Acanthamoeba spp. are free-living protozoa that are opportunistic pathogens for humans. Cysteine proteases of Acanthamoeba have been partially characterized, but their biochemical and functional properties are not clearly understood yet. In this study, we isolated a gene encoding cysteine protease of A. castellanii (AcCP) and its biochemical and functional properties were analyzed. Sequence analysis of AcCP suggests that this enzyme is a typical cathepsin L family cysteine protease, which shares similar structural characteristics with other cathepsin L-like enzymes. The recombinant AcCP showed enzymatic activity in acidic conditions with an optimum at pH 4.0. The recombinant enzyme effectively hydrolyzed human proteins including hemoglobin, albumin, immunoglobuins A and G, and fibronectin at acidic pH. AcCP mainly localized in lysosomal compartment and its expression was observed in both trophozoites and cysts. AcCP was also identified in cultured medium of A. castellanii. Considering to lysosomal localization, secretion or release by trophozoites and continuous expression in trophozoites and cysts, the enzyme could be a multifunctional enzyme that plays important biological functions for nutrition, development and pathogenicity of A. castellanii. These results also imply that AcCP can be a promising target for development of chemotherapeutic drug for Acanthamoeba infections.
Acanthamoeba castellanii ; Acanthamoeba ; Cathepsin L ; Cathepsins ; Cysteine Proteases ; Cysteine ; Fibronectins ; Genes, vif ; Humans ; Hydrogen-Ion Concentration ; Lysosomes ; Sequence Analysis ; Trophozoites ; Virulence

A 76-year-old female present to the emergency department with dysarthria, dizziness, dyspnea. The patient had hypertension and atrial fibrillation. Brain MRI revealed right cerebellar infarction. Transthoracic echocardiography showed a large round mass in the left atrium. Transesophageal echocardiography showed large complex echogenic round mass lesion attached on left atrial side of interatrial septum. Coronary angiogram revealed round movable mass lesion in left atrium with feeding arteries originated from right coronary artery. She underwent removal of mass and Maze operation, and pathologic finding was compatible with myxoma.
Aged ; Arteries ; Atrial Fibrillation ; Brain ; Coronary Vessels ; Dizziness ; Dysarthria ; Dyspnea ; Echocardiography ; Echocardiography, Transesophageal ; Emergencies ; Female ; Heart ; Heart Atria ; Heart Murmurs ; Humans ; Hypertension ; Infarction ; Myxoma

BACKGROUND: Real-time PCR for quantification of JAK2 V617F has recently been introduced and used to evaluate the importance of mutant allele burden in both diagnosis and disease progression in myeloproliferative diseases (MPDs). We evaluated the usefulness of JAK2 MutaScreen(TM) kit that uses a real-time semiquantitative PCR method and has been designed to screen JAK2 V617F mutant allele burden. METHODS: Forty MPD patients were included in this study. We screened JAK2 V617F and determined the mutant allele burden using JAK2 MutaScreen(TM) kit. The mutant allele burden was estimated by six-scaled standards of JAK2 V617F mutant allele (2%, 5%, 12.5%, 31%, 50%, and 78%). For evaluation of test performance, an allele-specific PCR (AS-PCR) was carried out in all samples by using Seeplex JAK2 Genotyping kit. We assessed the clinical differences in distinct disease entities of MPDs according to JAK2 V617F mutant allele burden. RESULTS: JAK2 V617F mutation was detected in 30 cases, including 10 of 11 cases (91%) of polycythemia vera (PV), 13 of 20 cases (65%) of essential thrombocythemia (ET), and 2 of 3 cases (67%) of chronic idiopathic myelofibrosis (CIMF). The concordance rate between the two tests was 95% (38/40). JAK2 V617F mutant allele burden was greater than 50% in 17 cases, and 10 of them (59%) were PV. In contrast, mutant allele burden was less than 50% in 13 cases and 11 of them (85%) were ET. CONCLUSIONS: JAK2 MutaScreen(TM) kit that utilizes a real-time semi-quantitative PCR method is a useful tool for diagnosing MPDs precisely. It can be used to assess the grade of mutant allele burden as well as to screen JAK2 V617F simultaneously.
Adult ; Aged ; *Alleles ; Amino Acid Substitution ; DNA Mutational Analysis ; Disease Progression ; Female ; Humans ; Janus Kinase 2/*genetics ; Male ; Middle Aged ; Mutation ; Myeloproliferative Disorders/*diagnosis/genetics ; Polymerase Chain Reaction/*methods ; Reagent Kits, Diagnostic

BACKGROUND AND OBJECTIVES: Increased QT dispersion (QTD) in patients with acute myocardial infarction (AMI) may be related with such adverse events as sudden cardiac death and ischemic heart failure. SUBJECTS AND METHODS: Two hundred eight patients (age : 62+/-10.4 years, 158 males), underwent diagnostic coronary angiography under the diagnosis of AMI between January and December 2001 at the Heart Center of Chonnam National University Hospital, and these patients were enrolled to evaluate the relationship between the QTD and myocardial injury and the complex coronary arterial lesion. RESULTS: A QTD of over 80 ms was observed in 89 patients (42.7%). There were in 61 patients with ST elevation myocardial infarction (STEMI, 68.5%) and 28 patients with non-ST elevation myocardial infarction (NSTEMI, 31.5%). There was no correlation between the QTD and such risk factors as hypertension, diabetes, gender, smoking, hyperlipidemia and family history. The level of CK-MB on admission was correlated with the QTD (112.5+/-98.1 U/L in the group with a QTD over 80 ms and 72.6+/-73.4 U/L in the group with a QTD under 80 ms, p<0.05). The ejection fraction measured by two dimensional echocardiography on admission showed correlation with the QTD (50.9+/-11.4% in the group with a QTD over 80 ms and 54.7+/-11.2% U/L in the group with a QTD under 80 ms, p<0.05). For the coronary angiographic findings, the lesion type, according to American College of Cardiology/American Heart Association classification, correlated with the QT dispersion (type B2 or C : 64.1% in the group with a QTD over 80 ms, 49.6% in the group with a QTD under 80 ms, p<0.05) CONCLUSION: There was significant correlation between the prolonged QTD and the severity of myocardial injury at admission, and the complex coronary arterial lesion in patients with AMI.
Classification ; Coronary Angiography ; Coronary Vessels* ; Death, Sudden, Cardiac ; Diagnosis ; Echocardiography ; Electrocardiography ; Heart ; Heart Failure ; Humans ; Hyperlipidemias ; Hypertension ; Jeollanam-do ; Myocardial Infarction* ; Risk Factors ; Smoke ; Smoking

BACKGROUND: This study was designed to evaluate the safety and clinical benefits of a glycoprotein IIb/IIIa receptor inhibitor, ReoPro(R) in the elderly patients with acute myocardial infarction (AMI) (>or=70 years of age) undergoing percutaneous coronary intervention (PCI). METHODS: AMI patients who underwent PCI with use of ReoPro(R) at Chonnam National University Hospital from Jan 2000 to Jan 2002 were divided into two groups: Group I (>or=70 years of age: 74 +/- 2.4 years, n=28) and Group II (<70 years of age: 56 +/- 8.0 years, n=122). Early and long-term clinical outcomes after PCI were analyzed in a retrospective fashion. RESULTS: As for risk factors and angiographic profiles, there were no differences between the two groups. Stenting was performed in 18 patients (64%) in group I and in 78 patients (63%) in group II. The incidence of gastrointestinal bleeding was 3 patients in group I and no patient in group II (p=0.005). At one-month evaluation, three cardiac deaths developed in group I, but no cardiac death in group II (p=0.005). During a period of 25 +/- 10.4 months of clinical follow-up, three cardiac deaths (11%) occurred in group I and 3 (2%) in group II, four AMIs (3%) in group II, and one stroke (0.8%) in group II. Target lesion revascularization (TLR) was performed in two patients (7%) in group I and in 24 patients (19%) in group II. No differences were found in the incidences of these variables between the two groups. CONCLUSION: ReoPro(R) in elderly patients with AMI undergoing PCI entailed higher bleeding complications and early mortality. However, it has comparable clinical effect in elderly patients to younger patients during long-term clinical follow-up.
Aged* ; Angioplasty ; Blood Platelets* ; Death ; Follow-Up Studies ; Glycoproteins* ; Hemorrhage ; Humans ; Incidence ; Jeollanam-do ; Mortality ; Myocardial Infarction* ; Percutaneous Coronary Intervention* ; Retrospective Studies ; Risk Factors ; Stents ; Stroke

BACKGROUND: Currently, the cardiopulmonary machine with non-pulsatile pumps, which are low in internal circuit pressure and cause little damage to blood cells, is widely used. However, a great number of experimental studies shows that pulsatile perfusions are more useful than non-pulsatile counterparts in many areas, such as hemodynamic, metabolism, organ functions, and micro-circulation. Yet, many concerns relating to pulsatile cardiopulmonary machines, such as high internal circuit pressure and blood cell damage, have long hindered the development of pulsatile cardiopulmonary machines. Against this backdrop, this study focuses on the safety and effectiveness of the pulsatile cardiopulmonary machines developed by a domestic research lab. MATERIAL AND METHOD: The dual-pulsatile cardiopulmonary bypass experiment with total extracorporeal circulation was conducted on six calves. Extracorporeal circulation was provided between superior/inferior vena cava and aorta. The membrane oxygenator, which was placed between the left and right pumps, was used for blood oxygenation. Circulation took four hours. Arterial blood gas analysis and blood tests were also conducted. Plasma hemoglobin levels were calculated, while pulse pressure and internal circuit pressure were carefully observed. Measurement was taken five times; once before the operation of the cardiopulmonary bypass, and after its operation it was taken every hour for four hours. RESULT: Through the arterial blood gas analysis, PCO2 and pH remained within normal levels. PO2 in arterial blood showed enough oxygenation of over 100 mmHg. The level of plasma hemoglobin, which had total cardiopulmonary circulation, steadily increased to 15.87+/-5.63 mg/dl after four hours passed, but remained below 20 mg/dl. There was no obvious abnormal findings in blood test. Systolic blood pressure which was at 97.5+/-5.7 mmHg during the pre-circulation contraction period, was maintained over 100 mmHg as time passed. Moreover, diastolic blood pressure was 72.2+/-7.7 mmHg during the expansion period and well kept at the appropriate level with time passing by. Average blood pressure which was 83+/-9.2 mmHg before circulation, increased as time passed, while pump flow was maintained over 3.3 L/min. Blood pressure fluctuation during total extracorporeal circulation showed a similar level of arterial blood pressure of pre-circulation heart. CONCLUSION: In the experiment mentioned above, pulsatile cardiopulmonary machines using the doual-pulsatile structure provided effective pulsatile blood flow with little damage in blood cells, showing excellence in the aspects of hematology and hemodynamic. Therefore, it is expected that the pulsatile cardiopulmonary machine, if it becomes a standard cardiopulmonary machine in all heart operations, will provide stable blood flow to end-organs.
Animals* ; Aorta ; Arterial Pressure ; Blood Cells ; Blood Gas Analysis ; Blood Pressure ; Cardiopulmonary Bypass* ; Extracorporeal Circulation ; Heart ; Hematologic Tests ; Hematology ; Hemodynamics ; Hydrogen-Ion Concentration ; Metabolism ; Models, Animal* ; Oxygen ; Oxygenators, Membrane ; Perfusion ; Plasma ; Pulsatile Flow

BACKGROUND: The preoperative coronary angiography is required in some patients underwent valvular surgery. The purpose of this study was to evaluate predictive factors for coronary lesion and to determine the indication for preoperative coronary angiography in Korean patients with valvular heart disease (VHD). METHODS: Two hundreds eight patients (55.5 +/- 12.8 years, 99 male) with VHD underwent preoperative coronary angiography between January 1996 and January 2003 were analyzed the variables of age, gender, previous or current angina, hypertension, diabetes mellitus, smoking, dyslipidemia, obesity and familial predisposition according to the presence of coronary lesion. RESULTS: The prevalence of significant coronary artery disease in VHD patients was 26.4% (55/208). The probability of coronary lesion was higher in male above 55 years and in female above 65 years than in younger patients. The predictive factors of coronary lesion were previous or current angina (p<0.05), dyslipidemia (p<0.01), hypertension (p<0.05), previous angina (p<0.05), diabetes (p<0.01) and smoking (p<0.01) by multiple logistic regression analysis. CONCLUSION: Previous or current angina and at least one of the four risk factors (hypertension, smoking, dyslipidemia and diabetes) are predictors of coronary lesion, especially in elderly Korean patients with VHD (older than 55 years in male and 65 years in female).
Aged ; Coronary Angiography* ; Coronary Artery Disease ; Coronary Disease ; Diabetes Mellitus ; Dyslipidemias ; Female ; Heart Valve Diseases* ; Humans ; Hypertension ; Logistic Models ; Male ; Obesity ; Prevalence ; Risk Factors ; Smoke ; Smoking