In order to evaluate the possible roles of secretory proteases in the pathogenesis of amoebic keratitis, we purified and characterized a serine protease secreted by Acanthamoeba lugdunensis KA/E2, isolated from a Korean keratitis patient. The ammonium sulfate-precipitated culture supernatant of the isolate was purified by sequential chromatography on CM-Sepharose, Sephacryl S-200, and mono Q-anion exchange column. The purified 33 kDa protease had a pH optimum of 8.5 and a temperature optimum of 55 degrees C. Phenylmethylsulfonylfluoride and 4- (2- Aminoethyl) -benzenesulfonyl-fluoride, both serine protease specific inhibitors, inhibited almost completely the activity of the 33 kDa protease whereas other classes of inhibitors did not affect its activity. The 33 kDa enzyme degraded various extracellular matrix proteins and serum proteins. Our results strongly suggest that the 33 kDa serine protease secreted from this keratopathogenic Acanthamoeba play important roles in the pathogenesis of amoebic keratitis, such as in corneal tissue invasion, immune evasion and nutrient uptake.
Acanthamoeba/*enzymology/isolation & purification/pathogenicity ; Acanthamoeba Keratitis/*parasitology ; Animals ; Cornea/parasitology ; Humans ; Hydrogen-Ion Concentration ; Korea ; Serine Endopeptidases/chemistry/*isolation & purification/*metabolism ; Substrate Specificity ; Temperature ; Virulence Factors

The reservoir hosts of Thelazia callipaeda were examined. The eyes of the 76 dogs raised at farm, 78 military dogs (shepherds), 96 cattle, and 105 pigs were investigated for the presence of eyeworm. Among them, six worms of T. callipaeda were collected from two dogs raised at farm (2.7%), and 188 worms from 26 shepherds (33.5%). No worms were recovered from the cattle or pigs. These results suggest that the dogs, especially the military dogs are serving as a reservoir host of T. callipaeda. in Korea.
Animals ; Cattle ; Disease Reservoirs/*veterinary ; Dog Diseases/epidemiology/*parasitology ; Dogs ; Eye/parasitology ; Female ; *Host-Parasite Interactions ; Korea/epidemiology ; Male ; Prevalence ; Spirurida Infections/epidemiology/parasitology/*veterinary ; Swine ; Thelazioidea/*isolation & purification

We attempted to identify parasite DNA in the biliary stones of humans via PCR and DNA sequencing. Genomic DNA was isolated from each of 15 common bile duct (CBD) stones and 5 gallbladder (GB) stones. The patients who had the CBD stones suffered from cholangitis, and the patients with GB stones showed acute cholecystitis, respectively. The 28S and 18S rDNA genes were amplified successfully from 3 and/or 1 common bile duct stone samples, and then cloned and sequenced. The 28S and 18S rDNA sequences were highly conserved among isolates. Identity of the obtained 28S D1 rDNA with that of Clonorchis sinensis was higher than 97.6%, and identity of the 18S rDNA with that of other Ascarididae was 97.9%. Almost no intra-specific variations were detected in the 28S and 18S rDNA with the exception of a few nucleotide variations, i.e., substitution and deletion. These findings suggest that C. sinensis and Ascaris lumbricoides may be related with the biliary stone formation and development.
Aged ; Aged, 80 and over ; Animals ; Ascaridida/genetics/isolation & purification ; Ascaris lumbricoides/genetics/isolation & purification ; Base Sequence ; Clonorchis sinensis/genetics/isolation & purification ; Common Bile Duct/*parasitology ; DNA, Helminth/*genetics ; DNA, Ribosomal/genetics ; Face/parasitology ; Female ; Gallbladder/parasitology ; Gallstones/*parasitology ; Helminths/genetics/*isolation & purification ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Polymerase Chain Reaction/*methods ; RNA, Ribosomal, 18S/genetics ; RNA, Ribosomal, 28S/genetics ; Sequence Alignment

PURPOSE: Troglitazone (TRO) is a peroxisome proliferator-activated receptor gamma (PPARgamma) agonist. TRO has antiproliferative activity on many kinds of cancer cells via G1 arrest. TRO also increases Cu2+/Zn2+-superoxide dismutase (CuZnSOD) and catalase. Cell cycle, and SOD and catalase may affect on radiation sensitivity. We investigated the effect of TRO on radiation sensitivity in cancer cells in vitro. MATERIALS AND METHODS: Three human cervix cancer cell lines (HeLa, Me180, and SiHa) were used. The protein expressions of SOD and catalase, and catalase activities were measured at 2-10 microM of TRO for 24 hours. Cell cycle was evaluated with flow cytometry. Reactive oxygen species (ROS) was measured using 2',7'-dichlorofluorescin diacetate. Cell survival by radiation was measured with clonogenic assay. RESULTS: By 5 microM TRO for 24 hours, the mRNA, protein expression and activity of catalase were increased in all three cell lines. G0-G1 phase cells were increased in HeLa and Me180 by 5 microM TRO for 24 hours, but those were not increased in SiHa. By pretreatment with 5 microM TRO radiation sensitivity was increased in HeLa and Me180, but it was decreased in SiHa. In Me180, with 2 microM TRO which increased catalase but not increased G0-G1 cells, radiosensitization was not observed. ROS produced by radiation was decreased with TRO. CONCLUSION: TRO increases radiation sensitivity through G0-G1 arrest or decreases radiation sensitivity through catalase-mediated ROS scavenging according to TRO dose or cell types. The change of radiation sensitivity by combined with TRO is not dependent on the PPARgamma expression level.
Catalase ; Cell Cycle ; Cell Line ; Cell Survival ; Cervix Uteri ; Chromans ; Female ; Flow Cytometry ; Fluoresceins ; Humans ; PPAR gamma ; Radiation Tolerance ; Reactive Oxygen Species ; RNA, Messenger ; Thiazolidinediones ; Uterine Cervical Neoplasms

Accessory renal artery (ARA) is a kind of developmental anomaly in renal artery. It is important in respect to clinical medicine, for example primary hypertension, renovascular disease, inferior vena caval obstruction, ureteral obstruction, occurrence of other vascular anomalies such as accessory renal veins, surgical importance and renal transplantation. However, up to few research of ARA was reported in dissection of cadavers. In our dissecting theater, 12 accessory renal arteries for 10 cadavers were found during dissection the 22 cadavers from 1995 to 1996. 1. Two cases were bilateral and 8 cases were unilateral accessory renal arteries. 2. Seven cases were left and 5 cases were right accessory renal arteries. 3. Two cases originated at the abdominal aorta between celiac trunk and superior mesenteric artery, 7 cases originated between superior mesenteric artery and inferior mesenteric artery, and 3 cases originated below inferior mesenteric artery. 4. Seven cases have no branches during their courses, 4 cases have 3 branches, and a case has 2 branches. 5. Seven cases entered into renal parenchyma through renal hilum, 5 cases entered into apical and arterosuperior segments, and 6 cases entered into inferior segment.
Aorta, Abdominal ; Cadaver ; Clinical Medicine ; Hypertension, Renovascular ; Kidney Transplantation ; Mesenteric Arteries ; Mesenteric Artery, Inferior ; Mesenteric Artery, Superior ; Renal Artery* ; Renal Veins ; Ureteral Obstruction

PURPOSE: The fibrates are ligands for peroxisome proliferator-activated receptor (PPAR) alpha and used clinically as hypolipidemic drugs. The fibrates are known to cause peroxisome proliferation, enhance superoxide dismutase (SOD) expression and catalase activity. The antioxidant actions of the fibrates may modify radiation sensitivity. Here, we investigated the change of the radiation sensitivity in two cervix cancer cell lines in combination with fenofibrate (FF). MATERIALS AND METHODS: Activity and protein expression of SOD were measured according to the concentration of FF. The mRNA expressions were measured by using real time reverse-transcription polymerase chain reaction. Combined cytotoxic effect of FF and radiation was measured by using clonogenic assay. RESULTS: In HeLa cells total SOD activity was increased with increasing FF doses up to 30 microM. In the other hand, the catalase activity was increased a little. As with activity the protein expression of SOD1 and SOD2 was increased with increasing doses of FF. The mRNAs of SOD1, SOD2, PPARalpha and PPARgamma were increased with increasing doses of FF. The reactive oxygen species (ROS) produced by radiation was decreased by preincubation with FF. The surviving fractions (SF) by combining FF and radiation was higher than those of radiation alone. In Me180 cells SOD and catalase activity were not increased with FF. Also, the mRNAs of SOD1, SOD2, and PPARalpha were not increased with FF. However, the mRNA of PPARgamma was increased with FF. CONCLUSION: FF can reduce radiation sensitivity by ROS scavenging via SOD induction in HeLa. SOD induction by FF is related with PPARalpha.
Catalase ; Cell Line ; Fenofibrate ; Fibric Acids ; Hand ; HeLa Cells ; Humans ; Hypolipidemic Agents ; Ligands ; Peroxisomes ; Polymerase Chain Reaction ; PPAR alpha ; PPAR gamma ; Radiation Tolerance ; Reactive Oxygen Species ; RNA, Messenger ; Superoxide Dismutase ; Superoxides ; Uterine Cervical Neoplasms

No abstract available.
Humans ; Infant, Newborn* ; Vitamin K Deficiency* ; Vitamin K* ; Vitamins*

Purpose: The aim of this study was to investigate the outcomes of endodontic treatment of mandibular second molar with C-shaped canal in elderly patients and related factors affecting the survival of the tooth. Materials and Methods: From 2010 to 2015, the survival rate following endodontic treatment was evaluated in elderly patients over 60 years who visited the Veterans Health Service Medical Center for endodontic treatment. The presence of C-shaped canals was confirmed using clinical records and radiographic features. Patientʼs age, sex, systemic diseases, tooth location, vitality, signs and symptoms, fractures, caries, apical radiolucency, canal shaping methods, sealer leakage, filling voids, and restoration of prosthesis were included in the analyses as confounding variables. The survival rate of teeth was analyzed using Kaplan–Meire analysis and the relationship between the survival rate and variables was analyzed using Simple and Multiple Cox regression analysis.Result: In total, 107 teeth in elderly patients had C-shaped canal. The survival rate of teeth that received endodontic treatment was 63.70%. None of the factors investigated significantly influenced the survival rate (P>0.05). Conclusion In elderly patients with C-shaped canal, the survival rate after root canal treatment was not significantly different from that of other mandibular molars.

PURPOSE: Kawasaki disease(KD) is an important acute febrile systemic vasculitis disease. Various cytokines have been studied to evaluate the pathogenesis of vascular injury. Some cytokines have been implicated for vascular injury, but there has been no direct evidence. We compared cytokine profiles of KD with Henoch-Schonlein purpura(HSP), and febrile infections. METHODS: We investigated the serum concentrations of six cytokines and intracellular TNF-alpha and IL-6 levels in T cells and monocytes with acute KD, HSP and infections. We determined serum cytokine concentrations using the cytometric bead array method and detected intracellular TNF-alpha and IL-6 levels using QuantiBRITE and dual color flow cytometry. RESULTS: Serum IFN-gamma concentrations were increased more in KD and infections than in HSP and normal controls. Serum TNF-alpha concentrations were higher in KD and infections than in HSP and normal controls. And TNF-alpha concentrations in KD were lower than in infections. Serum IL-10 concentrations were higher in KD and in infections than in HSP and normal controls. And IL-10 concentrations in KD were higher than in infections. Serum IL-6 concentrations were higher in KD and in infections than in HSP and normal controls. Serum IL-4 and IL-2 concentrations were lower and similar to those of normal controls, respectively. However, intracellular IL-6 levels in T cells were higher in KD than in infections and intracellular IL-6 levels in monocytes were higher in KD than in infections. CONCLUSION: These results suggest that the alterations of various cytokines in KD are related to fever rather than to vasculitis. KD showed higher IL-10 levels in serum and intracellular IL-6 levels in T cell and monocyte than HSP. This reveals that the pathogenesis of KD is different from that of HSP.
Cytokines ; Fever ; Flow Cytometry ; Interferons ; Interleukin-10 ; Interleukin-2 ; Interleukin-4 ; Interleukin-6 ; Monocytes ; Mucocutaneous Lymph Node Syndrome* ; Purpura ; Systemic Vasculitis ; T-Lymphocytes ; Tumor Necrosis Factor-alpha ; Vascular System Injuries ; Vasculitis

Objectives: ：This study was aimed to investigate the changes in metabolic syndrome (MetS) status and long-term impact of its components over a 10-year period in severe mental illness (SMI) patients in a national mental hospital. Methods: ：A total of 93 patients (schizophrenia=88, bipolar disorder=5) who met the diagnosis of Diagnostic and Statistical Manual of Mental Disorders, fourth edition, text revision (DSM-IV-TR) and participated in the MetS study in 2011 were included. MetS was defined by revised National Cholesterol Education Program-Adult Treatment Panel III (revised NCEP-ATP-III) guidelines. Results: ：The prevalence of MetS was significantly increased from 40.9% in 2011 to 60.2% in 2020. There were significant differences in admission status and hospitalization months, compared to the groups with and without MetS. Upon reviewing the changes over a decade, systolic blood pressure (SBP) was a significant factor in the group without MetS. In the group with MetS, SBP, waist circumference, and BMI (body mass index) were significant factors. Multiple logistic regression analysis revealed that hospitalization during follow-up periods [odds ratio (OR)=0.969, 95% confidence interval (CI): 0.948-0.991] and BMI (OR=1.426, 95% CI: 1.196-1.701) were significantly associated with MetS in subjects. Conclusion ：The prevalence of MetS in patients with SMI significantly increased over time. The admission status and hospitalization were also confirmed to be the significant values of MetS.