee, Deok-Chan
    • Journal of fish pathology
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    • v.22 no.3
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    • pp.263-273
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    • 2009
  • There is an increasing number of reports describing Streptococcus parauberis as an important pathogen of cultured olive flounder Paralichthys olivaceus and starry flounder Platichthys stellatus in Korea. We tried to determine the effects of water temperature (14${^{\circ}C}$ and 21${^{\circ}C}$) on the pathogenicity in Streptococcal disease caused by S. parauberis. We have challenged 180 olive flounder by i.p injection to $2.0{\times}10^{7}$ live cells/fish. Mortality was monitored for 21 days post challenge. And histopathological characterizations as infection degree, tissue degeneration and/or bacterial distribution were investigated with H&E stain and in situ hybridization technique. Fifty percent and 16.7% of mortality occurred within 21 days at 21${^{\circ}C}$ and 14${^{\circ}C}$ water temperature, respectively. In most cases, the typical symptoms of olive flounder infected with S. parauberis were darkness of the skin, lethargy, mild abdominal distension cause by ascites, splenomegaly, congested liver and internal organs paleness. The pericardial sac contained large amounts of cloudy fluid. Numerous whitish nodules, which were variable in size and often confluent, were randomly scattered throughout the myocardium. Especially, pericarditis and/or myocarditis was observed in all tested fishes after death. Positive in reaction with S. parauberis were found in all tissues in situ hybridization analysis. The relative numbers of S. parauberis in heart were much more than in liver, spleen, kidney and stomach. We evaluated that S. parauberis strain causes serious damage in the pericardium, shortness of breath and the blood disorder. Therefore, pericarditis and myocarditis caused by S. parauberis were closely related to mortality of olive flounder.

Isolation and Identification of Microorganisms Producing the Soy Protein-Hydrolyzing Enzyme from Traditional Mejus (전통메주로부터 대두단백질 가수분해효소 생산성 미생물의 분리 및 동정)

  • Kang, Min-Jung;Kim, Seong-Ho;Joo, Hyun-Kyu;Lee, Gap-Sang;Yim, Moo-Hyun
    • Applied Biological Chemistry
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    • v.43 no.2
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    • pp.86-94
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    • 2000
  • In order to develop the enzymatic hydrolysis system concerned with taste and flavor, strains having the high hydrolyzing activity on the soy protein were selected from some traditional Mejus. Two molds and one bacterium producing enzymes which were different in character of hydrolysis were isolated and identified. Leucine and azodye enzyme activities of both M4 and M5 were relatively high among in the isolated molds. And, leucine enzyme activity of B16 was the lowest in the isolated bacteria. These strains were isolated as microorganisms having a dissimilar hydrolysis pattern on the soy protein by enzymatic reactions. Mold M4 on the culture solid media was mycelium colors of white and its sclerotia colors were changed from white to black. According to the result of slide culture, radial conidial head, subclavate vesicle, conidia of subglobose, stipes of uncolored with smooth walls and metula and phialides were existed. Because M4 was taxonomically similar to the characteristics of Aspergillus oryzae (ahlburg) species, M4 was identified and named as Aspergillus oryzae M4.Mold M5 showed white and black mycelium on the MEA medium. Mold M5 colony exhibited grayish-green color and have long(7 mm) sporangiophores at slide culture. Sporangia became brownish-gray and the wall of larger sporangia was broken to form small collars, and smaller sporangia were fomed continually from large basal membrane. Columella is globose and hyaline, and sporangiospores are ellipsoidal of small diameter$(80\;{\mu}m)$. Because M5 was taxonomically similar to the Mucor circinelloides of zygomycetes, M5 was was identified and named as Mucor circinelloides M5. Bacteria B16 colony was opaque white, circular and lobate, and had rod shaped endospore. B16 was found positive in stain, catalase, ${\beta}-glucosidse$ and V-P tests. B16 was found to utilize D-fructose, ${\alpha}-D-glucose$, maltose, D-mannose, D-raffinose, stachyose and sucrose. By the morphological and physiological results, the characteristics of B16 was thought to correspond to that of Bacillus megaterium. However, fatty acid composition was similar to Paenibacillus marcerans, requiring further study for the definite identification. Accordingly, Bacteria B16 was provisionally classified and named as Bacillus megaterium B16.

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